Biomolecules

17 pages, 2534 KiB

Open AccessArticle

Citrus sudachi Peel Extract Suppresses Cell Proliferation and Promotes the Differentiation of Keratinocytes through Inhibition of the EGFR–ERK Signaling Pathway

by Shogo Abe, Misako Ueno, Mami Nishitani, Tetsuya Akamatsu, Takumi Sato, Marie Shimoda, Hiroki Kanaoka, Yoshitaka Nii, Hiroko Yamasaki and Keizo Yuasa

Biomolecules 2020, 10(10), 1468; https://doi.org/10.3390/biom10101468 - 21 Oct 2020

Cited by 8 | Viewed by 3400

Abstract

Citrus sudachi is a well-known fruit in Tokushima Prefecture, Japan, and its peels are rich in phytochemicals, including phenolic compounds. Although it is expected that the extract of the C. sudachi peel elicits various beneficial physiological activities, the effect on the skin has [...] Read more.

Citrus sudachi is a well-known fruit in Tokushima Prefecture, Japan, and its peels are rich in phytochemicals, including phenolic compounds. Although it is expected that the extract of the C. sudachi peel elicits various beneficial physiological activities, the effect on the skin has not been investigated. In this study, we report that the aqueous extract from the peel of C. sudachi suppresses cell proliferation of the immortalized human keratinocyte cell line, HaCaT, and primary normal human epidermal keratinocytes. The extract of C. sudachi peel suppressed epidermal growth factor (EGF)-induced EGF receptor activation and tumor necrosis factor (TNF)-α-induced extracellular regulated kinase (ERK) 1/2 activation, which suggests that the extract exerts its inhibitory effect through inhibition of both the EGF receptor (EGFR) and its downstream molecules. Additionally, the extract of C. sudachi peel potentiated calcium-induced keratinocyte differentiation. These results suggest that the extract of C. sudachi peel may have beneficial effects against skin diseases that are characterized by hyperproliferation of epidermal keratinocytes, such as those seen in psoriasis and in cutaneous squamous cell carcinoma. Full article

(This article belongs to the Special Issue MAPKs in Skin Health and Disease)

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22 pages, 2482 KiB

Open AccessArticle

Identification of Putative Non-Substrate-Based XT-I Inhibitors by Natural Product Library Screening

by Thanh-Diep Ly, Anika Kleine, Bastian Fischer, Vanessa Schmidt, Doris Hendig, Joachim Kuhn, Cornelius Knabbe and Isabel Faust

Biomolecules 2020, 10(10), 1467; https://doi.org/10.3390/biom10101467 - 21 Oct 2020

Cited by 10 | Viewed by 3067

Abstract

Fibroproliferative diseases are characterized by excessive accumulation of extracellular matrix (ECM) components leading to organ dysfunction. This process is characterized by an increase in myofibroblast content and enzyme activity of xylosyltransferase-I (XT-I), the initial enzyme in proteoglycan (PG) biosynthesis. Therefore, the inhibition of [...] Read more.

Fibroproliferative diseases are characterized by excessive accumulation of extracellular matrix (ECM) components leading to organ dysfunction. This process is characterized by an increase in myofibroblast content and enzyme activity of xylosyltransferase-I (XT-I), the initial enzyme in proteoglycan (PG) biosynthesis. Therefore, the inhibition of XT-I could be a promising treatment for fibrosis. We used a natural product-inspired compound library to identify non-substrate-based inhibitors of human XT-I by UPLC-MS/MS. We combined this cell-free approach with virtual and molecular biological analyses to confirm and prioritize the inhibitory potential of the compounds identified. The characterization for compound potency in TGF-β1-driven XYLT1 transcription regulation in primary dermal human fibroblasts (key cells in ECM remodeling) was addressed by gene expression analysis. Consequently, we identified amphotericin B and celastrol as new non-substrate-based XT-I protein inhibitors. Their XT-I inhibitory effects were mediated by an uncompetitive or a competitive inhibition mode, respectively. Both compounds reduced the cellular XYLT1 expression level and XT-I activity. We showed that these cellular inhibitor-mediated changes involve the TGF-β and microRNA-21 signaling pathway. The results of our study provide a strong rationale for the further optimization and future usage of the XT-I inhibitors identified as promising therapeutic agents of fibroproliferative diseases. Full article

(This article belongs to the Special Issue Enzymatic Inhibitors from Natural Sources: A Huge Collection of New Potential Drugs)

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16 pages, 2431 KiB

Open AccessArticle

Gene Expression Profiling of Multiple Histone Deacetylases (HDAC) and Its Correlation with NRF2-Mediated Redox Regulation in the Pathogenesis of Diabetic Foot Ulcers

by Rajan Teena, Umapathy Dhamodharan, Daoud Ali, Kesavan Rajesh and Kunka Mohanram Ramkumar

Biomolecules 2020, 10(10), 1466; https://doi.org/10.3390/biom10101466 - 21 Oct 2020

Cited by 21 | Viewed by 3863

Abstract

Nuclear factor erythroid-2-related factor 2 (Nrf2) is a protein of the leucine zipper family, which mitigates inflammation and employs cytoprotective effects. Attempting to unravel the epigenetic regulation of type 2 diabetes mellitus (T2DM) and diabetic foot ulcer (DFU), we profiled the expression of [...] Read more.

Nuclear factor erythroid-2-related factor 2 (Nrf2) is a protein of the leucine zipper family, which mitigates inflammation and employs cytoprotective effects. Attempting to unravel the epigenetic regulation of type 2 diabetes mellitus (T2DM) and diabetic foot ulcer (DFU), we profiled the expression of eleven isoform-specific histone deacetylases (HDACs) and correlated them with NRF2 and cytokines. This study recruited a total of 60 subjects and categorized into DFU patients (n = 20), T2DM patients (n = 20), and healthy controls (n = 20). The DFU patients were subcategorized into uninfected and infected DFU (n = 10 each). We observed a progressive decline in the expression of NRF2 and its downstream targets among T2DM and DFU subjects. The inflammatory markers IL-6 and TNF-α were significantly upregulated, whereas anti-inflammatory marker IL-10 was significantly downregulated in DFU. Of note, a significant upregulation of HDAC1, 3, 4, 11, SIRT3 and downregulation of HDAC2,8, SIRT1, SIRT2, SIRT3, SIRT7 among DFU patients were observed. The significant positive correlation between NRF2 and SIRT1 in DFU patients suggested the vital role of NRF2/SIRT1 in redox homeostasis and angiogenesis. In contrast, the significant negative correlation between NRF2 and HDAC1, 3 and 4, implied an imbalance in NRF2-HDAC1, 3, 4 circuit. Furthermore, a significant positive correlation was observed between HDAC4 and IL-6, and the negative correlation between SIRT1 and IL-6 suggested the pro-inflammatory role of HDAC4 and the anti-inflammatory role of SIRT1 in NRF2 signaling. In conclusion, the epigenetic changes such as upregulation of HDAC1, 3, 4, 11, SIRT3 and downregulation of HDAC2, 8, SIRT1, SIRT2, SIRT6, SIRT7 and their association with NRF2 as well as inflammatory markers are suggestive of their roles in pathophysiology of T2DM and DFU. Full article

(This article belongs to the Special Issue Role of Nrf2 in Disease: Novel Molecular Mechanisms and Therapeutic Approaches)

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19 pages, 821 KiB

Open AccessReview

Alpha-Synuclein in Alcohol Use Disorder, Connections with Parkinson’s Disease and Potential Therapeutic Role of 5’ Untranslated Region-Directed Small Molecules

by Catherine M. Cahill, Rozaleen Aleyadeh, Jin Gao, Changning Wang and Jack T. Rogers

Biomolecules 2020, 10(10), 1465; https://doi.org/10.3390/biom10101465 - 21 Oct 2020

Cited by 9 | Viewed by 3964

Abstract

Alpha-synuclein (α-Syn) is a 140-amino acid (aa) protein encoded by the Synuclein alpha SNCA gene. It is the synaptic protein associated with Parkinson’s disease (PD) and is the most highly expressed protein in the Lewy bodies associated with PD and other alpha synucleopathies, [...] Read more.

