Current Issues in Molecular Biology

15 pages, 1756 KiB

Open AccessArticle

Peroxiredoxins and Hypoxia-Inducible Factor-1α in Duodenal Tissue: Emerging Factors in the Pathophysiology of Pediatric Celiac Disease Patients

by Fadime Aydın Köse, Aysun Pabuccuoglu, Miray Karakoyun and Sema Aydogdu

Curr. Issues Mol. Biol. 2023, 45(2), 1779-1793; https://doi.org/10.3390/cimb45020114 - 20 Feb 2023

Cited by 2 | Viewed by 2360

Abstract

Celiac disease (CD) is an autoimmune enteropathy. Peroxiredoxins (PRDXs) are powerful antioxidant enzymes having an important role in significant cellular pathways including cell survival, apoptosis, and inflammation. This study aimed at investigating the expression levels of all PRDX isoforms (1–6) and their possible [...] Read more.

Celiac disease (CD) is an autoimmune enteropathy. Peroxiredoxins (PRDXs) are powerful antioxidant enzymes having an important role in significant cellular pathways including cell survival, apoptosis, and inflammation. This study aimed at investigating the expression levels of all PRDX isoforms (1–6) and their possible relationships with a transcription factor, HIF-1α, in the small intestinal tissue samples of pediatric CD patients. The study groups consisted of first-diagnosed CD patients (n = 7) and non-CD patients with functional gastrointestinal tract disorders as the controls (n = 7). The PRDXs and HIF-1α expression levels were determined by using real-time PCR and Western blotting in duodenal biopsy samples. It was observed that the mRNA and protein expression levels of PRDX 5 were significantly higher in the CD patients, whereas the PRDX 1, -2, and -4 expressions were decreased in each case compared to the control group. No significant differences were detected in the PRDX 3 and PRDX 6 expressions. The expression of HIF-1α was also significantly elevated in CD patients. These findings indicate, for the first time, that PRDXs, particularly PRDX 5, may play a significant role in the pathogenesis of CD. Furthermore, our results suggest that HIF-1α may upregulate PRDX-5 transcription in the duodenal tissue of CD. Full article

(This article belongs to the Special Issue Molecular Mechanisms and Regulation in Allergy and Immune Diseases, Immunodeficiencies)

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Protein expression levels of tTG and inflammatory proteins in the duodenal tissue of children with CD and the control group. (a) Protein expression of tTG, iNOS, p-NF-κB, and TNF-α were determined by WB analysis and quantified by densitometric analysis. Relative densitometric values of the (b) tTG protein expression; (c) iNOS protein expression; (d) p-NF-κB p65 protein expression; and (e) TNF-α protein expression levels. The β-actin signal was used to normalize the data, which were then expressed in arbitrary densitometric units. Data were expressed as mean ± SEM (n = 3). * p < 0.05 versus the control group (CD: celiac disease group). Full article ">Figure 2
Apoptotic cell death-related cleaved caspase-3 and p53 protein expression levels in the duodenal tissue of pediatric CD patients. (a) Protein expression of cleaved caspase-3 and p53 were determined by WB analysis and quantified by densitometric analysis. Relative densitometric values of the (b) p53 protein expression and (c) cleaved caspase-3 protein expression levels. The β-actin signal was used to normalize the data, which were then expressed in arbitrary densitometric units. Data were expressed as mean ± SEM (n = 3). * p < 0.05 versus the control group (CD: celiac disease group). Full article ">Figure 3
The relative protein and mRNA expression levels of HIF-1α in the duodenal tissue. (a) Protein expression of HIF-1α were determined by WB analysis. (b) Relative values of HIF-1α were quantified by densitometric analysis. The β-actin signal was used to normalize the data. Data were expressed as the mean ± SEM (n = 3). (c) The relative mRNA expressions were calculated with the 2−ΔΔCT method and normalized to GAPDH (n = 7/7). Data were expressed as the mean ± SEM. * p < 0.05 versus the control group (CD: celiac disease group). Full article ">Figure 4
Comparative graph of the relative PRDX (1–6) mRNA expression values. Data were expressed as the mean ± SEM (n = 7/7). * p < 0.05 versus the control group (CD: celiac disease group). Full article ">Figure 5
The protein expression levels of the PRDX isoforms in the duodenal biopsy samples. (a) Protein expression levels of PRDX -1, -2, -3, -4, -5, and -6 were determined by WB analysis. (b) Relative densitometric values of the protein expression levels of the PRDX isoforms. The β-actin signal was used to normalize the data, which were then expressed in arbitrary densitometric units. Data were expressed as the mean ± SEM (n = 3). * p < 0.05 versus the control group. Full article ">

17 pages, 808 KiB

Open AccessReview

Neurobiology and Applications of Inositol in Psychiatry: A Narrative Review

by Carmen Concerto, Cecilia Chiarenza, Antonio Di Francesco, Antimo Natale, Ivan Privitera, Alessandro Rodolico, Antonio Trovato, Andrea Aguglia, Francesco Fisicaro, Manuela Pennisi, Rita Bella, Antonino Petralia, Maria Salvina Signorelli and Giuseppe Lanza

Curr. Issues Mol. Biol. 2023, 45(2), 1762-1778; https://doi.org/10.3390/cimb45020113 - 20 Feb 2023

Cited by 14 | Viewed by 16367

Abstract

Inositol is a natural sugar-like compound, commonly present in many plants and foods. It is involved in several biochemical pathways, most of them controlling vital cellular mechanisms, such as cell development, signaling and nuclear processes, metabolic and endocrine modulation, cell growth, signal transduction, [...] Read more.

Inositol is a natural sugar-like compound, commonly present in many plants and foods. It is involved in several biochemical pathways, most of them controlling vital cellular mechanisms, such as cell development, signaling and nuclear processes, metabolic and endocrine modulation, cell growth, signal transduction, etc. In this narrative review, we focused on the role of inositol in human brain physiology and pathology, with the aim of providing an update on both potential applications and current limits in its use in psychiatric disorders. Overall, imaging and biomolecular studies have shown the role of inositol levels in the pathogenesis of mood disorders. However, when administered as monotherapy or in addition to conventional drugs, inositol did not seem to influence clinical outcomes in both mood and psychotic disorders. Conversely, more encouraging results have emerged for the treatment of panic disorders. We concluded that, despite its multifaceted neurobiological activities and some positive findings, to date, data on the efficacy of inositol in the treatment of psychiatric disorders are still controversial, partly due to the heterogeneity of supporting studies. Therefore, systematic use of inositol in routine clinical practice cannot be recommended yet, although further basic and translational research should be encouraged. Full article

(This article belongs to the Special Issue Food-Derived Bioactive Compounds in Health and Disease)

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21 pages, 11399 KiB

Open AccessArticle

Vaccine- and Breakthrough Infection-Elicited Pre-Omicron Immunity More Effectively Neutralizes Omicron BA.1, BA.2, BA.4 and BA.5 Than Pre-Omicron Infection Alone

by Eveline Santos da Silva, Jean-Yves Servais, Michel Kohnen, Victor Arendt, Georges Gilson, Therese Staub, Carole Seguin-Devaux and Danielle Perez-Bercoff

Curr. Issues Mol. Biol. 2023, 45(2), 1741-1761; https://doi.org/10.3390/cimb45020112 - 19 Feb 2023

Cited by 3 | Viewed by 2271

Abstract

Since the emergence of SARS-CoV-2 Omicron BA.1 and BA.2, several Omicron sublineages have emerged, supplanting their predecessors. Here we compared the neutralization of Omicron sublineages BA.1, BA.2, BA.4 and BA.5 by human sera collected from individuals who were infected with the ancestral B.1 [...] Read more.

Since the emergence of SARS-CoV-2 Omicron BA.1 and BA.2, several Omicron sublineages have emerged, supplanting their predecessors. Here we compared the neutralization of Omicron sublineages BA.1, BA.2, BA.4 and BA.5 by human sera collected from individuals who were infected with the ancestral B.1 (D614G) strain, who were vaccinated (3 doses) or with breakthrough infection with pre-Omicron strains (Gamma or Delta). All Omicron sublineages exhibited extensive escape from all sera when compared to the ancestral B.1 strain and to Delta, albeit to different levels depending on the origin of the sera. Convalescent sera were unable to neutralize BA.1, and partly neutralized BA.2, BA.4 and BA.5. Vaccinee sera partly neutralized BA.2, but BA.1, BA.4 and BA.5 evaded neutralizing antibodies (NAb). Some breakthrough infections (BTI) sera were non-neutralizing. Neutralizing BTI sera had similar neutralizing ability against all Omicron sublineages. Despite similar levels of anti-Spike and anti-Receptor Binding Domain (RBD) antibodies in all groups, BTI sera had the highest cross-neutralizing ability against all Omicron sublineages and convalescent sera were the least neutralizing. Antibody avidity inferred from the NT50:antibody titer ratio was highest in sera from BTI patients, underscoring qualitative differences in antibodies elicited by infection or vaccination. Together, these findings highlight the importance of vaccination to trigger highly cross-reactive antibodies that neutralize phylogenetically and antigenically distant strains, and suggest that immune imprinting by first generation vaccines may restrict, but not abolish, cross-neutralization. Full article

(This article belongs to the Special Issue Molecular and Real-World Evidence Research of Respiratory Diseases and Infections)

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21 pages, 4517 KiB

Open AccessArticle

Quantification of the Diversity in Gene Structures Using the Principles of Polarization Mapping

by Dmitry Zimnyakov, Marina Alonova, Anatoly Skripal, Sergey Dobdin and Valentina Feodorova

Curr. Issues Mol. Biol. 2023, 45(2), 1720-1740; https://doi.org/10.3390/cimb45020111 - 18 Feb 2023

Cited by 4 | Viewed by 1851

Abstract

Results of computational analysis and visualization of differences in gene structures using polarization coding are presented. A two-dimensional phase screen, where each element of which corresponds to a specific basic nucleotide (adenine, cytosine, guanine, or thymine), displays the analyzed nucleotide sequence. Readout of [...] Read more.

Results of computational analysis and visualization of differences in gene structures using polarization coding are presented. A two-dimensional phase screen, where each element of which corresponds to a specific basic nucleotide (adenine, cytosine, guanine, or thymine), displays the analyzed nucleotide sequence. Readout of the screen with a coherent beam characterized by a given polarization state forms a diffracted light field with a local polarization structure that is unique for the analyzed nucleotide sequence. This unique structure is described by spatial distributions of local values of the Stokes vector components. Analysis of these distributions allows the comparison of nucleotide sequences for different strains of pathogenic microorganisms and frequency analysis of the sequences. The possibilities of this polarization-based technique are illustrated by the model data obtained from a comparative analysis of the spike protein gene sequences for three different model variants (Wuhan, Delta, and Omicron) of the SARS-CoV-2 virus. Various modifications of polarization encoding and analysis of gene structures and a possibility for instrumental implementation of the proposed method are discussed. Full article

(This article belongs to the Section Bioinformatics and Systems Biology)

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8 pages, 3834 KiB

Open AccessCase Report

Recurrent PIK3CA H1047R-Mutated Congenital Infiltrative Facial Lipomatosis: A Case Report and Review of Literature

by Kei Shing Oh, Hisham F. Bahmad, Kalin Veselinov Stoyanov, Ibrahim H. Amjad and Carole Brathwaite

Curr. Issues Mol. Biol. 2023, 45(2), 1712-1719; https://doi.org/10.3390/cimb45020110 - 17 Feb 2023

Cited by 4 | Viewed by 2143

Abstract

Congenital infiltrating lipomatosis of the face (CILF) is a rare, congenital, nonhereditary facial overgrowth due to post-zygomatic activating mutations in PIK3CA gene. It is unilateral and involves hypertrophy of both the soft and hard tissue structures on the affected side of the face. [...] Read more.

Congenital infiltrating lipomatosis of the face (CILF) is a rare, congenital, nonhereditary facial overgrowth due to post-zygomatic activating mutations in PIK3CA gene. It is unilateral and involves hypertrophy of both the soft and hard tissue structures on the affected side of the face. This commonly results in early eruption of the teeth, hypertrophy of the facial bones, macroglossia, and proliferation of the parotid gland. Less than 80 cases of CILF have been reported in the literature so far. Treatment modalities include liposuction and surgical excision. However, since the hallmark of CILF is mutation in the PIK3CA gene, PI3K inhibitors may play a therapeutic role in CILF. We report a case of an 8-year-old boy with recurrent CILF of the scalp and nose, with PIK3CA H1047R mutation. We discuss the differential diagnoses, clinical outcomes, and management of this rare entity. Full article

(This article belongs to the Special Issue Linking Genomic Changes with Cancer in the NGS Era)

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19 pages, 4294 KiB

Open AccessArticle

Genome-Wide Identification and Expression Analysis of Calmodulin-Like Gene Family in Paspalums vaginatium Revealed Their Role in Response to Salt and Cold Stress

by Meizhen Yang, Jingjin Chen, Tingting Liu, Leilei Xiang and Biao-Feng Zhou

Curr. Issues Mol. Biol. 2023, 45(2), 1693-1711; https://doi.org/10.3390/cimb45020109 - 16 Feb 2023

Cited by 7 | Viewed by 2122

Abstract

The calmodulin-like (CML) family is an important calcium (Ca²⁺) sensor in plants and plays a pivotal role in the response to abiotic and biotic stresses. As one of the most salt-tolerant grass species, Paspalums vaginatum is resistant to multiple abiotic stresses, [...] Read more.

