International Journal of Molecular Sciences

3 pages, 169 KiB

Open AccessEditorial

Molecular Pathology, Diagnostics and Therapeutics: A Story of Success in 2022

by Stephen A. Bustin

Int. J. Mol. Sci. 2023, 24(5), 5063; https://doi.org/10.3390/ijms24055063 - 6 Mar 2023

Viewed by 1756

Molecular pathology, diagnostics and therapeutics are three closely related topics of critical importance in medical research and clinical practice [...] Full article

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

15 pages, 2786 KiB

Open AccessArticle

Loss of Blood-Brain Barrier Integrity in an In Vitro Model Subjected to Intermittent Hypoxia: Is Reversion Possible with a HIF-1α Pathway Inhibitor?

by Anne Cloé Voirin, Morgane Chatard, Anne Briançon-Marjollet, Jean Louis Pepin, Nathalie Perek and Frederic Roche

Int. J. Mol. Sci. 2023, 24(5), 5062; https://doi.org/10.3390/ijms24055062 - 6 Mar 2023

Cited by 4 | Viewed by 2314

Abstract

Several sleep-related breathing disorders provoke repeated hypoxia stresses, which potentially lead to neurological diseases, such as cognitive impairment. Nevertheless, consequences of repeated intermittent hypoxia on the blood-brain barrier (BBB) are less recognized. This study compared two methods of intermittent hypoxia induction on the [...] Read more.

Several sleep-related breathing disorders provoke repeated hypoxia stresses, which potentially lead to neurological diseases, such as cognitive impairment. Nevertheless, consequences of repeated intermittent hypoxia on the blood-brain barrier (BBB) are less recognized. This study compared two methods of intermittent hypoxia induction on the cerebral endothelium of the BBB: one using hydralazine and the other using a hypoxia chamber. These cycles were performed on an endothelial cell and astrocyte coculture model. Na-Fl permeability, tight junction protein, and ABC transporters (P-gp and MRP-1) content were evaluated with or without HIF-1 inhibitors YC-1. Our results demonstrated that hydralazine as well as intermittent physical hypoxia progressively altered BBB integrity, as shown by an increase in Na-Fl permeability. This alteration was accompanied by a decrease in concentration of tight junction proteins ZO-1 and claudin-5. In turn, microvascular endothelial cells up-regulated the expression of P-gp and MRP-1. An alteration was also found under hydralazine after the third cycle. On the other hand, the third intermittent hypoxia exposure showed a preservation of BBB characteristics. Furthermore, inhibition of HIF-1α with YC-1 prevented BBB dysfunction after hydralazine treatment. In the case of physical intermittent hypoxia, we observed an incomplete reversion suggesting that other biological mechanisms may be involved in BBB dysfunction. In conclusion, intermittent hypoxia led to an alteration of the BBB model with an adaptation observed after the third cycle. Full article

(This article belongs to the Special Issue Pathophysiological and Molecular Signaling Impacts of Chronic Hypoxia and Intermittent Hypoxia)

► Show Figures

Figure 1

36 pages, 3697 KiB

Open AccessArticle

Genetic Structure Analysis of 155 Transboundary and Local Populations of Cattle (Bos taurus, Bos indicus and Bos grunniens) Based on STR Markers

by Evgenia Solodneva, Gulnara Svishcheva, Rodion Smolnikov, Sergey Bazhenov, Evgenii Konorov, Vera Mukhina and Yurii Stolpovsky

Int. J. Mol. Sci. 2023, 24(5), 5061; https://doi.org/10.3390/ijms24055061 - 6 Mar 2023

Cited by 2 | Viewed by 2571

Abstract

Every week, 1–2 breeds of farm animals, including local cattle, disappear in the world. As the keepers of rare allelic variants, native breeds potentially expand the range of genetic solutions to possible problems of the future, which means that the study of the [...] Read more.

Every week, 1–2 breeds of farm animals, including local cattle, disappear in the world. As the keepers of rare allelic variants, native breeds potentially expand the range of genetic solutions to possible problems of the future, which means that the study of the genetic structure of these breeds is an urgent task. Providing nomadic herders with valuable resources necessary for life, domestic yaks have also become an important object of study. In order to determine the population genetic characteristics, and clarify the phylogenetic relationships of modern representatives of 155 cattle populations from different regions of the world, we collected a large set of STR data (10,250 individuals), including unique native cattle, 12 yak populations from Russia, Mongolia and Kyrgyzstan, as well as zebu breeds. Estimation of main population genetic parameters, phylogenetic analysis, principal component analysis and Bayesian cluster analysis allowed us to refine genetic structure and provided insights in relationships of native populations, transboundary breeds and populations of domestic yak. Our results can find practical application in conservation programs of endangered breeds, as well as become the basis for future fundamental research. Full article

(This article belongs to the Special Issue Bioinformatics of Gene Regulations and Structure - 2022)

► Show Figures

Figure 1

Figure 1
Phylogenetic tree constructed by the neighbor-joining algorithm using Da genetic distances. The circular range is represented by 7 main clusters (breed group): purple (Domestic yak), yellow (Zebu), light yellow (African), beige (Iberian), pink (European), light green (Asian) and blue (Creole). The root of the tree was selected automatically for better visualisation and represents the basic division between modern yaks and the rest. Full article ">Figure 2
Principal component analysis. Spatial representation of the genetic distances between the analysed breeds along the first two axes obtained by factor matching analysis based on microsatellite data. The values in parentheses on both axes represent the percentage contribution of each axis to the total inertia. The colours represent belonging to a breed group, as shown in the figure. The names of some breeds are given. (a) PCA results for 155 populations. (b) PCA results for a sample of predominantly European and Asian breeds from Cinisara to Alatau (according to the dendrogram). Full article ">Figure 3
The evaluation of the structure of each individual from the studied populations for K = 12. Each individual is represented by a separate column. Colour identification reflects the proportion of 12 estimated ancestral populations in the genome of a particular individual. (a) Structure of 10,250 individuals belonging to seven breed groups (European, Asian, Creole, Iberian, African, Zebu and Domestic yak). (b) Structure of individuals belonging to populations of the Asian breed group, indicating the country and 4 Russian breeds from the European breed group. Full article ">Figure 4
Cluster analysis of 155 cattle populations calculated from microsatellite data in the STRUCTURE program. Each breed is represented by a vertical column divided into K colours, according to the number of estimated ancestral populations. Size of the colour segment is proportional to the contribution of a particular ancestral population to the genome of the breed in question. The results for K = 4, 12 and 20 are presented. The values of K from 2 to 20 are shown in the additional <a href="#app1-ijms-24-05061" class="html-app">Figure S2</a>. Full article ">Figure 5
Genotype accumulation curve for all animals genotyped on 12 loci. The horizontal axis represents the number of loci randomly sampled without replacement up to <math display="inline"><semantics> <mrow> <mo>(</mo> <mi>n</mi> <mo>−</mo> <mn>1</mn> <mo>)</mo> </mrow> </semantics></math> loci; the vertical axis shows the number of multilocus genotypes observed, the number of unique multilocus genotypes in the dataset. The red dashed line represents 100% of the total observed multilocus genotypes. Full article ">

15 pages, 3164 KiB

Open AccessArticle

A Novel Wall-Associated Kinase TaWAK-5D600 Positively Participates in Defense against Sharp Eyespot and Fusarium Crown Rot in Wheat

by Haijun Qi, Xiuliang Zhu, Wenbiao Shen and Zengyan Zhang

Int. J. Mol. Sci. 2023, 24(5), 5060; https://doi.org/10.3390/ijms24055060 - 6 Mar 2023

Cited by 1 | Viewed by 2223

Abstract

Sharp eyespot and Fusarium crown rot, mainly caused by soil-borne fungi Rhizoctonia cerealis and Fusarium pseudograminearum, are destructive diseases of major cereal crops including wheat (Triticum aestivum). However, the mechanisms underlying wheat-resistant responses to the two pathogens are largely elusive. [...] Read more.

Sharp eyespot and Fusarium crown rot, mainly caused by soil-borne fungi Rhizoctonia cerealis and Fusarium pseudograminearum, are destructive diseases of major cereal crops including wheat (Triticum aestivum). However, the mechanisms underlying wheat-resistant responses to the two pathogens are largely elusive. In this study, we performed a genome-wide analysis of wall-associated kinase (WAK) family in wheat. As a result, a total of 140 TaWAK (not TaWAKL) candidate genes were identified from the wheat genome, each of which contains an N-terminal signal peptide, a galacturonan binding domain, an EGF-like domain, a calcium binding EGF domain (EGF-Ca), a transmembrane domain, and an intracellular Serine/Threonine protein kinase domain. By analyzing the RNA-sequencing data of wheat inoculated with R. cerealis and F. pseudograminearum, we found that transcript abundance of TaWAK-5D600 (TraesCS5D02G268600) on chromosome 5D was significantly upregulated, and that its upregulated transcript levels in response to both pathogens were higher compared with other TaWAK genes. Importantly, knock-down of TaWAK-5D600 transcript impaired wheat resistance against the fungal pathogens R. cerealis and F. pseudograminearum, and significantly repressed expression of defense-related genes in wheat, TaSERK1, TaMPK3, TaPR1, TaChitinase3, and TaChitinase4. Thus, this study proposes TaWAK-5D600 as a promising gene for improving wheat broad resistance to sharp eyespot and Fusarium crown rot (FCR) in wheat. Full article

(This article belongs to the Special Issue Molecular Insights into Plant-Biotic Interactions and Crop Yield)

► Show Figures

Figure 1

Figure 1
Chromosomal distribution of the typical TaWAK genes in wheat. The 140 TaWAK genes were unevenly distributed on 20 wheat chromosomes. The bar indicates the length of chromosome in megabases (MB). Full article ">Figure 2
The transcript profiles of 140 TaWAKs in the wheat RNA-sequencing (RNA-seq) data. (A) The transcript levels of 140 TaWAKs upon R. cerealis infection in the resistant recombinant inbred lines (RILs) derived from the cross ‘Shanhongmai’ × ‘Wenmai 6′. (B) The transcript profiles of 140 TaWAKs upon F. pseudograminearum infection. The RNA-seq data upon F. pseudograminearum infection were checked from the online RNA-seq data (<a href="http://www.wheat-expression.com/" target="_blank">http://www.wheat-expression.com/</a> accessed on 25 August 2022) [<a href="#B32-ijms-24-05060" class="html-bibr">32</a>]. Full article ">Figure 3
TaWAK-5D600 is involved in wheat responses to both sharp eyespot and Fusarium crown rot. (A) The transcript levels and fold change of the significantly upregulated 27 TaWAK genes in the RILs-R response to R. cerealis infection. (B) The transcript levels and fold change of the 27 R. cerealis induced TaWAK genes upon F. pseudograminearum infection in the online RNA-seq data (<a href="http://www.wheat-expression.com/" target="_blank">http://www.wheat-expression.com/</a> accessed on 25 August 2022) [<a href="#B32-ijms-24-05060" class="html-bibr">32</a>]. (C) Transcript levels of TaWAK-5D600 in sharp eyespot-resistant wheat line CI12633 at non-treatment and 4, 7, and 10 dpi with R. cerealis Rc207. (D) The transcript patterns of TaWAK-5D600 in FCR-mildly-resistant wheat line CI12633 at non-treatment and 1 and 2 dpi with F. pseudograminearum WHF220. TaWAK-5D600 transcript level at non-treatment is set to 1. TaActin gene was used as the internal control (t-test: * p < 0.05; ** p < 0.01). Full article ">Figure 4
Phylogenetic tree, conserved-domain, and gene-structure analyses of TaWAK-5D600. (A) A phylogenetic tree of TaWAK-5D600 and other 18 WAK proteins from different plants. The position of TaWAK-5D600 was indicated by a red blot. (B) Gene structure of TaWAK-5D600; black boxes represent exons and black lines indicate introns. (C) Schematic diagram of the TaWAK-5D600 protein. The conserved protein domains of TaWAK-5D600 were represented by different colored boxes. (D) Subcellular localization of TaWAK-5D600 in wheat protoplasts cells (bars = 20 μm). Full article ">Figure 5
Silencing of TaWAK-5D600-compromised wheat resistance both to sharp eyespot and Fusarium crown rot. (A) Typical BSMV symptoms on wheat leaves at 15 dpi infected with BSMV and transcripts of BSMV coat protein (CP) gene detecting by RT-PCR assays. (B) The silencing efficiency of TaWAK-5D600 detecting by RT-qPCR assay. The transcript level of TaWAK-5D600 in BSMV:GFP (control) wheat seedlings was set to 1. (C) Sharp eyespot symptoms on TaWAK-5D600-silenced and BSMV:GFP (control) wheat plants at 30 dpi with R. cerealis. (D) Disease indexes (DIs) of TaWAK-5D600-silenced and BSMV:GFP (control) wheat plants at 30 dpi with R. cerealis in two independent batches (t-test: ** p < 0.01). (E) Fusarium crown rot symptoms on TaWAK-5D600-silenced and control wheat plants at 30 dpi with F. pseudograminearum. (F) Disease index (DI) of TaWAK-5D600-silenced and control wheat plants at 30 dpi with F. pseudograminearum WHF220 in two independent batches (t-test: ** p < 0.01). Bars indicate SEs of the mean. Full article ">Figure 6
Transcript profiles of TaWAK-5D600 and defense-related genes in BSMV:GFP (control) and BSMV:TaWAK-5D600-infected wheat seedlings. Relative transcript abundances of TaWAK-5D600 and the tested genes TaSERK1, TaMPK3, TaPR1, TaChitinase3, and TaChitinase4 in BSMV: TaWAK5D600-infected CI12633 seedlings were quantified relative to those in BSMV:GFP (control) seedlings. Statistically significant differences were calculated based on three replications via a Student’s t-test (** p < 0.01). TaActin was used as an internal control. Full article ">

19 pages, 6634 KiB

Open AccessArticle

Ginsenoside Rb1 Improves Post-Cardiac Arrest Myocardial Stunning and Cerebral Outcomes by Regulating the Keap1/Nrf2 Pathway

by Long Chen, Na Geng, Taiwei Chen, Qingqing Xiao, Hengyuan Zhang, Huanhuan Huo, Lisheng Jiang, Qin Shao and Ben He

Int. J. Mol. Sci. 2023, 24(5), 5059; https://doi.org/10.3390/ijms24055059 - 6 Mar 2023

Cited by 8 | Viewed by 2402

Abstract

The prognosis of cardiac arrest (CA) is dismal despite the ongoing progress in cardiopulmonary resuscitation (CPR). ginsenoside Rb1 (Gn-Rb1) has been verified to be cardioprotective in cardiac remodeling and cardiac ischemia/reperfusion (I/R) injury, but its role is less known in CA. After 15 [...] Read more.