Alpha-synuclein (α-Syn) is a 140-amino acid (aa) protein encoded by the Synuclein alpha SNCA gene. It is the synaptic protein associated with Parkinson’s disease (PD) and is the most highly expressed protein in the Lewy bodies associated with PD and other alpha synucleopathies, including Lewy body dementia (LBD) and multiple system atrophy (MSA). Iron deposits are present in the core of Lewy bodies, and there are reports suggesting that divalent metal ions including Cu²⁺ and Fe²⁺ enhance the aggregation of α-Syn. Differential expression of α-Syn is associated with alcohol use disorder (AUD), and specific genetic variants contribute to the risk for alcoholism, including alcohol craving. Spliced variants of α-Syn, leading to the expression of several shorter forms which are more prone to aggregation, are associated with both PD and AUD, and common transcript variants may be able to predict at-risk populations for some movement disorders or subtypes of PD, including secondary Parkinsonism. Both PD and AUD are associated with liver and brain iron dyshomeostasis. Research over the past decade has shown that α-Syn has iron import functions with an ability to oxidize the Fe³⁺ form of iron to Fe²⁺ to facilitate its entry into cells. Our prior research has identified an iron-responsive element (IRE) in the 5’ untranslated region (5’UTR) of α-Syn mRNA, and we have used the α-Syn 5’UTR to screen for small molecules that modulate its expression in the H4 neuronal cell line. These screens have led us to identify several interesting small molecules capable of both decreasing and increasing α-Syn expression and that may have the potential, together with the recently described mesenchymal stem cell therapies, to normalize α-Syn expression in different regions of the alcoholic and PD brain. Full article

(This article belongs to the Special Issue Parkinson’s Disease and Associated Disorders)

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11 pages, 1190 KiB

Open AccessCommunication

Discovery of New Antibacterial Accramycins from a Genetic Variant of the Soil Bacterium, Streptomyces sp. MA37

by Fleurdeliz Maglangit, Yuting Zhang, Kwaku Kyeremeh and Hai Deng

Biomolecules 2020, 10(10), 1464; https://doi.org/10.3390/biom10101464 - 20 Oct 2020

Cited by 15 | Viewed by 3603

Abstract

Continued mining of natural products from the strain Streptomyces sp. MA37 in our laboratory led to the discovery of a minor specialized metabolite (SM) called accramycin A. Owing to its low yield (0.2 mg/L) in the wild type strain, we investigated the roles [...] Read more.

Continued mining of natural products from the strain Streptomyces sp. MA37 in our laboratory led to the discovery of a minor specialized metabolite (SM) called accramycin A. Owing to its low yield (0.2 mg/L) in the wild type strain, we investigated the roles of regulatory genes in the corresponding biosynthetic gene cluster (acc BGC) through gene inactivation with the aim of improving the titer of this compound. One of the resulting mutants (∆accJ) dramatically upregulated the production of accramycin A 1 by 330-fold (66 mg/L). Furthermore, ten new metabolites, accramycins B–K 2–11, were discovered, together with two known compounds, naphthacemycin B₁12 and fasamycin C 13 from the mutant extract. This suggested that accJ, annotated as multiple antibiotic resistance regulator (MarR), is a negative regulator gene in the accramycin biosynthesis. Compounds 1–13 inhibited the Gram-positive pathogens (Staphylococcus aureus, Enterococcus faecalis) and clinical isolates Enterococcus faecium (K59-68 and K60-39) and Staphylococcus haemolyticus with minimal inhibitory concentration (MIC) values in the range of 1.5–12.5 µg/mL. Remarkably, compounds 1–13 displayed superior activity against K60-39 (MIC = 3.1–6.3 µg/mL) compared to ampicillin (MIC = 25 µg/mL), and offered promising potential for the development of accramycin-based antibiotics that target multidrug-resistant Enterococcus clinical isolates. Our results highlight the importance of identifying the roles of regulatory genes in natural product discovery. Full article

(This article belongs to the Section Natural and Bio-derived Molecules)

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16 pages, 4305 KiB

Open AccessArticle

Standardized Fraction of Turbinaria ornata Alleviates Dextran Sulfate Sodium-Induced Chronic Colitis in C57BL/6 Mice via Upregulation of FOXP3⁺ Regulatory T Cells

by Na-Hyun Kim, Seon Min Lee, Yun Na Kim, You-Jin Jeon, Jeong-Doo Heo, Eun Ju Jeong and Jung-Rae Rho

Biomolecules 2020, 10(10), 1463; https://doi.org/10.3390/biom10101463 - 20 Oct 2020

Cited by 13 | Viewed by 2927

Abstract

Turbinaria ornata is a tropical brown algae (seaweed) known to have anti-inflammatory properties. In this study, we analyzed T. ornata extract (TOE) using liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) and nuclear magnetic resonance (NMR) and evaluated the in vivo efficacy [...] Read more.

Turbinaria ornata is a tropical brown algae (seaweed) known to have anti-inflammatory properties. In this study, we analyzed T. ornata extract (TOE) using liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) and nuclear magnetic resonance (NMR) and evaluated the in vivo efficacy of TOE against dextran sulfate sodium-induced chronic colitis in C57BL/6 mice. The bioactive fraction of TOE was administered orally daily for 6 weeks to mice under different treatments normal, colitis, and colitis + conventional drug (5-aminosalicylic acid, 5-ASA). Regarding clinical manifestation, the disease activity index and colon length of the colitis + TOE group were significantly reduced compared to those of the colitis group. The results of myeloperoxidase activity and histopathological examination showed similar results. Western blot analysis of colon tissues revealed that cyclooxygenase-2, tumor necrosis factor alpha (TNF-α), and phosphorylated signal transducer and activator of transcription-3 (p-STAT3) were significantly decreased in the colitis + 5-ASA group, whereas forkhead box P3 (FOXP3) was increased. qPCR results showed changes in T cell subsets; the administration of TOE upregulated regulatory T cell (Treg) expression, although T helper 17 cell (Th17) expression did not change significantly. Interestingly, the colitis + TOE group showed high levels of both Th1 and Th2 transcription factors, but the secreted cytokine interferon (IFN)-γ and interleukin (IL)-4 remained unchanged and somewhat reduced. Additionally, TNF-α gene expression was significantly reduced in the colitis + TOE group. IL-6 mRNA levels were also decreased, although not significantly. Four compounds were structurally elucidated using 1D- and 2D-NMR spectroscopy, and five compounds were fully identified or tentatively characterized using LC-QTOF-MS. In conclusion, TOE could alleviate chronic colitis via upregulation of Foxp3⁺ Treg cells and production of the anti-inflammatory cytokine IL-10, which directly inhibits macrophages and pro-inflammatory cytokine synthesis, leading to reduced colitis. Full article

(This article belongs to the Section Natural and Bio-derived Molecules)

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43 pages, 4071 KiB

Open AccessArticle

Synthesis and HPLC-ECD Study of Cytostatic Condensed O,N-Heterocycles Obtained from 3-Aminoflavanones

by Ádám Szappanos, Attila Mándi, Katalin Gulácsi, Erika Lisztes, Balázs István Tóth, Tamás Bíró, Anita Kónya-Ábrahám, Attila Kiss-Szikszai, Attila Bényei, Sándor Antus and Tibor Kurtán

Biomolecules 2020, 10(10), 1462; https://doi.org/10.3390/biom10101462 - 20 Oct 2020

Cited by 2 | Viewed by 3296

Abstract

Racemic chiral O,N-heterocycles containing 2-arylchroman or 2-aryl-2H-chromene subunit condensed with morpholine, thiazole, or pyrrole moieties at the C-3-C-4 bond were synthesized with various substitution patterns of the aryl group by the cyclization of cis- or trans-3-aminoflavanone analogues. The [...] Read more.