The calmodulin-like (CML) family is an important calcium (Ca²⁺) sensor in plants and plays a pivotal role in the response to abiotic and biotic stresses. As one of the most salt-tolerant grass species, Paspalums vaginatum is resistant to multiple abiotic stresses, such as salt, cold, and drought. However, investigations of PvCML proteins in P. vaginatum have been limited. Based on the recently published P. vaginatum genome, we identified forty-nine PvCMLs and performed a comprehensive bioinformatics analysis of PvCMLs. The main results showed that the PvCMLs were unevenly distributed on all chromosomes and that the expansion of PvCMLs was shaped by tandem and segmental duplications. In addition, cis-acting element analysis, expression profiles, and qRT–PCR analysis revealed that PvCMLs were involved in the response to salt and cold stress. Most interestingly, we found evidence of a tandem gene cluster that independently evolved in P. vaginatum and may participate in cold resistance. In summary, our work provides important insight into how grass species are resistant to abiotic stresses such as salt and cold and could be the basis of further gene function research on CMLs in P. vaginatum. Full article

(This article belongs to the Special Issue Functional Genomics and Comparative Genomics Analysis in Plants)

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Phylogenetic relationship of calmodulin-like (CML) proteins in P. vaginatum (green), A. thaliana (red), and rice (blue). An unrooted tree was built using the NJ method in MEGA with 1000 bootstrap replications. The bootstrap values of each node lower than 30 were not shown. Full article ">Figure 2
The conserved motif composition and gene structure of CMLs proteins in P. vaginatum. (A) phylogenetic relationship of PvCMLs. (B) The conserved EF-hand motifs of PvCMLs. (C) Gene structure of PvCMLs. Pink boxes represent untranslated regions (UTR), green boxes represent coding regions (CDS), and black lines represent introns. Full article ">Figure 3
Chromosomal distribution and synteny analysis in the genomes of P. vaginatum, Oryza sativa, and A. thaliana. (A) Paralogous genes of PvCMLs in P. vaginatum. (B) Orthologous CML genes between P. vaginatum and rice. (C) Orthologous CML genes between P. vaginatum and A. thaliana and between P. vaginatum and rice. Red lines indicate orthologous gene pairs. Full article ">Figure 4
Bar plot of average Ka/Ks ratios for homologous CML gene pairs between rice and P. vaginatum. The different colors indicate eight subfamily groups identified in phylogenetic analysis. The error bar is shown in the figure. Full article ">Figure 5
Prediction of cis-acting elements contained in PvCMLs. Analysis of cis-elements was carried out by the PlantCARE database. The analysis showed the presence of 14 cis-elements. The different colors represent the different numbers of cis-elements, with red indicating a high number of cis-elements, while white represents a low number of cis-elements. Values in the boxes represent the number of cis-regulatory elements. Full article ">Figure 6
Expression pattern of PvCML genes under salt and cold stress conditions. (A). Heatmap of log2–transformed RPKM of PvCML genes after 400 nM NaCl treatment. The colors changing from blue to red indicate the expression level transition from low to high. (B). Volcano plot displaying the differentially expressed genes after salt treatment. The blue dots represent genes differentially expressed (adjusted p < 0.05) in the 400 nM NaCl treatment group vs. the normal control group. The red dots represent PvCMLs. (C). Heatmap of log2–transformed RPKM of PvCML genes after 6 °C cold treatment. The colors changing from blue to red indicate the expression level transition from low to high. (D). Volcano plot displaying the differentially expressed genes after 6 °C cold treatment. The blue dots represent genes differentially expressed (adjusted p < 0.05) in the 6 °C treatment group vs. the normal control group. The red dots represent PvCMLs. Full article ">Figure 7
Gene expression analysis by RT–qPCR for the PvCMLs under salt stress conditions. The four-week-old seashore paspalum cultivars (sea spray) were treated with 200 mM NaCl, and three leaves were collected for each sample at 0 h, 2 h, and 24 h following treatment. Three biological replicates were performed for this analysis. The bar plot indicates the relative mRNA expression of each PvCML gene at each treatment time. Full article ">Figure 8
Gene expression analysis by RT–qPCR for the PvCMLs under cold stress conditions. The four-week-old seashore paspalum cultivars (sea spray) were treated with 4 °C, and three leaves were collected for each sample at 0 h, 2 h, and 24 h following treatment. Three biological replicates were performed for this analysis. The bar plot indicates the relative mRNA expression of each PvCML gene at each treatment time. Full article ">

12 pages, 2250 KiB

Open AccessArticle

Effect of C-Type Natriuretic Peptide (CNP) on Spermatozoa Maturation in Adult Rat Epididymis

by Hu Zhao, Yuejin Yu, Chunlei Mei, Tianyu Zhang, Yafei Kang, Na Li and Donghui Huang

Curr. Issues Mol. Biol. 2023, 45(2), 1681-1692; https://doi.org/10.3390/cimb45020108 - 16 Feb 2023

Cited by 3 | Viewed by 2439

Abstract

C-type natriuretic peptide (CNP) is highly expressed in male reproductive tissues, such as the epididymis. The aim of this study is to explore the role of CNP in the maturation of rat epididymal spermatozoa. First, the expression levels of CNP and its specific [...] Read more.

C-type natriuretic peptide (CNP) is highly expressed in male reproductive tissues, such as the epididymis. The aim of this study is to explore the role of CNP in the maturation of rat epididymal spermatozoa. First, the expression levels of CNP and its specific natriuretic peptide receptor-B (NPR-B) were detected in various tissues of rats and epididymis at different stages after birth. Then a castrated rat model was established to analyze the relationship between testosterone and CNP/NPR-B expression in the epididymis. Finally, CNP and different inhibitors (NPR-B inhibitors, cGMP inhibitors) were used to incubate epididymal sperm in vitro to examine sperm mobility and expression of sperm maturation-related factors. The results showed CNP/NPR-B mRNAs were expressed in all tissues of rats, but were extremely highly expressed in male genital ducts (seminal vesicle, prostate and epididymis). The expression of CNP/NPR-B in epididymis was the highest at birth and the fifth week after birth. In the epididymis, CNP/NPR-B were highly expressed in the caput and located in the epididymal epithelial cells. After castration, the expression of CNP/NPR-B decreased sharply and was restored quickly after testosterone supplementation. In vitro, CNP could significantly promote the acquisition of epididymal sperm motility through the NPR-B/cGMP pathway and induce the expression of sperm maturation-related factors (such as Bin1b, Catsper 1, Dnah17, Fertilin). This study shows that CNP plays a role in epididymal sperm maturation. The mechanism of CNP is to promote the acquisition of epididymal sperm fluidity through the NPR-B/cGMP signaling pathway and also to regulate sperm maturation-related genes. Moreover, the expression of CNP/NPR-B was regulated by testosterone. Full article

(This article belongs to the Special Issue Molecular Research in Reproductive Biology)

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The expression of CNP/NPR-B in rat. (A,B) The expression of CNP (A) and NPR-B (B) mRNA in different rat tissues. (C,D) The expression of CNP (C) and NPR-B (D) mRNA in epididymis at different stages after birth. Full article ">Figure 2
The expression of CNP/NPR-B in the different segments of epididymis. (A) Immunohistochemistry shows the localization of CNP and NPR-B in different segments of the epididymis and prostate. a–c: caput of epididymis; d–f: corpse of epididymis; g–i: cauda of epididymis; j–l: prostate. (B,C) The level of CNP (B) and NPR-B (C) mRNA in different segments of epididymis detected by RT- PCR. (D,E) The content of CNP (D) and cGMP (E) in different segments of epididymal fluid by ELISA. (* p < 0.05, ** p < 0.01, *** p < 0.01, NS: not significant). Full article ">Figure 3
The expression of CNP/NPR−B in the castrated rat model. The red line represents the content of testosterone in the serum, while the blue line represents the expression of CNP (A) or NPR−B (B) mRNA in the epididymis. Full article ">Figure 4
The effect of CNP on maturation of the epididymis sperm in vitro. (A) Spermatozoa immunofluorescence in the different segments of the epididymis detected by laser confocal photography (600×). (B) Sperm motilities at different segments of epididymis detected at 0, 2, 4, 6 and 8 h after incubation with CNP. (C) Sperm motility at the caput of the epididymis after incubation with either CNP, 8-Br-cGMP, KT5823, or NPR-B antagonist. (D) Expression of maturation-related genes in the epididymal caput sperm after incubation with CNP for 6 h. (* p < 0.05, ** p < 0.01). Full article ">Figure 5
The role of CNP in sperm maturation. Testosterone secreted by Leydig cells could promote the expression of CNP/NPR-B in rat epididymal epithelium, especially in the caput. CNP not only promotes the acquisition of epididymal sperm mobility via the NPR-B/cGMP signal pathway but also stimulates sperm mature-related genes (such as Bin1b, Catsper 1, Dnah17), thus playing a role in sperm maturation in epididymis. Full article ">

26 pages, 5532 KiB

Open AccessArticle

Molecular Aspects of Hypoxic Stress Effects in Chronic Ethanol Exposure of Neuronal Cells

by Simona Isabelle Stoica, Gelu Onose, Ioana Madalina Pitica, Ana Iulia Neagu, Gabriela Ion, Lilia Matei, Laura Denisa Dragu, Lacramioara-Elena Radu, Mihaela Chivu-Economescu, Laura Georgiana Necula, Aurelian Anghelescu, Carmen Cristina Diaconu, Constantin Munteanu and Coralia Bleotu

Curr. Issues Mol. Biol. 2023, 45(2), 1655-1680; https://doi.org/10.3390/cimb45020107 - 16 Feb 2023

Cited by 6 | Viewed by 2972

Abstract

Experimental models of a clinical, pathophysiological context are used to understand molecular mechanisms and develop novel therapies. Previous studies revealed better outcomes for spinal cord injury chronic ethanol-consuming patients. This study evaluated cellular and molecular changes in a model mimicking spinal cord injury [...] Read more.

Experimental models of a clinical, pathophysiological context are used to understand molecular mechanisms and develop novel therapies. Previous studies revealed better outcomes for spinal cord injury chronic ethanol-consuming patients. This study evaluated cellular and molecular changes in a model mimicking spinal cord injury (hypoxic stress induced by treatment with deferoxamine or cobalt chloride) in chronic ethanol-consuming patients (ethanol-exposed neural cultures (SK-N-SH)) in order to explain the clinical paradigm of better outcomes for spinal cord injury chronic ethanol-consuming patients. The results show that long-term ethanol exposure has a cytotoxic effect, inducing apoptosis. At 24 h after the induction of hypoxic stress (by deferoxamine or cobalt chloride treatments), reduced ROS in long-term ethanol-exposed SK-N-SH cells was observed, which might be due to an adaptation to stressful conditions. In addition, the HIF-1α protein level was increased after hypoxic treatment of long-term ethanol-exposed cells, inducing fluctuations in its target metabolic enzymes proportionally with treatment intensity. The wound healing assay demonstrated that the cells recovered after stress conditions, showing that the ethanol-exposed cells that passed the acute step had the same proliferation profile as the cells unexposed to ethanol. Deferoxamine-treated cells displayed higher proliferative activity than the control cells in the proliferation–migration assay, emphasizing the neuroprotective effect. Cells have overcome the critical point of the alcohol-induced traumatic impact and adapted to ethanol (a chronic phenomenon), sustaining the regeneration process. However, further experiments are needed to ensure recovery efficiency is more effective in chronic ethanol exposure. Full article

(This article belongs to the Special Issue Molecular Research on Oxidative Stress and Health)

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11 pages, 1796 KiB

Open AccessCommunication

Stability of Dengue 2 Nonstructural Glycoprotein 1 (NS1) Is Affected by the Nature of Basic Residue at Position NS1-324

by Eva Ogire, Chaker El-Kalamouni, Philippe Desprès and Marjolaine Roche

Curr. Issues Mol. Biol. 2023, 45(2), 1644-1654; https://doi.org/10.3390/cimb45020106 - 14 Feb 2023

Cited by 2 | Viewed by 2143

Abstract

Dengue is the most prevalent mosquito-borne viral disease. It is caused by the infection of any of the four dengue virus (DENV) serotypes DENV-1 to DENV-4. The DENV non-structural glycoprotein 1 (NS1) plays an important role in virus replication and the immunopathogenesis of [...] Read more.