The prognosis of cardiac arrest (CA) is dismal despite the ongoing progress in cardiopulmonary resuscitation (CPR). ginsenoside Rb1 (Gn-Rb1) has been verified to be cardioprotective in cardiac remodeling and cardiac ischemia/reperfusion (I/R) injury, but its role is less known in CA. After 15 min of potassium chloride-induced CA, male C57BL/6 mice were resuscitated. Gn-Rb1 was blindly randomized to mice after 20 s of CPR. We assessed the cardiac systolic function before CA and 3 h after CPR. Mortality rates, neurological outcome, mitochondrial homeostasis, and the levels of oxidative stress were evaluated. We found that Gn-Rb1 improved the long-term survival during the post-resuscitation period but did not affect the ROSC rate. Further mechanistic investigations revealed that Gn-Rb1 ameliorated CA/CPR-induced mitochondrial destabilization and oxidative stress, partially via the activation of Keap1/Nrf2 axis. Gn-Rb1 improved the neurological outcome after resuscitation partially by balancing the oxidative stress and suppressing apoptosis. In sum, Gn-Rb1 protects against post-CA myocardial stunning and cerebral outcomes via the induction of the Nrf2 signaling pathway, which may offer a new insight into therapeutic strategies for CA. Full article

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

► Show Figures

Figure 1

Figure 1
Flow chart of the experimental groups. (a) Schematic of the experimental workflow. min: minute; h: hour; min: minute; CA: cardiac arrest; CPR: cardiopulmonary resuscitation; KCl: potassium chloride; ROSC: return of spontaneous circulation; echo: echocardiography; Rb1: ginsenoside Rb1; (b) flow chart of the experimental groups. sham: mice underwent a sham surgery without CA/CPR; vehicle: mice subjected to CA/CPR without Rb1; n: number of mice; EF: ejection fraction; FS: fractional shortening; CO: cardiac output; DHE: dihydroethidium; 4-HNE: 4 hydroxynonenal; NT: nitrotyrosine; TUNEL: terminal dUTP nick-end labeling; NADH: NADH dehydrogenase; and TEM: transmission electron microscopy. Full article ">Figure 2
Gn-Rb1 treatment attenuated myocardial dysfunction and improved survival rate after CA/CPR. (a) Structure of ginsenoside Rb1. (b) Return of spontaneous circulation (ROSC) rate (n = 15 per group). (c) Time for ROSC (n = 15 per group). (d–g) Cardiac dysfunction measured by echocardiography (n = 9–15 per group). (h) Kaplan–Meier curve demonstrating survival following CA/CPR (n = 10 in sham, n = 20 in CA, and CA + Rb1 group). * p < 0.05; and NS = not significant. LVEF: left ventricular ejection fraction; LVFS: left ventricular fractional shortening; CO: cardiac output; CA: cardiac arrest; and Rb1: ginsenoside Rb1; h: hour. Full article ">Figure 3
Gn-Rb1 attenuated oxidative stress in the heart following CA/CPR. (a,b) Representative immunofluorescence staining images and quantitative result of dihydroethidium (DHE), performed to assess myocardial ROS accumulation (n = 3 per group). (c,d) Representative immunohistochemical staining images and quantitative result of nitrotyrosine (NT), performed to assess myocardial nitrotyrosine production in different groups (n = 3 per group). (e,f) Representative immunohistochemical staining images and quantitative result of 4 hydroxynonenal (4-HNE), performed to assess myocardial lipid peroxidation (n = 3 per group). (g,h) Western blot analysis of antioxidant proteins SOD2 and oxidative markers gp91phox in different cardiac homogenates (n = 6 per group). (i,j) Western blot analysis of subunits of NADH dehydrogenase in different cardiac homogenates (n = 6 per group). (k) NADH dehydrogenase was assessed using an ELISA assay 3 h following CA/CPR (n = 6 per group). CA: cardiac arrest; CPR: cardiopulmonary resuscitation; Rb1: ginsenoside Rb1. * p < 0.05; and NS = not significant. Full article ">Figure 4
Gn-Rb1 improves mitochondrial homeostasis and energy metabolism in the heart following CA/CPR. (a) Transmission electron microscopy was performed to observe the ultrastructure of post-CA myocardial tissues (n = 3 per group). (b,c) Representative Western blots and quantitative analysis of total p-Drp1 (Ser 616) and p-Drp1 (Ser 637), as well as GAPDH protein expression (n = 6 per group). (d,e) Representative Western blots and quantitative analysis of mitochondrial Drp1, Fis1, Mfn2, Opa1, and VDAC1 protein expression (n = 6 per group). (f) The total ATP levels in different group were determined by Luciferase assay (n = 6 per group). (g) The membrane potential in isolated mitochondria was assessed by Luciferase assay (n = 6 per group). CA: cardiac arrest; CPR: cardiopulmonary resuscitation; Rb1: ginsenoside Rb1. * p < 0.05; and NS = not significant. Full article ">Figure 5
Gn-Rb1 activates the Keap1/Nrf2 signaling pathway in the heart following CA/CPR. (a,b) Representative Western blots and quantitative analysis of Keap1, nuclear and cytosolic Nrf2, HO-1, and NQO1 in the myocardium for 3 h, following CA/CPR in different groups (n = 6). (c,d) NRCMs were transfected with small interfering RNA targeting Nrf2 and exposed to hypoxia for 12 h, and then treated as indicated during reoxygenation for 3 h. Representative Western blots and quantitative analysis of HO-1 and NQO1 were assessed (n = 3). H/R: hypoxia/reoxygenation; CA: cardiac arrest; CPR: cardiopulmonary resuscitation; and Rb1: ginsenoside Rb1. * p < 0.05; and NS = not significant. Full article ">Figure 6
Gene knockdown of Nrf2 attenuates the ameliorative effect of ginsenoside Rb1 on oxidative stress after H/R. (a–c) The mRNA and protein expression of Nrf2 were assessed 48 h after Nrf2 siRNA or NC siRNA transfection in NRCM. (d,e) Representative fluorescent images of DHE, DCFH-DA, and MitoSOX red in the presence of Nrf2 siRNA or NC siRNA with or without Gn-Rb1 treatment (10 μM) in NRCM after H/R insult (n = 3). (f,g) Western blot analysis of antioxidant proteins SOD2 and oxidative markers gp91 in different groups. (h) NADH dehydrogenase activity was assessed using an ELISA assay in different groups (n = 3 per group). (i,j) Western blot analysis of subunits of NADH dehydrogenase in different groups (n = 3 per group). H/R: hypoxia/reoxygenation; DHE: dihydroethidium; ROS: Reactive oxygen species; and Rb1: ginsenoside Rb1. # p < 0.05 vs. Sham; * p < 0.05 vs. NC+Rb1+H/R; and NS = not significant. Full article ">Figure 7
Gene knockdown of Nrf2 attenuates the ameliorative effect of ginsenoside Rb1 on mitochondrial injury and metabolic destabilization after H/R. (a–d) Representative fluorescent images of Rhod-2 and JC-1 in the presence of Nrf2 siRNA or NC siRNA with or without Rb1 treatment (10 μM) in NRCMs after H/R insult (n = 3). (e,f) Representative Western blots and quantitative analysis of mitochondrial Drp1, Fis1, Mfn2, Opa1, and VDAC1 protein expression (n = 3). (g,h) Representative Western blots and quantitative analysis of total p-Drp1 (Ser 616), p-Drp1 (Ser 637), and GAPDH protein expression (n = 3). H/R: hypoxia/reoxygenation; Rb1: ginsenoside Rb1. # p < 0.05 vs. Sham; * p < 0.05 vs. NC+Rb1+H/R. Full article ">Figure 8
Gn-Rb1 treatment improved neurological outcomes after CA/CPR. (a) The neurological function score was assessed in the surviving mice within 72 h at indicated time points after CA/CPR. (b,c) Representative immunofluorescence staining images of dihydroethidium (DHE) performed to assess brain tissue ROS accumulation (n = 3 per group). (d,e) Western blot analysis of antioxidant proteins SOD2 and oxidative markers gp91 in different groups (n = 6 per group). (f,g) Representative immunofluorescence staining images of TUNEL performed to assess the number of TUNEL-positive cells in the brain tissue (n = 3 per group). (h,i) Representative Western blots and quantitative analysis of Bax, Bcl-2, and cleaved caspase-3 in different groups (n = 6 per group). DHE: dihydroethidium; CA: cardiac arrest; CPR: cardiopulmonary resuscitation; and Rb1: ginsenoside Rb1. * p < 0.05; and NS = not significant. Full article ">

12 pages, 4787 KiB

Open AccessCommunication

Transcriptome Sequencing Reveals the Mechanism behind Chemically Induced Oral Mucositis in a 3D Cell Culture Model

by Maria Lambros, Jonathan Moreno, Qinqin Fei, Cyrus Parsa, Robert Orlando and Lindsey Van Haute

Int. J. Mol. Sci. 2023, 24(5), 5058; https://doi.org/10.3390/ijms24055058 - 6 Mar 2023

Cited by 1 | Viewed by 2057

Abstract

Oral mucositis is a common side effect of cancer treatment, and in particular of treatment with the mTORC1 inhibitor everolimus. Current treatment methods are not efficient enough and a better understanding of the causes and mechanisms behind oral mucositis is necessary to find [...] Read more.

Oral mucositis is a common side effect of cancer treatment, and in particular of treatment with the mTORC1 inhibitor everolimus. Current treatment methods are not efficient enough and a better understanding of the causes and mechanisms behind oral mucositis is necessary to find potential therapeutic targets. Here, we treated an organotypic 3D oral mucosal tissue model consisting of human keratinocytes grown on top of human fibroblasts with a high or low dose of everolimus for 40 or 60 h and investigated (1) the effect of everolimus on microscopic sections of the 3D cell culture for evidence of morphologic changes and (2) changes in the transcriptome by high throughput RNA-Seq analysis. We show that the most affected pathways are cornification, cytokine expression, glycolysis, and cell proliferation and we provide further details. This study provides a good resource towards a better understanding of the development of oral mucositis. It gives a detailed overview of the different molecular pathways that are involved in mucositis. This in turn provides information about potential therapeutic targets, which is an important step towards preventing or managing this common side effect of cancer treatment. Full article

(This article belongs to the Special Issue Design, Discovery and Development of Modulators Targeting Protein-Protein Interactions)

► Show Figures

Figure 1

15 pages, 4958 KiB

Open AccessArticle

The CCCH-Type Zinc-Finger Protein GhC3H20 Enhances Salt Stress Tolerance in Arabidopsis thaliana and Cotton through ABA Signal Transduction Pathway

by Qi Zhang, Jingjing Zhang, Fei Wei, Xiaokang Fu, Hengling Wei, Jianhua Lu, Liang Ma and Hantao Wang

Int. J. Mol. Sci. 2023, 24(5), 5057; https://doi.org/10.3390/ijms24055057 - 6 Mar 2023

Cited by 6 | Viewed by 2083

Abstract

The CCCH zinc-finger protein contains a typical C3H-type motif widely existing in plants, and it plays an important role in plant growth, development, and stress responses. In this study, a CCCH zinc-finger gene, GhC3H20, was isolated and thoroughly characterized to regulate salt [...] Read more.