Racemic chiral O,N-heterocycles containing 2-arylchroman or 2-aryl-2H-chromene subunit condensed with morpholine, thiazole, or pyrrole moieties at the C-3-C-4 bond were synthesized with various substitution patterns of the aryl group by the cyclization of cis- or trans-3-aminoflavanone analogues. The 3-aminoflavanone precursors were obtained in a Neber rearrangement of oxime tosylates of flavanones, which provided the trans diastereomer as the major product and enabled the isolation of both the cis- and trans-diastereomers. The cis- and trans-aminoflavanones were utilized to prepare three diastereomers of 5-aryl-chromeno[4,3-b][1,4]oxazines. Antiproliferative activity of the condensed heterocycles and precursors was evaluated against A2780 and WM35 cancer cell lines. For a 3-(N-chloroacetylamino)-flavan-4-ol derivative, showing structural analogy with acyclic acid ceramidase inhibitors, 0.15 μM, 3.50 μM, and 6.06 μM IC₅₀ values were measured against A2780, WM35, and HaCat cell lines, and apoptotic mechanism was confirmed. Low micromolar IC₅₀ values down to 2.14 μM were identified for the thiazole- and pyrrole-condensed 2H-chromene derivatives. Enantiomers of the condensed heterocycles were separated by HPLC using chiral stationary phase, HPLC-ECD spectra were recorded and TDDFT-ECD calculations were performed to determine the absolute configuration and solution conformation. Characteristic ECD transitions of the separated enantiomers were correlated with the absolute configuration and effect of substitution pattern on the HPLC elution order was determined. Full article

(This article belongs to the Section Natural and Bio-derived Molecules)

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20 pages, 1718 KiB

Open AccessReview

Understanding the Pathogenesis of Spondyloarthritis

by Aigul Sharip and Jeannette Kunz

Biomolecules 2020, 10(10), 1461; https://doi.org/10.3390/biom10101461 - 20 Oct 2020

Cited by 69 | Viewed by 10079

Abstract

Spondyloarthritis comprises a group of inflammatory diseases of the joints and spine, with various clinical manifestations. The group includes ankylosing spondylitis, reactive arthritis, psoriatic arthritis, arthritis associated with inflammatory bowel disease, and undifferentiated spondyloarthritis. The exact etiology and pathogenesis of spondyloarthritis are still [...] Read more.

Spondyloarthritis comprises a group of inflammatory diseases of the joints and spine, with various clinical manifestations. The group includes ankylosing spondylitis, reactive arthritis, psoriatic arthritis, arthritis associated with inflammatory bowel disease, and undifferentiated spondyloarthritis. The exact etiology and pathogenesis of spondyloarthritis are still unknown, but five hypotheses explaining the pathogenesis exist. These hypotheses suggest that spondyloarthritis is caused by arthritogenic peptides, an unfolded protein response, HLA-B*27 homodimer formation, malfunctioning endoplasmic reticulum aminopeptidases, and, last but not least, gut inflammation and dysbiosis. Here we discuss the five hypotheses and the evidence supporting each. In all of these hypotheses, HLA-B*27 plays a central role. It is likely that a combination of these hypotheses, with HLA-B*27 taking center stage, will eventually explain the development of spondyloarthritis in predisposed individuals. Full article

(This article belongs to the Special Issue Molecular Basis of Autoimmunity Diseases)

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16 pages, 3013 KiB

Open AccessArticle

Predicting Deep Learning Based Multi-Omics Parallel Integration Survival Subtypes in Lung Cancer Using Reverse Phase Protein Array Data

by Satoshi Takahashi, Ken Asada, Ken Takasawa, Ryo Shimoyama, Akira Sakai, Amina Bolatkan, Norio Shinkai, Kazuma Kobayashi, Masaaki Komatsu, Syuzo Kaneko, Jun Sese and Ryuji Hamamoto

Biomolecules 2020, 10(10), 1460; https://doi.org/10.3390/biom10101460 - 19 Oct 2020

Cited by 47 | Viewed by 5728

Abstract

Mortality attributed to lung cancer accounts for a large fraction of cancer deaths worldwide. With increasing mortality figures, the accurate prediction of prognosis has become essential. In recent years, multi-omics analysis has emerged as a useful survival prediction tool. However, the methodology relevant [...] Read more.

Mortality attributed to lung cancer accounts for a large fraction of cancer deaths worldwide. With increasing mortality figures, the accurate prediction of prognosis has become essential. In recent years, multi-omics analysis has emerged as a useful survival prediction tool. However, the methodology relevant to multi-omics analysis has not yet been fully established and further improvements are required for clinical applications. In this study, we developed a novel method to accurately predict the survival of patients with lung cancer using multi-omics data. With unsupervised learning techniques, survival-associated subtypes in non-small cell lung cancer were first detected using the multi-omics datasets from six categories in The Cancer Genome Atlas (TCGA). The new subtypes, referred to as integration survival subtypes, clearly divided patients into longer and shorter-surviving groups (log-rank test: p = 0.003) and we confirmed that this is independent of histopathological classification (Chi-square test of independence: p = 0.94). Next, an attempt was made to detect the integration survival subtypes using only one categorical dataset. Our machine learning model that was only trained on the reverse phase protein array (RPPA) could accurately predict the integration survival subtypes (AUC = 0.99). The predicted subtypes could also distinguish between high and low risk patients (log-rank test: p = 0.012). Overall, this study explores novel potentials of multi-omics analysis to accurately predict the prognosis of patients with lung cancer. Full article

(This article belongs to the Special Issue Application of Artificial Intelligence for Medical Research)

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25 pages, 5803 KiB

Open AccessArticle

Analysis of Amygdalin in Various Matrices Using Electrospray Ionization and Flowing Atmospheric-Pressure Afterglow Mass Spectrometry

by Maria Guć, Sandra Rutecka and Grzegorz Schroeder

Biomolecules 2020, 10(10), 1459; https://doi.org/10.3390/biom10101459 - 19 Oct 2020

Cited by 6 | Viewed by 3448

Abstract

Amygdalin is a natural cyanogenic compound that plants produce in the fight against insects and herbivores. Excessive amounts of amygdalin by animals and humans can potentially lead to fatal intoxication. However, studies confirm that amygdalin has antitumor properties, including the ability to inhibit [...] Read more.

Amygdalin is a natural cyanogenic compound that plants produce in the fight against insects and herbivores. Excessive amounts of amygdalin by animals and humans can potentially lead to fatal intoxication. However, studies confirm that amygdalin has antitumor properties, including the ability to inhibit the proliferation of cancer cells and to induce their apoptosis. The analysis of amygdalin in various matrices is an important analytical problem today. The publication presents the methodology of direct determination of amygdalin in water, sewage, and biological materials using electrospray ionization mass spectrometry (ESI-MS) and a new analytical method using flowing atmospheric-pressure afterglow mass spectrometry (FAPA-MS). The methods of analyte pre-concentration using a magnetic, molecularly imprinted polymer (mag-MIP) and the influence of interferents on the recorded spectra were discussed. Analytical parameters in ESI-MS and FAPA-MS methods were established. The linearity range was 4.5 µg L⁻¹–45 mg L⁻¹ in positive mode ESI-MS and FAPA-MS. The limit of detection (LOD) for ESI-MS was 0.101 ± 0.003 µg L⁻¹ and the limit of quantification (LOQ) was 0.303 ± 0.009 µg L⁻¹. In FAPA-MS, the LOD was 0.050 ± 0.002 µg L⁻¹ and the LOQ was 0.150 ± 0.006 µg L⁻¹. The content of amygdalin in various matrices was determined. Full article

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22 pages, 4242 KiB

Open AccessArticle

Robust Cell-Free Expression of Sub-Pathological and Pathological Huntingtin Exon-1 for NMR Studies. General Approaches for the Isotopic Labeling of Low-Complexity Proteins

by Anna Morató, Carlos A. Elena-Real, Matija Popovic, Aurélie Fournet, Karen Zhang, Frédéric Allemand, Nathalie Sibille, Annika Urbanek and Pau Bernadó

Biomolecules 2020, 10(10), 1458; https://doi.org/10.3390/biom10101458 - 19 Oct 2020

Cited by 8 | Viewed by 2990

Abstract

The high-resolution structural study of huntingtin exon-1 (HttEx1) has long been hampered by its intrinsic properties. In addition to being prone to aggregate, HttEx1 contains low-complexity regions (LCRs) and is intrinsically disordered, ruling out several standard structural biology approaches. Here, we use a [...] Read more.