Dengue is the most prevalent mosquito-borne viral disease. It is caused by the infection of any of the four dengue virus (DENV) serotypes DENV-1 to DENV-4. The DENV non-structural glycoprotein 1 (NS1) plays an important role in virus replication and the immunopathogenesis of virus infection. The NS1 protein has been identified as both a cell-associated homodimer and a soluble secreted lipoprotein nanoparticle. The nature of the residues at positions NS1-272 and NS1-324 in the β-ladder domain may have an effect on the biological behaviors of DENV-2 NS1 protein in human hepatoma Huh7 cells. The stability of the NS1 protein from the Reunion 2018 DENV-2 strain was affected by the presence of lysine residues at positions 272 and 324. In the present study, we evaluated the impact of mutations into lysine at positions 272 and 324 on recombinant NS1 protein from the DES-14 DENV-2 strain bearing arginine residue on these two positions. The DES-14 NS1 protein mutant bearing a lysine at position 324 was deficient in protein stability and secretion compared to wild-type protein. The defect in the DES-14 NS1 protein mutant was associated to oxidative stress and pro-inflammatory cytokine activation in Huh7 cells. The ubiquitin-proteasome proteolytic pathway might play a key role in the stability of DENV-2 protein bearing a lysine residue at position 324. Full article

(This article belongs to the Collection Feature Papers in Current Issues in Molecular Biology)

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Structural features of DENV-2 NS1 protein. Tridimensional structure prediction of DES-14 NS1 protein (Genbank access number MG189962) and the three mutants DES-14 NS1-(K272, K324), NS1-(K272), and NS1-(K324) was performed by modeling on PHYRE2 protein recognition server (<a href="http://www.sbg.bio.ic.ac.uk/~phyre2/html/page.cgi?id=index" target="_blank">http://www.sbg.bio.ic.ac.uk/~phyre2/html/page.cgi?id=index</a>; accessed on 23 December 2022). The predicted structures were analyzed with PyMOL 2.5, a program for interactive visualization of tridimensional proteins. The colored dots indicate the positions of residues NS1-272 and NS1-324 in the 3D structure of NS1 protein. Full article ">Figure 2
Effects of amino-acid substitutions R272K and R324K on DENV-2 rNS1 expression. Huh7 cells were transfected for 24 h with plasmids expressing DENV-2 rNS1wt or rNS1-(K272), rNS1-(K324), and rNS1-(K272, K324) mutants. In (A), immunoblot assay was performed on non-heated RIPA cell lysates (intracellular proteins) and cell supernatants (extracellular proteins) using anti-NS1 antibody D/2/D6/B7 (anti-NS1 Ab). The intracellular NS1 dimer is indicated as (rNS1)2. The β-actin protein served as a protein loading control. The estimated apparent molecular weights of (rNS1)2 and β-actin are indicated. The basic residues at the positions NS1-272 and NS1-324 are shown at the top. In (B), signal intensity was quantified using ImageJ software to evaluate the amount of rNS1 mutants relative to rNS1wt. The results are the mean (± SEM) of 10 (intracellular rNS1) or 4 (secreted rNS1) replicates (open circles). p-values were determined on the comparison of rNS1-(K324) with rNS1-(K272) mutant (**** p < 0.0001; * p < 0.05). Full article ">Figure 3
Effect of proteasome inhibition on DENV-2 rNS1 expression. Huh7 cells were transfected for 24 h with plasmids expressing DENV-2 rNS1wt or rNS1-(K272), rNS1-(K324), and rNS1-(K272, K324) mutants or mock-transfected (control) in presence (+) or absence of 10 µM of the proteasome inhibitor MG132 in 0.1% DMSO for 6 h. In (A), immunoblot assays were performed on RIPA cell lysates using anti-NS1 antibody. Cell lysates were analyzed before (top) and after (bottom) heat-denaturation in the presence of a reducing agent. The mouse anti-NS1 antibody D/2/D6/B7 (anti-NS1 mAb) was used to detect heat-labile dimer rNS1 which is indicated as (NS1)2. The rabbit anti-NS1 antibody PA5-33207 (anti-NS1 pAb) was used to detect monomeric form of rNS1 which is indicated as (rNS1)m. The β-actin protein served as a protein loading control. The estimated apparent molecular weights of (rNS1)2 and (rNS1)m are indicated. In (B), signal intensity was quantified using ImageJ software to calculate the fold-increase between the amounts of rNS1 expressed with or without MG132 (control). The results are the mean (± SEM) of 3 replicates (open circles). p-values were determined on the comparison of different rNS1 (* p < 0.05; n.s.: not significant). Full article ">Figure 4
ROS production in Huh7 cells expressing the rNS1-(K24) mutant. Huh7 cells were transfected for 24 h with plasmids expressing DENV-2 rNS1wt, rNS1-(K272), or rNS1-(K324). In (A), the production of total intracellular reactive oxygen species (ROS) was measured using DCFH-DA dye. Intracellular ROS in samples was expressed as a fold change relative to mock-transfected cells. In (B), antioxidant enzyme superoxide dismutase (SOD-1) mRNA levels were quantified by RT-qPCR. Results are expressed as the fold induction of transcripts in samples relative to those in mock-transfected cells. The results are the mean (± SEM) at least seven (A) and three (B) replicates (open circles). p-values were determined on the comparison of rNS1 mutants versus rNS1wt and rNS1-(K324) mutant versus rNS1-(K272) mutant (n.s.: not significant). Full article ">Figure 5
Pro-inflammatory cytokine activation in Huh7 cells expressing the rNS1-(K324) mutant. Huh7 cells were transfected for 24 h with plasmids expressing rNS1wt or the rNS1-(K272) and rNS1-(K324) mutants. The IL-1β (A) and IL-6 (B) transcripts were quantified by RT-qPCR. Results are expressed as the fold induction of transcripts in samples relative to those in mock-transfected cells. The results are the mean (± SEM) at least three replicates (open circles). p-values were determined on the comparison of rNS1 mutants versus rNS1wt. Full article ">Figure 6
Expression of DES-14 NS1 protein mutant bearing a lysine residue at position 324 (DES-14 NS1-(K324) mutant) is associated to oxidative stress and inflammatory in human hepatoma Huh7 cells. Full article ">

17 pages, 40206 KiB

Open AccessArticle

Possible Mechanisms of the Neuroprotective Actions of Date Palm Fruits Aqueous Extracts against Valproic Acid-Induced Autism in Rats

by Abdelaziz M. Hussein, Seham Ahmed Mahmoud, Khalid Mohammed Elazab, Ahmed F. Abouelnaga, Marwa Abass, Ahmed A. H. Mosa, Mennatullah A. M. Hussein and Mohamed E. G. Elsayed

Curr. Issues Mol. Biol. 2023, 45(2), 1627-1643; https://doi.org/10.3390/cimb45020105 - 14 Feb 2023

Cited by 8 | Viewed by 2818

Abstract

The current study aimed to determine how palm date aqueous fruit extracts (AFE) affected the autistic-like behaviors brought on by valproic acid (VPA) injection, as well as any potential contributions from Sirt-1, oxidative stress, apoptosis, and autophagy. The pregnant Sprague Dawley females were [...] Read more.

The current study aimed to determine how palm date aqueous fruit extracts (AFE) affected the autistic-like behaviors brought on by valproic acid (VPA) injection, as well as any potential contributions from Sirt-1, oxidative stress, apoptosis, and autophagy. The pregnant Sprague Dawley females were treated with VPA at 12.5th gestation day and pregnant females and their offspring were treated with AFE orally at doses of 4 mg/Kg by gastric gavage for 45 days after birth. The elevated plus-T maze, water maze, and rotarod tests were used to examine autism-like behaviors. At the end of the study, the expression of Nrf2, heme oxygenase (HO-1), Sirt-1, caspase-3 (a marker of apoptosis), LC3 (a marker of autophagy), and NFκB (inflammatory cytokines) were evaluated along with the oxidative stress in brain tissues and the histological changes in the cerebellum and hippocampus. The neurobehavioral assessments significantly declined due to VPA, which also significantly increased oxidative stress in the brain tissues and significantly decreased Nrf2 and HO-1 expression. Additionally, VPA administration caused significant increase in the expression of caspase-3 in the cerebellar cortex, not in the hippocampus; LC3 and NFκB in the hippocampus, not in the cerebellar cortex; and significant reduction in the expression of Sirt-1 in the hippocampus, not in the cerebellum. On the other hand, AFE treatment significantly improved the neurobehavioral changes as well as it improved significantly the oxidative stress and the expression of LC3, NFκB, NrF2, HO-1, and Sirt-1 in the cerebellum and hippocampus. Conclusions: AFE administration might improve the autistic-like symptoms induced by VPA in rats via attenuation of the oxidative stress, upregulation of Nrf2 and HO-1, Sirt-1 and LC3 expression with downregulation of caspase-3, and NFκB expression in the cerebellum and hippocampus. Full article

(This article belongs to the Topic Autism: Molecular Bases, Diagnosis and Therapies)

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14 pages, 2747 KiB

Open AccessArticle

Neuroprotective Effects of Geopung-Chunghyuldan Based on Its Salvianolic Acid B Content Using an In Vivo Stroke Model

by Han-Gyul Lee, Seungwon Kwon, Sang-Kwan Moon, Seung-Yeon Cho, Seong-Uk Park, Woo-Sang Jung, Jung-Mi Park, Chang-Nam Ko and Ki-Ho Cho

Curr. Issues Mol. Biol. 2023, 45(2), 1613-1626; https://doi.org/10.3390/cimb45020104 - 13 Feb 2023

Cited by 6 | Viewed by 2407

Abstract

Background: Geopung-Chunghyuldan (GCD) has neuroprotective properties. Salviae miltiorrhizae Radix plays an essential role in GCD’s effect. The Salviae miltiorrhizae Radix marker compound is salvianolic acid B; however, its content is not uniform among samples. This study aimed to evaluate the neuroprotective effects of [...] Read more.

Background: Geopung-Chunghyuldan (GCD) has neuroprotective properties. Salviae miltiorrhizae Radix plays an essential role in GCD’s effect. The Salviae miltiorrhizae Radix marker compound is salvianolic acid B; however, its content is not uniform among samples. This study aimed to evaluate the neuroprotective effects of GCD based on salvianolic acid B content. Methods: The neuroprotective effects of GCD based on the salvianolic acid B content were evaluated by measuring infarct volume 24 h after permanent middle cerebral artery occlusion in an in vivo stroke model. For the experimental group, each GCD was administered immediately before surgery. The control groups were administered distilled water and aspirin (30 mg/kg) in the same way. The salvianolic acid B content in five types of Salviae Miltiorrhizae Radix (two Chinese and three Korean regions) based on different cultivation regions was analyzed by high-performance liquid chromatography. Results: Three samples met the Korean and Chinese Pharmacopeia standards for salvianolic acid B. However, two samples did not. GCDs with high salvianolic acid B showed marked neuroprotective effects compared to the control groups, whereas GCDs with low salvianolic acid B did not. Conclusions: The salvianolic acid B content of Salviae miltiorrhizae Radix affects the neuroprotection effect of GCD. Stable, raw Salviae miltiorrhizae Radix is essential for GCD homogenization. Full article

(This article belongs to the Special Issue Pathophysiology and Molecular Mechanisms of Acute Stroke)