The CCCH zinc-finger protein contains a typical C3H-type motif widely existing in plants, and it plays an important role in plant growth, development, and stress responses. In this study, a CCCH zinc-finger gene, GhC3H20, was isolated and thoroughly characterized to regulate salt stress in cotton and Arabidopsis. The expression of GhC3H20 was up-regulated under salt, drought, and ABA treatments. GUS activity was detected in the root, stem, leaves, and flowers of ProGhC3H20::GUS transgenic Arabidopsis. Compared with the control, the GUS activity of ProGhC3H20::GUS transgenic Arabidopsis seedlings under NaCl treatment was stronger. Through the genetic transformation of Arabidopsis, three transgenic lines of 35S-GhC3H20 were obtained. Under NaCl and mannitol treatments, the roots of the transgenic lines were significantly longer than those of the wild-type (WT) Arabidopsis. The leaves of the WT turned yellow and wilted under high-concentration salt treatment at the seedling stage, while the leaves of the transgenic Arabidopsis lines did not. Further investigation showed that compared with the WT, the content of catalase (CAT) in the leaves of the transgenic lines was significantly higher. Therefore, compared with the WT, overexpression of GhC3H20 enhanced the salt stress tolerance of transgenic Arabidopsis. A virus-induced gene silencing (VIGS) experiment showed that compared with the control, the leaves of pYL156-GhC3H20 plants were wilted and dehydrated. The content of chlorophyll in pYL156-GhC3H20 leaves was significantly lower than those of the control. Therefore, silencing of GhC3H20 reduced salt stress tolerance in cotton. Two interacting proteins (GhPP2CA and GhHAB1) of GhC3H20 have been identified through a yeast two-hybrid assay. The expression levels of PP2CA and HAB1 in transgenic Arabidopsis were higher than those in the WT, and pYL156-GhC3H20 had expression levels lower than those in the control. GhPP2CA and GhHAB1 are the key genes involved in the ABA signaling pathway. Taken together, our findings demonstrate that GhC3H20 may interact with GhPP2CA and GhHAB1 to participate in the ABA signaling pathway to enhance salt stress tolerance in cotton. Full article

(This article belongs to the Special Issue Genes Function and Mechanism Identification in Plant Stress Resistance)

► Show Figures

Figure 1

17 pages, 3816 KiB

Open AccessArticle

Ceria Nanoparticles Alleviated Osteoarthritis through Attenuating Senescence and Senescence-Associated Secretory Phenotype in Synoviocytes

by Xunshan Ren, Huangming Zhuang, Fuze Jiang, Yuelong Zhang and Panghu Zhou

Int. J. Mol. Sci. 2023, 24(5), 5056; https://doi.org/10.3390/ijms24055056 - 6 Mar 2023

Cited by 5 | Viewed by 2496

Abstract

Accumulation of senescent cells is the prominent risk factor for osteoarthritis (OA), accelerating the progression of OA through a senescence-associated secretory phenotype (SASP). Recent studies emphasized the existence of senescent synoviocytes in OA and the therapeutic effect of removing senescent synoviocytes. Ceria nanoparticles [...] Read more.

Accumulation of senescent cells is the prominent risk factor for osteoarthritis (OA), accelerating the progression of OA through a senescence-associated secretory phenotype (SASP). Recent studies emphasized the existence of senescent synoviocytes in OA and the therapeutic effect of removing senescent synoviocytes. Ceria nanoparticles (CeNP) have exhibited therapeutic effects in multiple age-related diseases due to their unique capability of ROS scavenging. However, the role of CeNP in OA remains unknown. Our results revealed that CeNP could inhibit the expression of senescence and SASP biomarkers in multiple passaged and hydrogen-peroxide-treated synoviocytes by removing ROS. In vivo, the concentration of ROS in the synovial tissue was remarkably suppressed after the intra-articular injection of CeNP. Likewise, CeNP reduced the expression of senescence and SASP biomarkers as determined by immunohistochemistry analysis. The mechanistic study showed that CeNP inactivated the NFκB pathway in senescent synoviocytes. Finally, safranin O–fast green staining showed milder destruction of articular cartilage in the CeNP-treated group compared with the OA group. Overall, our study suggested that CeNP attenuated senescence and protected cartilage from degeneration via scavenging ROS and inactivating the NFκB signaling pathway. This study has potentially significant implications in the field of OA as it provides a novel strategy for OA treatment. Full article

(This article belongs to the Special Issue Novel Strategies in the Development of New Anti-inflammatory Drug Substances and Therapies)

► Show Figures

Figure 1

Figure 1
Characterization of CeNP. (a) Representative TEM images of CeNP and magnified views of the black box. (b) The hydrodynamic size of CeNP. (c) Representative EDS spectrum of CeNP. (d) The zeta potential of CeNP. UV–visible spectra (e) and appearance (f) of CeNP in aqueous solution at room temperature for one week. Full article ">Figure 2
ROS scavenging capacity of CeNP. Inhibition rate of superoxide anion (a), hydrogen peroxide (b), DPPH (c), and OH·(d) in different CeNP concentrations (n = 3). Data represent the mean ± SEM from three independent experiments, NS, not significant; ** p < 0.01 vs. 0 μg/mL. Full article ">Figure 3
Cytotoxicity and cellular uptake of CeNP in synoviocytes. (a) Appearance of synoviocytes under the light microscope. (b) Immunofluorescence staining of vimentin in synoviocytes. (c) CeNP uptake capacity of synoviocytes and mean intake number of CeNP (n = 3). (d) Synoviocyte viability detected by the CCK-8 assay after treatment with different concentrations of CeNP for 24 h (n = 3). Data represent the mean ± SEM from three independent experiments, NS, not significant; ** p < 0.01 vs. NC. Full article ">Figure 4
CeNP attenuated H2O2-elicited senescence and inhibited SASP in synoviocytes. (a) Representative fluorescence imaging of intracellular ROS and mean fluorescence intensity in NC, H2O2 and H2O2 + CeNP group (n = 3). (b) SA-β-Gal staining and quantification of SA-β-Gal positive rate in NC, H2O2 and H2O2 + CeNP groups (n = 3). (c–e) RT-qPCR analysis of P16, P21, iNOS, COX2, MMP3, ADAMTS5, IL-6 and TNFα in NC, H2O2 and H2O2 + CeNP groups (n = 3). (f–h) Western blot results of iNOS, COX2, ADAMTS5 and MMP3 in NC, H2O2 and H2O2 + CeNP groups (n = 3). (i,j) Elisa assays of IL-6 and TNFα in NC, H2O2 and H2O2 + CeNP groups (n = 3). Data represent the mean ± SEM from three independent experiments, ** p < 0.01 vs. NC; # p < 0.05 vs. H2O2 group; ## p < 0.01 vs. H2O2 group. Full article ">Figure 5
CeNP attenuated senescence and inhibited SASP in multiple passaged synoviocytes. (a) Representative fluorescence imaging of intracellular ROS and mean fluorescence intensity in NC, MP, and MP with CeNP treatment synoviocytes (n = 3). (b) SA-β-Gal staining and quantification of SA-β-Gal positive rate in NC, MP, and MP with CeNP treatment synoviocytes (n = 3). (c–e) RT-qPCR analysis of P16, P21, iNOS, COX2, MMP3, ADAMTS5, IL-6 and TNFα in NC, MP, and MP with CeNP treatment synoviocytes (n = 3). (f–h) Western blot results of iNOS, COX2, ADAMTS5 and MMP3 in NC, MP, and MP with CeNP treatment synoviocytes (n = 3). (i,j) Elisa assays of IL-6 and TNFα in NC, MP, and MP with CeNP treatment synoviocytes (n = 3). Data represent the mean ± SEM from three independent experiments, ** p < 0.01 vs. NC; ## p < 0.01 vs. MP. Full article ">Figure 6
CeNP inhibited the activation of the NFκB pathway in senescent synoviocytes. (a) The protein levels of p-p65, p65, p-IκBα, and IκBα were detected by western blot. (b–d) Quantitative analysis of the results. * p < 0.05 vs. NC; ** p < 0.01 vs. NC; # p < 0.05 vs. MP; ## p < 0.01 vs. MP. Full article ">Figure 7
CeNP scavenged ROS and attenuated senescence of synoviocytes in vivo. (a) DHE staining and mean fluorescence intensity of ROS in the synovium (n = 5). (b–e) Immunohistochemical staining and quantitative results of P16, P21, iNOS and COX2 in the synovium (n = 5). ** p < 0.01 vs. sham; # p < 0.05 vs. ACLT; ## p < 0.01 vs. ACLT. Full article ">Figure 8
CeNP suppressed SASP, inactivated the NFκB signaling pathway and protected cartilage in vivo. (a–d) Immunohistochemical staining and quantitative results of ADAMTS5, MMP3, IL-6 and TNFα (n = 5). (e,f) Immunohistochemical staining and quantitative results of p65 and p-p65 (n = 5). (g) Representative pictures of HE, safranin O–fast green staining of the cartilage and OARSI score in each group (n = 5). * p < 0.05 vs. sham; ** p < 0.01 vs. sham; # p < 0.05 vs. ACLT; ## p < 0.01 vs. ACLT. Full article ">

19 pages, 1299 KiB

Open AccessArticle

Cannabidiol as a Promising Therapeutic Option in IC/BPS: In Vitro Evaluation of Its Protective Effects against Inflammation and Oxidative Stress

by Tadeja Kuret, Mateja Erdani Kreft, Rok Romih and Peter Veranič

Int. J. Mol. Sci. 2023, 24(5), 5055; https://doi.org/10.3390/ijms24055055 - 6 Mar 2023

Cited by 7 | Viewed by 2657

Abstract

Several animal studies have described the potential effect of cannabidiol (CBD) in alleviating the symptoms of interstitial cystitis/bladder pain syndrome (IC/BPS), a chronic inflammatory disease of the urinary bladder. However, the effects of CBD, its mechanism of action, and modulation of downstream signaling [...] Read more.

Several animal studies have described the potential effect of cannabidiol (CBD) in alleviating the symptoms of interstitial cystitis/bladder pain syndrome (IC/BPS), a chronic inflammatory disease of the urinary bladder. However, the effects of CBD, its mechanism of action, and modulation of downstream signaling pathways in urothelial cells, the main effector cells in IC/BPS, have not been fully elucidated yet. Here, we investigated the effect of CBD against inflammation and oxidative stress in an in vitro model of IC/BPS comprised of TNFα-stimulated human urothelial cells SV-HUC1. Our results show that CBD treatment of urothelial cells significantly decreased TNFα-upregulated mRNA and protein expression of IL1α, IL8, CXCL1, and CXCL10, as well as attenuated NFκB phosphorylation. In addition, CBD treatment also diminished TNFα-driven cellular reactive oxygen species generation (ROS), by increasing the expression of the redox-sensitive transcription factor Nrf2, the antioxidant enzymes superoxide dismutase 1 and 2, and hem oxygenase 1. CBD-mediated effects in urothelial cells may occur by the activation of the PPARγ receptor since inhibition of PPARγ resulted in significantly diminished anti-inflammatory and antioxidant effects of CBD. Our observations provide new insights into the therapeutic potential of CBD through modulation of PPARγ/Nrf2/NFκB signaling pathways, which could be further exploited in the treatment of IC/BPS. Full article

(This article belongs to the Special Issue Inflammation in Health and Disease: New Insights and Therapeutic Avenues 3.0)

► Show Figures

Figure 1

12 pages, 2982 KiB

Open AccessArticle

Properties of a Single Amino Acid Residue in the Third Transmembrane Domain Determine the Kinetics of Ambient Light-Sensitive Channelrhodopsin

by Akito Hatakeyama, Eriko Sugano, Tatsuki Sayama, Yoshito Watanabe, Tomoya Suzuki, Kitako Tabata, Yuka Endo, Tetsuya Sakajiri, Tomokazu Fukuda, Taku Ozaki and Hiroshi Tomita

Int. J. Mol. Sci. 2023, 24(5), 5054; https://doi.org/10.3390/ijms24055054 - 6 Mar 2023

Cited by 1 | Viewed by 1625

Abstract

Channelrhodopsins have been utilized in gene therapy to restore vision in patients with retinitis pigmentosa and their channel kinetics are an important factor to consider in such applications. We investigated the channel kinetics of ComV1 variants with different amino acid residues at the [...] Read more.

Channelrhodopsins have been utilized in gene therapy to restore vision in patients with retinitis pigmentosa and their channel kinetics are an important factor to consider in such applications. We investigated the channel kinetics of ComV1 variants with different amino acid residues at the 172nd position. Patch clamp methods were used to record the photocurrents induced by stimuli from diodes in HEK293 cells transfected with plasmid vectors. The channel kinetics (τon and τoff) were considerably altered by the replacement of the 172nd amino acid and was dependent on the amino acid characteristics. The size of amino acids at this position correlated with τon and decay, whereas the solubility correlated with τon and τoff. Molecular dynamic simulation indicated that the ion tunnel constructed by H172, E121, and R306 widened due to H172A variant, whereas the interaction between A172 and the surrounding amino acids weakened compared with H172. The bottleneck radius of the ion gate constructed with the 172nd amino acid affected the photocurrent and channel kinetics. The 172nd amino acid in ComV1 is a key residue for determining channel kinetics as its properties alter the radius of the ion gate. Our findings can be used to improve the channel kinetics of channelrhodopsins. Full article

(This article belongs to the Special Issue Retinal Diseases and Cell Signaling)