The high-resolution structural study of huntingtin exon-1 (HttEx1) has long been hampered by its intrinsic properties. In addition to being prone to aggregate, HttEx1 contains low-complexity regions (LCRs) and is intrinsically disordered, ruling out several standard structural biology approaches. Here, we use a cell-free (CF) protein expression system to robustly and rapidly synthesize (sub-) pathological HttEx1. The open nature of the CF reaction allows the application of different isotopic labeling schemes, making HttEx1 amenable for nuclear magnetic resonance studies. While uniform and selective labeling facilitate the sequential assignment of HttEx1, combining CF expression with nonsense suppression allows the site-specific incorporation of a single labeled residue, making possible the detailed investigation of the LCRs. To optimize CF suppression yields, we analyze the expression and suppression kinetics, revealing that high concentrations of loaded suppressor tRNA have a negative impact on the final reaction yield. The optimized CF protein expression and suppression system is very versatile and well suited to produce challenging proteins with LCRs in order to enable the characterization of their structure and dynamics. Full article

(This article belongs to the Section Molecular Structure and Dynamics)

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32 pages, 2420 KiB

Open AccessEditor’s ChoiceReview

Updating Phospholipase A₂ Biology

by Makoto Murakami, Hiroyasu Sato and Yoshitaka Taketomi

Biomolecules 2020, 10(10), 1457; https://doi.org/10.3390/biom10101457 - 19 Oct 2020

Cited by 127 | Viewed by 12097

Abstract

The phospholipase A₂ (PLA₂) superfamily contains more than 50 enzymes in mammals that are subdivided into several distinct families on a structural and biochemical basis. In principle, PLA₂ has the capacity to hydrolyze the sn-2 position of glycerophospholipids [...] Read more.

The phospholipase A₂ (PLA₂) superfamily contains more than 50 enzymes in mammals that are subdivided into several distinct families on a structural and biochemical basis. In principle, PLA₂ has the capacity to hydrolyze the sn-2 position of glycerophospholipids to release fatty acids and lysophospholipids, yet several enzymes in this superfamily catalyze other reactions rather than or in addition to the PLA₂ reaction. PLA₂ enzymes play crucial roles in not only the production of lipid mediators, but also membrane remodeling, bioenergetics, and body surface barrier, thereby participating in a number of biological events. Accordingly, disturbance of PLA₂-regulated lipid metabolism is often associated with various diseases. This review updates the current state of understanding of the classification, enzymatic properties, and biological functions of various enzymes belonging to the PLA₂ superfamily, focusing particularly on the novel roles of PLA₂s in vivo. Full article

(This article belongs to the Special Issue Phospholipases: From Structure to Biological Function)

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17 pages, 3022 KiB

Open AccessArticle

Anti-Inflammatory Properties of Injectable Betamethasone-Loaded Tyramine-Modified Gellan Gum/Silk Fibroin Hydrogels

by Isabel Matos Oliveira, Cristiana Gonçalves, Myeong Eun Shin, Sumi Lee, Rui Luis Reis, Gilson Khang and Joaquim Miguel Oliveira

Biomolecules 2020, 10(10), 1456; https://doi.org/10.3390/biom10101456 - 17 Oct 2020

Cited by 30 | Viewed by 3791

Abstract

Rheumatoid arthritis is a rheumatic disease for which a healing treatment does not presently exist. Silk fibroin has been extensively studied for use in drug delivery systems due to its uniqueness, versatility and strong clinical track record in medicine. However, in general, natural [...] Read more.

Rheumatoid arthritis is a rheumatic disease for which a healing treatment does not presently exist. Silk fibroin has been extensively studied for use in drug delivery systems due to its uniqueness, versatility and strong clinical track record in medicine. However, in general, natural polymeric materials are not mechanically stable enough, and have high rates of biodegradation. Thus, synthetic materials such as gellan gum can be used to produce composite structures with biological signals to promote tissue-specific interactions while providing the desired mechanical properties. In this work, we aimed to produce hydrogels of tyramine-modified gellan gum with silk fibroin (Ty–GG/SF) via horseradish peroxidase (HRP), with encapsulated betamethasone, to improve the biocompatibility and mechanical properties, and further increase therapeutic efficacy to treat rheumatoid arthritis (RA). The Ty–GG/SF hydrogels presented a β-sheet secondary structure, with gelation time around 2–5 min, good resistance to enzymatic degradation, a suitable injectability profile, viscoelastic capacity with a significant solid component and a betamethasone-controlled release profile over time. In vitro studies showed that Ty–GG/SF hydrogels did not produce a deleterious effect on cellular metabolic activity, morphology or proliferation. Furthermore, Ty–GG/SF hydrogels with encapsulated betamethasone revealed greater therapeutic efficacy than the drug applied alone. Therefore, this strategy can provide an improvement in therapeutic efficacy when compared to the traditional use of drugs for the treatment of rheumatoid arthritis. Full article

(This article belongs to the Special Issue Recent Advances in Silk Fibroin)

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23 pages, 4405 KiB

Open AccessFeature PaperArticle

Metabolic Effects of Selective Deletion of Group VIA Phospholipase A₂ from Macrophages or Pancreatic Islet Beta-Cells

by John Turk, Haowei Song, Mary Wohltmann, Cheryl Frankfater, Xiaoyong Lei and Sasanka Ramanadham

Biomolecules 2020, 10(10), 1455; https://doi.org/10.3390/biom10101455 - 17 Oct 2020

Cited by 8 | Viewed by 2757

Abstract

To examine the role of group VIA phospholipase A₂ (iPLA₂β) in specific cell lineages in insulin secretion and insulin action, we prepared mice with a selective iPLA₂β deficiency in cells of myelomonocytic lineage, including macrophages (MØ-iPLA₂β-KO), [...] Read more.

To examine the role of group VIA phospholipase A₂ (iPLA₂β) in specific cell lineages in insulin secretion and insulin action, we prepared mice with a selective iPLA₂β deficiency in cells of myelomonocytic lineage, including macrophages (MØ-iPLA₂β-KO), or in insulin-secreting β-cells (β-Cell-iPLA₂β-KO), respectively. MØ-iPLA₂β-KO mice exhibited normal glucose tolerance when fed standard chow and better glucose tolerance than floxed-iPLA₂β control mice after consuming a high-fat diet (HFD). MØ-iPLA₂β-KO mice exhibited normal glucose-stimulated insulin secretion (GSIS) in vivo and from isolated islets ex vivo compared to controls. Male MØ-iPLA₂β-KO mice exhibited enhanced insulin responsivity vs. controls after a prolonged HFD. In contrast, β-cell-iPLA₂β-KO mice exhibited impaired glucose tolerance when fed standard chow, and glucose tolerance deteriorated further when introduced to a HFD. β-Cell-iPLA₂β-KO mice exhibited impaired GSIS in vivo and from isolated islets ex vivo vs. controls. β-Cell-iPLA₂β-KO mice also exhibited an enhanced insulin responsivity compared to controls. These findings suggest that MØ iPLA₂β participates in HFD-induced deterioration in glucose tolerance and that this mainly reflects an effect on insulin responsivity rather than on insulin secretion. In contrast, β-cell iPLA₂β plays a role in GSIS and also appears to confer some protection against deterioration in β-cell functions induced by a HFD. Full article

(This article belongs to the Special Issue Phospholipases: From Structure to Biological Function)

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18 pages, 1788 KiB

Open AccessArticle

Hypothermic Effect of Acute Citral Treatment during LPS-induced Systemic Inflammation in Obese Mice: Reduction of Serum TNF-α and Leptin Levels

by Maycon T. Emílio-Silva, Vinicius P. Rodrigues, Gabriela Bueno, Rie Ohara, Marina G. Martins, José A. C. Horta-Júnior, Luiz G. S. Branco, Lúcia R. M. Rocha and Clélia A. Hiruma-Lima

Biomolecules 2020, 10(10), 1454; https://doi.org/10.3390/biom10101454 - 17 Oct 2020

Cited by 14 | Viewed by 4337

Abstract

Citral is a mixture of monoterpenes present in the essential oil of several plants, such as Cymbopogon citratus and Zingiber officinale, possessing anti-inflammatory, anti-ulcerogenic, and antipyretic actions. We investigated the action of citral on body temperature (Tb) and inflammatory signaling in eutrophic [...] Read more.