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Results of high-performance liquid chromatography chromatogram for the salvianolic acid B in five types of Salviae mitiorrhizae Radix. (A) Standardized salvianolic acid B. (B) Salvianolic acid B in C1. (C) Salvianolic acid B in C2. (D) Salvianolic acid B in K1. (E) Salvianolic acid B in K2. (F) Salvianolic acid B in K3. K1, K2, and K3 showed high peak values similar to that of the standardized salvianolic acid B. C1 and C2 showed low peak values. C, Chinese Salviae miltiorrhizae Radix; K, Korean Salviae miltiorrhizae Radix. Full article ">Figure 2
Effects of experimental drug A60+B(C1) 30 mg/kg on ischemic brain injury after permanent middle cerebral artery occlusion (pMCAO). Infarct volume was measured 24 h after pMCAO. DW and all drugs were administered just before pMCAO. (A) Individual infarct volume values for each group (n = 13–17 mice in each group). (B Upper) Representative photographs of infarcted brain slices from DW- and drug-treated mice. (B Lower) There was no significant difference in infarct volume among groups. DW, distilled water; ASA, Aspirin®; A, Ethanol extract of Coptidis Rhizoma, Phellodendri Cortex, Scutellariae Radix, Gardeniae Fructus, and Rhei Rhizoma; B(C1), Ethanol extract of 2018 Chinese Salviae miltiorrhizae Radix and Notoginseng Radix. Data represent means ± standard deviation. Full article ">Figure 3
Effects of experimental drug A60+B(C2) 30 mg/kg on ischemic brain injury after permanent middle cerebral artery occlusion (pMCAO). Infarct volume was measured 24 h after pMCAO. DW and all drugs were administered just before pMCAO. (A) Individual infarct volume values for each group (n = 9–17 mice in each group). (B Upper) Representative photographs of infarcted brain slices from DW- and drug-treated mice. (B Lower) No significant difference in infarct volume was observed among groups. DW, distilled water; ASA, Aspirin®; A, Ethanol extract of Coptidis Rhizoma, Phellodendri Cortex, Scutellariae Radix, Gardeniae Fructus, and Rhei Rhizoma; B(C2), Ethanol extract of 2019 Chinese Salviae miltiorrhizae Radix and Notoginseng Radix. Data represent means ± standard deviation. Full article ">Figure 4
Effects of experimental drug A60+B(K1) 30 mg/kg on ischemic brain injury after permanent middle cerebral artery occlusion (pMCAO). Infarct volume was measured 24 h after pMCAO. DW and all drugs were administered just before pMCAO. (A) Individual infarct volume values for each group (n = 15–19 mice for each group). (B Upper) Representative photographs of infarcted brain slices from DW- and drug-treated mice. (B Lower) The group of experimental drug A+BK1 exhibited a significant decrease in infarct volume after pMCAO compared to groups DW and A60. DW, distilled water; ASA, Aspirin®; A, Ethanol extract of Coptidis Rhizoma, Phellodendri Cortex, Scutellariae Radix, Gardeniae Fructus, and Rhei Rhizoma; B(K1), Ethanol extract of 2019 Yeongyang Korean Salviae miltiorrhizae Radix and Notoginseng Radix. Data represent means ± standard deviation. * p < 0.05 vs. DW; ** p < 0.01 vs. A60 by analysis of variance followed by Tukey’s test. Full article ">Figure 5
Effects of experimental drug A60+B(K2) 30 mg/kg on ischemic brain injury after permanent middle cerebral artery occlusion (pMCAO). Infarct volume was measured 24 h after pMCAO. DW and all drugs were administered just before pMCAO. (A) Individual infarct volume values for each group (n = 15–19 mice for each group). (B Upper) Representative photographs of infarcted brain slices from DW- and drug-treated mice. (B Lower) The group of experimental drug A+BK2 exhibited a significant decrease in infarct volume after pMCAO compared to groups DW and A60. DW, distilled water; ASA, Aspirin®; A, Ethanol extract of Coptidis Rhizoma, Phellodendri Cortex, Scutellariae Radix, Gardeniae Fructus, and Rhei Rhizoma; B(K2), Ethanol extract of 2019 Jangheung Korean Salviae miltiorrhizae Radix and Notoginseng Radix. Data represent means ± standard deviation. * p < 0.05 vs. DW; ** p < 0.01 vs. A60 by analysis of variance followed by Tukey’s test. Full article ">Figure 6
Effects of experimental drug A60+B(K3) 30 mg/kg on ischemic brain injury after permanent middle cerebral artery occlusion (pMCAO). Infarct volume was measured 24 h after pMCAO. DW and all drugs were administered just before pMCAO. (A) Individual infarct volume values for each group (n = 15–19 mice for each group). (B Upper) Representative photographs of infarcted brain slices from DW- and drug-treated mice. (B Lower) The group administered the experimental drug A+BK3 exhibited a significant decrease in infarct volume after pMCAO compared to the A60 group. DW, distilled water; ASA, Aspirin®; A, Ethanol extract of Coptidis Rhizoma, Phellodendri Cortex, Scutellariae Radix, Gardeniae Fructus, and Rhei Rhizoma; B(K3), Ethanol extract of 2020 Korean Salviae miltiorrhizae Radix and Notoginseng Radix. Data represent means ± standard deviation. * p < 0.05 vs. A60 by analysis of variance followed by Tukey’s test. Full article ">

12 pages, 3071 KiB

Open AccessArticle

Isodorsmanin A Prevents Inflammatory Response in LPS-Stimulated Macrophages by Inhibiting the JNK and NF-κB Signaling Pathways

by You Chul Chung, Ami Lee, Jin Ah Ryuk and Youn-Hwan Hwang

Curr. Issues Mol. Biol. 2023, 45(2), 1601-1612; https://doi.org/10.3390/cimb45020103 - 13 Feb 2023

Cited by 4 | Viewed by 1963

Abstract

Natural and synthetic chalcones exhibit anti-inflammatory, antitumoral, antibacterial, antifungal, antimalarial, and antitubercular activities. Isodorsmanin A (IDA), a chalcone, is a well-known constituent of the dried seeds of Psoralea corylifolia L. (PC). Although other constituents of PC have been widely investigated, there are no [...] Read more.

Natural and synthetic chalcones exhibit anti-inflammatory, antitumoral, antibacterial, antifungal, antimalarial, and antitubercular activities. Isodorsmanin A (IDA), a chalcone, is a well-known constituent of the dried seeds of Psoralea corylifolia L. (PC). Although other constituents of PC have been widely investigated, there are no studies on the biological properties of IDA. In this study, we focused on the anti-inflammatory effects of IDA and evaluated its effects on lipopolysaccharide (LPS)-stimulated macrophages. The results showed that IDA suppressed the production of inflammatory mediators (nitric oxide [NO] and prostaglandin E₂ [PGE₂]) and proinflammatory cytokines (tumor necrosis factor-α [TNF-α], interleukin-6 [IL-6], and interleukin-1β [IL-1β]) without cytotoxicity. In addition, it downregulated the mRNA levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) within the treatment concentrations. In our mechanistic studies, IDA inhibited the phosphorylation of the c-Jun N-terminal kinase (JNK), mitogen-activated protein kinase (MAPK), and protected the nuclear factor of the kappa light polypeptide gene enhancer in the B-cells’ inhibitor, alpha (IκB-α), from degradation, thus preventing the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells’ (NF-κB) transcription factor. Our results suggest that IDA is a promising compound for attenuating excessive inflammatory responses. Full article

(This article belongs to the Topic Nitrite and Nitric Oxide in Life)

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14 pages, 3978 KiB

Open AccessArticle

Hesperetin Induces Autophagy and Delayed Apoptosis by Modulating the AMPK/Akt/mTOR Pathway in Human Leukemia Cells In Vitro

by Ching-Yeh Lin, Ya-Hui Chen and Ying-Chih Huang

Curr. Issues Mol. Biol. 2023, 45(2), 1587-1600; https://doi.org/10.3390/cimb45020102 - 13 Feb 2023

Cited by 10 | Viewed by 2897

Abstract

Background: Hesperetin has been reported to have anticancer properties. However, the molecular mechanisms underlying its action on leukemia cells remain unclear. This in vitro study evaluated the possible mechanisms of hesperetin in leukemia cells (HL-60 and U937). Methods: Cell viability was evaluated using [...] Read more.

Background: Hesperetin has been reported to have anticancer properties. However, the molecular mechanisms underlying its action on leukemia cells remain unclear. This in vitro study evaluated the possible mechanisms of hesperetin in leukemia cells (HL-60 and U937). Methods: Cell viability was evaluated using a cell counting kit-8 (CCK-8) assay. Apoptosis and autophagy assays were conducted through annexin V/PI staining and acidic vesicular organelle (AVO) staining. Cell cycle analysis was conducted through propidium iodide (PI) and flow cytometry. The expression of proteins related to apoptosis and autophagy, including cleaved-PARP-1, Bcl-2, Bax, LC3-I/II, Beclin-1, Atg5, p62, phospho-AMPK, AMPK, phospho-mTOR, mTOR, phospho-Akt, and Akt, in human leukemia cells were evaluated using Western blotting. Results: Hesperetin dose-dependently inhibited leukemia cell viability. However, we found a low degree of apoptosis and cell cycle arrest induced by hesperetin in U937 cells. These findings imply the presence of additional mechanisms modulating hesperetin-induced cell death. Next, we evaluated autophagy, the possible mechanism modulating cell death or survival, to clarify the underlying mechanism of hesperetin-induced cell death. Hesperetin also dose-dependently increased the ratio of LC3II/I, Atg5, and Beclin 1 and decreased p62. Moreover, 3-methyladenine (3-MA) and bafilomycin A1 (Baf-A1) inhibited hesperetin-induced autophagy. We suggest that hesperetin can protect cancer cells during the transient period and may extend survival. Furthermore, a decrease in p-mTOR and p-Akt expression and an increase in p-AMPK expression were observed. Collectively, these findings suggest that hesperetin induces autophagy by modulating the AMPK/Akt/mTOR pathway. Conclusion: Hesperetin promoted cell death in the human leukemic cell line U937 by inducing a low degree of slight apoptosis, cell cycle arrest, and autophagy. It is therefore a potential adjuvant to antileukemia therapy and may be combined with other chemotherapeutic drugs to reduce chemoresistance and side effects. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Leukemia)

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17 pages, 4382 KiB

Open AccessArticle

Analysis of the Properties of 44 ABC Transporter Genes from Biocontrol Agent Trichoderma asperellum ACCC30536 and Their Responses to Pathogenic Alternaria alternata Toxin Stress

by Hua-Ying Du, Yu-Zhou Zhang, Kuo Liu, Pei-Wen Gu, Shuang Cao, Xiang Gao, Zhi-Ying Wang, Zhi-Hua Liu and Ze-Yang Yu

Curr. Issues Mol. Biol. 2023, 45(2), 1570-1586; https://doi.org/10.3390/cimb45020101 - 12 Feb 2023

Cited by 7 | Viewed by 2025

Abstract

ATP-binding cassette (ABC) transporters are involved in transporting multiple substrates, such as toxins, and may be important for the survival of Trichoderma when encountering biotic toxins. In this study, genome searching revealed that there are 44 ABC transporters encoded in the genome of [...] Read more.

ATP-binding cassette (ABC) transporters are involved in transporting multiple substrates, such as toxins, and may be important for the survival of Trichoderma when encountering biotic toxins. In this study, genome searching revealed that there are 44 ABC transporters encoded in the genome of Trichoderma asperellum. These ABC transporters were divided into six types based on three-dimensional (3D) structure prediction, of which four, represented by 39 ABCs, are involved in transport and the remaining two, represented by 5 ABCs, are involved in regulating translation. The characteristics of nucleotide-binding domain (NBD) are important in the identification of ABC proteins. Even though the 3D structures of the 79 NBDs in the 44 ABCs are similar, multiple sequence alignment showed they can be divided into three classes. In total, 794 motifs were found in the promoter regions of the 44 ABC genes, of which 541 were cis-regulators related to stress responses. To characterize how their ABCs respond when T. asperellum interact with fungi or plants, T. asperellum was cultivated in either minimal media (MM) control, C-hungry, N-hungry, or poplar medium (PdPap) to simulate normal conditions, competition with pathogens, interaction with pathogens, and interaction with plants, respectively. The results show that 17 of 39 transport ABCs are highly expressed in at least one condition, whereas four of the five translation-regulating ABCs are highly expressed in at least one condition. Of these 21 highly expressed ABCs, 6 were chosen for RT-qPCR expression under the toxin stress of phytopathogen Alternaria alternata, and the results show ABC01, ABC04, ABC05, and ABC31 were highly expressed and may be involved in pathogen interaction and detoxifying toxins from A. alternata. Full article

(This article belongs to the Special Issue Microbial Engineering: Gene Expression Regulation and Its Application)

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Alignment of the 79 NBDs within the 44 ABCs. NBDs were predicted using NCBI BlastP, and amino acid sequences that differ from previous studies are highlighted in yellow. Numbers in names indicate the ABC protein number. The B with the shadow in the names represent the NBD and the position of the peptide. Full article ">Figure 2
Phylogenetic analysis of the 44 ABC proteins. The phylogenetic tree was constructed using the maximum-likelihood method (model: LG + G + I + F) with 1000 bootstrap replicates in the MEGA 7.0 program. The positions of multidomains were drawn in the TBtools program. Full article ">Figure 3
The predicted three-dimensional structures of the 44 ABC transporters in T. asperellum. The prediction was conducted using Clustal Omega (<a href="http://www.ebi.ac.uk/services/proteins" target="_blank">http://www.ebi.ac.uk/services/proteins</a>, accessed on 13 March 2021). The number at the top right corner shows the protein number. The first line below the structure shows the order of conserved domains (A–E represents domains belonging to the ABC-membrane, ABC-ATPase, ABC2-membrane, ABC-tran-2, and acyl-transfer-3 superfamilies, respectively). The second line below the structure shows the multidomain type. The third line below the structure shows the protein molecular weight (kDa). Full article ">Figure 4
Regulatory element-binding motifs in the promoters of the 44 ABC transporter genes in Trichoderma asperellum. Full article ">Figure 5
Heatmap of expression of 44 ABC transporters under four different conditions based on RNA-Seq. C-hungry = MM with 0.5% (w/v) ammonium sulfate, N-hungry = MM with 0.5% (w/v) glucose, and PdPap = variable carbon source in MM containing either 1% (w/v) root powder, 1% (w/v) stem powder, or 1% (w/v) leaf powder from PdPap. The heatmap of genes value = log2(RPKM), where red means high expression and blue means low expression, and the genes are clustered according to their expression. Full article ">Figure 6
Transcription analysis of six ABC genes. Expression of ABC protein in Trichoderma asperellum cultured in MM medium (Control) or toxin-induced medium, MM with 10% fermentation filtrate of Alternaria alternata (Treatment). Differential expression = Treatment − Control. Error bars represent standard deviation. (A): ABC01; (B): ABC02; (C): ABC04; (D): ABC05; (E): ABC30; (F): ABC31 (* p < 0.05, ** p < 0.01, n = 3). Full article ">

2 pages, 174 KiB

Open AccessEditorial

Editorial for Special Issue: “Effects of Nanoparticles on Living Organisms”

by Yoshitaka Miyamoto

Curr. Issues Mol. Biol. 2023, 45(2), 1568-1569; https://doi.org/10.3390/cimb45020100 - 10 Feb 2023

Viewed by 1247

Abstract

This Special Issue provides an overview of the “Effects of Nanoparticles on Living Organisms” [...] Full article

(This article belongs to the Special Issue Effects of Nanoparticles on Living Organisms)

32 pages, 1679 KiB

Open AccessReview

Advances in Molecular Regulation of Prostate Cancer Cells by Top Natural Products of Malaysia

by Jose M. Prieto and Mohd Mukrish Mohd Hanafi

Curr. Issues Mol. Biol. 2023, 45(2), 1536-1567; https://doi.org/10.3390/cimb45020099 - 9 Feb 2023

Cited by 4 | Viewed by 4482

Abstract

Prostate cancer (PCa) remains both a global health burden and a scientific challenge. We present a review of the molecular targets driving current drug discovery to fight this disease. Moreover, the preventable nature of most PCa cases represents an opportunity for phytochemicals as [...] Read more.