► Show Figures

Figure 1

Figure 1
Comparison of functions between ComV1 and H172R. (a) The structure of ComV1 predicted using a bioinformatic tool. The protein is indicated by the green ribbon. H172 is located near the ion tunnel. (b,c) The fluorescence of 4′,6-diamidino-2-phenylindole (DAPI) (blue) and ComV1-Venus or H172R-Venus (green) introduced into HEK293 cells. The scale bar indicates 5 µm. Both genes are mainly expressed on the plasma membrane. (d,e) The typical waveforms of ComV1 (d) and H172R (e) were evoked with a 505-nm light-emitting diode adjusted to 1 µW/mm2 for 1 s. (f–i) Comparison of photocurrents (f), decays at 505 nm (g), τon (h), and τoff (i) between ComV1 and H172R. Data are expressed as mean ± standard error of mean (SEM) (ComV1 n = 18–19, H172R n = 8). The data were compared using two-way analysis of variance (ANOVA) with Bonferroni post-test (** p < 0.01, *** p < 0.001). Full article ">Figure 2
Electrophysiological profiles of ComV1 and the H172G, H172A, H172K, and H172Y mutants. (a) The typical waveforms evoked with 505-nm light adjusted to 1 µW/mm2 for 1 s. (b–e) Comparison of τon (b), τoff (c), photocurrents (d), and decays (e) between ComV1 and its mutants. τon, τoff, and decays at 505 nm (<a href="#app1-ijms-24-05054" class="html-app">Supplementary Material File Figure S1</a>). Photocurrents were evoked with a light-emitting diode adjusted to 1 µW/mm2 for 1 s. The data are shown as mean ± SEM (ComV1 n = 18–19, H172G n = 8, H172A n = 8, H172K n = 8, H172Y n = 8), and were analyzed using Dunnett’s multiple comparison test vs. ComV1 (* p < 0.05, ** p < 0.01). All data measured using light of various wavelengths and the results of statistical analysis are shown in <a href="#app1-ijms-24-05054" class="html-app">Supplementary File Table S1 and Supplementary Files Table S2–S4</a>, respectively. (f) The fluorescence of channelrhodopsin-Venus (green) and DAPI (blue) in each mutant. Full article ">Figure 3
Correlation between properties of the 172nd amino acid and channel function of ComV1 and mutants. (a–d) Correlation between the size of the 172nd amino acid residue and channel properties, peak currents (a), τon (b), τoff (c), and decay (d) at 505 nm. (e–h) Correlation between the solubility of the 172nd amino acid residue and channel properties, peak current (e), τon (f), τoff (g), and decay (h). All data were analyzed with Pearson correlation coefficient (* p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant). Full article ">Figure 4
Structures of ComV1 and its mutants analyzed with molecular dynamic simulation. (a–e), The simulated structure around the 172nd residue in ComV1 (a), H172G (b), H172A (c), H172K (d), and H172Y (e) at an early excited state. The protein and the estimated ion gating pathway are shown as green and yellow ribbons, respectively. The figure in the frame is the structure near the 172nd residue observed vertically from the ion tunnel. The first structural models were built with homology modeling using the structure of PDBID: 7E71 as the template. Full article ">Figure 5
Correlation analysis between the ion tunnel radius and the channel properties. (a–d) Correlation between the radius of the bottleneck of the ion gating pathway and peak currents (a), τon (b), τoff (c), and decays (d) at 505 nm. All data were analyzed with Pearson correlation coefficient (* p < 0.05, *** p < 0.001). Full article ">

17 pages, 1409 KiB

Open AccessReview

Female Reproductive Aging and Oxidative Stress: Mesenchymal Stem Cell Conditioned Medium as a Promising Antioxidant

by Kihae Ra, Se Chang Park and Byeong Chun Lee

Int. J. Mol. Sci. 2023, 24(5), 5053; https://doi.org/10.3390/ijms24055053 - 6 Mar 2023

Cited by 13 | Viewed by 3330

Abstract

The recent tendency to delay pregnancy has increased the incidence of age-related infertility, as female reproductive competence decreases with aging. Along with aging, a lowered capacity of antioxidant defense causes a loss of normal function in the ovaries and uterus due to oxidative [...] Read more.

The recent tendency to delay pregnancy has increased the incidence of age-related infertility, as female reproductive competence decreases with aging. Along with aging, a lowered capacity of antioxidant defense causes a loss of normal function in the ovaries and uterus due to oxidative damage. Therefore, advancements have been made in assisted reproduction to resolve infertility caused by reproductive aging and oxidative stress, following an emphasis on their use. The application of mesenchymal stem cells (MSCs) with intensive antioxidative properties has been extensively validated as a regenerative therapy, and proceeding from original cell therapy, the therapeutic effects of stem cell conditioned medium (CM) containing paracrine factors secreted during cell culture have been reported to be as effective as that of direct treatment of source cells. In this review, we summarized the current understanding of female reproductive aging and oxidative stress and present MSC-CM, which could be developed as a promising antioxidant intervention for assisted reproductive technology. Full article

(This article belongs to the Special Issue Advanced Research on Reproductive System Disease 2022)

► Show Figures

Figure 1

16 pages, 41816 KiB

Open AccessArticle

Finding miRNA–RNA Network Biomarkers for Predicting Metastasis and Prognosis in Cancer

by Seokwoo Lee, Myounghoon Cho, Byungkyu Park and Kyungsook Han

Int. J. Mol. Sci. 2023, 24(5), 5052; https://doi.org/10.3390/ijms24055052 - 6 Mar 2023

Viewed by 1785

Abstract

Despite remarkable progress in cancer research and treatment over the past decades, cancer ranks as a leading cause of death worldwide. In particular, metastasis is the major cause of cancer deaths. After an extensive analysis of miRNAs and RNAs in tumor tissue samples, [...] Read more.

Despite remarkable progress in cancer research and treatment over the past decades, cancer ranks as a leading cause of death worldwide. In particular, metastasis is the major cause of cancer deaths. After an extensive analysis of miRNAs and RNAs in tumor tissue samples, we derived miRNA–RNA pairs with substantially different correlations from those in normal tissue samples. Using the differential miRNA–RNA correlations, we constructed models for predicting metastasis. A comparison of our model to other models with the same data sets of solid cancer showed that our model is much better than the others in both lymph node metastasis and distant metastasis. The miRNA–RNA correlations were also used in finding prognostic network biomarkers in cancer patients. The results of our study showed that miRNA–RNA correlations and networks consisting of miRNA–RNA pairs were more powerful in predicting prognosis as well as metastasis. Our method and the biomarkers obtained using the method will be useful for predicting metastasis and prognosis, which in turn will help select treatment options for cancer patients and targets of anti-cancer drug discovery. Full article

(This article belongs to the Special Issue Network Medicine in Human Diseases)

► Show Figures

Figure 1

Figure 1
ROC curves of three types of features in predicting metastasis of COAD in independent testing. (A) Average ROC curves in predicting lymph node metastasis of COAD. (B) Average ROC curves in predicting distant metastasis of COAD. <math display="inline"><semantics> <mo>Δ</mo> </semantics></math>PCC: <math display="inline"><semantics> <mo>Δ</mo> </semantics></math>PCC of miRNA–RNA pairs, Exp: Expressions of genes involved in miRNA–RNA pairs, Exp191: Expressions of 191 metastasis-predictive genes [<a href="#B19-ijms-24-05052" class="html-bibr">19</a>]. The curves are the average ROC curves of 10 runs. The gray part indicates the error range of the ROC curves. Full article ">Figure 2
Kaplan–Meier plots comparing the survival rates of two groups of cancer patients with respect to a miRNA–RNA pair. (A) The survival rates of two groups of LUAD patients with respect to MIR3125_OR1F1. The larger <math display="inline"><semantics> <mrow> <mo>|</mo> <mo>Δ</mo> <mi>PCC</mi> <mo>|</mo> </mrow> </semantics></math> values of the MIR3125_OR1F1 pair are associated with the shorter survival rates of LUAD patients. (B) The survival rates of two groups of PRAD patients with respect to MIR5087_EZR-AS1. The larger <math display="inline"><semantics> <mrow> <mo>|</mo> <mo>Δ</mo> <mi>PCC</mi> <mo>|</mo> </mrow> </semantics></math> values of the MIR5087_EZR-AS1 pair are associated with the longer survival rates of PRAD patients. The risk tables below the Kaplan–Meier plots show the numbers at risk of each group over five years. Full article ">Figure 3
(A) Network_MIR645 for prognosis of LUAD, which consists of 3 mRNAs (white ellipse), 6 lncRNAs (grey ellipse), and 2 pseudogenes (pink ellipse) connected to 1 miRNA (shown as green circle). (B) Network_MIR4666A for prognosis of PRAD. It consists of 2 mRNAs, 2 lncRNAs, and 3 pseudogenes connected with miRNA. The Kaplan–Meier plots compare the survival rates of two groups of risk scores, which were defined using Equation (<a href="#FD4-ijms-24-05052" class="html-disp-formula">4</a>). Full article ">Figure 4
Network of miRNA–RNA correlations in PRAD, which consists of 5036 edges between 4121 nodes (125 miRNAs, 2330 mRNAs, 1169 lncRNAs, and 479 pseudogenes). Subnetworks enclosed with rounded boxes are potential prognostic network biomarkers found in our study. Full article ">Figure 5
(A) Distribution of the p-values derived from the log–rank test with respect to subnetworks, miRNA–RNA pairs, and individual genes. (B) Distribution of the C-index values with respect to subnetworks, miRNA–RNA pairs, and individual genes. Full article ">Figure 6
The overall framework of our method. (A) Deriving miRNA–RNA pairs based on <math display="inline"><semantics> <mo>Δ</mo> </semantics></math>PCC from RNA–seq and clinical data. (B) Constructing prediction two models for predicting metastasis (LNM and DM) based on differential correlations between miRNAs and their target RNAs and predicting metastasis using the models. (C) Identifying prognostic network biomarkers from miRNA–RNA pairs. Full article ">

16 pages, 2253 KiB

Open AccessArticle

Gene Expression Analysis of Immune Regulatory Genes in Circulating Tumour Cells and Peripheral Blood Mononuclear Cells in Patients with Colorectal Carcinoma

by Sharmin Aktar, Faysal Bin Hamid, Sujani Madhurika Kodagoda Gamage, Tracie Cheng, Nahal Pakneshan, Cu Tai Lu, Farhadul Islam, Vinod Gopalan and Alfred King-yin Lam

Int. J. Mol. Sci. 2023, 24(5), 5051; https://doi.org/10.3390/ijms24055051 - 6 Mar 2023

Cited by 3 | Viewed by 2522 | Correction

Abstract

Information regarding genetic alterations of driver cancer genes in circulating tumour cells (CTCs) and their surrounding immune microenvironment nowadays can be employed as a real-time monitoring platform for translational applications such as patient response to therapeutic targets, including immunotherapy. This study aimed to [...] Read more.

Information regarding genetic alterations of driver cancer genes in circulating tumour cells (CTCs) and their surrounding immune microenvironment nowadays can be employed as a real-time monitoring platform for translational applications such as patient response to therapeutic targets, including immunotherapy. This study aimed to investigate the expression profiling of these genes along with immunotherapeutic target molecules in CTCs and peripheral blood mononuclear cells (PBMCs) in patients with colorectal carcinoma (CRC). Expression of p53, APC, KRAS, c-Myc, and immunotherapeutic target molecules PD-L1, CTLA-4, and CD47 in CTCs and PBMCs were analysed by qPCR. Their expression in high versus low CTC-positive patients with CRC was compared and clinicopathological correlations between these patient groups were analysed. CTCs were detected in 61% (38 of 62) of patients with CRC. The presence of higher numbers of CTCs was significantly correlated with advanced cancer stages (p = 0.045) and the subtypes of adenocarcinoma (conventional vs. mucinous, p = 0.019), while being weakly correlated with tumour size (p = 0.051). Patients with lower numbers of CTCs had higher expression of KRAS. Higher KRAS expression in CTCs was negatively correlated with tumour perforation (p = 0.029), lymph node status (p = 0.037), distant metastasis (p = 0.046) and overall staging (p = 0.004). CTLA-4 was highly expressed in both CTCs and PBMCs. In addition, CTLA-4 expression was positively correlated with KRAS (r = 0.6878, p = 0.002) in the enriched CTC fraction. Dysregulation of KRAS in CTCs might evade the immune system by altering the expression of CTLA-4, providing new insights into the selection of therapeutic targets at the onset of the disease. Monitoring CTCs counts, as well as gene expression profiling of PBMCs, can be helpful in predicting tumour progression, patient outcome and treatment. Full article

(This article belongs to the Special Issue Heterogeneity and Precision Oncology in Circulating Tumor Cells)

► Show Figures

Figure 1

Figure 1
Enumeration of circulating tumour cells (CTCs) and numbers of different subpopulation of CTCs in patients with colorectal carcinoma (CRC). The figure depicts: (A) representative images of CTCs detected from patients with CRC captured using an Olympus Fluoview FV1000 Confocal Microscope (scale bar: 20 µm); (B) a comparison of the number of different subpopulations of CTCs detected in patients with CRC; and (C) the number of populations recruited in different groups based on the range of CTC counts, along with healthy donors. (HD, healthy donor; N, No CTC; L, low CTC-positive group; H, high CTC-positive group). Full article ">Figure 2
Relative fold change values (2–ΔΔCt) of p53, APC, KRAS, c-Myc and CD47, CTLA-4 in CTC-enriched fraction from patients with CRC and from healthy donors (HDs) (n = 6). All the values are plotted as a scatter plot with the median. Line indicates the normal fold change value. ** p < 0.005, * p < 0.05. Full article ">Figure 3
Comparison of the gene expression of oncogenes (KRAS, c-Myc), tumour suppressor genes (p53, APC) and immune checkpoint molecules (CTLA-4, CD47) between high versus low CTC-positive groups in CTCs and PBMCs from patients with colorectal carcinoma (CRC). Data are depicted as scatter plots interleaved with bar plots, indicating min. to max. value. All the values are plotted as mean ± SEM. The dashed line indicates the normal fold change value. The PCR data were shown on a log2 scale and analysed by unpaired two-way ANOVA (Bonferroni’s multiple comparison test). Comparisons were considered significant at p ≤ 0.05. Full article ">Figure 4
Correlation between KRAS gene and CTLA-4 expression in CTCs. (A) The mRNA expression level of CTLA-4 correlated positively with KRAS (r = 0.6878, p = 0.0002). r; coefficient correlation value (Spearman’s rank test). (B) The mRNA expression levels of CTLA-4 in CTCs and PBMCs from patients with CRC according to the KRAS mutation status of the primary tumour. Full article ">Figure 5
Relationship of the gene expressions (P53, APC, KRAS, c-Myc, CD47 and CTLA-4) between high and low CTC groups in CTCs and PBMCs, in patients with CRC. Full article ">Figure 6
Heat map depicting the mRNA expression level and percentage of positive expression of the tumour suppressor genes p53 and APC; the oncogenes, KRAS and c-Myc; and the immune-regulatory molecules CD47 and CTLA-4 in CTCs and PBMCs among individual patients. The values were calculated from the log2 value of the relative quantification of each gene. The colour indicates the expression level for each gene. Red fields represent downregulated genes; blue fields represent upregulated genes; crossed-out fields represent no expression. Full article ">

17 pages, 3031 KiB

Open AccessArticle

Identification and Functional Analysis of Two Mitoferrins, CsMIT1 and CsMIT2, Participating in Iron Homeostasis in Cucumber

by Karolina Małas and Katarzyna Kabała

Int. J. Mol. Sci. 2023, 24(5), 5050; https://doi.org/10.3390/ijms24055050 - 6 Mar 2023

Viewed by 1667

Abstract

Mitochondria are one of the major iron sinks in plant cells. Mitochondrial iron accumulation involves the action of ferric reductase oxidases (FRO) and carriers located in the inner mitochondrial membrane. It has been suggested that among these transporters, mitoferrins (mitochondrial iron transporters, MITs) [...] Read more.