Citral is a mixture of monoterpenes present in the essential oil of several plants, such as Cymbopogon citratus and Zingiber officinale, possessing anti-inflammatory, anti-ulcerogenic, and antipyretic actions. We investigated the action of citral on body temperature (Tb) and inflammatory signaling in eutrophic and obese mice during Systemic Inflammation (SI) induced by Lipopolysaccharide (LPS). Thus, we assessed the effect of citral (25, 100, and 300 mg/kg) and ibuprofen in LPS-induced SI in Swiss male mice fed a standard diet (SD) or high-fat diet (HFD) for 12 weeks. Following SI induction, we measured Tb and collected the serum, hypothalamus, and gastric mucosa for biochemical measurements. Acute treatment with citral decreased the Tb of both SD and HFD-fed animals. Citral (300 mg/kg) treatment caused a significantly lower Tb variation in HFD-fed animals than in those fed the SD. Citral reduced peripheral levels of tumor necrosis factor (TNF)-α in SD and HFD mice and decreased serum leptin concentration in HFD mice 90 min after the LPS challenge. Furthermore, citral also reduced interleukin (IL)-6 levels in the hypothalamus of obese mice. In summary, citral effectively reduced Tb during SI by reducing inflammatory mediators with a distinct action profile in HFD mice when compared with SD. Full article

(This article belongs to the Collection Bioactive Lipids in Inflammation, Diabetes and Cancer)

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22 pages, 1142 KiB

Open AccessEditor’s ChoiceReview

Ubiquitomics: An Overview and Future

by George Vere, Rachel Kealy, Benedikt M. Kessler and Adan Pinto-Fernandez

Biomolecules 2020, 10(10), 1453; https://doi.org/10.3390/biom10101453 - 17 Oct 2020

Cited by 26 | Viewed by 6867

Abstract

Covalent attachment of ubiquitin, a small globular polypeptide, to protein substrates is a key post-translational modification that determines the fate, function, and turnover of most cellular proteins. Ubiquitin modification exists as mono- or polyubiquitin chains involving multiple ways how ubiquitin C-termini are connected [...] Read more.

Covalent attachment of ubiquitin, a small globular polypeptide, to protein substrates is a key post-translational modification that determines the fate, function, and turnover of most cellular proteins. Ubiquitin modification exists as mono- or polyubiquitin chains involving multiple ways how ubiquitin C-termini are connected to lysine, perhaps other amino acid side chains, and N-termini of proteins, often including branching of the ubiquitin chains. Understanding this enormous complexity in protein ubiquitination, the so-called ‘ubiquitin code’, in combination with the

\sim

1000 enzymes involved in controlling ubiquitin recognition, conjugation, and deconjugation, calls for novel developments in analytical techniques. Here, we review different headways in the field mainly driven by mass spectrometry and chemical biology, referred to as “ubiquitomics”, aiming to understand this system’s biological diversity. Full article

(This article belongs to the Special Issue Looking Back and Ahead: Emerging Concepts in Ubiquitin and UBLs)

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23 pages, 6204 KiB

Open AccessFeature PaperArticle

Calmodulin Directly Interacts with the Cx43 Carboxyl-Terminus and Cytoplasmic Loop Containing Three ODDD-Linked Mutants (M147T, R148Q, and T154A) that Retain α-Helical Structure, but Exhibit Loss-of-Function and Cellular Trafficking Defects

by Li Zheng, Sylvie Chenavas, Fabien Kieken, Andrew Trease, Sarah Brownell, Asokan Anbanandam, Paul L. Sorgen and Gaelle Spagnol

Biomolecules 2020, 10(10), 1452; https://doi.org/10.3390/biom10101452 - 17 Oct 2020

Cited by 7 | Viewed by 3336

Abstract

The autosomal-dominant pleiotropic disorder called oculodentodigital dysplasia (ODDD) is caused by mutations in the gap junction protein Cx43. Of the 73 mutations identified to date, over one-third are localized in the cytoplasmic loop (Cx43CL) domain. Here, we determined the mechanism by which three [...] Read more.

The autosomal-dominant pleiotropic disorder called oculodentodigital dysplasia (ODDD) is caused by mutations in the gap junction protein Cx43. Of the 73 mutations identified to date, over one-third are localized in the cytoplasmic loop (Cx43CL) domain. Here, we determined the mechanism by which three ODDD mutations (M147T, R148Q, and T154A), all of which localize within the predicted 1-5-10 calmodulin-binding motif of the Cx43CL, manifest the disease. Nuclear magnetic resonance (NMR) and circular dichroism revealed that the three ODDD mutations had little-to-no effect on the ability of the Cx43CL to form α-helical structure as well as bind calmodulin. Combination of microscopy and a dye-transfer assay uncovered these mutations increased the intracellular level of Cx43 and those that trafficked to the plasma membrane did not form functional channels. NMR also identify that CaM can directly interact with the Cx43CT domain. The Cx43CT residues involved in the CaM interaction overlap with tyrosines phosphorylated by Pyk2 and Src. In vitro and in cyto data provide evidence that the importance of the CaM interaction with the Cx43CT may lie in restricting Pyk2 and Src phosphorylation, and their subsequent downstream effects. Full article

(This article belongs to the Special Issue Connexins, Innexins, and Pannexins: From Biology to Clinical Targets)

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17 pages, 2307 KiB

Open AccessArticle

Suppression of Inflammation Delays Hair Cell Regeneration and Functional Recovery Following Lateral Line Damage in Zebrafish Larvae

by Ru Zhang, Xiaopeng Liu, Yajuan Li, Ming Wang, Lin Chen and Bing Hu

Biomolecules 2020, 10(10), 1451; https://doi.org/10.3390/biom10101451 - 16 Oct 2020

Cited by 5 | Viewed by 3753

Abstract

Cochlear hair cells in human beings cannot regenerate after loss; however, those in fish and other lower species can. Recently, the role of inflammation in hair cell regeneration has been attracting the attention of scientists. In the present study, we investigated how suppression [...] Read more.

Cochlear hair cells in human beings cannot regenerate after loss; however, those in fish and other lower species can. Recently, the role of inflammation in hair cell regeneration has been attracting the attention of scientists. In the present study, we investigated how suppression of inflammatory factors affects hair cell regeneration and the functional recovery of regenerated hair cells in zebrafish. We killed hair cells in the lateral line of zebrafish larvae with CuSO₄ to induce an inflammatory response and coapplied BRS-28, an anti-inflammatory agent to suppress the inflammation. The recovery of the hair cell number and rheotaxis was slower when CuSO₄ and BRS-28 were coapplied than when CuSO₄ was applied alone. The recovery of hair cell count lagged behind that of the calcium imaging signal during the regeneration. The calcium imaging signal in the neuromasts in the inflammation-inhibited group was weaker than that in the noninflammation-inhibited group at the early stage of regeneration, although it returned to normal at the late stage. Our study demonstrates that suppressing inflammation by BRS-28 delays hair cell regeneration and functional recovery when hair cells are damaged. We suspect that BRS-28 inhibits pro-inflammatory factors and thereby reduces the migration of macrophages to delay the regeneration of hair cells. Full article

(This article belongs to the Special Issue Fish as Simple Models for Human Disease and Drug Screen)

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24 pages, 9254 KiB

Open AccessArticle

Nonhistone Proteins HMGB1 and HMGB2 Differentially Modulate the Response of Human Embryonic Stem Cells and the Progenitor Cells to the Anticancer Drug Etoposide

by Alireza Jian Bagherpoor, Martin Kučírek, Radek Fedr, Soodabeh Abbasi Sani and Michal Štros

Biomolecules 2020, 10(10), 1450; https://doi.org/10.3390/biom10101450 - 15 Oct 2020

Cited by 5 | Viewed by 3148

Abstract

HMGB1 and HMGB2 proteins are abundantly expressed in human embryonic stem cells
(hESCs) and hESC-derived progenitor cells (neuroectodermal cells, hNECs), though their functional
roles in pluripotency and the mechanisms underlying their dierentiation in response to the anticancer
drug etoposide remain to be elucidated. [...] Read more.

HMGB1 and HMGB2 proteins are abundantly expressed in human embryonic stem cells
(hESCs) and hESC-derived progenitor cells (neuroectodermal cells, hNECs), though their functional
roles in pluripotency and the mechanisms underlying their dierentiation in response to the anticancer
drug etoposide remain to be elucidated. Here, we show that HMGB1 and/or HMGB2 knockdown
(KD) by shRNA in hESCs did not aect the cell stemness/pluripotency regardless of etoposide
treatments, while in hESC-derived neuroectodermal cells, treatment resulted in dierential eects on
cell survival and the generation of rosette structures. The objective of this work was to determine
whether HMGB1/2 proteins could modulate the sensitivity of hESCs and hESC-derived progenitor
cells (hNECs) to etoposide. We observed that HMGB1 KD knockdown (KD) and, to a lesser extent,
HMGB2 KD enhanced the sensitivity of hESCs to etoposide. Enhanced accumulation of 53BP1 on
telomeres was detected by confocal microscopy in both untreated and etoposide-treated HMGB1
KD hESCs and hNECs, indicating that the loss of HMGB1 could destabilize telomeres. On the other
hand, decreased accumulation of 53BP1 on telomeres in etoposide-treated HMGB2 KD hESCs
(but not in HMGB2 KD hNECs) suggested that the loss of HMGB2 promoted the stability of telomeres.
Etoposide treatment of hESCs resulted in a significant enhancement of telomerase activity, with
the highest increase observed in the HMGB2 KD cells. Interestingly, no changes in telomerase activity
were found in etoposide-treated control hNECs, but HMGB2 KD (unlike HMGB1 KD) markedly
decreased telomerase activity in these cells. Changes in telomerase activity in the etoposide-treated
HMGB2 KD hESCs or hNECs coincided with the appearance of DNA damage markers and could
already be observed before the onset of apoptosis. Collectively, we have demonstrated that HMGB1
or HMGB2 dierentially modulate the impact of etoposide treatment on human embryonic stem cells
and their progenitor cells, suggesting possible strategies for the enhancement of the ecacy of this
anticancer drug. Full article

(This article belongs to the Special Issue HMG Proteins from Molecules to Disease)

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18 pages, 700 KiB

Open AccessReview

Interface of Phospholipase Activity, Immune Cell Function, and Atherosclerosis

by Robert M. Schilke, Cassidy M. R. Blackburn, Temitayo T. Bamgbose and Matthew D. Woolard

Biomolecules 2020, 10(10), 1449; https://doi.org/10.3390/biom10101449 - 15 Oct 2020

Cited by 13 | Viewed by 5767

Abstract

Phospholipases are a family of lipid-altering enzymes that can either reduce or increase bioactive lipid levels. Bioactive lipids elicit signaling responses, activate transcription factors, promote G-coupled-protein activity, and modulate membrane fluidity, which mediates cellular function. Phospholipases and the bioactive lipids they produce are [...] Read more.

Phospholipases are a family of lipid-altering enzymes that can either reduce or increase bioactive lipid levels. Bioactive lipids elicit signaling responses, activate transcription factors, promote G-coupled-protein activity, and modulate membrane fluidity, which mediates cellular function. Phospholipases and the bioactive lipids they produce are important regulators of immune cell activity, dictating both pro-inflammatory and pro-resolving activity. During atherosclerosis, pro-inflammatory and pro-resolving activities govern atherosclerosis progression and regression, respectively. This review will look at the interface of phospholipase activity, immune cell function, and atherosclerosis. Full article

(This article belongs to the Special Issue Phospholipases: From Structure to Biological Function)

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19 pages, 2814 KiB

Open AccessArticle

Assessment of Fibrinogen Macromolecules Interaction with Red Blood Cells Membrane by Means of Laser Aggregometry, Flow Cytometry, and Optical Tweezers Combined with Microfluidics

by Alexey N. Semenov, Andrei E. Lugovtsov, Evgeny A. Shirshin, Boris P. Yakimov, Petr B. Ermolinskiy, Polina Y. Bikmulina, Denis S. Kudryavtsev, Peter S. Timashev, Alexei V. Muravyov, Christian Wagner, Sehyun Shin and Alexander V. Priezzhev

Biomolecules 2020, 10(10), 1448; https://doi.org/10.3390/biom10101448 - 15 Oct 2020

Cited by 19 | Viewed by 4207

Abstract

An elevated concentration of fibrinogen in blood is a significant risk factor during many pathological diseases, as it leads to an increase in red blood cells (RBC) aggregation, resulting in hemorheological disorders. Despite the biomedical importance, the mechanisms of fibrinogen-induced RBC aggregation are [...] Read more.

An elevated concentration of fibrinogen in blood is a significant risk factor during many pathological diseases, as it leads to an increase in red blood cells (RBC) aggregation, resulting in hemorheological disorders. Despite the biomedical importance, the mechanisms of fibrinogen-induced RBC aggregation are still debatable. One of the discussed models is the non-specific adsorption of fibrinogen macromolecules onto the RBC membrane, leading to the cells bridging in aggregates. However, recent works point to the specific character of the interaction between fibrinogen and the RBC membrane. Fibrinogen is the major physiological ligand of glycoproteins receptors IIbIIIa (GPIIbIIIa or αIIββ3 or CD41/CD61). Inhibitors of GPIIbIIIa are widely used in clinics for the treatment of various cardiovascular diseases as antiplatelets agents preventing the platelets’ aggregation. However, the effects of GPIIbIIIa inhibition on RBC aggregation are not sufficiently well studied. The objective of the present work was the complex multimodal in vitro study of the interaction between fibrinogen and the RBC membrane, revealing the role of GPIIbIIIa in the specificity of binding of fibrinogen by the RBC membrane and its involvement in the cells’ aggregation process. We demonstrate that GPIIbIIIa inhibition leads to a significant decrease in the adsorption of fibrinogen macromolecules onto the membrane, resulting in the reduction of RBC aggregation. We show that the mechanisms underlying these effects are governed by a decrease in the bridging components of RBC aggregation forces. Full article

(This article belongs to the Special Issue Biochemical and Biophysical Properties of Red Blood Cells in Disease)

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18 pages, 4222 KiB

Open AccessEditor’s ChoiceReview

Genetic Alterations in the INK4a/ARF Locus: Effects on Melanoma Development and Progression

by Zizhen Ming, Su Yin Lim and Helen Rizos

Biomolecules 2020, 10(10), 1447; https://doi.org/10.3390/biom10101447 - 15 Oct 2020

Cited by 28 | Viewed by 4498

Abstract

Genetic alterations in the INK4a/ARF (or CDKN2A) locus have been reported in many cancer types, including melanoma; head and neck squamous cell carcinomas; lung, breast, and pancreatic cancers. In melanoma, loss of function CDKN2A alterations have been identified in approximately 50% of [...] Read more.

Genetic alterations in the INK4a/ARF (or CDKN2A) locus have been reported in many cancer types, including melanoma; head and neck squamous cell carcinomas; lung, breast, and pancreatic cancers. In melanoma, loss of function CDKN2A alterations have been identified in approximately 50% of primary melanomas, in over 75% of metastatic melanomas, and in the germline of 40% of families with a predisposition to cutaneous melanoma. The CDKN2A locus encodes two critical tumor suppressor proteins, the cyclin-dependent kinase inhibitor p16^INK4a and the p53 regulator p14^ARF. The majority of CDKN2A alterations in melanoma selectively target p16^INK4a or affect the coding sequence of both p16^INK4a and p14^ARF. There is also a subset of less common somatic and germline INK4a/ARF alterations that affect p14^ARF, while not altering the syntenic p16^INK4a coding regions. In this review, we describe the frequency and types of somatic alterations affecting the CDKN2A locus in melanoma and germline CDKN2A alterations in familial melanoma, and their functional consequences in melanoma development. We discuss the clinical implications of CDKN2A inactivating alterations and their influence on treatment response and resistance. Full article

(This article belongs to the Special Issue Deciphering alternative functions of the INK4a/ARF locus)

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13 pages, 8427 KiB

Open AccessBrief Report

TRPA1 Expression in Synovial Sarcoma May Support Neural Origin

by Francesco De Logu, Filippo Ugolini, Chiara Caporalini, Annarita Palomba, Sara Simi, Francesca Portelli, Domenico Andrea Campanacci, Giovanni Beltrami, Daniela Massi and Romina Nassini

Biomolecules 2020, 10(10), 1446; https://doi.org/10.3390/biom10101446 - 15 Oct 2020

Cited by 7 | Viewed by 2905

Abstract

Synovial sarcoma (SS) is a malignant mesenchymal soft tissue neoplasm. Despite its name, the cells of origin are not synovial cells, but rather neural, myogenic, or multipotent mesenchymal stem cells have been proposed as possible cells originators. Unlike other sarcomas, an unusual presentation [...] Read more.

Synovial sarcoma (SS) is a malignant mesenchymal soft tissue neoplasm. Despite its name, the cells of origin are not synovial cells, but rather neural, myogenic, or multipotent mesenchymal stem cells have been proposed as possible cells originators. Unlike other sarcomas, an unusual presentation of long-term pain at the tumor site has been documented, but the exact mechanisms have not been fully clarified yet. The transient receptor potential ankyrin 1 (TRPA1) is a nonselective cation channel mainly expressed in primary sensory neurons, where it functions as a pain sensor. TRPA1 have also been described in multiple non-excitable cells, including those derived from neural crest stem cells such as glial cells and, in particular, Schwann cell oligodendrocytes and astrocytes. We evaluated TRPA1 expression in SS. We selected a cohort of 41 SSs, and by immunohistochemistry, we studied TRPA1 expression. TRPA1 was found in 92.6% of cases. Triple TRPA1/pS100/SOX10 and TRPA1/SLUG/SNAIL staining strongly supports a neural origin of SS. TRPA1 positivity was also observed in a subset of cases negative with pS100, SOX10 and/or SLUG/SNAIL, and these divergent phenotypes may reflect a process of tumor plasticity and dedifferentiation of neural-derived SSs. Given the functional diversity of TRPA1 and its expression in neuronal and non-neuronal multipotent neural crest stem cells, it remains to be determined whether TRPA1 expression in SSs neoplastic cells plays a role in the molecular mechanism associated with premonitory pain symptoms and tumor progression. Full article

(This article belongs to the Special Issue TRP Channels in Cancer Pathophysiology and Therapy)

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16 pages, 817 KiB

Open AccessFeature PaperEditor’s ChoiceReview

Metabolic Functions of G Protein-Coupled Receptors in Hepatocytes—Potential Applications for Diabetes and NAFLD

by Takefumi Kimura, Sai P. Pydi, Jonathan Pham and Naoki Tanaka

Biomolecules 2020, 10(10), 1445; https://doi.org/10.3390/biom10101445 - 15 Oct 2020

Cited by 27 | Viewed by 9465

Abstract

G protein-coupled receptors (GPCRs) are cell surface receptors that mediate the function of extracellular ligands. Understanding how GPCRs work at the molecular level has important therapeutic implications, as 30–40% of the drugs currently in clinical use mediate therapeutic effects by acting on GPCRs. [...] Read more.

G protein-coupled receptors (GPCRs) are cell surface receptors that mediate the function of extracellular ligands. Understanding how GPCRs work at the molecular level has important therapeutic implications, as 30–40% of the drugs currently in clinical use mediate therapeutic effects by acting on GPCRs. Like many other cell types, liver function is regulated by GPCRs. More than 50 different GPCRs are predicted to be expressed in the mouse liver. However, knowledge of how GPCRs regulate liver metabolism is limited. A better understanding of the metabolic role of GPCRs in hepatocytes, the dominant constituent cells of the liver, could lead to the development of novel drugs that are clinically useful for the treatment of various metabolic diseases, including type 2 diabetes, nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). In this review, we describe the functions of multiple GPCRs expressed in hepatocytes and their role in metabolic processes. Full article

(This article belongs to the Special Issue Molecular Pathology and Therapeutics in Non-alcoholic Fatty Liver Disease)

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11 pages, 3308 KiB

Open AccessArticle

Preventive Effect of Muscone against Cisplatin Nephrotoxicity in LLC-PK1 Cells

by Hung Manh Phung, Sullim Lee, Ji Hye Hwang and Ki Sung Kang

Biomolecules 2020, 10(10), 1444; https://doi.org/10.3390/biom10101444 - 15 Oct 2020

Cited by 11 | Viewed by 4808

Abstract

Cisplatin, one of the most common antitumor agents, is widely applied to treat various cancerous diseases and is included in the World Health Organization Model List of Essential Medicines. Cisplatin therapy is used to treat 10–20% of all cancerous cases, and its cure [...] Read more.

Cisplatin, one of the most common antitumor agents, is widely applied to treat various cancerous diseases and is included in the World Health Organization Model List of Essential Medicines. Cisplatin therapy is used to treat 10–20% of all cancerous cases, and its cure rate is especially high in testicular cancer (over 90%). However, a major side effect of this anticancer drug is nephrotoxicity, limiting treatment effect and reducing the quality of life in cancer patients. Muscone, an odoriferous constituent of musk, was confirmed to inhibit cisplatin-induced LLC-PK1 kidney proximal tubule cell death in a dose-dependent manner. In term of renal protective mechanism, muscone inhibited cisplatin oxidative toxicity by decreasing reactive oxygen species (ROS) level and stimulating HO-1 expression. Muscone also exerted anti-inflammation effect through inhibition of p38 phosphorylation. Furthermore, muscone mitigated cisplatin-induced apoptosis in LLC-PK1 cells via both intrinsic and extrinsic pathways by inhibiting pro-apoptotic protein Bax expression, and cleaved caspase-3, 7, and 8; and increase of anti-apoptotic protein Bcl-2 level. In addition, the anti-apoptotic effect of muscone also was enhanced by preventing p53 expression and its phosphorylation. Our study showed that muscone may be a potential protective agent against cisplatin-induced nephrotoxicity. Full article

(This article belongs to the Special Issue Phytochemical Constituents of Medicinal Plants for the Treatment of Chronic Inflammation)

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20 pages, 11431 KiB

Open AccessFeature PaperArticle

Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes

by Akina Au, Qing Shao, Kyra K. White, Sergiu A. Lucaciu, Jessica L. Esseltine, Kevin Barr and Dale W. Laird

Biomolecules 2020, 10(10), 1443; https://doi.org/10.3390/biom10101443 - 14 Oct 2020

Cited by 12 | Viewed by 3003

Abstract

When considering connexin expression and regulation, the epidermis of the skin is one of the most complex tissues found in mammals even though it largely contains a single cell type, the keratinocyte. In the rodent epidermis, up to 9 connexin family members have [...] Read more.

When considering connexin expression and regulation, the epidermis of the skin is one of the most complex tissues found in mammals even though it largely contains a single cell type, the keratinocyte. In the rodent epidermis, up to 9 connexin family members have been detected at the mRNA level. Many of these connexins are temporally and spatially regulated in coordination with keratinocyte progenitor cell differentiation and migration from the stratum basale to form the stratum spinosum and stratum granulosum layers before finally forming the stratum corneum. Cx43 is the principal connexin found in basal keratinocytes and to a lesser degree found in keratinocytes that have begun to differentiate where Cx26, Cx30 and Cx31 become prevalent. Here we show that the CRISPR-Cas9 ablation of Cx43 reduces overall gap junction coupling in monolayer cultures of rat epidermal keratinocytes (REKs) and dysregulates the differentiation of REKs when grown in organotypic cultures. Natively found in differentiated keratinocytes, Cx31 readily assembles into gap junctions when expressed in REKs where it can extensively co-assemble into the same gap junctions with co-expressed Cx30. Time-lapse imaging indicated that many Cx31 gap junctions are mobile within the plasma membrane undergoing both fusion and fission events. Finally, the persistence of pre-existing Cx31 gap junctions in the presence of the protein trafficking blocker, brefeldin A, is longer than that found for Cx43 gap junctions indicating that it has a distinctly different life expectancy in REKs. Collectively, this study highlights the importance of Cx43 in rodent keratinocyte differentiation and suggests that Cx31 acquires life-cycle properties that are distinct from Cx43. Full article

(This article belongs to the Special Issue Connexins, Innexins, and Pannexins: From Biology to Clinical Targets)

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14 pages, 1328 KiB

Open AccessReview

Decrypting UFMylation: How Proteins Are Modified with UFM1

by Sayanika Banerjee, Manoj Kumar and Reuven Wiener

Biomolecules 2020, 10(10), 1442; https://doi.org/10.3390/biom10101442 - 14 Oct 2020

Cited by 40 | Viewed by 8244

Abstract

Besides ubiquitin (Ub), humans have a set of ubiquitin-like proteins (UBLs) that can also covalently modify target proteins. To date, less is known about UBLs than Ub and even less is known about the UBL called ubiquitin-fold modifier 1 (UFM1). Currently, our understanding [...] Read more.

Besides ubiquitin (Ub), humans have a set of ubiquitin-like proteins (UBLs) that can also covalently modify target proteins. To date, less is known about UBLs than Ub and even less is known about the UBL called ubiquitin-fold modifier 1 (UFM1). Currently, our understanding of protein modification by UFM1 (UFMylation) is like a jigsaw puzzle with many missing pieces, and in some cases it is not even clear whether these pieces of data are in the right place. Here we review the current data on UFM1 from structural biology to biochemistry and cell biology. We believe that the physiological significance of protein modification by UFM1 is currently underestimated and there is more to it than meets the eye. Full article

(This article belongs to the Special Issue Ubiquitin-Like Modifiers and Their Diverse Impact on Cell Signaling)

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16 pages, 1559 KiB

Open AccessFeature PaperArticle

GluN2 Subunit-Dependent Redox Modulation of NMDA Receptor Activation by Homocysteine

by Dmitry A. Sibarov, Sergei I. Boikov, Tatiana V. Karelina and Sergei M. Antonov

Biomolecules 2020, 10(10), 1441; https://doi.org/10.3390/biom10101441 - 14 Oct 2020

Cited by 10 | Viewed by 2892

Abstract

Homocysteine (HCY) molecule combines distinct pharmacological properties as an agonist of N-methyl-d-aspartate receptors (NMDARs) and a reducing agent. Whereas NMDAR activation by HCY was elucidated, whether the redox modulation contributes to its action is unclear. Here, using patch-clamp recording and [...] Read more.

Homocysteine (HCY) molecule combines distinct pharmacological properties as an agonist of N-methyl-d-aspartate receptors (NMDARs) and a reducing agent. Whereas NMDAR activation by HCY was elucidated, whether the redox modulation contributes to its action is unclear. Here, using patch-clamp recording and imaging of intracellular Ca²⁺, we study dithiothreitol (DTT) effects on currents and Ca²⁺ responses activated by HCY through native NMDARs and recombinant diheteromeric GluN1/2A, GluN1/2B, and GluN1/2C receptors. Within a wide range (1–800 μM) of [HCY]s, the concentration–activation relationships for recombinant NMDARs revealed a biphasicness. The high-affinity component obtained between 1 and 100 µM [HCY]s corresponding to the NMDAR activation was not affected by 1 mM DTT. The low-affinity phase observed at [HCY]s above 200 μM probably originated from thiol-dependent redox modulation of NMDARs. The reduction of NMDAR disulfide bonds by either 1 mM DTT or 1 mM HCY decreased GluN1/2A currents activated by HCY. In contrast, HCY-elicited GluN1/2B currents were enhanced due to the remarkable weakening of GluN1/2B desensitization. In fact, cleaving NMDAR disulfide bonds in neurons reversed the HCY-induced Ca²⁺ accumulation, making it dependent on GluN2B- rather than GluN2A-containing NMDARs. Thus, estimated concentrations for the HCY redox effects exceed those in the plasma during intermediate hyperhomocysteinemia but may occur during severe hyperhomocysteinemia. Full article

(This article belongs to the Special Issue Homocysteine: Biochemistry, Molecular Biology, and Role in Disease)

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18 pages, 2637 KiB

Open AccessArticle

Erinacine C Activates Transcription from a Consensus ETS DNA Binding Site in Astrocytic Cells in Addition to NGF Induction

by Monique Rascher, Kathrin Wittstein, Barbara Winter, Zeljka Rupcic, Alexandra Wolf-Asseburg, Marc Stadler and Reinhard W. Köster

Biomolecules 2020, 10(10), 1440; https://doi.org/10.3390/biom10101440 - 14 Oct 2020

Cited by 8 | Viewed by 3651

Abstract

Medicinal mushrooms of the genus Hericium are known to produce secondary metabolites with homeostatic properties for the central nervous system. We and others have recently demonstrated that among these metabolites cyathane diterpenoids and in particular erinacine C possess potent neurotrophin inducing properties in [...] Read more.

Medicinal mushrooms of the genus Hericium are known to produce secondary metabolites with homeostatic properties for the central nervous system. We and others have recently demonstrated that among these metabolites cyathane diterpenoids and in particular erinacine C possess potent neurotrophin inducing properties in astrocytic cells. Yet, the signaling events downstream of erinacine C induced neurotrophin acitivity in neural-like adrenal phaeochromocytoma cells (PC12) cells have remained elusive. Similar, signaling events activated by erinacine C in astrocytic cells are unknown. Using a combination of genetic and pharmacological inhibitors we show that erinacine C induced neurotrophic activity mediates PC12 cell differentiation via the TrkA receptor and likely its associated PLCγ-, PI3K-, and MAPK/ERK pathways. Furthermore, a small library of transcriptional activation reporters revealed that erinacine C induces transcriptional activation mediated by DNA consensus binding sites of selected conserved transcription factor families. Among these, transcription is activated from an ETS consensus in a concentration dependent manner. Interestingly, induced ETS-consensus transcription occurs in parallel and independent of neurotrophin induction. This finding helps to explain the many pleiotropic functions of cyathane diterpenoids. Moreover, our studies provide genetic access to cyathane diterpenoid functions in astrocytic cells and help to mechanistically understand the action of cyathanes in glial cells. Full article

(This article belongs to the Section Natural and Bio-derived Molecules)

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26 pages, 1456 KiB

Open AccessEditor’s ChoiceReview

Microglia in Alzheimer’s Disease in the Context of Tau Pathology

by Juan Ramón Perea, Marta Bolós and Jesús Avila

Biomolecules 2020, 10(10), 1439; https://doi.org/10.3390/biom10101439 - 14 Oct 2020

Cited by 68 | Viewed by 12694

Abstract

Microglia are the cells that comprise the innate immune system in the brain. First described more than a century ago, these cells were initially assigned a secondary role in the central nervous system (CNS) with respect to the protagonists, neurons. However, the latest [...] Read more.

Microglia are the cells that comprise the innate immune system in the brain. First described more than a century ago, these cells were initially assigned a secondary role in the central nervous system (CNS) with respect to the protagonists, neurons. However, the latest advances have revealed the complexity and importance of microglia in neurodegenerative conditions such as Alzheimer’s disease (AD), the most common form of dementia associated with aging. This pathology is characterized by the accumulation of amyloid-β peptide (Aβ), which forms senile plaques in the neocortex, as well as by the aggregation of hyperphosphorylated tau protein, a process that leads to the development of neurofibrillary tangles (NFTs). Over the past few years, efforts have been focused on studying the interaction between Aβ and microglia, together with the ability of the latter to decrease the levels of this peptide. Given that most clinical trials following this strategy have failed, current endeavors focus on deciphering the molecular mechanisms that trigger the tau-induced inflammatory response of microglia. In this review, we summarize the most recent studies on the physiological and pathological functions of tau protein and microglia. In addition, we analyze the impact of microglial AD-risk genes (APOE, TREM2, and CD33) in tau pathology, and we discuss the role of extracellular soluble tau in neuroinflammation. Full article

(This article belongs to the Special Issue Microglia in Neurodegeneration)

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Biomolecules, Volume 10, Issue 10 (October 2020) – 115 articles

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