Prostate cancer (PCa) remains both a global health burden and a scientific challenge. We present a review of the molecular targets driving current drug discovery to fight this disease. Moreover, the preventable nature of most PCa cases represents an opportunity for phytochemicals as chemopreventive when adequately integrated into nutritional interventions. With a renovated interest in natural remedies as a commodity and their essential role in cancer drug discovery, Malaysia is looking towards capitalizing on its mega biodiversity, which includes the oldest rainforest in the world and an estimated 1200 medicinal plants. We here explore whether the list of top Malay plants prioritized by the Malaysian government may fulfill the potential of becoming newer, sustainable sources of prostate cancer chemotherapy. These include Andrographis paniculate, Centella asiatica, Clinacanthus nutans, Eurycoma longifolia, Ficus deltoidea, Hibiscus sabdariffa, Marantodes pumilum (syn. Labisia pumila), Morinda citrifolia, Orthosiphon aristatus, and Phyllanthus niruri. Our review highlights the importance of resistance factors such as Smac/DIABLO in cancer progression, the role of the CXCL12/CXCR4 axis in cancer metastasis, and the regulation of PCa cells by some promising terpenes (andrographolide, Asiatic acid, rosmarinic acid), flavonoids (isovitexin, gossypin, sinensetin), and alkylresorcinols (labisiaquinones) among others. Full article

(This article belongs to the Special Issue Advances in Molecular Pathogenesis Regulation in Cancer)

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Chemical structure of andrographolide. Full article ">Figure 2
Chemical structure of Asiatic acid, also known as dammarolic acid. Full article ">Figure 3
Chemical structure of vitexin (left) and isovitexin (right). Full article ">Figure 4
Chemical structure of gossypin. Full article ">Figure 5
Chemical structure of cytotoxic alkylresorcinols from M. pumilum. 1-O-methyl-6-acetoxy-5-(pentadec-10Z-enyl)resorcinol (a), labisiaquinone A (b), labisiaquinone B (c), and 1-O-methyl-6-acetoxy-5-pentadecylresorcinol (d). Full article ">Figure 6
Chemical structure of cytotoxic saccharide fatty acid esters (a), asperulosidic acid (b), adamnacanthal (c), and escopoletin (d) present in M. citrifolia L. Full article ">Figure 7
Chemical structure of rosmarinic acid (a) and sinensetin (b) present in O. aristatus. Full article ">

17 pages, 726 KiB

Open AccessReview

The Quest for Neurodegenerative Disease Treatment—Focusing on Alzheimer’s Disease Personalised Diets

by Matei Palimariciuc, Ioana-Miruna Balmus, Bogdan Gireadă, Alin Ciobica, Roxana Chiriță, Alin-Constantin Iordache, Mihai Apostu and Romeo Petru Dobrin

Curr. Issues Mol. Biol. 2023, 45(2), 1519-1535; https://doi.org/10.3390/cimb45020098 - 9 Feb 2023

Cited by 5 | Viewed by 3044

Abstract

Dementia represents a clinical syndrome characterised by progressive decline in memory, language, visuospatial and executive function, personality, and behaviour, causing loss of abilities to perform instrumental or essential activities of daily living. The most common cause of dementia is Alzheimer’s disease (AD), which [...] Read more.

Dementia represents a clinical syndrome characterised by progressive decline in memory, language, visuospatial and executive function, personality, and behaviour, causing loss of abilities to perform instrumental or essential activities of daily living. The most common cause of dementia is Alzheimer’s disease (AD), which accounts for up to 80% of all dementia cases. Despite that extensive studies regarding the etiology and risk factors have been performed in recent decades, and how the current knowledge about AD pathophysiology significantly improved with the recent advances in science and technology, little is still known about its treatment options. In this controverted context, a nutritional approach could be a promising way to formulate improved AD management strategies and to further analyse possible treatment strategy options based on personalised diets, as Nutritional Psychiatry is currently gaining relevance in neuropsychiatric disease treatment. Based on the current knowledge of AD pathophysiology, as well as based on the repeatedly documented anti-inflammatory and antioxidant potential of different functional foods, we aimed to find, describe, and correlate several dietary compounds that could be useful in formulating a nutritional approach in AD management. We performed a screening for relevant studies on the main scientific databases using keywords such as “Alzheimer’s disease”, “dementia”, “treatment”, “medication”, “treatment alternatives”, “vitamin E”, “nutrition”, “selenium”, “Ginkgo biloba”, “antioxidants”, “medicinal plants”, and “traditional medicine” in combinations. Results: nutrients could be a key component in the physiologic and anatomic development of the brain. Several nutrients have been studied in the pursuit of the mechanism triggered by the pathology of AD: vitamin D, fatty acids, selenium, as well as neuroprotective plant extracts (i.e., Ginkgo biloba, Panax ginseng, Curcuma longa), suggesting that the nutritional patterns could modulate the cognitive status and provide neuroprotection. The multifactorial origin of AD development and progression could suggest that nutrition could greatly contribute to the complex pathological picture. The identification of adequate nutritional interventions and the not yet fully understood nutrient activity in AD could be the next steps in finding several innovative treatment options for neurodegenerative disorders. Full article

(This article belongs to the Special Issue Aging, Age-Related Changes in the Brain and the Progression of Alzheimer’s Disease)

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19 pages, 1918 KiB

Open AccessArticle

Changes in the Expression Profile of Pyroptosis-Related Genes in Senescent Retinal Pigment Epithelial Cells after Lutein Treatment

by Barbara Strzalka-Mrozik, Marcel Madej, Natalia Kurowska, Celina Kruszniewska-Rajs, Magdalena Kimsa-Dudek, Jolanta Adamska and Joanna Magdalena Gola

Curr. Issues Mol. Biol. 2023, 45(2), 1500-1518; https://doi.org/10.3390/cimb45020097 - 9 Feb 2023

Cited by 7 | Viewed by 2648

Abstract

Retinal pigment epithelium (RPE) is a specialized structure essential for proper vision, which is constantly exposed to oxidative damage. With aging, this damage accumulates within the RPE cells, causing various diseases, including age-related macular degeneration (AMD). Numerous antioxidant substances are used to prevent [...] Read more.

Retinal pigment epithelium (RPE) is a specialized structure essential for proper vision, which is constantly exposed to oxidative damage. With aging, this damage accumulates within the RPE cells, causing various diseases, including age-related macular degeneration (AMD). Numerous antioxidant substances are used to prevent this process in humans, including lutein. This study aims to determine the differences in the expression patterns of pyroptosis genes in senescent human retinal pigment epithelial cell line ARPE-19 exposed to lutein. Changes in the expression of pyroptosis-related genes were assessed by oligonucleotide microarrays, and the results were validated by real-time RT-qPCR. The microarray analysis showed seven transcripts were differentially expressed both in the H₂O₂-treated cells versus the controls and in the lutein/H₂O₂-treated cells compared to the H₂O₂-treated cells (FC > 2.0). Depending on the used lutein, H₂O₂, or co-treatment of ARPE-19 cells, statistically significant differences in the expression of TXNIP, CXCL8, BAX, and CASP1 genes were confirmed by the RT-qPCR (p < 0.05). A STRING database analysis showed that the proteins encoded by the analyzed genes form a strong interaction network (p < 0.001). These data indicate that lutein modulates the expression level of pyroptosis-related genes, which may be useful for the development of new methods preventing pyroptosis pathway activation in the future. Full article

(This article belongs to the Collection Application of Natural and Pseudo Natural Products in Drug Discovery and Development)

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Cell viability of the ARPE-19 cells that were treated with (a) H2O2 for 30, 60, and 120 min., as well as (b) treated with lutein for 24 h. Each bar represents the mean ± standard deviation (SD), Dunnett’s test, * p < 0.05 versus control (C). Sample size: five wells and three technical replicates for each concetration of H202 and eight wells and three technical replicates for each concetration of lutein. Full article ">Figure 2
Non-treated and H2O2-treated ARPE-19 cells stained for senescence-associated β-galactosidase activity. The cells were treated with H2O2 at a concentration of 400 μM for 60 min. The arrows indicate blue SA-β-gal-positive cells. Full article ">Figure 3
Percentage of SA-β-gal-positive cells in non-treated (C) and H2O2-treated ARPE-19 cells (H). The bars present mean ± standard deviation (SD); Student’s t-test; * p < 0.05. The percent of SA-β-gal-positive cells was counted in ten microscopic fields. Full article ">Figure 4
Changes in the mRNA levels of (a) TP53, (b) CDKN1A and (c) CDKN1B genes in non-treated (C) and H2O2-treated (H) ARPE-19 cells. The box and whisker plots present the mean ± standard error (SE) and standard deviation (SD) of copy numbers per 1 μg of the total RNA; Student’s t-test; * p < 0.05 versus C. Sample size: three biological and three technical replicates for each test group. Full article ">Figure 5
Fluorescence signal intensity maps specific to 128 ID mRNA pyroptosis-related genes specific to the control ARPE--19 cells (C), lutein-treated ARPE-19 cells (L), H2O2--treated ARPE-19 cells (H), and lutein-- and H2O2-treated ARPE-19 cells (LH). Blue—indicates the lowest values and red—indicates the highest values of fluorescence signals. Full article ">Figure 6
Changes in the mRNA levels of (a) TXNIP, (b) CXCL8, (c) BCL2, (d) BAX, (e) CASP1, and (f) CASP9 genes in senescent retinal pigment epithelial cells after treatment with H2O2 (H), lutein (L), co-treated with lutein and H2O2 (LH), and non-treated cells (C). The box and whisker plots present the mean ± standard error (SE) and standard deviation (SD) of copy numbers per 1 μg of the total RNA; Tukey’s post hoc test; * p < 0.05 versus C; # p < 0.05 versus H; ^ p < 0.05 versus LH. Sample size—three biological and three technical replicates for each test group. Full article ">Figure 7
Protein–protein interaction network generated using the STRING database. STRING database—Search Tool for the Retrieval of Interacting Genes/Proteins. Full article ">

17 pages, 4930 KiB

Open AccessArticle

Therapeutic Effects of Aloe saponaria against Ulcerative Colitis Induced by Dextran Sulfate Sodium

by Do Yeong Kweon, Hee Jin Song, Ji Eun Kim, You Jeong Jin, Yu Jeong Roh, Ayun Seol, Ju Min Park, Eun Suk Lee, Won Sik Choi and Dae Youn Hwang

Curr. Issues Mol. Biol. 2023, 45(2), 1483-1499; https://doi.org/10.3390/cimb45020096 - 9 Feb 2023

Cited by 4 | Viewed by 2898

Abstract

Aloe vera (A. vera) has been studied as a treatment option for ulcerative colitis (UC), but there is a lack of scientific evidence showing whether treatment with Aloe saponaria (A. saponaria) can also be beneficial. To investigate the therapeutic [...] Read more.

Aloe vera (A. vera) has been studied as a treatment option for ulcerative colitis (UC), but there is a lack of scientific evidence showing whether treatment with Aloe saponaria (A. saponaria) can also be beneficial. To investigate the therapeutic potential of A. saponaria as a treatment for UC, clinical symptoms, histopathological characteristics of the colon, inflammatory response, and toxicity were analyzed in dextran sulfate sodium (DSS)-induced UC mice after administration of aqueous extracts of A. saponaria (AAS) for 7 days. The total polyphenol and tannin content of AAS was 272 µg/g and 163 µg/g, respectively. AAS exhibited significant antioxidant activity. Several clinical symptoms, including body weight, colon length, and hematochezia, remarkably improved in the DSS+AAS treated group compared to the DSS+Vehicle-treated group. In addition, similar improvements were detected in the histopathological characteristics and mucin-secreting ability in the colon of DSS-induced UC mice after the administration of AAS. The levels of infiltrated inflammatory cells and cytokine expression were significantly decreased in a dose-dependent manner in the colon of the DSS+AAS-treated group. These alterations in inflammatory response were accompanied by a significant recovery of the protein kinase C/extracellular signal-regulated kinase (PKC/ERK) and phosphatidylinositol-3-kinase/serine-threonine protein kinase (PI3K/Akt) signaling pathways. However, the levels of key markers for hepatotoxicity and nephrotoxicity consistently remained between those of the DSS+AAS-treated and the No groups. Therefore, the results of the present study provide novel evidence that AAS may improve the clinical symptoms and attenuate the inflammatory response in DSS-induced UC mice and does not have any significant hepatotoxicity or nephrotoxicity. Full article

(This article belongs to the Special Issue Bioactives and Inflammation)

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12 pages, 2139 KiB

Open AccessCommunication

Proof-of-Concept Analysis of B Cell Receptor Repertoire in COVID-19 Patients Undergoing ECMO by Single-Cell V(D)J and Gene Expression Sequencing

by Alessia Gallo, Nicola Cuscino, Claudia Carcione, Rosalia Busà, Pier Giulio Conaldi and Matteo Bulati

Curr. Issues Mol. Biol. 2023, 45(2), 1471-1482; https://doi.org/10.3390/cimb45020095 - 9 Feb 2023

Viewed by 2608

Abstract

SARS-CoV-2, which causes COVID-19, has altered human activities all over the world and has become a global hazard to public health. Despite considerable advancements in pandemic containment techniques, in which vaccination played a key role, COVID-19 remains a global threat, particularly for frail [...] Read more.