Mitochondria are one of the major iron sinks in plant cells. Mitochondrial iron accumulation involves the action of ferric reductase oxidases (FRO) and carriers located in the inner mitochondrial membrane. It has been suggested that among these transporters, mitoferrins (mitochondrial iron transporters, MITs) belonging to the mitochondrial carrier family (MCF) function as mitochondrial iron importers. In this study, two cucumber proteins, CsMIT1 and CsMIT2, with high homology to Arabidopsis, rice and yeast MITs were identified and characterized. CsMIT1 and CsMIT2 were expressed in all organs of the two-week-old seedlings. Under Fe-limited conditions as well as Fe excess, the mRNA levels of CsMIT1 and CsMIT2 were altered, suggesting their regulation by iron availability. Analyses using Arabidopsis protoplasts confirmed the mitochondrial localization of cucumber mitoferrins. Expression of CsMIT1 and CsMIT2 restored the growth of the Δmrs3Δmrs4 mutant (defective in mitochondrial Fe transport), but not in mutants sensitive to other heavy metals. Moreover, the altered cytosolic and mitochondrial Fe concentrations, observed in the Δmrs3Δmrs4 strain, were recovered almost to the levels of WT yeast by expressing CsMIT1 or CsMIT2. These results indicate that cucumber proteins are involved in the iron transport from the cytoplasm to the mitochondria. Full article

(This article belongs to the Special Issue Iron and Sulfur in Plants 3.0)

► Show Figures

Figure 1

Figure 1
In silico analysis of MIT proteins from yeast, human, fish, and monocotyledonous and dicotyledonous plants. Sequence alignment of rice (OsMIT1), Arabidopsis (AtMIT1 and AtMIT2), cucumber (CsMIT1 and CsMIT2), human (HsMfrn1), yeast (ScMRS3, ScMRS4) and O. niloticus (TMfrn1) generated using Clustal O. Grey highlights represent the characteristic sequence motif of MCs (P-x-[DE]-x-[LIVAT]-[KR]-x-[LRK]-[LIVMFY]-(20-30 residues)-[DE]-G-x-x-x-x-[WYF]-[KR]-G). Transmembrane helices predicted for these proteins are depicted by H1-H6. Putative substrate contact sites are highlighted in pink, and the residues involved in iron transport are marked in red boxes. Green boxes represent the conserved motifs forming salt bridges on the cytosolic (dark green) and matrix (light green) side. Full article ">Figure 2
CsMIT1 and CsMIT2 gene expression in different organs of cucumber plants grown under various iron treatments. (a) The expression of CsMIT1 and CsMIT2 in the vegetative organs of two-week-old cucumber seedlings grown in control medium. (b) The expression of CsMIT1 in the roots and leaves of two-week-old cucumber seedlings grown in control medium or subjected to Fe deficiency (-Fe) and Fe excess (+Fe) for 24 h or two weeks. (c) The expression of CsMIT2 in the roots and leaves of two-week-old cucumber seedlings grown in control medium or subjected to Fe deficiency (-Fe) and Fe excess (+Fe) for 24 h or two weeks. For real-time analysis, expression of individual genes was calculated relative to the reference gene CsCACS according to the ΔΔCT method. Results shown are the means of three biological replicates; error bars represent standard error (+/-SE). Different letters represent statistically significant differences (p < 0.05; ANOVA with Tukey’s correction). Full article ">Figure 3
Subcellular localization of CsMIT1 and CsMIT2 in A. thaliana protoplasts. DIC—transmission images of the protoplasts expressing CsMIT1-GFP or CsMIT2-GFP, GFP—GFP fluorescence of the protoplast, MitoTracker—fluorescence of mitochondria specific marker. The scale corresponds to 10 μm. Full article ">Figure 4
Effect of CsMIT1 and CsMIT2 expression in yeast. (a) Representative of serial dilutions corresponding to OD of 0.2; 0.02 and 0.002 of WT and Δmrs3Δmrs4 cells transformed with empty vector or vector carrying CsMIT1 or/and CsMIT2 placed onto control SC/Glu-His medium or SC/Glu-His low-Fe medium containing 0.1 mM BPS. (b) Localization of CsMIT1 and CsMIT2 in Δmrs3Δmrs4 cells. DIC—transmission images of the cells expressing CsMIT1-GFP or CsMIT2-GFP, GFP—GFP fluorescence of the cell, MitoTracker—fluorescence of mitochondria specific marker. The scale corresponds to 5 μm. (c) Representative of serial dilutions corresponding to OD of 0.2; 0.02 and 0.002 of WT and yeast mutants sensitive to various heavy metals. Cells were transformed with empty vector or vector carrying CsMIT1 or CsMIT2 placed onto control SC/Glu-His medium or SC/Glu-His medium supplemented with suitable metal. Full article ">Figure 5
Cytosolic and mitochondrial Fe content in yeast expressing CsMIT1 and CsMIT2. (a) Western blot analysis of CsMIT1-GFP and CsMIT2-GFP (left) or c-GDO-FLAG (right) proteins in yeast Δmrs3Δmrs4 transformants using the antibodies against GFP and FLAG, respectively. (b) c-GDO activity in WT and Δmrs3Δmrs4 cells expressing c-GDO-FLAG and either empty vector, CsMIT1-GFP or CsMIT2-GFP. c-GDO activity is expressed as nanomoles of substrate converted per minute per mg of protein. Results shown are the means of four biological replicates; error bars represent standard error (+/-SE). Different letters represent statistically significant differences (p < 0.05; ANOVA with Tukey’s correction). (c) Mitochondrial Fe content in WT and Δmrs3Δmrs4 cells expressing either empty vector, CsMIT1-GFP or CsMIT2-GFP. Results as shown are expressed as the mean Fe level of three biological replicates (nmol mg-1 protein); error bars represent standard error (+/-SE). Different letters represent statistically significant differences (p < 0.05; ANOVA with Tukey’s correction). Full article ">Figure 6
Possible role of mitoferrins in cucumber cells and their gene expression under different iron availability. Full article ">

15 pages, 2771 KiB

Open AccessArticle

Effect of Chitosan-Diosgenin Combination on Wound Healing

by Lubomir Petrov, Olya Stoilova, Georgi Pramatarov, Hristiyana Kanzova, Elina Tsvetanova, Madlena Andreeva, Almira Georgieva, Dimitrinka Atanasova, Stanislav Philipov and Albena Alexandrova

Int. J. Mol. Sci. 2023, 24(5), 5049; https://doi.org/10.3390/ijms24055049 - 6 Mar 2023

Cited by 3 | Viewed by 1977

Abstract

The difficult-to-heal wounds continue to be a problem for modern medicine. Chitosan and diosgenin possess anti-inflammatory and antioxidant effects making them relevant substances for wound treatment. That is why this work aimed to study the effect of the combined application of chitosan and [...] Read more.

The difficult-to-heal wounds continue to be a problem for modern medicine. Chitosan and diosgenin possess anti-inflammatory and antioxidant effects making them relevant substances for wound treatment. That is why this work aimed to study the effect of the combined application of chitosan and diosgenin on a mouse skin wound model. For the purpose, wounds (6 mm diameter) were made on mice’s backs and were treated for 9 days with one of the following: 50% ethanol (control), polyethylene glycol (PEG) in 50% ethanol, chitosan and PEG in 50% ethanol (Chs), diosgenin and PEG in 50% ethanol (Dg) and chitosan, diosgenin and PEG in 50% ethanol (ChsDg). Before the first treatment and on the 3rd, 6th and 9th days, the wounds were photographed and their area was determined. On the 9th day, animals were euthanized and wounds’ tissues were excised for histological analysis. In addition, the lipid peroxidation (LPO), protein oxidation (POx) and total glutathione (tGSH) levels were measured. The results showed that ChsDg had the most pronounced overall effect on wound area reduction, followed by Chs and PEG. Moreover, the application of ChsDg maintained high levels of tGSH in wound tissues, compared to other substances. It was shown that all tested substances, except ethanol, reduced POx comparable to intact skin levels. Therefore, the combined application of chitosan and diosgenin is a very promising and effective medication for wound healing. Full article

(This article belongs to the Special Issue The Emerging Role of Polymeric Materials in Pharmaceutical Designs and Applications)

► Show Figures

Figure 1

16 pages, 4051 KiB

Open AccessArticle

Discovering the Biological Significance and Therapeutic Potential of miR-29b-3p in Triple-Negative Breast Cancer

by Ancuta Jurj, Oana Zanoaga, Lajos Raduly, Vlad Morhan, Zsofia Papi, Cristina Ciocan, Laura-Ancuta Pop, Ioana Berindan-Neagoe and Cornelia Braicu

Int. J. Mol. Sci. 2023, 24(5), 5048; https://doi.org/10.3390/ijms24055048 - 6 Mar 2023

Cited by 2 | Viewed by 2235

Abstract

The lack of estrogen or progesterone receptors and absence of HER2 amplification/overexpression in triple-negative breast cancer (TNBC) restricts therapeutic options used in clinical management. MicroRNAs (miRNAs) are small, non-coding transcripts which affect important cellular mechanisms by regulating gene expression at the post-transcriptional level. [...] Read more.

The lack of estrogen or progesterone receptors and absence of HER2 amplification/overexpression in triple-negative breast cancer (TNBC) restricts therapeutic options used in clinical management. MicroRNAs (miRNAs) are small, non-coding transcripts which affect important cellular mechanisms by regulating gene expression at the post-transcriptional level. Among this class, attention was focused on miR-29b-3p with a high profile in TNBC and correlated with the overall survival rates, as TCGA data revealed. This study aims to investigate the implication of the miR-29b-3p inhibitor in TNBC cell lines by identifying a potential therapeutic transcript, improving the clinical outcomes of this disease. The experiments were performed on two TNBC cell lines (MDA-MB-231 and BT549) as in vitro models. An established dose of 50 nM was used for all functional assays performed on the miR-29b-3p inhibitor. A decreased level of miR-29b-3p determined a significant reduction in cell proliferation and colony-forming capacity. At the same time, the changes occurring at the molecular and cellular levels were highlighted. We observed that, when inhibiting the expression level of miR-29b-3p, processes such as apoptosis and autophagy were activated. Further, microarray data revealed that the miRNA expression pattern was altered after miR-29b-3p inhibition, pointing out 8 overexpressed and 11 downregulated miRNAs specific for BT549 cells and 33 upregulated and 10 downregulated miRNAs that were specific for MDA-MB-231 cells. As a common signature for both cell lines, three transcripts were observed, two downregulated, miR-29b-3p and miR-29a, and one upregulated, miR-1229-5p. According to DIANA miRPath, the main predicted targets are related to ECM (extracellular matrix) receptor interaction and TP53 signaling. An additional validation step through qRT-PCR was performed, which showed an upregulation of MCL1 and TGFB1. By inhibiting the expression level of miR-29b-3p, it was shown that complex regulatory pathways targeted this transcript in TNBC cells. Full article

(This article belongs to the Special Issue Breast Cancer Mechanistic Insights and Targeted Therapies)

► Show Figures

Figure 1

Figure 1
The expression level of miR-29b-3p in TNBC (TCGA dataset) and breast cancer cell lines. (A) The expression level of miR-29b-3p in TNBC versus adjacent normal tissue according to TCGA patient’s cohort. (B) A detailed view of miR-29b-3p expression level in TNBC patients according to TCGA patient’s cohort (TT: tumor tissue, TN: normal adjacent tissue). (C) The overall survival rate for miR-29b-3p in TNBC patients, according to data available from the online application KM Plotter developed based on TCGA data. Patients with expression above the median are indicated in the red line, and patients with expression below the median in the black line compared by the log-rank test, HR: hazard ratio (D) The evaluation of miR-29b-3p expression levels in triple-negative breast cancer cell lines (Hs578T, BT549 and MDA-MB-231), compared to the expression level for this transcript in normal epithelial breast cell line, FR2, based on ΔΔCt method and U6 for normalization (* p < 0.05, **** p < 0.0001). Full article ">Figure 2
The inhibition of miR-29b-3p reduces cell viability and the capacity for colony formation. (A) Using MTT assay, it was observed that the downregulation of miR-29b-3p reduces cell viability rate in both TNBC cell lines (B). The inhibition of miR-29b-3p reduced the capacity of colony assay formation. Colony formation was observed in both TNBC cell lines transfected with miR-29b-3p inhibitor and NC (negative control) inhibitor. Data are presented as % of NC inhibitor group, NC is considered as 100%; data presented as mean ± SD, n = 3; Student’s t-test was considered statistically significant for * p < 0.05, ** p < 0.01, *** p < 0.001 when compared to the NC inhibitor group. Full article ">Figure 3
miR-29b-3p inhibits mitochondrial activity and activates autophagy and apoptosis in both TNBC cell lines. (A) The inhibition of miR-29b-3p reduces the mitochondrial activity in transfected TNBC cell lines, shown through TMRE/Hoechst double staining (active mitochondria are stained in red; cell nuclei are stained in blue); (B) The evaluation of autophagic vacuoles using MDC/PI double staining. Thus, more autophagic vacuoles can be observed in both TNBC cell lines transfected with miR-29b-3p inhibitor compared to the NC inhibitor group. In the MDA-MB-231 cell line, the presence of cell nuclei stained with PI can be observed, suggesting the activation of apoptotic processes to a late phase or necrosis. (C) The evaluation of apoptotic cells was evaluated by using Annexin V-FITC double staining. Post-transfection with miR-29b-3p inhibitor, apoptotic cells in both TNBC cell lines are significantly increased, suggesting that cells undergo apoptosis. An increased number of cells were found in the early and late phases of apoptosis and the necrotic phases. Images were visualized under the inverted fluorescent microscope, IX71 Olympus (20X magnification). Data were analyzed with GraphPad Prism 8 software, using Student’s t-test (* p < 0.05, ** p < 0.01, **** p < 0.0001) based on manual counting of the cells. Full article ">Figure 4
The altered miRNA pattern in TNBC cell lines as an effect of miR-29b-3p inhibition versus NC inhibitor group. Hierarchical clustering of miRNA expression in (A) BT549 and (B) MDA-MB-231 cell lines represented as heat-map, a fold change > ±1.25 and significantly expressed p < 0.05. (C) The Venn diagram used for upregulated miRNAs and (D) The Venn diagram used for downregulated miRNAs as an effect of miR-29b-3p inhibitor on both TNBC cell lines, generated by using Venny software; (E) heatmap representation of the common up- and downregulated miRs in both TNBC cell lines by highlighted the main biological processes using DIANA-miRPath software. Full article ">Figure 5
The network interaction between mRNA–miRNA is generated by using miRNet software. (A) BT549 cell line. (B) MDA-MB-231 cell line. Network generated with miRNet online tool. Full article ">Figure 6
The relative expression level of miR-29b-3p in both transfected TNBC cell lines using qRT-PCR, based on ΔΔCt method (miR-29 inhibitor versus NC inhibitor group). Data were analyzed with GraphPad Prism 8 software, using Student’s t-test (** p < 0.01 and *** p < 0.001). Full article ">Figure 7
The relative expression level of selected genes according to miRNet diagram using qRT-PCR based on ΔΔCt method (miR-29 inhibitor versus NC inhibitor group). In both TNBC cell lines, BT549 and MDA-MB-231, transfected with miR-29b-3p inhibitor for 48 h, the following genes, MCL1, BCL2, TP53, CASP3, TGFB1, TGFBR2, were analyzed. Data were analyzed with GraphPad Prism 8 software, using Student’s t-test (* p < 0.05, ** p < 0.01). Full article ">Figure 8
The quantification of IL6 protein in both TNBC cell lines, BT549 and MBA-MB-231, transfected with miR-29b-3p inhibitor for 48 h. Data were analyzed with GraphPad Prism 8 software, using Student’s t-test (* p < 0.05, ** p < 0.01). Full article ">

25 pages, 4116 KiB

Open AccessArticle

Parental Preconception and Pre-Hatch Exposure to a Developmental Insult Alters Offspring’s Gene Expression and Epigenetic Regulations: An Avian Model

by Issam Rimawi, Gadi Turgeman, Nataly Avital-Cohen, Israel Rozenboim and Joseph Yanai

Int. J. Mol. Sci. 2023, 24(5), 5047; https://doi.org/10.3390/ijms24055047 - 6 Mar 2023

Cited by 2 | Viewed by 2155

Abstract

Parental exposure to insults was initially considered safe if stopped before conception. In the present investigation, paternal or maternal preconception exposure to the neuroteratogen chlorpyrifos was investigated in a well-controlled avian model (Fayoumi) and compared to pre-hatch exposure focusing on molecular [...] Read more.

Parental exposure to insults was initially considered safe if stopped before conception. In the present investigation, paternal or maternal preconception exposure to the neuroteratogen chlorpyrifos was investigated in a well-controlled avian model (Fayoumi) and compared to pre-hatch exposure focusing on molecular alterations. The investigation included the analysis of several neurogenesis, neurotransmission, epigenetic and microRNA genes. A significant decrease in the vesicular acetylcholine transporter (SLC18A3) expression was detected in the female offspring in the three investigated models: paternal (57.7%, p < 0.05), maternal (36%, p < 0.05) and pre-hatch (35.6%, p < 0.05). Paternal exposure to chlorpyrifos also led to a significant increase in brain-derived neurotrophic factor (BDNF) gene expression mainly in the female offspring (27.6%, p < 0.005), while its targeting microRNA, miR-10a, was similarly decreased in both female (50.5%, p < 0.05) and male (56%, p < 0.05) offspring. Doublecortin’s (DCX) targeting microRNA, miR-29a, was decreased in the offspring after maternal preconception exposure to chlorpyrifos (39.8%, p < 0.05). Finally, pre-hatch exposure to chlorpyrifos led to a significant increase in protein kinase C beta (PKCß; 44.1%, p < 0.05), methyl-CpG-binding domain protein 2 (MBD2; 44%, p < 0.01) and 3 (MBD3; 33%, p < 0.05) genes expression in the offspring. Although extensive studies are required to establish a mechanism–phenotype relationship, it should be noted that the current investigation does not include phenotype assessment in the offspring. Full article

(This article belongs to the Special Issue Regulation and Function of Adult Neurogenesis)

► Show Figures

Graphical abstract

17 pages, 1513 KiB

Open AccessReview

The Functions of TRIM56 in Antiviral Innate Immunity and Tumorigenesis

by Lin Fu, Xiaotong Zhou, Qian Jiao and Xi Chen

Int. J. Mol. Sci. 2023, 24(5), 5046; https://doi.org/10.3390/ijms24055046 - 6 Mar 2023

Cited by 14 | Viewed by 2773

Abstract

As a member of the TRIM (tripartite motif) protein family, TRIM56 can function as an E3 ubiquitin ligase. In addition, TRIM56 has been shown to possess deubiquitinase activity and the ability to bind RNA. This adds to the complexity of the regulatory mechanism [...] Read more.

As a member of the TRIM (tripartite motif) protein family, TRIM56 can function as an E3 ubiquitin ligase. In addition, TRIM56 has been shown to possess deubiquitinase activity and the ability to bind RNA. This adds to the complexity of the regulatory mechanism of TRIM56. TRIM56 was initially found to be able to regulate the innate immune response. In recent years, its role in direct antiviral and tumor development has also attracted the interest of researchers, but there is no systematic review on TRIM56. Here, we first summarize the structural features and expression of TRIM56. Then, we review the functions of TRIM56 in TLR and cGAS-STING pathways of innate immune response, the mechanisms and structural specificity of TRIM56 against different types of viruses, and the dual roles of TRIM56 in tumorigenesis. Finally, we discuss the future research directions regarding TRIM56. Full article

(This article belongs to the Section Molecular Immunology)

► Show Figures

Figure 1

Figure 1
Schematic representation of the domains of TRIM56. TRIM56 has three structural domains, an N-terminal RING domain (red), a B-box domain (green), and a coiled-coil domain (blue). The human TRIM56 transcript is 755 aa long. Full article ">Figure 2
Regulatory network of TRIM56 in innate immunity. (A) TRIM56 in the TLR signaling pathway. TRIM56 catalyzes the M1-type ubiquitination modification of TAK1 and thus the interaction between TAK1 and IKKα. TRIM56 interacts with TRIF to positively regulate the TLR3-mediated interferon pathway in an E3-independent manner. (B) TRIM56 in the cGAS-STING signaling pathway. TRIM56 induces the Lys335 monoubiquitination of cGAS, resulting in a significant increase in cGAMP production. TRIM56 catalyzes the formation of the K63-linked ubiquitination of STING. This modification induces STING dimerization, which recruits TBK1 and induces IFN-1β. TRIM56 synthesizes a ubiquitin chain that binds to NEMO and mediates the ubiquitination of NEMO to activate IKKβ. P, phosphate; Ub, ubiquitin. Full article ">Figure 3
TRIM56 is involved in the development of cancer (see also <a href="#ijms-24-05046-t002" class="html-table">Table 2</a>). TRIM56 plays a dual role in tumors. The red color in the figure represents the cancer-promoting function and the blue color represents the cancer-suppressing function. TRIM56 promotes the development of glioma, breast cancer, and Kaposi’s sarcoma, but is an oncogenic repressor in ovarian cancer, multiple myeloma, lung adenocarcinoma, hepatocellular carcinoma, and leukemia. TRIM56 affects multiple signaling pathways, including the TLR3-TRIF pathway, the ELF4-IRF4 pathway, the Wnt pathway, and the NF-κB pathway. TRIM56 mediates the ubiquitin degradation of key proteins, such as Vimentin, FXR1, DVL2, and SAP18. TRIM56 can stabilize some key proteins via K63 ubiquitination (ERα) and deubiquitination (cIAP1 and FOXM1). Ub, ubiquitin. Full article ">Figure 4
TRIM56 is aberrantly expressed in several types of tumors. By analyzing the data in the TCGA database, we found that the expression levels of TRIM56 were significantly low in lung squamous cell carcinoma (LUSC), uterine corpus endometrial carcinoma (UCEC), and uterine carcinosarcoma (UCS), and significantly high in pancreatic adenocarcinoma (PAAD), glioblastoma (GBM), lower-grade glioma (LGG), and thymoma (THYM). Data were analyzed using GEPIA 2.0. Full article ">

17 pages, 837 KiB

Open AccessReview

Impacts of Environmental Pollution on Brain Tumorigenesis

by Cristina Pagano, Giovanna Navarra, Laura Coppola, Beatrice Savarese, Giorgio Avilia, Antonella Giarra, Giovanni Pagano, Alessandra Marano, Marco Trifuoggi, Maurizio Bifulco and Chiara Laezza

Int. J. Mol. Sci. 2023, 24(5), 5045; https://doi.org/10.3390/ijms24055045 - 6 Mar 2023

Cited by 6 | Viewed by 4815

Abstract

Pollutants consist of several components, known as direct or indirect mutagens, that can be associated with the risk of tumorigenesis. The increased incidence of brain tumors, observed more frequently in industrialized countries, has generated a deeper interest in examining different pollutants that could [...] Read more.

Pollutants consist of several components, known as direct or indirect mutagens, that can be associated with the risk of tumorigenesis. The increased incidence of brain tumors, observed more frequently in industrialized countries, has generated a deeper interest in examining different pollutants that could be found in food, air, or water supply. These compounds, due to their chemical nature, alter the activity of biological molecules naturally found in the body. The bioaccumulation leads to harmful effects for humans, increasing the risk of the onset of several pathologies, including cancer. Environmental components often combine with other risk factors, such as the individual genetic component, which increases the chance of developing cancer. The objective of this review is to discuss the impact of environmental carcinogens on modulating the risk of brain tumorigenesis, focusing our attention on certain categories of pollutants and their sources. Full article

(This article belongs to the Special Issue Latest Review Papers in Molecular Oncology 2023)

► Show Figures

Figure 1

20 pages, 12533 KiB

Open AccessArticle

Integrating Multi-Omics Analysis Reveals the Regulatory Mechanisms of White–Violet Mutant Flowers in Grape Hyacinth (Muscari latifolium)

by Junren Ma, Zhi Li and Yali Liu

Int. J. Mol. Sci. 2023, 24(5), 5044; https://doi.org/10.3390/ijms24055044 - 6 Mar 2023

Cited by 2 | Viewed by 2014

Abstract

Grape hyacinth (Muscari spp.) is a famous bulbous blue flower; however, few bicolor varieties are available in the market. Therefore, the discovery of bicolor varieties and understanding of their mechanisms are crucial to the breeding of new varieties. In this study, we [...] Read more.