SARS-CoV-2, which causes COVID-19, has altered human activities all over the world and has become a global hazard to public health. Despite considerable advancements in pandemic containment techniques, in which vaccination played a key role, COVID-19 remains a global threat, particularly for frail patients and unvaccinated individuals, who may be more susceptible to developing ARDS. Several studies reported that patients with COVID-19-related ARDS who were treated with ECMO had a similar survival rate to those with COVID-19-unrelated ARDS. In order to shed light on the potential mechanisms underlying the COVID-19 infection, we conducted this proof-of-concept study using single-cell V(D)J and gene expression sequencing of B cells to examine the dynamic changes in the transcriptomic BCR repertoire present in patients with COVID-19 at various stages. We compared a recovered and a deceased COVID-19 patient supported by ECMO with one COVID-19-recovered patient who did not receive ECMO treatment and one healthy subject who had never been infected previously. Our analysis revealed a downregulation of FXYD, HLA-DRB1, and RPS20 in memory B cells; MTATP8 and HLA-DQA1 in naïve cells; RPS4Y1 in activated B cells; and IGHV3-73 in plasma cells in COVID-19 patients. We further described an increased ratio of IgA + IgG to IgD + IgM, suggestive of an intensive memory antibody response, in the COVID ECMO D patient. Finally, we assessed a V(D)J rearrangement of heavy chain IgHV3, IGHJ4, and IGHD3/IGHD2 families in COVID-19 patients regardless of the severity of the disease. Full article

(This article belongs to the Special Issue Next-Generation Sequencing (NGS) Technique and Personalized Medicine)

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The antigen-binding domain of immunoglobulins is composed of two polypeptide chains, namely heavy and light chains. The exons that encode the antigen-binding domains of the heavy chain are assembled from V (variable), D (diversity), and J (joining) gene segments by a process defined as “cut-and-paste” DNA rearrangements. This process, named V(D)J recombination, selects a pair of segments, introduces double-strand breaks next to each segment, deletes the intermediate DNA, and ligates the segments together. Rearrangements take place in a well-ordered way, with D-to-J joining proceeding before a V segment is joined to the rearranged D–J segments. The rearrangement of the light chain is identical except for the absence of D gene fragments. Full article ">Figure 2
Features of B cell subsets. (A) Violin plots show the expression distribution of canonical cell markers in B cell subsets. (B) Proportion of B cell clusters in each sample. (C) t-SNE projection of B cells from all four samples. Each dot corresponds to a single cell, colored according to cell clusters. (D) Heat map representation of differentially expressed genes of B cell populations in the COVID ECMO S, COVID ECMO D, COVID R, and the healthy control. Genes with a maximum adjusted p-value of 0.01 and an absolute value of log2 (fold change) >0.5 were considered to be differentially expressed genes. Full article ">Figure 3
Features of B cell subpopulations. (A) Heat map representation of differentially expressed genes of B cell subpopulations (naïve cells, activated B cells, memory B cells, and plasma cells) of the COVID ECMO S, COVID ECMO D, COVID R, and healthy control. Genes with a maximum adjusted p-value of 0.01 and an absolute value of log2 (fold change) >0.5 were considered to be differentially expressed genes. (B) The graphs show the sequencing results of highly deregulated mRNA in the COVID ECMO S, COVID ECMO D, COVID R, and healthy control. Full article ">Figure 4
(A) t-SNE plot of B cells colored by immunoglobulin heavy chain expression. (B) The proportion of different immunoglobulin isotypes in each sample. (C) The pie plot shows the proportion of clonally expanded B cells in the COVID ECMO S, COVID ECMO D, and COVID R patients and in the healthy control. (D) The distribution of immunoglobulin heavy chain (IGH) CDR3 length. (E) The bubble chart shows the usage frequency of V–J gene combination in each sample. Full article ">

28 pages, 1156 KiB

Open AccessReview

Multiple Sclerosis: Inflammatory and Neuroglial Aspects

by Giulio Papiri, Giordano D’Andreamatteo, Gabriella Cacchiò, Sonila Alia, Mauro Silvestrini, Cristina Paci, Simona Luzzi and Arianna Vignini

Curr. Issues Mol. Biol. 2023, 45(2), 1443-1470; https://doi.org/10.3390/cimb45020094 - 8 Feb 2023

Cited by 36 | Viewed by 5501

Abstract

Multiple sclerosis (MS) represents the most common acquired demyelinating disorder of the central nervous system (CNS). Its pathogenesis, in parallel with the well-established role of mechanisms pertaining to autoimmunity, involves several key functions of immune, glial and nerve cells. The disease’s natural history [...] Read more.

Multiple sclerosis (MS) represents the most common acquired demyelinating disorder of the central nervous system (CNS). Its pathogenesis, in parallel with the well-established role of mechanisms pertaining to autoimmunity, involves several key functions of immune, glial and nerve cells. The disease’s natural history is complex, heterogeneous and may evolve over a relapsing-remitting (RRMS) or progressive (PPMS/SPMS) course. Acute inflammation, driven by infiltration of peripheral cells in the CNS, is thought to be the most relevant process during the earliest phases and in RRMS, while disruption in glial and neural cells of pathways pertaining to energy metabolism, survival cascades, synaptic and ionic homeostasis are thought to be mostly relevant in long-standing disease, such as in progressive forms. In this complex scenario, many mechanisms originally thought to be distinctive of neurodegenerative disorders are being increasingly recognized as crucial from the beginning of the disease. The present review aims at highlighting mechanisms in common between MS, autoimmune diseases and biology of neurodegenerative disorders. In fact, there is an unmet need to explore new targets that might be involved as master regulators of autoimmunity, inflammation and survival of nerve cells. Full article

(This article belongs to the Special Issue Molecular Mechanisms in Demyelinating Disorders of the Central Nervous System)

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21 pages, 6442 KiB

Open AccessArticle

Synthesis, Molecular Docking, and Dynamic Simulation Targeting Main Protease (Mpro) of New, Thiazole Clubbed Pyridine Scaffolds as Potential COVID-19 Inhibitors

by Adel Alghamdi, Amr S. Abouzied, Abdulwahab Alamri, Sirajudheen Anwar, Mukhtar Ansari, Ibrahim Khadra, Yasser H. Zaki and Sobhi M. Gomha

Curr. Issues Mol. Biol. 2023, 45(2), 1422-1442; https://doi.org/10.3390/cimb45020093 - 7 Feb 2023

Cited by 42 | Viewed by 3329

Abstract

Many biological activities of pyridine and thiazole derivatives have been reported, including antiviral activity and, more recently, as COVID-19 inhibitors. Thus, in this paper, we designed, synthesized, and characterized a novel series of N-aminothiazole-hydrazineethyl-pyridines, beginning with a N′-(1-(pyridine-3-yl)ethylidene)hydrazinecarbothiohydrazide derivative and various [...] Read more.

Many biological activities of pyridine and thiazole derivatives have been reported, including antiviral activity and, more recently, as COVID-19 inhibitors. Thus, in this paper, we designed, synthesized, and characterized a novel series of N-aminothiazole-hydrazineethyl-pyridines, beginning with a N′-(1-(pyridine-3-yl)ethylidene)hydrazinecarbothiohydrazide derivative and various hydrazonoyl chlorides and phenacyl bromides. Their Schiff bases were prepared from the condensation of N-aminothiazole derivatives with 4-methoxybenzaldehyde. FTIR, MS, NMR, and elemental studies were used to identify new products. The binding energy for non-bonding interactions between the ligand (studied compounds) and receptor was determined using molecular docking against the SARS-CoV-2 main protease (PDB code: 6LU7). Finally, the best docked pose with highest binding energy (8a = −8.6 kcal/mol) was selected for further molecular dynamics (MD) simulation studies to verify the outcomes and comprehend the thermodynamic properties of the binding. Through additional in vitro and in vivo research on the newly synthesized chemicals, it is envisaged that the achieved results will represent a significant advancement in the fight against COVID-19. Full article

(This article belongs to the Special Issue Drug Development and Repositioning Methodology on COVID-19)

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15 pages, 18625 KiB

Open AccessArticle

Urinary Proteome Differences in Patients with Type 2 Diabetes Pre and Post Liraglutide Treatment

by Mohamed Rafiullah, Hicham Benabdelkamel, Afshan Masood, Aishah A. Ekhzaimy, Mohthash Musambil, Salini Scaria Joy and Assim A. Alfadda

Curr. Issues Mol. Biol. 2023, 45(2), 1407-1421; https://doi.org/10.3390/cimb45020092 - 6 Feb 2023

Cited by 8 | Viewed by 2401

Abstract

Diabetes mellitus is a chronic multisystem disease with a high global prevalence. The glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide is known to lower glucose levels and reduce weight. However, the mechanisms underlying the benefits of liraglutide treatment in patients with type 2 diabetes [...] Read more.

Diabetes mellitus is a chronic multisystem disease with a high global prevalence. The glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide is known to lower glucose levels and reduce weight. However, the mechanisms underlying the benefits of liraglutide treatment in patients with type 2 diabetes mellitus (T2DM) remain unclear. Twelve male patients with T2DM (pre and post liraglutide treatment) and HbA1c between 8% and 11% were recruited. In the present study, a two-dimensional difference gel electrophoresis (2D-DIGE) matrix-assisted laser desorption/ionization-time of flight (MALDI TOF) mass spectrometric approach combined with bioinformatics and network pathway analysis was used to explore the urine proteomic profile. The mean age of the patients was 52.4 ± 7.5 years. After treatment with liraglutide, a statistically significant change (p < 0.006) was observed in HbA1c with no significant changes in body weight or markers of dyslipidemia. Two-dimensional difference gel electrophoresis identified significant changes (≥1.5-fold change, ANOVA, p ≤ 0.05) in 32 proteins (4 down- and 28 upregulated) in liraglutide post treatment compared to the pre-treatment state. Albumin, serotransferrin, metallothionein-2 (MT-2), and keratins K1 and K10 were found to be upregulated after liraglutide treatment. The patients showed significant improvement in glycemic control after the 12-week treatment with liraglutide. The renoprotective effect of liraglutide may be linked to the increased urinary abundance of MT-2 and the decreased abundance of zinc alpha 2-glycoprotein (ZAG) and Alpha-1 antitrypsin (α1-AT). More studies are needed to elucidate the molecular mechanisms behind the renoprotective effects of liraglutide. Full article

(This article belongs to the Special Issue Natural Products as Potential Sources of Antidiabetic Compounds)

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Representative fluorescent protein profile of a two-dimensional difference in gel electrophoresis (2D-DIGE) containing urine sample from post-treatment with liraglutide labeled with Cy3 (A), pre-treatment labeled with Cy5 (B), a pooled internal control labeled with Cy2 (C), and a merged image of PRE/POST (D). Full article ">Figure 2
2D-DIGE numbered spots indicate proteins with differential abundance (defined as fold-change ≥ 1.5, p ≤ 0.05) between pre-treated and post-treated samples successfully identified with matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) mass spectrometry (MS). MW, protein molecular weight; pI, isoelectric point. Full article ">Figure 3
Principal component analysis of the proteomic dataset. Pink dots denote the 12 liraglutide pre-treated urine samples, and blue dots indicate the 12 liraglutide post-treated samples. Together, these explained 60.95% of the selected spots’ variability values. Colored dots and numbers represent gels and spots, respectively. Full article ">Figure 4
The differentially expressed proteins’ most enriched interaction network in both the pre- and post-treatment phases. Protein nodes with a blue or red halo denote upregulation or downregulation, respectively. Protein nodes without a halo are proposed by the STRING database and indicate potential targets that were functionally coordinated with the differentially expressed proteins. The solid black line displays co-expression; the green line indicates the gene neighborhood; the dark-blue line indicates gene co-occurrence; the purple line shows experimentally determined protein interactions. Interaction network of differentially expressed proteins in the pretreatment group compared to the post-treatment group. Full article ">Figure 5
Comparative representation (%) of reported proteins divided into groups based on their cellular components (A), molecular functions (B), and biological processes (C). Full article ">

11 pages, 5176 KiB

Open AccessArticle

Chloride Intracellular Channel Protein 1 Expression and Angiogenic Profile of Liver Metastasis of Digestive Origin

by Amalia Raluca Ceausu, Alexandru Ciolofan, Alexandru Blidisel, Andrei Alexandru Cosma, Pusa Nela Gaje and Octavian Cretu

Curr. Issues Mol. Biol. 2023, 45(2), 1396-1406; https://doi.org/10.3390/cimb45020091 - 6 Feb 2023

Viewed by 1808

Abstract

Chloride intracellular channel 1 (CLIC1) is involved in cell migration and metastasis. The histological growth patterns of liver metastasis are as follows: desmoplastic (d-HGP), replacement (r-HGP), pushing (p-HGP), and mixed. The aim of this study was to evaluate the relation between HGP, angiogenesis, [...] Read more.

Chloride intracellular channel 1 (CLIC1) is involved in cell migration and metastasis. The histological growth patterns of liver metastasis are as follows: desmoplastic (d-HGP), replacement (r-HGP), pushing (p-HGP), and mixed. The aim of this study was to evaluate the relation between HGP, angiogenesis, and CLIC1 expression. Materials and Methods: A total of 40 cases of primary tumors and their LM: d-HGP (12 cases), r-HGP (13 cases), and p-HGP (15 cases), were evaluated through simple and double immunostaining. CLIC1 assessment was conducted as follows: scores of 0 (less than 10% of positive cells), 1 (10–30%), 2 (30–50%), or 3 (more than 50%) were assigned. Heterogeneous CLIC1 expression was found. CLIC1 in primary tumors correlated with grade G for all cases of LM with a p-HGP (p = 0.004). The CLIC1 score for LMs with an r-HGP correlated with grade G of the corresponding primary tumor (p = 0.027). CLIC1 and CD34+/Ki67+ vessels (p = 0.006) correlated in primary tumors. CLIC1 in primary tumors correlated with CD34+/Ki67+ vessels of LMs with a d HGP (p = 0.024). Conclusions: The CLIC1 score may have prognostic value, mainly for LMs with a p-HGP and r-HGP, and therapeutic value for LMs with a d-HGP. Full article

(This article belongs to the Special Issue Molecular-Based Approaches in Therapy for Gastrointestinal Cancers)

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CD 34 immunoexpression, poorly differentiated colon adenocarcinomas, intratumoral vessels, ×200 magnification (A,B), and corresponding metastasis (p-HGP), ×200 magnification (C). Full article ">Figure 2
CD34 immunoexpression in moderately differentiated colon adenocarcinoma, ×200 magnification (A), and corresponding liver metastasis, replacement HGP, ×100 magnification (B). Full article ">Figure 3
CLIC1 immunoexpression, colorectal adenocarcinoma, score of 2, ×400 magnification (A), and LM, desmoplastic HGP, score of 2, ×200 magnification (B). Full article ">Figure 4
CLIC1 immunoexpression, gastric adenocarcinoma, score of 2, ×400 magnification (A), and corresponding LM, pushing HGP, score of 3, ×200 magnification (B). CLIC1 expression intensity in the cytoplasm (purple arrow <img alt="Cimb 45 00091 i001" src="/cimb/cimb-45-00091/article_deploy/html/images/cimb-45-00091-i001.png"/>) increased in the nucleus (blue arrow <img alt="Cimb 45 00091 i002" src="/cimb/cimb-45-00091/article_deploy/html/images/cimb-45-00091-i002.png"/>) of the cell. Full article ">Figure 5
CLIC1 immunoexpression, pancreatic adenocarcinoma, score of 3, ×200 magnification (A) and corresponding LM, replacement HGP, score of 3, ×400 magnification (B). Full article ">Figure 6
The overall survival in CLIC1 positive cases of LM, divided in three groups (A = d-HGP; B = p-HGP and C = r-HGP). Full article ">Figure 7
CLIC1 alteration from TCGA database in gastric adenocarcinoma (A), pancreatic adenocarcinoma (B) and colorectal adenocarcinoma (C). CLIC1 expression and overall survival interrelation (D). Full article ">

9 pages, 606 KiB

Open AccessArticle

In Silico Integrated Analysis of Genomic, Transcriptomic, and Proteomic Data Reveals QTL-Specific Genes for Bacterial Canker Resistance in Tomato (Solanum lycopersicum L.)

by Ibrahim Celik

Curr. Issues Mol. Biol. 2023, 45(2), 1387-1395; https://doi.org/10.3390/cimb45020090 - 6 Feb 2023

Cited by 2 | Viewed by 1965

Abstract

Bacterial canker of tomato, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), is a devasting disease that leads to significant yield losses. Although QTLs originating from three wild species (Solanum arcanum, S. habrochaites, and S. pimpinellifolium) were identified, [...] Read more.

Bacterial canker of tomato, caused by Clavibacter michiganensis subsp. michiganensis (Cmm), is a devasting disease that leads to significant yield losses. Although QTLs originating from three wild species (Solanum arcanum, S. habrochaites, and S. pimpinellifolium) were identified, none of the QTLs was annotated for candidate gene identification. In the present study, a QTL-based physical map was constructed to reveal the meta-QTLs for Cmm resistance. As a result, seven major QTLs were mapped. Functional annotation of QTLs revealed 48 candidate genes. Additionally, experimentally validated Cmm resistance-related genes based on transcriptomic and proteomic studies were mapped in the genome and 25 genes were found to be located in the QTL regions. The present study is the first report to construct a physical map for Cmm resistance QTLs and identify QTL-specific candidate genes. The candidate genes identified in the present study are valuable targets for fine mapping and developing markers for marker-assisted selection in tomatoes for Cmm resistance breeding. Full article

(This article belongs to the Special Issue Functional Genomics and Comparative Genomics Analysis in Plants)

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14 pages, 1549 KiB

Open AccessReview

Carbonic Anhydrase II Activators in Osteopetrosis Treatment: A Review

by Zikra Alkhayal, Zakia Shinwari, Ameera Gaafar and Ayodele Alaiya

Curr. Issues Mol. Biol. 2023, 45(2), 1373-1386; https://doi.org/10.3390/cimb45020089 - 6 Feb 2023

Cited by 7 | Viewed by 3270

Abstract

Osteopetrosis is a rare hereditary illness generated by failure in osteoclasts resulting in elevated bone densities. Patients with osteopetrosis possess several complications, like dental caries, earlier teeth loss, delayed eruption, malformed crowns and roots, and lamina dura thickening. Since deficiency of carbonic anhydrase [...] Read more.

Osteopetrosis is a rare hereditary illness generated by failure in osteoclasts resulting in elevated bone densities. Patients with osteopetrosis possess several complications, like dental caries, earlier teeth loss, delayed eruption, malformed crowns and roots, and lamina dura thickening. Since deficiency of carbonic anhydrase II is a major cause behind osteopetrosis, carbonic anhydrase II activators have a large number of applications in osteopetrosis treatment. There is a lack of a comprehensive review on osteopetrosis, pathogenesis of dental abnormalities, and the role of carbonic anhydrase II activators in osteopetrosis treatment. To address this research gap, the authros perfomed a comprehensive review on osteopetrosis and its types, pathogenesis of dental abnormalities, and the role of carbonic anhydrase II activators in osteopetrosis treatment. A brief introduction to the pathogenesis of dental abnormalities and regeneration is provided in this survey. A discussion of types of osteopetrosis depending on genetic inheritance, such as autosomal dominant, autosomal recessive, and X-linked inheritance osteopetrosis, is presented in this survey. The paper also focuses on the importance of carbonic anhydrase II activators as a potential drug therapy for dental osteopetrosis. In addition, a brief note on the role of azole and fluconazole in treating osteopetrosis is given. Finally, future directions involving gene therapy for dental osteopetrosis are described. Full article

(This article belongs to the Collection Feature Papers in Current Issues in Molecular Biology)

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24 pages, 6647 KiB

Open AccessArticle

Development and Optimization of Label-Free Quantitative Proteomics under Different Crossing Periods of Bottle Gourd

by Anurag Malik, Virender Singh Mor, Himani Punia, D. S. Duhan, Jayanti Tokas, Axay Bhuker, Mohammed Nasser Alyemeni and Awais Shakoor

Curr. Issues Mol. Biol. 2023, 45(2), 1349-1372; https://doi.org/10.3390/cimb45020088 - 6 Feb 2023

Cited by 3 | Viewed by 2561

Abstract

Bottle gourd, a common vegetable in the human diet, has been valued for its medicinal and energetic properties. In this experiment, the time-resolved analysis of the changes in the proteins’ electrophoretic patterning of the seed development at different crossing periods was studied in [...] Read more.

Bottle gourd, a common vegetable in the human diet, has been valued for its medicinal and energetic properties. In this experiment, the time-resolved analysis of the changes in the proteins’ electrophoretic patterning of the seed development at different crossing periods was studied in bottle gourd using label-free quantitative proteomics. Hybrid HBGH-35 had the highest observed protein levels at the 4th week of the crossing period (F₄) compared to the parental lines, viz. G-2 (M) and Pusa Naveen (F). The crossing period is significantly correlated with grain filling and reserve accumulation. The observed protein expression profile after storage was related to seed maturation and grain filling in bottle gourds. A total of 2517 proteins were identified in differentially treated bottle gourd fruits, and 372 proteins were differentially expressed between different crossing periods. Proteins related to carbohydrate and energy metabolism, anthocyanin biosynthesis, cell stress response, and fruit firmness were characterized and quantified. Some proteins were involved in the development, while others were engaged in desiccation and the early grain-filling stage. F4 was distinguished by an increase in the accumulation of low molecular weight proteins and enzymes such as amylase, a serine protease, and trypsin inhibitors. The seed vigor also followed similar patterns of differential expression of seed storage proteins. Our findings defined a new window during seed production, which showed that at F₄, maximum photosynthetic assimilates accumulated, resulting in an enhanced source–sink relationship and improved seed production. Our study attempts to observe the protein expression profiling pattern under different crossing periods using label-free quantitative proteomics in bottle gourd. It will facilitate future detailed investigation of the protein associated with quality traits and the agronomic importance of bottle gourd through selective breeding programs. Full article

(This article belongs to the Special Issue Genetic Sight: Plant Traits during Postharvest)

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16 pages, 2238 KiB

Open AccessArticle

Ozoile Reduces the LPS-Induced Inflammatory Response in Colonic Epithelial Cells and THP-1 Monocytes

by Maria Paola Bertuccio, Valentina Rizzo, Salvatore Arena, Alessandra Trainito, Angela Simona Montalto, Daniela Caccamo, Monica Currò, Carmelo Romeo and Pietro Impellizzeri

Curr. Issues Mol. Biol. 2023, 45(2), 1333-1348; https://doi.org/10.3390/cimb45020087 - 5 Feb 2023

Cited by 12 | Viewed by 2997

Abstract

Inappropriate activation of immune functions in intestinal epithelial cells can lead to inflammation that is characterized also by infiltration into intestinal tissue of monocytes/macrophages. Current therapies for intestinal inflammation include anti-inflammatory, immunosuppressive and biological drugs. Ozoile (stable ozonides) has been reported to exert [...] Read more.

Inappropriate activation of immune functions in intestinal epithelial cells can lead to inflammation that is characterized also by infiltration into intestinal tissue of monocytes/macrophages. Current therapies for intestinal inflammation include anti-inflammatory, immunosuppressive and biological drugs. Ozoile (stable ozonides) has been reported to exert anti-inflammatory effects. However, ozonated oil has been used mainly for topical applications and no data are available about its effects on intestinal cells or immune cells. In this study, we evaluated Ozoile effects on human HT-29 colonic cells and THP-1 monocytic cells stimulated with LPS to induce inflammation. HT-29 and THP-1 cells were treated with LPS in the presence/absence of Ozoile for 4 h. Biomarkers of inflammation, some members of tight junctions and the adhesion molecule ICAM were assessed by qRT-PCR. Protein expression was analyzed by Western blotting. The release of TNF-α and IL-1β was measured by ELISA. In HT-29, Ozoile inhibited LPS-induced expression of TNF-α, IL-1β, ZO-1, CLDN1, NOS2 and MMP-2 and increased the expression of Nrf2 and SOD2 antioxidant proteins. In THP-1 cells, the LPS induction of TNF-α, IL-1β and ICAM was counteracted by Ozoile treatment. Our in vitro results demonstrate the effectiveness of Ozoile in reducing the inflammatory response in intestinal and monocytic cells. Further in vivo studies are necessary to confirm its possible use for intestinal inflammatory conditions. Full article

(This article belongs to the Special Issue Bioactives and Inflammation)

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Figure 1

Figure 1
Effects of Ozoile (OZ) on HT-29 (A) and THP-1 (B) cell viability in the presence or absence of LPS (1 µg/mL). Different concentrations of Ozoile (0.1, 0.5, 1 and 2 mg/mL) were added to the culture medium 30 min before LPS treatment (1 µg/mL for 4 h), and then cell viability was assessed by the MTT test. Results are expressed as percentages relative to untreated cells. Data are means ± SEM from five independent experiments. Full article ">Figure 2
Effects of Ozoile (OZ) on cytokine gene expression (A) and release (B,C) in HT-29 cells stimulated with LPS. (A) HT-29 cells were treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL) 30 min before LPS treatment (1 µg/mL for 4 h). The mRNA transcript levels of TNF-α and IL-1β were analyzed by real-time PCR. (B,C) Cytokine release into culture medium was measured by ELISA. Data are expressed as means ± SEM from three independent experiments. * p < 0.05, ** p < 0.01 and *** p < 0.001 indicate significant differences vs. control cells. § p < 0.05, §§ p < 0.01 and §§§ p < 0.001 indicate significant differences vs. LPS-treated cells. Full article ">Figure 3
Effects of Ozoile (OZ) on IL-4 gene expression in HT-29 cells stimulated with LPS. HT-29 cells were treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL) 30 min before LPS exposure (1 µg/mL for 4 h). The mRNA transcript levels were analyzed by real-time PCR. Data are expressed as means ± SEM from three independent experiments. * p < 0.05 indicates a significant difference vs. control cells. § p < 0.05 indicates a significant difference vs. LPS-treated cells. Full article ">Figure 4
Effects of Ozoile (OZ) on ZO-1 and CLDN1 gene expression in HT-29 cells stimulated with LPS. HT-29 cells were treated with 1 mg/mL of Ozoile 30 min before LPS treatment (1 µg/mL for 4 h). The mRNA transcript levels were analyzed by real-time PCR. Data are expressed as means ± SEM from three independent experiments. * p < 0.05 and *** p < 0.001 indicate significant differences vs. control cells. §§ p < 0.01 and §§§ p <0.001 indicate significant differences vs. LPS-treated cells. Full article ">Figure 5
Effects of Ozoile (OZ) on NOS2 (A) and MMP2 (B) gene expression in HT-29 cells stimulated with LPS. HT-29 cells were treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL) 30 min before LPS treatment (1 µg/mL for 4 h). The mRNA transcript levels of NOS2 and MMP2 were analyzed by real-time PCR. Data are expressed as means ± SEM from three independent experiments. *** p < 0.001 indicates a significant difference vs. control cells. §§§ p < 0.001 indicates a significant difference vs. LPS-treated cells. Full article ">Figure 6
Evaluation of NOS2, MMP2, SOD2 (A) and Nrf2 (E) protein expression by Western blotting in HT-29 cells treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL), added 30 min before LPS treatment (1 µg/mL for 4 h). Densitometric analysis of NOS2 (B), MMP2 (C) and SOD2 (D) after normalization against β-actin and of Nrf2 (F) after normalization against laminin. The results are expressed as means ± SEM from three independent experiments. * p < 0.05 and ** p < 0.01 indicate significant differences vs. control cells. Full article ">Figure 7
Effects of Ozoile (OZ) on TNF-α and IL-1β gene expression (A) and release (B,C) in THP-1 cells stimulated with LPS. (A) THP-1 cells were treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL) before LPS treatment (1 µg/mL for 4 h). The mRNA transcript levels were analyzed by real-time PCR. (B,C) Cytokine release into the culture medium was measured by ELISA. Data are expressed as means ± SEM from three independent experiments. ** p < 0.01 and *** p < 0.001 indicate significant differences vs. control cells. §§§ p < 0.001 indicates a significant difference vs. LPS-treated cells. Full article ">Figure 8
Effects of Ozoile (OZ) on IL-4 gene expression in THP-1 cells stimulated with LPS. THP-1 cells were treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL) 30 min before LPS exposure (1 µg/mL for 4 h). The mRNA transcript levels were analyzed by real-time PCR. Data are expressed as means ± SEM from three independent experiments. ** p < 0.01 indicates a significant difference vs. control cells. § p < 0.05 and §§§ p < 0.001 indicate significant differences vs. LPS-treated cells. Full article ">Figure 9
Effects of Ozoile (OZ) on ICAM gene expression in THP-1 cells stimulated with LPS. THP-1 cells were treated with different concentrations of Ozoile (0.1, 0.5 and 1 mg/mL) before LPS treatment (1 µg/mL for 4 h). The mRNA transcript levels were analyzed by real-time PCR. Data are expressed as means ± SEM from three independent experiments. *** p < 0.001 indicates a significant difference vs. control cells. § p < 0.05 and §§ p < 0.01 indicate significant differences vs. LPS-treated cells. Full article ">

19 pages, 6027 KiB

Open AccessArticle

Network Pharmacological Analysis of a New Herbal Combination Targeting Hyperlipidemia and Efficacy Validation In Vitro

by Tae-Hyoung Kim, Ga-Ram Yu, Hyuck Kim, Jai-Eun Kim, Dong-Woo Lim and Won-Hwan Park

Curr. Issues Mol. Biol. 2023, 45(2), 1314-1332; https://doi.org/10.3390/cimb45020086 - 4 Feb 2023

Cited by 8 | Viewed by 3425

Abstract

The network pharmacology (NP) approach is a valuable novel methodology for understanding the complex pharmacological mechanisms of medicinal herbs. In addition, various in silico analysis techniques combined with the NP can improve the understanding of various issues used in natural product research. This [...] Read more.

The network pharmacology (NP) approach is a valuable novel methodology for understanding the complex pharmacological mechanisms of medicinal herbs. In addition, various in silico analysis techniques combined with the NP can improve the understanding of various issues used in natural product research. This study assessed the therapeutic effects of Arum ternata (AT), Poria cocos (PC), and Zingiber officinale (ZO) on hyperlipidemia after network pharmacologic analysis. A protein–protein interaction (PPI) network of forty-one key targets was analyzed to discover core functional clusters of the herbal compounds. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and gene ontology (GO) term enrichment analysis identified significant categories of hypolipidemic mechanisms. The STITCH database indicated a high connection with several statin drugs, deduced by the similarity in targets. AT, PC, and ZO regulated the genes related to the energy metabolism and lipogenesis in HepG2 cells loaded with free fatty acids (FFAs). Furthermore, the mixture of three herbs had a combinational effect. The herbal combination exerted superior efficacy compared to a single herb, particularly in regulating acetyl-CoA carboxylase (ACC) and carnitine palmitoyltransferase 1 (CPT-1). In conclusion, the network pharmacologic approach was used to assess potential targets of the herbal combination for treatment. Experimental data from FFA-induced HepG2 cells suggested that the combination of AT, PC, and ZO might attenuate hyperlipidemia and its associated hepatic steatosis. Full article

(This article belongs to the Special Issue Bioactive Compounds of Natural Products on Metabolic Disorders and Complications)

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Figure 1

Figure 1
Venn diagrams of the compounds and target genes from the herbal combination. (A) Distribution of potential compounds from PC, AT, and ZO. (B) Target of potential form PC, AT, and ZO (C) Hyperlipidemia-related target genes from two web databases. (D) Dyslipidemia-related common target genes from both web databases and TCMSP. Full article ">Figure 2
(A) Full protein–protein interaction network of 41 hyperlipidemia-related target genes from the herbal combination. (B,C) Functional cluster of full PPI network created from MCODE of Cytoscape. Full article ">Figure 3
Visualization of the compounds–targets–pathways network constructed with the components of the herbal combination. Full article ">Figure 4
Visualization of the KEGG pathways and targets of the herbal combination analyzed by ClueGO and CluePedia. (A) Visualization of significantly enriched KEGG pathways and their genes (created with ClueGO) (B) Cerebral layout of significant KEGG pathways and their genes by their cellular compartment (created with CluePedia). Full article ">Figure 5
Bubble plot visualization of enrichment analysis (BP terms and KEGG pathways) from two target clusters from the herbal combination. Significant (A) BP terms and (B) KEGG pathways of target cluster 1. Significant (C) BP terms and (D) KEGG pathways of target cluster 2. Full article ">Figure 6
Visualization of the target–chemical interaction of targets from cluster 1 using the STITCH database. Full article ">Figure 7
AT, PC, ZO, and MIX stimulate ACC, AMPK, and AKT phosphorylation in FFA-induced hepatic steatosis and activate CPT-1 related to fatty acid oxidation. HepG2 cells were incubated in the absence or presence of FFAs (1 mM) with AT, PC, ZO, and MIX for 24 h. (A) HepG2 cells were treated with various concentrations (0–50 µg/mL) of AT, PC, ZO, and MIX for 24 h. The results are presented as means ± SDs of the percentages determined by three independent experiments versus the non-treated controls. (B) HepG2 cells were co-treated with each sample and FFAs for 24 h. Western blot analysis shows the effect on the phosphorylation of ACC and AMPK and AKT protein expression related to the energy metabolism and lipogenesis in FFA-induced hepatic steatosis HepG2 cells. The band intensities were measured by densitometry and normalized versus the intensities of the total forms and β-actin. The results are presented as the means ± SDs of three independent experiments. # p < 0.05 versus FFA-treated controls, and * p < 0.05, ** p < 0.01 versus FFA-treated HepG2 cells. Full article ">Figure 8
AT, PC, ZO, and MIX ameliorate lipogenesis in FFA-induced hepatic steatosis HepG2 cells. HepG2 cells were incubated in the absence or presence of FFAs (1 mM) with AT, PC, ZO, and MIX for six or 48 h. (A) Relative expressions of CEBPA, PPARγ, ACC, and HMGCR genes were determined by qPCR. (B) The impact on lipid accumulation was evaluated by measuring Oil Red O staining and comparing the microscopic images. Representative images (100×) of lipid accumulation via Oil Red O staining in HepG2 cells under different conditions. The results are presented as the means ± SDs of three independent experiments. # p < 0.05, ## p < 0.01 versus the FFA-treated controls, and * p < 0.05, ** p < 0.01 versus FFA-treated HepG2 cells. Full article ">

8 pages, 2599 KiB

Open AccessArticle

Prophylaxis of Antifungal Drugs against Systemic Fungemia induced by Oral Candidiasis in Mice

by Kazunori Ninomiya, Hiroki Katagiri, Hajime Hara, Kayoko Fukui, Maiko Haga-Tsujimura, Ken Nakahara and Kenjirou Nakamura

Curr. Issues Mol. Biol. 2023, 45(2), 1306-1313; https://doi.org/10.3390/cimb45020085 - 4 Feb 2023

Cited by 1 | Viewed by 1764

Abstract

Oral mucositis is highly prevalent among the elderly, for whom oral care is often difficult. Oral mucositis, such as candidiasis, can induce systemic fungemia. Antifungal prophylaxis may be useful in such cases to prevent systemic fungemia; however, studies on this are limited. The [...] Read more.

Oral mucositis is highly prevalent among the elderly, for whom oral care is often difficult. Oral mucositis, such as candidiasis, can induce systemic fungemia. Antifungal prophylaxis may be useful in such cases to prevent systemic fungemia; however, studies on this are limited. The objective of this study was to demonstrate the effectiveness of antifungal prophylaxis to prevent systemic Candida dissemination compared to oral care using a mice model. Oral candidiasis was induced using chemotherapy and inoculation with C. albicans in 8-week-old male mice. Group A was given oral care, Group B was orally administered an antifungal drug, Group C was intravenously administered an antifungal drug, and Group D was used as the negative control group. Macroscopic features of the tongue surface, colony forming units (CFU) on the tongue, and blood culture for C. albicans were evaluated. CFU was significantly higher in Group A than in Groups B and C. The oral care group, but not the groups administered antifungal agents, showed significantly higher positive numbers of animals with C. albicans in the blood as compared to the control group, indicating the effectiveness of antifungal prophylaxis over oral care. Antifungal prophylaxis may be an option for the prevention of systemic fungemia in individuals with difficulty in oral care. Full article

(This article belongs to the Special Issue Advances in Research on Molecular Oral Microorganisms)

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Figure 1

Figure 1
Experimental schedule. Full article ">Figure 2
Macroscopic pictures of the tongue. (a) Group A: The presence of clinical oral candidiasis and food residue (circle). (b) Group B: A decrease in white patches on the tongue was observed compared to Group A. (c) Group C: The oral mucositis was decreased as compared to Group A and Group B. (d) Group D: The tongue of the negative control group. Full article ">Figure 3
Colony forming Unit (CFU) counts in the tongue. The CFU counts were obtained in Group A. Between Groups A and B, Groups A and C were statistically significant (** p < 0.01). There was no significant difference between Groups B and C. Full article ">Figure 4
Positive results of multiplex PCR for blood cultures. The 615 bp band specifically identifies Candida albicans among Candida spp. Lane −: negative control (buffer), lane +: positive control (C. albicans). Full article ">Figure 5
Results of nested PCR. Lane (a): negative control (buffer), lane (b): positive control (C. albicans); the lane to the right indicates positive (+) or negative (−). All specimens from the blood, the tongue, the liver, and the kidney were negative for C. albicans, indicating that there was no contamination of the experimental environment and experimental animals. Full article ">

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Curr. Issues Mol. Biol., Volume 45, Issue 2 (February 2023) – 63 articles

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