Grape hyacinth (Muscari spp.) is a famous bulbous blue flower; however, few bicolor varieties are available in the market. Therefore, the discovery of bicolor varieties and understanding of their mechanisms are crucial to the breeding of new varieties. In this study, we report a significant bicolor mutant with white upper and violet lower portions, with both parts belonging to a single raceme. Ionomics showed that pH and metal element contents were not responsible for the bicolor formation. Targeted metabolomics illustrated that the content of the 24 color-related compounds was significantly lower in the upper part than that in the lower part. Moreover, full-length transcriptomics combined with second-generation transcriptomics revealed 12,237 differentially expressed genes in which anthocyanin synthesis gene expression of the upper part was noted to be significantly lower than that of the lower part. Transcription factor differential expression analysis was used to describe the presence of a pair of MaMYB113a/b sequences, with low levels of expression in the upper part and high expression in the lower part. Furthermore, tobacco transformation confirmed that overexpression of MaMYB113a/b can promote anthocyanin accumulation in tobacco leaves. Accordingly, the differential expression of MaMYB113a/b contributes the formation of a bicolor mutant in Muscari latifolium. Full article

(This article belongs to the Section Molecular Plant Sciences)

► Show Figures

Figure 1

21 pages, 14902 KiB

Open AccessReview

Polyoxometalates Impact as Anticancer Agents

by Fátima Carvalho and Manuel Aureliano

Int. J. Mol. Sci. 2023, 24(5), 5043; https://doi.org/10.3390/ijms24055043 - 6 Mar 2023

Cited by 27 | Viewed by 5276

Abstract

Polyoxometalates (POMs) are oxoanions of transition metal ions, such as V, Mo, W, Nb, and Pd, forming a variety of structures with a wide range of applications. Herein, we analyzed recent studies on the effects of polyoxometalates as anticancer agents, particularly their effects [...] Read more.

Polyoxometalates (POMs) are oxoanions of transition metal ions, such as V, Mo, W, Nb, and Pd, forming a variety of structures with a wide range of applications. Herein, we analyzed recent studies on the effects of polyoxometalates as anticancer agents, particularly their effects on the cell cycle. To this end, a literature search was carried out between March and June 2022, using the keywords “polyoxometalates” and “cell cycle”. The effects of POMs on selected cell lines can be diverse, such as their effects in the cell cycle, protein expression, mitochondrial effects, reactive oxygen species (ROS) production, cell death and cell viability. The present study focused on cell viability and cell cycle arrest. Cell viability was analyzed by dividing the POMs into sections according to the constituent compound, namely polyoxovanadates (POVs), polyoxomolybdates (POMos), polyoxopaladates (POPds) and polyoxotungstates (POTs). When comparing and sorting the IC₅₀ values in ascending order, we obtained first POVs, then POTs, POPds and, finally, POMos. When comparing clinically approved drugs and POMs, better results of POMs in relation to drugs were observed in many cases, since the dose required to have an inhibitory concentration of 50% is 2 to 200 times less, depending on the POMs, highlighting that these compounds could become in the future an alternative to existing drugs in cancer therapy. Full article

(This article belongs to the Special Issue Novel Biosystems in Toxicology and Pharmacology)

► Show Figures

Figure 1

35 pages, 1530 KiB

Open AccessReview

Role of Dopamine in the Heart in Health and Disease

by Joachim Neumann, Britt Hofmann, Stefan Dhein and Ulrich Gergs

Int. J. Mol. Sci. 2023, 24(5), 5042; https://doi.org/10.3390/ijms24055042 - 6 Mar 2023

Cited by 18 | Viewed by 8393

Abstract

Dopamine has effects on the mammalian heart. These effects can include an increase in the force of contraction, and an elevation of the beating rate and the constriction of coronary arteries. Depending on the species studied, positive inotropic effects were strong, very modest, [...] Read more.

Dopamine has effects on the mammalian heart. These effects can include an increase in the force of contraction, and an elevation of the beating rate and the constriction of coronary arteries. Depending on the species studied, positive inotropic effects were strong, very modest, or absent, or even negative inotropic effects occurred. We can discern five dopamine receptors. In addition, the signal transduction by dopamine receptors and the regulation of the expression of cardiac dopamine receptors will be of interest to us, because this might be a tempting area of drug development. Dopamine acts in a species-dependent fashion on these cardiac dopamine receptors, but also on cardiac adrenergic receptors. We will discuss the utility of drugs that are currently available as tools to understand cardiac dopamine receptors. The molecule dopamine itself is present in the mammalian heart. Therefore, cardiac dopamine might act as an autocrine or paracrine compound in the mammalian heart. Dopamine itself might cause cardiac diseases. Moreover, the cardiac function of dopamine and the expression of dopamine receptors in the heart can be altered in diseases such as sepsis. Various drugs for cardiac and non-cardiac diseases are currently in the clinic that are, at least in part, agonists or antagonists at dopamine receptors. We define the research needs in order to understand dopamine receptors in the heart better. All in all, an update on the role of dopamine receptors in the human heart appears to be clinically relevant, and is thus presented here. Full article

(This article belongs to the Collection Feature Paper Collection in Biochemistry)

► Show Figures

Figure 1

Figure 1
Dopamine might stimulate D1-dopamine receptors in the sarcolemma. This would activate the stimulatory GTP-binding protein (Gs), and thereby, the activity of adenylyl cyclases (AC) would be enhanced. AC will catalyze the conversion of ATP to cAMP. This cAMP can activate cAMP-dependent protein kinases (PKA), leading to an increase in the phosphorylation states of regulatory proteins. For instance, phosphorylation of L-type Ca2+ channels (LTCC) will lead to more of an influx of Ca2+ from the extracellular space. This Ca2+ will facilitate the release of Ca2+ through the phosphorylated ryanodine receptor (RYR) into the cytosol. The cytosolic Ca2+ activates a Ca2+ calmodulin-dependent protein kinase (CaMKII) that can phosphorylate phospholamban (PLB), the inhibitory protein of the sarcoplasmic reticulum Ca2+ ATPase (SERCA). Possibly, dopamine can also be synthesized in cardiomyocytes. Phenylalanine is oxidized by phenylalanine hydroxylase to tyrosine. Tyrosine is in due course oxidized by tyrosine hydroxylase to L-DOPA, and aromatic L-amino acid decarboxylase (AADC) forms dopamine. Dopamine can be oxidized by dopamine-β-hydroxylase (DBH) to noradrenaline. Catechol-O-methyl transferase (COMT) forms 3-methoxytyramine. Monoamine oxidase A (MAO-A) will oxidize dopamine to dihydroxyphenylacetaldehyde (DOPAL) and aldehyde dehydrogenase (ALDH) to dihydroxyphenylacetic acid (DOPAC), and finally, COMT will form homovanillic acid (HVA). Dopamine, at least in nerve cells, can be transported by a vesicular monoamine transporter (VMAT2) into vesicles. These vesicles can release dopamine to the extracellular space. Dopamine can enter the cell via a dopamine transporter (DAT) or via other transporters (OCT, organic cation transporter; PMAT, plasma membrane monoamine transporter). Additionally, the D1-dopamine receptor can also be stimulated (see <a href="#ijms-24-05042-t001" class="html-table">Table 1</a>A) by fenoldopam, SKF38393, bromocriptine, pergolide, and possibly epinine. Epinine is formed from ibopamine through the activity of esterases. The D1-dopamine receptor will be blocked by haloperidol, but also by SCH23390 (see <a href="#ijms-24-05042-t001" class="html-table">Table 1</a>B). Full article ">Figure 2
Speculative mechanisms of D1-dopamine receptor-mediated signal transduction in the heart. These pathways are known in the central nervous system, but they need to be elucidated in the human heart. Protein phosphorylations can be reversed via the action of protein phosphatases, such as PP1, PP2A, or PP2B. The so-called “dopamine- and cAMP-regulated phosphoproteins with an apparent weight of 32 kDa” (DARPP32), protein phosphatase-1-inhibitor-1 (I-1) protein phosphatase-1-inhibitor-2 (I-2), can inhibit PP1 and thus amplify the function of the D1-dopamine receptor. The inhibitory actions of I-1 and DARPP32 on PP1 are amplified after their phosphorylation by cAMP-dependent protein kinase (PKA). I-1 and DAPRP32 are dephosphorylated, and thus they are inactivated by PP2A and PP2B. PP2A might increase the insertion of D1-dopamine receptors (D1-R) into the sarcolemma. I-2 can be regulated in its activity to inhibit PP1 by the kinases shown here: casein kinase II (CamKII), glycogen synthase kinase 3 (GSK-3), a kinase abbreviated as PFTAIRE kinase (PFTAIRE), by ERK, and a Cdc25C-associated kinase 1 (Cdc25C a K). I-1 is phosphorylated, and its activity is altered by Cdk1, CdK5, protein kinase C (PKC), and a mitogen-activated kinase (MAPK). See text for further details. Full article ">

10 pages, 252 KiB

Open AccessReview

Ovarian Cancer and Glutamine Metabolism

by Zacharias Fasoulakis, Antonios Koutras, Thomas Ntounis, Ioannis Prokopakis, Paraskevas Perros, Athanasios Chionis, Ioakeim Sapantzoglou, Alexandros Katrachouras, Kyriakos Konis, Athina A. Samara, Asimina Valsamaki, Vasileios-Chrysovalantis Palios, Panagiotis Symeonidis, Konstantinos Nikolettos, Athanasios Pagkalos, Sotirios Sotiriou, Marianna Theodora, Panos Antsaklis, Georgios Daskalakis and Emmanuel N. Kontomanolis

Int. J. Mol. Sci. 2023, 24(5), 5041; https://doi.org/10.3390/ijms24055041 - 6 Mar 2023

Cited by 9 | Viewed by 2843

Abstract

Cancer cells are known to have a distinct metabolic profile and to exhibit significant changes in a variety of metabolic mechanisms compared to normal cells, particularly glycolysis and glutaminolysis, in order to cover their increased energy requirements. There is mounting evidence that there [...] Read more.

Cancer cells are known to have a distinct metabolic profile and to exhibit significant changes in a variety of metabolic mechanisms compared to normal cells, particularly glycolysis and glutaminolysis, in order to cover their increased energy requirements. There is mounting evidence that there is a link between glutamine metabolism and the proliferation of cancer cells, demonstrating that glutamine metabolism is a vital mechanism for all cellular processes, including the development of cancer. Detailed knowledge regarding its degree of engagement in numerous biological processes across distinct cancer types is still lacking, despite the fact that such knowledge is necessary for comprehending the differentiating characteristics of many forms of cancer. This review aims to examine data on glutamine metabolism and ovarian cancer and identify possible therapeutic targets for ovarian cancer treatment. Full article

(This article belongs to the Special Issue Molecular and Cellular Mechanisms of Gynecologic Malignancies, Benign Diseases, and Their Medications)

14 pages, 1159 KiB

Open AccessReview

Sepsis-Associated Muscle Wasting: A Comprehensive Review from Bench to Bedside

by Ikumi Yoshihara, Yutaka Kondo, Ken Okamoto and Hiroshi Tanaka

Int. J. Mol. Sci. 2023, 24(5), 5040; https://doi.org/10.3390/ijms24055040 - 6 Mar 2023

Cited by 8 | Viewed by 5502

Abstract

Sepsis-associated muscle wasting (SAMW) is characterized by decreased muscle mass, reduced muscle fiber size, and decreased muscle strength, resulting in persistent physical disability accompanied by sepsis. Systemic inflammatory cytokines are the main cause of SAMW, which occurs in 40–70% of patients with sepsis. [...] Read more.

Sepsis-associated muscle wasting (SAMW) is characterized by decreased muscle mass, reduced muscle fiber size, and decreased muscle strength, resulting in persistent physical disability accompanied by sepsis. Systemic inflammatory cytokines are the main cause of SAMW, which occurs in 40–70% of patients with sepsis. The pathways associated with the ubiquitin–proteasome and autophagy systems are particularly activated in the muscle tissues during sepsis and may lead to muscle wasting. Additionally, expression of muscle atrophy-related genes Atrogin-1 and MuRF-1 are seemingly increased via the ubiquitin–proteasome pathway. In clinical settings, electrical muscular stimulation, physiotherapy, early mobilization, and nutritional support are used for patients with sepsis to prevent or treat SAMW. However, there are no pharmacological treatments for SAMW, and the underlying mechanisms are still unknown. Therefore, research is urgently required in this field. Full article

(This article belongs to the Special Issue Muscle Atrophy: From Bench to Bedside)

► Show Figures

Figure 1

16 pages, 4699 KiB

Open AccessArticle

Effects of Terahertz Radiation on the Aggregation of Alzheimer’s Aβ42 Peptide

by Lei Wang, Yuanyuan Cheng, Wenxia Wang, Jinwu Zhao, Yinsong Wang, Xumei Zhang, Meng Wang, Tianhe Shan and Mingxia He

Int. J. Mol. Sci. 2023, 24(5), 5039; https://doi.org/10.3390/ijms24055039 - 6 Mar 2023

Cited by 3 | Viewed by 2348

Abstract

The pathophysiology of Alzheimer’s disease is thought to be directly linked to the abnormal aggregation of β-amyloid (Aβ) in the nervous system as a common neurodegenerative disease. Consequently, researchers in many areas are actively looking for factors that affect Aβ aggregation. Numerous investigations [...] Read more.

The pathophysiology of Alzheimer’s disease is thought to be directly linked to the abnormal aggregation of β-amyloid (Aβ) in the nervous system as a common neurodegenerative disease. Consequently, researchers in many areas are actively looking for factors that affect Aβ aggregation. Numerous investigations have demonstrated that, in addition to chemical induction of Aβ aggregation, electromagnetic radiation may also affect Aβ aggregation. Terahertz waves are an emerging form of non-ionizing radiation that has the potential to affect the secondary bonding networks of biological systems, which in turn could affect the course of biochemical reactions by altering the conformation of biological macromolecules. As the primary radiation target in this investigation, the in vitro modeled Aβ42 aggregation system was examined using fluorescence spectrophotometry, supplemented by cellular simulations and transmission electron microscopy, to see how it responded to 3.1 THz radiation in various aggregation phases. The results demonstrated that in the nucleation aggregation stage, 3.1 THz electromagnetic waves promote Aβ42 monomer aggregation and that this promoting effect gradually diminishes with the exacerbation of the degree of aggregation. However, by the stage of oligomer aggregation into the original fiber, 3.1 THz electromagnetic waves exhibited an inhibitory effect. This leads us to the conclusion that terahertz radiation has an impact on the stability of the Aβ42 secondary structure, which in turn affects how Aβ42 molecules are recognized during the aggregation process and causes a seemingly aberrant biochemical response. Molecular dynamics simulation was employed to support the theory based on the aforementioned experimental observations and inferences. Full article

(This article belongs to the Special Issue Alzheimer’s Disease: Role and Structure of Soluble Oligomers)

► Show Figures

Figure 1

Figure 1
(a) Relative fluorescence intensities of experimental groups exposed to different irradiation times. (b) Relative fluorescence values of the experimental monomeric exposed group expressed as relative intensities. (c) Comparison of the relative fluorescence readings for the radiation-exposed groups at various incubation times. *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001. Full article ">Figure 2
Aβ42 transmission electron microscope images. (a) Aβ42 monomer’s morphology after 2 hours of aggregation. (b) Aβ42 monomer’s morphology after 2 hours of aggregation under terahertz radiation. (c) Aβ42 oligomer’s morphology after 2 hours of aggregation. (d) Aβ42 oligomer’s morphology after 2 hours of aggregation under terahertz radiation. All scale lengths are 300 nm. Full article ">Figure 3
Cell viability under various interventional conditions. (a) The viability of PC12 cells exposed to terahertz radiation directly in the absence of Aβ42. (b) The viability of PC12 cells subjected to terahertz radiation in a mixed culture containing monomeric Aβ42. (c) The viability of PC12 cells that survived after being exposed to terahertz radiation in combination with oligomeric Aβ42. M, Aβ42 monomers; O, Aβ42 oligomers; M E, Monomer exposed to radiation; O E, Oligomer exposed to radiation. **: p < 0.01, ****: p < 0.0001. Full article ">Figure 4
Live-cell staining under various interventional conditions. (a–c) Fluorescence images of PC12 cells without any intervention; (d–f) fluorescence images of PC12 cells after co-incubation with Aβ42; and (g–i) fluorescence images of PC12 cells following co-incubation with Aβ42 and terahertz radiation exposure. Full article ">Figure 5
(a) Aβ42 monomer molecular dynamics trajectories. (b) Aβ42 oligomer molecular dynamics trajectories. Full article ">Figure 6
(a) Aβ42 monomer RMSD in 20 ns without an applied electric field. (b) Aβ42 monomer RMSD in 20 ns in a terahertz electric field. (c) RMSD of the hydrophobic fragment of the Aβ42 oligomer in 20 ns without an applied electric field. (d) Hydrophobic Aβ42 oligomer fragment RMSD in 20 ns in a terahertz electric field. Full article ">Figure 7
(a) Aβ42 monomer secondary structures in 20 ns without an applied electric field. (b) Aβ42 monomer secondary structures in a terahertz electric field in 20 ns. (c) Hydrophobic fragment of Aβ42 oligomer secondary structures in 20 ns without an applied electric field. (d) A hydrophobic fragment of the Aβ42 oligomer’s secondary structures in a terahertz electric field in 20 ns. Structure = α-Helix + β-Sheet + β-Bridge + Turn. Full article ">Figure 8
(a) The number of hydrogen bonds formed in 20 ns between the Aβ42 monomer and water molecules in the absence of an electric field. (b) The number of hydrogen bonds formed in 20 ns between the Aβ42 monomer and water molecules under a terahertz electric field. (c) The number of hydrogen bonds formed in 20 ns between the Aβ42 oligomer and water molecules in the absence of an electric field. (d) The number of hydrogen bonds formed in 20 ns between the Aβ42 oligomer and water molecules under a terahertz electric field. Full article ">Figure 9
(a) SASA of the Aβ42 monomer in 20 ns. (b) SASA of the Aβ42 monomer in 20 ns under a terahertz electric field. (c) SASA of the Aβ42 oligomer in 20 ns. (d) SASA of the Aβ42 oligomer in 20 ns under a terahertz electric field. Full article ">Figure 10
Schematic representation of the irradiation system. (a) a terahertz waves generator; (b) OAP 1; (c) OAP 2; (d) a plane mirror; and (e) an opaque chamber. Full article ">Figure 11
(a) The initial structure of the Aβ42 monomer simulation. (b) The initial structure of the Aβ42 oligomer simulation. The different colors represent the various amino acids. Full article ">

3 pages, 185 KiB

Open AccessEditorial

Genetic Variation in Transcription Factor Binding Sites

by Gabriel Santpere

Int. J. Mol. Sci. 2023, 24(5), 5038; https://doi.org/10.3390/ijms24055038 - 6 Mar 2023

Cited by 1 | Viewed by 1560

Abstract

The interaction between transcription factors (TFs) and DNA is the core process that determines the state of a cell’s transcriptome [...] Full article

(This article belongs to the Special Issue The Role of Genetic Variation on Transcription Factor Binding Sites in Evolution and Disease)

17 pages, 2248 KiB

Open AccessArticle

[4+2]-Cycloaddition to 5-Methylidene-Hydantoins and 5-Methylidene-2-Thiohydantoins in the Synthesis of Spiro-2-Chalcogenimidazolones

by Dmitry E. Shybanov, Maxim E. Kukushkin, Yanislav S. Hrytseniuk, Yuri K. Grishin, Vitaly A. Roznyatovsky, Viktor A. Tafeenko, Dmitry A. Skvortsov, Nikolai V. Zyk and Elena K. Beloglazkina

Int. J. Mol. Sci. 2023, 24(5), 5037; https://doi.org/10.3390/ijms24055037 - 6 Mar 2023

Cited by 4 | Viewed by 1777

Abstract

Novel hydantion and thiohydantoin-based spiro-compounds were prepared via theDiels–Alder reactions between 5-methylidene-hydantoins or 5-methylidene-2-thiohydantoins and 1,3-dienes (cyclopentadiene, cyclohexadiene, 2,3-dimethylbutadiene, isoprene). It was shown that the cycloaddition reactions proceed regioselectively and stereoselectively with the formation of exo-isomers in the reactions with cyclic dienes andthe [...] Read more.

Novel hydantion and thiohydantoin-based spiro-compounds were prepared via theDiels–Alder reactions between 5-methylidene-hydantoins or 5-methylidene-2-thiohydantoins and 1,3-dienes (cyclopentadiene, cyclohexadiene, 2,3-dimethylbutadiene, isoprene). It was shown that the cycloaddition reactions proceed regioselectively and stereoselectively with the formation of exo-isomers in the reactions with cyclic dienes andthe less sterically hindered products in the reactions with isoprene. Reactions of methylideneimidazolones with cyclopentadiene proceed viaco-heating the reactants; reactions with cyclohexadiene, 2,3-dimethylbutadiene, and isoprene require catalysis by Lewis acids. It was demonstrated that ZnI₂ is an effective catalyst in the Diels–Alder reactions of methylidenethiohydantoins with non-activated dienes. The possibility of alkylation and acylation of the obtained spiro-hydantoinsat the N(1)nitrogen atoms with PhCH₂Cl or Boc₂O and the alkylation of the spiro-thiohydantoinsat the S atoms with MeI or PhCH₂Cl in high yields have been demonstrated. The preparativetransformation of spiro-thiohydantoins into corresponding spiro-hydantoinsin mild conditions by treating with 35% aqueous H₂O₂ or nitrile oxide has been carried out. The obtained compounds show moderate cytotoxicity in the MTT test on MCF7, A549, HEK293T, and VA13 cell lines. Some of the tested compounds demonstrated some antibacterial effect against Escherichia coli (E. coli) BW25113 DTC-pDualrep2 but were almost inactive against E. coli BW25113 LPTD-pDualrep2. Full article

(This article belongs to the Special Issue Development and Synthesis of Biologically Active Compounds)

► Show Figures

Figure 1

15 pages, 51753 KiB

Open AccessArticle

Fabrication of Silane-Grafted Cellulose Nanocrystals and Their Effects on the Structural, Thermal, Mechanical, and Hysteretic Behavior of Thermoplastic Polyurethane

by Xuenan Sun, Xinze Yang, Jiajing Zhang, Bin Shang, Pei Lyu, Chunhua Zhang, Xin Liu and Liangjun Xia

Int. J. Mol. Sci. 2023, 24(5), 5036; https://doi.org/10.3390/ijms24055036 - 6 Mar 2023

Cited by 6 | Viewed by 1785

Abstract

Reinforcement of polymer nanocomposites can be achieved by the selection of the appropriate fabrication method, surface modification, and orientation of the filler. Herein, we present a nonsolvent-induced phase separation method with ternary solvents to prepare thermoplastic polyurethane (TPU) composite films with excellent mechanical [...] Read more.

Reinforcement of polymer nanocomposites can be achieved by the selection of the appropriate fabrication method, surface modification, and orientation of the filler. Herein, we present a nonsolvent-induced phase separation method with ternary solvents to prepare thermoplastic polyurethane (TPU) composite films with excellent mechanical properties using 3-Glycidyloxypropyltrimethoxysilane-modified cellulose nanocrystals (GLCNCs). ATR-IR and SEM analyses of the GLCNCs confirmed that GL was successfully coated on the surface of the nanocrystals. The incorporation of GLCNCs in TPU resulted in the enhancement of the tensile strain and toughness of pure TPU owing to the enhanced interfacial interactions between them. The GLCNC–TPU composite film had tensile strain and toughness values of 1740.42% and 90.01 MJ/m³, respectively. Additionally, GLCNC–TPU exhibited a good elastic recovery rate. CNCs were readily aligned along the fiber axis after the spinning and drawing of the composites into fibers, which further improved the mechanical properties of the composites. The stress, strain, and toughness of the GLCNC–TPU composite fiber increased by 72.60%, 10.25%, and 103.61%, respectively, compared to those of the pure TPU film. This study demonstrates a facile and effective strategy for fabricating mechanically enhanced TPU composites. Full article

(This article belongs to the Special Issue Recent Advances in Cellulose Chemistry)

► Show Figures

Figure 1

14 pages, 3136 KiB

Open AccessReview

Role of Omega-Hydroxy Ceramides in Epidermis: Biosynthesis, Barrier Integrity and Analyzing Method

by Fei Ge, Keyan Sun, Zhenlin Hu and Xin Dong

Int. J. Mol. Sci. 2023, 24(5), 5035; https://doi.org/10.3390/ijms24055035 - 6 Mar 2023

Cited by 5 | Viewed by 2840

Abstract

Attached to the outer surface of the corneocyte lipid envelope (CLE), omega-hydroxy ceramides (ω-OH-Cer) link to involucrin and function as lipid components of the stratum corneum (SC). The integrity of the skin barrier is highly dependent on the lipid components of SC, especially [...] Read more.

Attached to the outer surface of the corneocyte lipid envelope (CLE), omega-hydroxy ceramides (ω-OH-Cer) link to involucrin and function as lipid components of the stratum corneum (SC). The integrity of the skin barrier is highly dependent on the lipid components of SC, especially on ω-OH-Cer. Synthetic ω-OH-Cer supplementation has been utilized in clinical practice for epidermal barrier injury and related surgeries. However, the mechanism discussion and analyzing methods are not keeping pace with its clinical application. Though mass spectrometry (MS) is the primary choice for biomolecular analysis, method modifications for ω-OH-Cer identification are lacking in progress. Therefore, finding conclusions on ω-OH-Cer biological function, as well as on its identification, means it is vital to remind further researchers of how the following work should be done. This review summarizes the important role of ω-OH-Cer in epidermal barrier functions and the forming mechanism of ω-OH-Cer. Recent identification methods for ω-OH-Cer are also discussed, which could provide new inspirations for study on both ω-OH-Cer and skin care development. Full article

(This article belongs to the Section Molecular Neurobiology)

► Show Figures

Figure 1

20 pages, 1439 KiB

Open AccessReview

Neutrophil Extracellular Traps in Airway Diseases: Pathological Roles and Therapeutic Implications

by Ara Jo and Dae Woo Kim

Int. J. Mol. Sci. 2023, 24(5), 5034; https://doi.org/10.3390/ijms24055034 - 6 Mar 2023

Cited by 20 | Viewed by 5756

Abstract

Neutrophils are important effector cells of the innate immune response that fight pathogens by phagocytosis and degranulation. Neutrophil extracellular traps (NETs) are released into the extracellular space to defend against invading pathogens. Although NETs play a defensive role against pathogens, excessive NETs can [...] Read more.

Neutrophils are important effector cells of the innate immune response that fight pathogens by phagocytosis and degranulation. Neutrophil extracellular traps (NETs) are released into the extracellular space to defend against invading pathogens. Although NETs play a defensive role against pathogens, excessive NETs can contribute to the pathogenesis of airway diseases. NETs are known to be directly cytotoxic to the lung epithelium and endothelium, highly involved in acute lung injury, and implicated in disease severity and exacerbation. This review describes the role of NET formation in airway diseases, including chronic rhinosinusitis, and suggests that targeting NETs could be a therapeutic strategy for airway diseases. Full article

(This article belongs to the Special Issue Chronic Rhinosinusitis: Aetiology, Immunology and Treatment)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Int. J. Mol. Sci., Volume 24, Issue 5 (March-1 2023) – 810 articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI