International Journal of Molecular Sciences

2472 KiB

Open AccessArticle

Exploring the Role of Different Neonatal Nutrition Regimens during the First Week of Life by Urinary GC-MS Metabolomics

by Angelica Dessì, Antonio Murgia, Rocco Agostino, Maria Grazia Pattumelli, Andrea Schirru, Paola Scano, Vassilios Fanos and Pierluigi Caboni

Int. J. Mol. Sci. 2016, 17(2), 265; https://doi.org/10.3390/ijms17020265 - 22 Feb 2016

Cited by 45 | Viewed by 6743

Abstract

In this study, a gas-chromatography mass spectrometry (GC-MS) metabolomics study was applied to examine urine metabolite profiles of different classes of neonates under different nutrition regimens. The study population included 35 neonates, exclusively either breastfed or formula milk fed, in a seven-day timeframe. [...] Read more.

In this study, a gas-chromatography mass spectrometry (GC-MS) metabolomics study was applied to examine urine metabolite profiles of different classes of neonates under different nutrition regimens. The study population included 35 neonates, exclusively either breastfed or formula milk fed, in a seven-day timeframe. Urine samples were collected from intrauterine growth restriction (IUGR), large for gestational age (LGA), and appropriate gestational age (AGA) neonates. At birth, IUGR and LGA neonates showed similarities in their urine metabolite profiles that differed from AGA. When neonates started milk feeding, their metabolite excretion profile was strongly characterized by the different diet regimens. After three days of formula milk nutrition, urine had higher levels of glucose, galactose, glycine and myo-inositol, while up-regulated aconitic acid, aminomalonic acid and adipic acid were found in breast milk fed neonates. At seven days, neonates fed with formula milk shared higher levels of pseudouridine with IUGR and LGA at birth. Breastfed neonates shared up-regulated pyroglutamic acid, citric acid, and homoserine, with AGA at birth. The role of most important metabolites is herein discussed. Full article

(This article belongs to the Section Biochemistry)

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Gas-chromatography mass spectrometry (GC-MS) chromatograms of urine samples collected at the first day of life. Full article ">Figure 2
Orthogonal partial least square discriminant analysis (OPLS-DA) of appropriate gestational age (AGA) vs. intrauterine growth restriction (IUGR) + large for gestational age (LGA) neonates at birth, R2Y = 0.71, Q2Y = 0.36. Score plot, triangles represent the AGA class, filled and empty circles LGA and IUGR, respectively. Separation of classes is maximized through the predictive component (x-axis), intraclass variability is described through the orthogonal component (y-axis). Full article ">Figure 3
OPLS-DA of breast milk (BM) vs. formula milk (FM) fed neonates at T2, R2Y = 0.81 and Q2Y = 0.69. Score plot, triangles represent the AGA class, filled and empty circles LGA and IUGR, respectively. Black and grey colours represent BM and FM feeding, respectively. Separation of classes is maximized through the predictive component (x-axis), intraclass variability is described through the orthogonal component (y-axis). Full article ">Figure 4
Comparison between the most discriminant metabolites. Box plots display the metabolite quantitative variation in each class. Full article ">Figure 5
OPLS-DA of BM vs. FM neonates at T3, R2Y = 0.88 and Q2Y = 0.61. Score plot, triangles represent the AGA class, filled and empty circles LGA and IUGR, respectively. Black and grey colours represent BM and FM feeding, respectively. Full article ">Figure 6
Comparison between most discriminant metabolites. Box plots display the metabolite quantitative variation in each class. Full article ">

2125 KiB

Open AccessArticle

Benzyl Isothiocyanate Inhibits Prostate Cancer Development in the Transgenic Adenocarcinoma Mouse Prostate (TRAMP) Model, Which Is Associated with the Induction of Cell Cycle G1 Arrest

by Han Jin Cho, Do Young Lim, Gyoo Taik Kwon, Ji Hee Kim, Zunnan Huang, Hyerim Song, Yoon Sin Oh, Young-Hee Kang, Ki Won Lee, Zigang Dong and Jung Han Yoon Park

Int. J. Mol. Sci. 2016, 17(2), 264; https://doi.org/10.3390/ijms17020264 - 22 Feb 2016

Cited by 21 | Viewed by 7105

Abstract

Benzyl isothiocyanate (BITC) is a hydrolysis product of glucotropaeolin, a compound found in cruciferous vegetables, and has been shown to have anti-tumor properties. In the present study, we investigated whether BITC inhibits the development of prostate cancer in the transgenic adenocarcinoma mouse prostate [...] Read more.

Benzyl isothiocyanate (BITC) is a hydrolysis product of glucotropaeolin, a compound found in cruciferous vegetables, and has been shown to have anti-tumor properties. In the present study, we investigated whether BITC inhibits the development of prostate cancer in the transgenic adenocarcinoma mouse prostate (TRAMP) mice. Five-week old, male TRAMP mice and their nontransgenic littermates were gavage-fed with 0, 5, or 10 mg/kg of BITC every day for 19 weeks. The weight of the genitourinary tract increased markedly in TRAMP mice and this increase was suppressed significantly by BITC feeding. H and E staining of the dorsolateral lobes of the prostate demonstrated that well-differentiated carcinoma (WDC) was a predominant feature in the TRAMP mice. The number of lobes with WDC was reduced by BITC feeding while that of lobes with prostatic intraepithelial neoplasia was increased. BITC feeding reduced the number of cells expressing Ki67 (a proliferation marker), cyclin A, cyclin D1, and cyclin-dependent kinase (CDK)2 in the prostatic tissue. In vitro cell culture results revealed that BITC decreased DNA synthesis, as well as CDK2 and CDK4 activity in TRAMP-C2 mouse prostate cancer cells. These results indicate that inhibition of cell cycle progression contributes to the inhibition of prostate cancer development in TRAMP mice treated with BITC. Full article

(This article belongs to the Special Issue The Mechanism of Action of Food Components in Disease Prevention)

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BITC administration delays prostate cancer development in TRAMP mice. Male TRAMP mice and their nontransgenic littermates at five weeks of age were randomly divided into control and BITC-treatment groups and gavage-fed with 0 (vehicle), 5, or 10 mg/kg of BITC every day. At 24 weeks of age, all mice were sacrificed, the GU tracts were excised from the mice and weighed. (A) Body weights and (B) the GU tract weights; (C) Representative photographs of the H and E stained DP from each group (×200); (D) The incidence of the prostatic intraepithelial neoplasia and adenocarcinoma of the prostate in TRAMP mice. Data represents the mean ± SEM (n = 5). Means without a common letter differ, p < 0.05. GU tract, genitourinary tract; PIN, prostatic intraepithelial neoplasia; WDC, well-differentiated carcinoma. Scale bar, 100 μm. Full article ">Figure 2
BITC administration reduces the expression of Ki67, CDK2, cyclin A, and cyclin D1 in the prostate epithelium. The prostate sections were immunohistochemically stained using a Ki67, CDK2, CDK4, cyclin A, cyclin D1, or p21 antibody. (A) Representative photographs of DAB-stained tissue specimens; (B) The number of immune-positive cells were counted and expressed as a percentage of total cells; (C) The staining intensity was quantified and the control groups (0 mg/kg BITC-fed TRAMP mice) were set as 100%. Data represents the mean ± SEM (n = 5). Means without a common letter differ, p < 0.05. Scale bar, 100 μm. Full article ">Figure 3
BITC inhibits cell proliferation and induces G1 cell cycle arrest in TRAMP-C2 cells. TRAMP-C2 or DU145 cells were plated in 24-well plates at 5 × 104 cells/well. Twenty-four hours after plating, the monolayers were serum-deprived in DMEM/F12 containing 1% charcoal-stripped FBS for 6 h. (A) TRAMP-C2 cells were treated with 0, 5, 10, or 20 µmol/L BITC for one, two, and three days; (B) DU145 human prostate cancer cells were treated with 0, 5, or 10 µmol/L BITC for one day. (A,B) Viable cell numbers were estimated by MTT assay. Each bar represents the mean ± SEM (n = 6); (C) TRAMP-C2 cells were treated with 0, 5, 10, or 20 µmol/L BITC in the presence of [3H]thymidine for 3 h. Each bar represents the mean ± SEM (n = 6); (D) TRAMP-C2 cells were incubated for 3 h in serum-deprivation medium containing BITC (0 or 20 µmol/L). The nuclei were stained with propidium iodide and the cell cycle was analyzed via flow cytometry. Each bar represents the mean ± SEM (n = 5). (A–C) Means without a common letter differ, p < 0.05; (D) * p < 0.05 as compared with the control group. Full article ">Figure 4
BITC reduces the expression of cyclin A and cyclin D1 and inhibits the activity of CDK2 and CDK4 in TRAMP-C2 mouse prostate cancer cells. Cells were plated in 100 mm dishes at 1 × 106 cells/dish. Twenty-four h after plating, the monolayers were serum-deprived in serum-deprivation medium for 6 h. Cells were then incubated for 3 h with serum-deprivation medium containing various concentrations of BITC (0, 5, 10, or 20 µmol/L); (A) The expression of CDK2, CDK4, cyclin A, and cyclin D1 were estimated by immunoblotting. The relative intensity of each band (normalized with its own β-actin) is shown above each band; (B,C) Cell lysates were incubated with an anti-CDK2 (B) or an anti-CDK4 (C) antibody and the immune complexes were precipitated with protein A sepharose. For the in vitro kinase assay, immunoprecipitated proteins were incubated with a substrate (B: Histone H1, C: retinoblastoma protein (Rb)) and [γ-32P]ATP. Each sample was subjected to SDS-PAGE and the gel was dried. The bands were visualized by autoradiography. For Western blot analysis, immunoprecipitated proteins were analyzed by Western blotting with an anti-CDK2 (B) or an anti-CDK4 (C) antibody. The relative intensity of each band was quantified and the control (0 μmol/L BITC) levels were set at 100%. Data denotes the mean ± SEM (n = 3). Means without a common letter differ, p < 0.05. Full article ">Figure 5
Proposed mechanisms by which BITC inhibits prostate cancer development. In vitro cell culture results (blue arrow) revealed that BITC treatment decreases the expression of CDK2, CDK4, cyclin A, and cyclin D1, as well as the activity of CDK2 and CDK4, thereby resulting in the induction of G1 cell cycle arrest. In vivo results (red arrow) revealed that BITC administration decreases the expression of CDK2, cyclin A, cyclin D1, and the number of Ki67 (a proliferation marker)-positive cells in the prostatic tissue, resulting in decreases in prostate cancer development. Full article ">

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Open AccessReview

Intracellular Delivery of Proteins with Cell-Penetrating Peptides for Therapeutic Uses in Human Disease

by Ana Dinca, Wei-Ming Chien and Michael T. Chin

Int. J. Mol. Sci. 2016, 17(2), 263; https://doi.org/10.3390/ijms17020263 - 22 Feb 2016

Cited by 123 | Viewed by 9695

Abstract

Protein therapy exhibits several advantages over small molecule drugs and is increasingly being developed for the treatment of disorders ranging from single enzyme deficiencies to cancer. Cell-penetrating peptides (CPPs), a group of small peptides capable of promoting transport of molecular cargo across the [...] Read more.

Protein therapy exhibits several advantages over small molecule drugs and is increasingly being developed for the treatment of disorders ranging from single enzyme deficiencies to cancer. Cell-penetrating peptides (CPPs), a group of small peptides capable of promoting transport of molecular cargo across the plasma membrane, have become important tools in promoting the cellular uptake of exogenously delivered proteins. Although the molecular mechanisms of uptake are not firmly established, CPPs have been empirically shown to promote uptake of various molecules, including large proteins over 100 kiloDaltons (kDa). Recombinant proteins that include a CPP tag to promote intracellular delivery show promise as therapeutic agents with encouraging success rates in both animal and human trials. This review highlights recent advances in protein-CPP therapy and discusses optimization strategies and potential detrimental effects. Full article

(This article belongs to the Special Issue Cell-Penetrating Peptides)

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Open AccessArticle

Identification and Evolution of Functional Alleles of the Previously Described Pollen Specific Myrosinase Pseudogene AtTGG6 in Arabidopsis thaliana

by Lili Fu, Bingying Han, Deguan Tan, Meng Wang, Mei Ding and Jiaming Zhang

Int. J. Mol. Sci. 2016, 17(2), 262; https://doi.org/10.3390/ijms17020262 - 22 Feb 2016

Cited by 6 | Viewed by 5036

Abstract

Myrosinases are β-thioglucoside glucohydrolases and serve as defense mechanisms against insect pests and pathogens by producing toxic compounds. AtTGG6 in Arabidopsis thaliana was previously reported to be a myrosinase pseudogene but specifically expressed in pollen. However, we found that AlTGG6, an ortholog [...] Read more.

Myrosinases are β-thioglucoside glucohydrolases and serve as defense mechanisms against insect pests and pathogens by producing toxic compounds. AtTGG6 in Arabidopsis thaliana was previously reported to be a myrosinase pseudogene but specifically expressed in pollen. However, we found that AlTGG6, an ortholog to AtTGG6 in A. lyrata (an outcrossing relative of A. thaliana) was functional, suggesting that functional AtTGG6 alleles may still exist in A. thaliana. AtTGG6 alleles in 29 A. thaliana ecotypes were cloned and sequenced. Results indicate that ten alleles were functional and encoded Myr II type myrosinase of 512 amino acids, and myrosinase activity was confirmed by overexpressing AtTGG6 in Pichia pastoris. However, the 19 other ecotypes had disabled alleles with highly polymorphic frame-shift mutations and diversified sequences. Thirteen frame-shift mutation types were identified, which occurred independently many times in the evolutionary history within a few thousand years. The functional allele was expressed specifically in pollen similar to the disabled alleles but at a higher expression level, suggesting its role in defense of pollen against insect pests such as pollen beetles. However, the defense function may have become less critical after A. thaliana evolved to self-fertilization, and thus resulted in loss of function in most ecotypes. Full article

(This article belongs to the Special Issue Gene–Environment Interactions)

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Chromosomal locations and phylogenetic analysis of Myr II members in Arabidopsis lyrata and A. thaliana. (A) Chromosomal locations of Myr II genes in A. thaliana; (B) Chromosomal locations of Myr II genes in A. lyrata; (C) Phylogeny inferred from cDNAs of Myr II members in A. thaliana and A. lyrata, using Myr I myrosinases AtTGG1 (At5g26000), AtTGG2 (At5g25980), and AtTGG3 (At5g48375) as outgroups. Note: AtTGG4 (At1g47600) and AtTGG5 (At1g51470) cDNA sequences were from ecotype Col-0, AtTGG6 (At1g51490) sequence was from ecotype Tsu-1 (GenBank accession number KU301834). AlTGG cDNA sequences were obtained by annotating a genomic scaffold (NW_003302555) in A. lyrata, and partially confirmed by sequencing the PCR-amplified cDNAs. The sequences were deposited in the GenBank database under accession numbers KU301856, KU301857, KU301858, and KU301859 for AlTGG4, AlTGG5, AlTGG6, and AlTGG45, respectively. Full article ">Figure 2
Myrosinase activity of recombinant AtTGG6 from Arabidopsis ecotype Tsu-1. (A) Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of recombinant protein overexpressed in Pichia pastoris GS115; Lane 1, molecular weight marker, the mass of each band in kilodalton (kD) is shown to the left; Lane 2, purified recombinant AtTGG6; (B) Myrosinase activity of purified AtTGG6 as visualized using a glucose test reagent; AtTGG6 + Vc, AtTGG6 (50 ng) plus 0.8 mM ascorbic acid (Vc); AtTGG6 − Vc, AtTGG6 (50 ng) only, Vc not added; CK, AtTGG6 (50 ng) disabled by heating at 95 °C for 5 min before use. Full article ">Figure 3
Phylogenetic analysis of AtTGG6 genomic (A) and cDNA (B) sequences by Maximum Likelihood method. The evolutionary history was inferred based on the Tamura–Nei model [<a href="#B23-ijms-17-00262" class="html-bibr">23</a>]. The bootstrap values over 50% are shown next to the branches. Initial tree(s) for the heuristic search were obtained automatically. The tree is drawn to scale, with branch lengths measured in million years (MYA) and substitutions per site (SPS). Evolutionary analyses were conducted in MEGA6 [<a href="#B24-ijms-17-00262" class="html-bibr">24</a>]. Functional alleles are highlighted in green or blue. The two functional alleles in Tsu-1 and Mr-0 which are clustered different in genomic and cDNA phylogenies are highlighted in blue. Full article ">Figure 4
Expression pattern of functional (A–H) and disabled (I–L) alleles of AtTGG6 in Arabidopsis thaliana. Functional Prom::GUS and disabled Prom::GUS were transformed into Col-0, and stained with X-gluxuronide solution. (A) Rosette; (B) stem; (C) leaves; and (D) root of transgenic plant with functional Prom::GUS; (E) a flower bud opened with a needle showing no GUS staining; (F) a flower one day before flowering opened with a needle showing the early expression; (G) a fully opened flower showing the high expression level; (H) an anther showing predominant expression in pollen; (I) a flower bud of disabled Prom::GUS opened with a needle showing no GUS staining; (J) a flower of disabled Prom::GUS one day before flowering showing weak expression; (K) a fully opened flower of disabled Prom::GUS showing expression at moderate level; (L) an anther showing disabled AtTGG6 predominantly transcribed in pollen. Full article ">

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Open AccessArticle

Cloning and Characterization of a Flavonoid 3′-Hydroxylase Gene from Tea Plant (Camellia sinensis)

by Tian-Shan Zhou, Rui Zhou, You-Ben Yu, Yao Xiao, Dong-Hua Li, Bin Xiao, Oliver Yu and Ya-Jun Yang

Int. J. Mol. Sci. 2016, 17(2), 261; https://doi.org/10.3390/ijms17020261 - 22 Feb 2016

Cited by 38 | Viewed by 8211

Abstract

Tea leaves contain abundant flavan-3-ols, which include dihydroxylated and trihydroxylated catechins. Flavonoid 3′-hydroxylase (F3′H: EC 1.14.13.21) is one of the enzymes in the establishment of the hydroxylation pattern. A gene encoding F3′H, designated as CsF3′H, was isolated from Camellia sinensis [...] Read more.

Tea leaves contain abundant flavan-3-ols, which include dihydroxylated and trihydroxylated catechins. Flavonoid 3′-hydroxylase (F3′H: EC 1.14.13.21) is one of the enzymes in the establishment of the hydroxylation pattern. A gene encoding F3′H, designated as CsF3′H, was isolated from Camellia sinensis with a homology-based cloning technique and deposited in the GenBank (GenBank ID: KT180309). Bioinformatic analysis revealed that CsF3′H was highly homologous with the characterized F3′Hs from other plant species. Four conserved cytochrome P450-featured motifs and three F3′H-specific conserved motifs were discovered in the protein sequence of CsF3′H. Enzymatic analysis of the heterologously expressed CsF3′H in yeast demonstrated that tea F3′H catalyzed the 3′-hydroxylation of naringenin, dihydrokaempferol and kaempferol. Apparent K_m values for these substrates were 17.08, 143.64 and 68.06 μM, and their apparent V_max values were 0.98, 0.19 and 0.44 pM·min⁻¹, respectively. Transcription level of CsF3′H in the new shoots, during tea seed germination was measured, along with that of other key genes for flavonoid biosynthesis using real-time PCR technique. The changes in 3′,4′-flavan-3-ols, 3′,4′,5′-flavan-3-ols and flavan-3-ols, were consistent with the expression level of CsF3′H and other related genes in the leaves. In the study of nitrogen supply for the tea plant growth, our results showed the expression level of CsF3′H and all other tested genes increased in response to nitrogen depletion after 12 days of treatment, in agreement with a corresponding increase in 3′,4′-catechins, 3′,4′,5′-catechins and flavan 3-ols content in the leaves. All these results suggest the importance of CsF3′H in the biosynthesis of 3′,4′-catechins, 3′,4′,5′-catechins and flavan 3-ols in tea leaves. Full article

(This article belongs to the Special Issue Molecular Research in Plant Secondary Metabolism 2015)

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8877 KiB

Open AccessArticle

Chalcone-Induced Apoptosis through Caspase-Dependent Intrinsic Pathways in Human Hepatocellular Carcinoma Cells

by Rodrigo Ramirez-Tagle, Carlos A. Escobar, Valentina Romero, Ignacio Montorfano, Ricardo Armisén, Vincenzo Borgna, Emanuel Jeldes, Luis Pizarro, Felipe Simon and Cesar Echeverria

Int. J. Mol. Sci. 2016, 17(2), 260; https://doi.org/10.3390/ijms17020260 - 22 Feb 2016

Cited by 33 | Viewed by 6994

Abstract

Hepatocellular carcinoma (HCC) is one of the most commonly diagnosed cancers worldwide. Chemoprevention of HCC can be achieved through the use of natural or synthetic compounds that reverse, suppress or prevent the development of cancer progression. In this study, we investigated the antiproliferative [...] Read more.

Hepatocellular carcinoma (HCC) is one of the most commonly diagnosed cancers worldwide. Chemoprevention of HCC can be achieved through the use of natural or synthetic compounds that reverse, suppress or prevent the development of cancer progression. In this study, we investigated the antiproliferative effects and the mechanism of action of two compounds, 2,3,4′-trimethoxy-2′-hydroxy-chalcone (CH1) and 3′-bromo-3,4-dimethoxy-chalcone (CH2), over human hepatoma cells (HepG2 and Huh-7) and cultured mouse hepatocytes (HepM). Cytotoxic effects were observed over the HepG2 and Huh-7, and no effects were observed over the HepM. For HepG2 cells, treated separately with each chalcone, typical apoptotic laddering and nuclear condensation were observed. Additionally, the caspases and Bcl-2 family proteins activation by using Western blotting and immunocytochemistry were studied. Caspase-8 was not activated, but caspase-3 and -9 were both activated by chalcones in HepG2 cells. Chalcones also induced reactive oxygen species (ROS) accumulation after 4, 8 and 24 h of treatment in HepG2 cells. These results suggest that apoptosis in HepG2 was induced through: (i) a caspase-dependent intrinsic pathway; and (ii) by alterations in the cellular levels of Bcl-2 family proteins, and also, that the chalcone moiety could be a potent candidate as novel anticancer agents acting on human hepatomas. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Human Liver Diseases)

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2410 KiB

Open AccessArticle

Synthetic Human TLR9-LRR11 Peptide Attenuates TLR9 Signaling by Binding to and thus Decreasing Internalization of CpG Oligodeoxynucleotides

by Xichun Pan, Bin Li, Mei Kuang, Xin Liu, Yanyan Cen, Rongxin Qin, Guofu Ding, Jiang Zheng and Hong Zhou

Int. J. Mol. Sci. 2016, 17(2), 242; https://doi.org/10.3390/ijms17020242 - 22 Feb 2016

Cited by 30 | Viewed by 6042

Abstract

Toll-like receptor (TLR) 9 is an endosomal receptor recognizing bacterial DNA/CpG-containing oligodeoxynucleotides (CpG ODN). Blocking CpG ODN/TLR9 activity represents a strategy for therapeutic prevention of immune system overactivation. Herein, we report that a synthetic peptide (SP) representing the leucine-rich repeat 11 subdomain of [...] Read more.

Toll-like receptor (TLR) 9 is an endosomal receptor recognizing bacterial DNA/CpG-containing oligodeoxynucleotides (CpG ODN). Blocking CpG ODN/TLR9 activity represents a strategy for therapeutic prevention of immune system overactivation. Herein, we report that a synthetic peptide (SP) representing the leucine-rich repeat 11 subdomain of the human TLR9 extracellular domain could attenuate CpG ODN/TLR9 activity in RAW264.7 cells by binding to CpG ODN and decreasing its internalization. Our results demonstrate that preincubation with SP specifically inhibited CpG ODN- but not lipopolysaccharide (LPS)- and lipopeptide (PAM3CSK4)-stimulated TNF-α and IL-6 release. Preincubation of SP with CpG ODN dose-dependently decreased TLR9-driven phosphorylation of IκBα and ERK and activation of NF-κB/p65. Moreover, SP dose-dependently decreased FAM-labeled CpG ODN internalization, whereas non-labeled CpG ODN reversed the inhibition. The K_D value of SP-CpG ODN binding was within the micromolar range. Our results demonstrated that SP was a specific inhibitor of CpG ODN/TLR9 activity via binding to CpG ODN, leading to reduced ODN internalization and decreased activation of subsequent pathways within cells. Thus, SP could be used as a potential CpG ODN antagonist to block TLR9 signaling. Full article

(This article belongs to the Section Biochemistry)

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3778 KiB

Open AccessArticle

Transcriptome-Based Identification of Differently Expressed Genes from Xanthomonas oryzae pv. oryzae Strains Exhibiting Different Virulence in Rice Varieties

by Tae-Hwan Noh, Eun-Sung Song, Hong-Il Kim, Mi-Hyung Kang and Young-Jin Park

Int. J. Mol. Sci. 2016, 17(2), 259; https://doi.org/10.3390/ijms17020259 - 19 Feb 2016

Cited by 6 | Viewed by 6383

Abstract

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB) in rice (Oryza sativa L.). In this study, we investigated the genome-wide transcription patterns of two Xoo strains (KACC10331 and HB1009), which showed different virulence patterns against eight rice cultivars, including [...] Read more.

Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB) in rice (Oryza sativa L.). In this study, we investigated the genome-wide transcription patterns of two Xoo strains (KACC10331 and HB1009), which showed different virulence patterns against eight rice cultivars, including IRBB21 (carrying Xa21). In total, 743 genes showed a significant change (p-value < 0.001 in t-tests) in their mRNA expression levels in the HB1009 (K3a race) strain compared with the Xoo KACC10331 strain (K1 race). Among them, four remarkably enriched GO terms, DNA binding, transposition, cellular nitrogen compound metabolic process, and cellular macromolecule metabolic process, were identified in the upregulated genes. In addition, the expression of 44 genes was considerably higher (log2 fold changes > 2) in the HB1009 (K3a race) strain than in the Xoo KACC10331 (K1 race) strain. Furthermore, 13 and 12 genes involved in hypersensitive response and pathogenicity (hrp) and two-component regulatory systems (TCSs), respectively, were upregulated in the HB1009 (K3a race) strain compared with the Xoo KACC10331 (K1 race) strain, which we determined using either quantitative real-time PCR analysis or next-generation RNA sequencing. These results will be helpful to improve our understanding of Xoo and to gain a better insight into the Xoo–rice interactions. Full article

(This article belongs to the Special Issue Microbial Genomics and Metabolomics)

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Virulence assay (A) and lesion length (B) of rice varieties inoculated with two Xanthomonas oryzae pv. oryzae strains (K1 and K3a race). Asterisks represent statistically significant differences relative to a susceptible rice cultivar (IR24) (paired, two-tailed Student’s t test, ** p-value < 0.01). Full article ">Figure 2
Total number of R-seq reads from each Xanthomonas oryzae pv. oryzae strain library and mapped reads to the Xanthomonas oryzae pv. oryzae KACC10331 genome. Full article ">Figure 3
Functional categorization of the upregulated and downregulated genes in Xanthomonas oryzae pv. oryzae HB1009 (K3a race) based on the GO annotation. Full article ">Figure 4
The expression patterns of hrp genes of the Xanthomonas oryzae pv. oryzae KACC10331 (K1 race) and HB1009 (K3a race) strains. The gene expression levels (arbitrary units) of hpaB (A); hrpD5 (B); hpaA (C); hrcS (D); hrcR (E); hrcQ (F); hpaP (G); hrpB2 (H); hrpB8 (I); and hpa2 (J) were normalized using 16S RNA as an internal reference. Gene expression levels were quantified by real-time RT-PCR. Full article ">Figure 5
The expression patterns of TCS and rax genes of the Xanthomonas oryzae pv. oryzae KACC10331 (K1 race) and HB1009 (K3a race) strains. The gene expression levels (arbitrary units) of regR (A); XOO0519 (B); XOO3710 (C); colS (D); XOO3875 (E); raxP (F); raxR (G); raxB (H); raxA (I); and raxST (J) were normalized using 16S RNA as an internal reference. Gene expression levels were quantified by real-time RT-PCR. Full article ">

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Open AccessArticle

In Silico Screening Identifies a Novel Potential PARP1 Inhibitor Targeting Synthetic Lethality in Cancer Treatment

by Jian Li, Nan Zhou, Peiling Cai and Jinku Bao

Int. J. Mol. Sci. 2016, 17(2), 258; https://doi.org/10.3390/ijms17020258 - 19 Feb 2016

Cited by 25 | Viewed by 6986

Abstract

Synthetic lethality describes situations in which defects in two different genes or pathways together result in cell death. This concept has been applied to drug development for cancer treatment, as represented by Poly (ADP-ribose) polymerase (PARPs) inhibitors. In the current study, we performed [...] Read more.

Synthetic lethality describes situations in which defects in two different genes or pathways together result in cell death. This concept has been applied to drug development for cancer treatment, as represented by Poly (ADP-ribose) polymerase (PARPs) inhibitors. In the current study, we performed a computational screening to discover new PARP inhibitors. Among the 11,247 compounds analyzed, one natural product, ZINC67913374, stood out by its superior performance in the simulation analyses. Compared with the FDA approved PARP1 inhibitor, olaparib, our results demonstrated that the ZINC67913374 compound achieved a better grid score (−86.8) and amber score (−51.42). Molecular dynamics simulations suggested that the PARP1-ZINC67913374 complex was more stable than olaparib. The binding free energy for ZINC67913374 was −177.28 kJ/mol while that of olaparib was −159.16 kJ/mol. These results indicated ZINC67913374 bound to PARP1 with a higher affinity, which suggest ZINC67913374 has promising potential for cancer drug development. Full article

(This article belongs to the Special Issue Molecule- and Cell-Targeted Drug Design and Screening and Identification of Natural Product)

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ROC evaluation of grid and amber scoring functions. Corresponding AUC values for each ROC curve are labeled above the line. Color code: Red—amber scoring function; Blue—grid scoring function; Gray—random. Full article ">Figure 2
Binding modes of inhibitors towards PARP1 at the binding site. (a) Binding mode of olaparib; (b) binding mode of ZINC67913374. The surface of PARP1 is presented as gray with 70% transparency. Inhibitors are shown as yellow stick. Corresponding residues of PRAP1 forming hydrogen bonds with ligands are displayed as tan stick. Color code for elements: tan—C of PARP1; yellow—C of inhibitor; blue—N; red—O; green—H. Full article ">Figure 3
Backbone RMSD of PARP1®C inhibitor complexes. Black line denotes RMSD of the olaparib system while red line represents the PARP1-ZINC67913374 complex. Full article ">Figure 4
Superimposed 2D interaction diagrams of olaparib (background) and ZINC67913374 (foreground) with PARP1. Ball and stick denotes ligands. Corresponding PARP1 residues are shown as wires. Full article ">Figure 5
RMSF plots of backbone atoms for PARP1®Cinhibitor systems. Black line is for olaparib and red line is the PARP1-ZINC67913374 system. Full article ">Figure 6
Binding free energy decomposition on a per-residue basis for olaparib- and ZINC67913374-PARP1 complexes. Full article ">

975 KiB

Open AccessArticle

Deletion of Phytochelatin Synthase Modulates the Metal Accumulation Pattern of Cadmium Exposed C. elegans

by Yona J. Essig, Samuel M. Webb and Stephen R. Stürzenbaum

Int. J. Mol. Sci. 2016, 17(2), 257; https://doi.org/10.3390/ijms17020257 - 19 Feb 2016

Cited by 15 | Viewed by 5361

Abstract

Environmental metal pollution is a growing health risk to flora and fauna. It is therefore important to fully elucidate metal detoxification pathways. Phytochelatin synthase (PCS), an enzyme involved in the biosynthesis of phytochelatins (PCs), plays an important role in cadmium detoxification. The PCS [...] Read more.

Environmental metal pollution is a growing health risk to flora and fauna. It is therefore important to fully elucidate metal detoxification pathways. Phytochelatin synthase (PCS), an enzyme involved in the biosynthesis of phytochelatins (PCs), plays an important role in cadmium detoxification. The PCS and PCs are however not restricted to plants, but are also present in some lower metazoans. The model nematode Caenorhabditis elegans, for example, contains a fully functional phytochelatin synthase and phytochelatin pathway. By means of a transgenic nematode strain expressing a pcs-1 promoter-tagged GFP (pcs-1::GFP) and a pcs-1 specific qPCR assay, further evidence is presented that the expression of the C. elegans phytochelatin synthase gene (pcs-1) is transcriptionally non-responsive to a chronic (48 h) insult of high levels of zinc (500 μM) or acute (3 h) exposures to high levels of cadmium (300 μM). However, the accumulation of cadmium, but not zinc, is dependent on the pcs-1 status of the nematode. Synchrotron based X-ray fluorescence imaging uncovered that the cadmium body burden increased significantly in the pcs-1(tm1748) knockout allele. Taken together, this suggests that whilst the transcription of pcs-1 may not be mediated by an exposure zinc or cadmium, it is nevertheless an integral part of the cadmium detoxification pathway in C. elegans. Full article

(This article belongs to the Special Issue Metal Metabolism in Animals)

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The transcriptional activity of the pcs-1 promoter. The expression pattern of the pcs-1 promoter was visualized in Ppcs-1::GFP transgenic nematodes (synchronized at L4 stage) by means of confocal microscopy (Leica DMIRE2) (A); fluorescence in the head and tail region was quantified using ImageJ (B); Note, the fluorescence intensity, though universally higher in the pharyngeal intestinal valve compared to the intensity in the rectal valve, were statistically indistinguishable in nematodes raised under control conditions or nematodes exposed to 500 μM zinc for 48 h or 300 μM cadmium for 3 h. The error bars represent the standard error of the mean (±SEM, biological repeats n = 9). A quantitative RT-PCR confirmed the transcriptional invariance of pcs-1 in wild-type nematodes challenged with metals (C). The error bars denote ±SEM (technical repeats n = 3, biological repeats n = 3). Statistical analyses were performed using a one way ANOVA). Note, Ppcs-1::GFP fluorescence signal intensity or pcs-1 transcription did not differ (p > 0.05) in control and metal exposed nematodes. Full article ">Figure 2
Accumulation of metals in the nematode body. X-ray fluorescence imaging (XFI) was utilized to assess the distribution and accumulation of zinc and cadmium in the body of C. elegans raised either on control plates or transferred to plates supplemented with 500 μM zinc for 48 h or 300 μM cadmium for 3 h (A); Note, base-line levels of zinc (but not cadmium) were observed in nematodes raised on control plates. All quantitative analyses of metal load within the body of nematodes were performed using the MicroAnalysis Toolkit (A,B). A highly significant difference (p = 0.002) was apparent in nematodes exposed to zinc (500 μM zinc for 48 h), however no significant difference was observed between the strains (wild-type and the pcs-1(tm1748) mutant). Likewise, exposure to cadmium resulted in a highly significant (p = 0.002) increase in metal load in wild-type and pcs-1(tm1748). However, strain specific differences were also observed, where the pcs-1(tm1748) accumulated significantly (p = 0.002) more cadmium than the respective wild-type (B). Note: whilst zinc could be measured in nematodes raised on control plates, the cadmium signal was below the detection limit. Statistical analyses were performed using a factorial ANOVA. Note, the pixel densities differ because Cd and Zn quantifications were performed at different beamlines (Cd: 14-3; Zn 2-3), different incident energies (Cd: 3.575 keV; Zn 10 keV), and different spot sizes (Cd: 5 × 5 μM; Zn 2 × 2 μM). ** denotes p ≤ 0.01. Full article ">

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Open AccessReview

Recent Advances in Disease Modeling and Drug Discovery for Diabetes Mellitus Using Induced Pluripotent Stem Cells

by Mohammed Kawser Hossain, Ahmed Abdal Dayem, Jihae Han, Subbroto Kumar Saha, Gwang-Mo Yang, Hye Yeon Choi and Ssang-Goo Cho

Int. J. Mol. Sci. 2016, 17(2), 256; https://doi.org/10.3390/ijms17020256 - 19 Feb 2016

Cited by 27 | Viewed by 9911

Abstract

Diabetes mellitus (DM) is a widespread metabolic disease with a progressive incidence of morbidity and mortality worldwide. Despite extensive research, treatment options for diabetic patients remains limited. Although significant challenges remain, induced pluripotent stem cells (iPSCs) have the capacity to differentiate into any [...] Read more.

Diabetes mellitus (DM) is a widespread metabolic disease with a progressive incidence of morbidity and mortality worldwide. Despite extensive research, treatment options for diabetic patients remains limited. Although significant challenges remain, induced pluripotent stem cells (iPSCs) have the capacity to differentiate into any cell type, including insulin-secreting pancreatic β cells, highlighting its potential as a treatment option for DM. Several iPSC lines have recently been derived from both diabetic and healthy donors. Using different reprogramming techniques, iPSCs were differentiated into insulin-secreting pancreatic βcells. Furthermore, diabetes patient-derived iPSCs (DiPSCs) are increasingly being used as a platform to perform cell-based drug screening in order to develop DiPSC-based cell therapies against DM. Toxicity and teratogenicity assays based on iPSC-derived cells can also provide additional information on safety before advancing drugs to clinical trials. In this review, we summarize recent advances in the development of techniques for differentiation of iPSCs or DiPSCs into insulin-secreting pancreatic β cells, their applications in drug screening, and their role in complementing and replacing animal testing in clinical use. Advances in iPSC technologies will provide new knowledge needed to develop patient-specific iPSC-based diabetic therapies. Full article

(This article belongs to the Special Issue Pluripotent Stem Cell Technology for Disease Modeling, Drug Discovery and Regenerative Medicine)

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Schematic presentation of generation of iPSCs (induced pluripotent stem cells) from healthy and diabetic patients and their application in the patient-specific iPSC-based diabetic therapy. Footprint-free iPSCs can be generated from healthy individual- or diabetic patient-derived somatic cells using reprogramming DNA-, RNA-, protein-, miRNA-, or small molecule-mediated reprogramming system. DiPSCs (iPSCs derived from diabetic patients) can be further differentiated into insulin-secreting pancreatic β cells for cell-based diabetic drug screening or for transplantation into diabetic patients for cell therapy. DiPSCs can also be repaired by gene correction and then be differentiated into functional insulin-secreting pancreatic β cells, which can be transplanted into a specific diabetic patient. For disease modeling, DiPSCs can be differentiated into insulin-secreting pancreatic β cells for drug screening or pathogenesis studies to develop the compounds or therapies to treat specific type of diabetes. Full article ">Figure 2
Schematic diagram depicting the various pancreatic β cell differentiation protocols for healthy iPSCs (A) and/or DiPSCs (B). The iPSCs and DiPSC can be differentiated into insulin-secreting functional β cells through the stages, embryoid body (EB), definitive endoderm (DE), pancreatic gut tube (PGT), pancreatic progenitor (PP), posterior fore gut (PFG), multi-lineage progenitor (MP), spontaneous differentiation (SD), progenitor expansion (PE), pancreatic differentiation (PD), NKX6-1+/C-peptide+ EN cells, stem cell-derived β (SC-β) cells, and/or pancreatic β-cells using specific transcription factors and small molecules. The following transcription factors, small molecules, and specific differentiation markers were used for the pancreatic β cell differentiation; EGF (epidermal growth factor), bFGF (basic fibroblast growth factor), NA butyrate (sodium butyrate), CHIR99021 (aGSK3β inhibitor), KGF (keratinocyte growth factor), RA (retinoic acid), SANT1 (a sonic hedgehog pathway antagonist), CMRL-supplement, PdBu (phorbol 12,13-dibutyrate; a PKC activator), LDN (LDN193189; a BMP pathway inhibitor), T3 (triiodothyronine; a thyroid hormone), Alk5i (ALK5 receptor inhibitor), FGF10 (fibroblast growth factor 10), CYC (cyclopamine; a Hh signaling pathway inhibitor), ILV (indolactam V; a PKC activator), SST (somatostatin; somatotropin (growth hormone) release–inhibiting hormone), GCG (glucagon), INS (insulin), HGF (hepatocyte growth factor), DAPT (a γ-secretase (NOTCH signaling pathway) inhibitor), GLP-1 (glucagon-like peptide 1), LP1 (synaptic membrane fractions), Nico (nicotinamide), FOXA2 (forkhead box protein A2), PAX4 (paired homeobox transcription factor 4), PAX 6 (paired homeobox transcription factor 6), NGN3 (neurogenin 3), HNF (hepatocyte nuclear factor), PDX1 (pancreatic and duodenal homeobox 1); NKX6.1 (NK6 homeobox transcription factor related, locus 1), SOX17 (SRY-box 17), and NKX2.2 (NK2 homeobox transcription factor related, locus 2). Full article ">

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Open AccessArticle

Prenylated Flavonoids from Cudrania tricuspidata Suppress Lipopolysaccharide-Induced Neuroinflammatory Activities in BV2 Microglial Cells

by Dong-Cheol Kim, Chi-Su Yoon, Tran Hong Quang, Wonmin Ko, Jong-Su Kim, Hyuncheol Oh and Youn-Chul Kim

Int. J. Mol. Sci. 2016, 17(2), 255; https://doi.org/10.3390/ijms17020255 - 19 Feb 2016

Cited by 28 | Viewed by 6394

Abstract

In Korea and China, Cudrania tricuspidata Bureau (Moraceae) is an important traditional medicinal plant used to treat lumbago, hemoptysis, and contusions. The C. tricuspidata methanol extract suppressed both production of NO and PGE₂ in BV2 microglial cells. Cudraflavanone D (1), isolated from [...] Read more.

In Korea and China, Cudrania tricuspidata Bureau (Moraceae) is an important traditional medicinal plant used to treat lumbago, hemoptysis, and contusions. The C. tricuspidata methanol extract suppressed both production of NO and PGE₂ in BV2 microglial cells. Cudraflavanone D (1), isolated from this extract, remarkably suppressed the protein expression of inducible NO synthase and cyclooxygenase-2, and decreased the levels of NO and PGE₂ in BV2 microglial cells exposed to lipopolysaccharide. Cudraflavanone D (1) also decreased IL-6, TNF-α, IL-12, and IL-1β production, blocked nuclear translocation of NF-κB heterodimers (p50 and p65) by interrupting the degradation and phosphorylation of inhibitor of IκB-α, and inhibited NF-κB binding. In addition, cudraflavanone D (1) suppressed the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 MAPK pathways. This study indicated that cudraflavanone D (1) can be a potential drug candidate for the cure of neuroinflammation. Full article

(This article belongs to the Special Issue Plant-Derived Pharmaceuticals by Molecular Farming 2016)

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The structures of compounds 1–7. Full article ">Figure 2
The effects of compounds 1–7 on nitrite production in BV2 microglial cells stimulated with LPS. The cells were pre-treated for 3 h with the indicated concentrations of compounds 1–7 and then stimulated for 24 h with LPS (1 μg/mL). The concentrations of nitrite were determined as described in the Materials and Methods section. The data represent the mean values ± SD of three experiments. * p < 0.05, as compared with cells treated with LPS only. Full article ">Figure 3
The effects of cudraflavanone D (1) on TNF-α (A), IL-1β (B), IL-12 (C), and IL-6 (D) mRNA expression in BV2 microglial cells stimulated with LPS. Cells were pre-treated for 3 h with the indicated concentrations of cudraflavanone D (1) and then stimulated for 12 h with LPS (1 μg/mL). The concentrations of TNF-α (A), IL-1β (B), IL-12 (C), and IL-6 (D) were determined as described in Materials and Methods. RNA quantification was performed as described in Materials and Methods and representative blots of three independent experiments are shown. The data represent the mean values of three experiments ± SD. * p < 0.05, as compared with the cells treated with LPS only. Full article ">Figure 3 Cont.
The effects of cudraflavanone D (1) on TNF-α (A), IL-1β (B), IL-12 (C), and IL-6 (D) mRNA expression in BV2 microglial cells stimulated with LPS. Cells were pre-treated for 3 h with the indicated concentrations of cudraflavanone D (1) and then stimulated for 12 h with LPS (1 μg/mL). The concentrations of TNF-α (A), IL-1β (B), IL-12 (C), and IL-6 (D) were determined as described in Materials and Methods. RNA quantification was performed as described in Materials and Methods and representative blots of three independent experiments are shown. The data represent the mean values of three experiments ± SD. * p < 0.05, as compared with the cells treated with LPS only. Full article ">Figure 4
(A) The effects of cudraflavanone D (1) on protein expression of iNOS and COX-2 (B) in BV2 microglial cells stimulated with LPS. Cells were pre-treated for 3 h with the indicated concentrations of cudraflavanone D (1) and then stimulated for 24 h with LPS (1 μg/mL). The concentrations of iNOS and COX-2 (B) were determined as described in Materials and Methods. Western blot analyses were performed as described in Materials and Methods and representative blots of three independent experiments are shown. Band intensity was quantified by densitometry and normalized to β-actin; the values are presented below each band. Relative data represent the mean values of three experiments ± SD. * p < 0.05, as compared to the cells treated with LPS only. Full article ">Figure 5
The effects of cudraflavanone D (1) on IκB-α phosphorylation and degradation (A), NF-κB activation (B,C), NF-κB localization (D), and NF-κB DNA binding activity (E) in BV2 microglial cells. Cells were pre-treated for 3 h with the indicated concentrations of cudraflavanone D (1), and then stimulated for 1 h with LPS (1 μg/mL). Western blot analyses of IκB-α and phosphorylated (p)-IκB-α in the cytoplasm (A), and NF-κB in the cytoplasm (B) and nucleus (C), and immunofluorescent analysis (E), were performed as described in Materials and Methods. Band intensity was quantified by densitometry and normalized to β-actin and PCNA, and the values are presented below each band. Relative data represent the mean values of three experiments ± SD. * p < 0.05, as compared to the cells treated with LPS only. Full article ">Figure 6
The effects of cudraflavanone D (1) on ERK, JNK, and p38 MAPK protein expression and phosphorylation. Cells were pre-treated for 3 h with the indicated concentrations of cudraflavanone D (1) and stimulated for 1 h with LPS (1 μg/mL) (A–C). The levels of (A) phosphorylated-ERK (p-ERK), (B) phosphorylated-JNK (p-JNK), and (C) phosphorylated-p38 MAPK (p-p38 MAPK) were determined by Western blotting. Representative blots from three independent experiments are shown. Band intensity was quantified by densitometry and normalized to β-actin; the values are presented below each band. Relative data represent the mean values of three experiments. Full article ">

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Open AccessArticle

Effect of the Solvent Temperatures on Dynamics of Serine Protease Proteinase K

by Peng Sang, Qiong Yang, Xing Du, Nan Yang, Li-Quan Yang, Xing-Lai Ji, Yun-Xin Fu, Zhao-Hui Meng and Shu-Qun Liu

Int. J. Mol. Sci. 2016, 17(2), 254; https://doi.org/10.3390/ijms17020254 - 19 Feb 2016

Cited by 21 | Viewed by 6022

Abstract

To obtain detailed information about the effect of the solvent temperatures on protein dynamics, multiple long molecular dynamics (MD) simulations of serine protease proteinase K with the solute and solvent coupled to different temperatures (either 300 or 180 K) have been performed. Comparative [...] Read more.

To obtain detailed information about the effect of the solvent temperatures on protein dynamics, multiple long molecular dynamics (MD) simulations of serine protease proteinase K with the solute and solvent coupled to different temperatures (either 300 or 180 K) have been performed. Comparative analyses demonstrate that the internal flexibility and mobility of proteinase K are strongly dependent on the solvent temperatures but weakly on the protein temperatures. The constructed free energy landscapes (FELs) at the high solvent temperatures exhibit a more rugged surface, broader spanning range, and higher minimum free energy level than do those at the low solvent temperatures. Comparison between the dynamic hydrogen bond (HB) numbers reveals that the high solvent temperatures intensify the competitive HB interactions between water molecules and protein surface atoms, and this in turn exacerbates the competitive HB interactions between protein internal atoms, thus enhancing the conformational flexibility and facilitating the collective motions of the protein. A refined FEL model was proposed to explain the role of the solvent mobility in facilitating the cascade amplification of microscopic motions of atoms and atomic groups into the global collective motions of the protein. Full article

(This article belongs to the Section Physical Chemistry, Theoretical and Computational Chemistry)

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Flexibility profiles of proteinase K calculated from the joined trajectories at the four combined temperatures. (A) Per-residue Cα RMSF profiles as a function of residue number. SSEs were marked along the horizontal axis with spirals and arrows representing α (or 3/10) helices and β strands, respectively; (B) Cα RMSF values as a function of distance from the protein core to the protein surface. In both (A) and (B), the black, red, green, and blue lines are profiles corresponding to the combined temperatures P180/S180, P180/S300, P300/S180, and P300/S300, respectively. Full article ">Figure 2
Eigenvalues as a function of eigenvector index derived from ED analyses of the joined trajectories at the four combined temperatures. Only eigenvalues of the first 30 eigenvectors (total is 837) are shown. Black line: P180/S180; red line: P180/S300; green line: P300/S180; blue line: P300/S300. Full article ">Figure 3
Projection extremes of the first eigenvector obtained from ED analyses of the joined trajectories at the four combined temperatures: (A) P180/S180; (B) P300/S180; (C) P180/S300; and (D) P300/S300. The linear interpolations between the two extremes are colored from blue to red to highlight conformational differences between these two states but do not represent the transition pathway. Full article ">Figure 4
Eigenvector projections and their properties obtained from combined ED analysis of the merged MD trajectories of P300/S180 and P180/S300. (A) Projections of the merged trajectory (P300/S180: 0–84 ns; P180/S300: 84–168 ns) onto the combined eigenvectors 1–4 and 30; (B) Distributions of the corresponding eigenvector projections; (C) Average values of the first 30 eigenvector projections as a function of eigenvector index; (D) Mean square displacement (MSD) values of the first 30 eigenvector projections as a function of eigenvector index. The projection properties (average and MSD values) were calculated separately from the two equal halves of a combined eigenvector projection that correspond to the P300/S180 and P180/S300 parts, respectively. Full article ">Figure 5
Projections of the joined MD trajectories onto the two-dimensional conformational subspace spanned by the first two eigenvectors: (A) P180/S180; (B) P300/S180; (C) P180/S300; and (D) P300/S300. Full article ">Figure 6
Constructed FELs of the proteinase K-solvent system at the four combined temperatures. (A–D) FELs as a function of two-dimensional conformational subspace at the combined temperatures: (A) P180/S180; (B) P300/S180; (C) P180/S300; and (D) P300/S300. The color bar represents the free energy value in unit of kJ/mol; (E,F) One-dimensional FELs along the projection of the eigenvectors (E) 1 and (F) 2. Black line: P180/S180; red line: P180/S300; green line: P300/S180; blue line: P300/S300. Full article ">Figure 7
A proposed FEL model to explain the effect of the solvent mobility on protein dynamics. This model is represented as a hierarchical organization of free energy wells (i.e., the smallest tier-2 wells are within the relatively large tier-1 wells, and the tier-1 wells are within the largest tier-0 wells), which dictates the hierarchical dynamics of the protein (i.e., different structural components feature different amplitude and timescale of the fluctuations). The tier-2 and tier-1 substates and tier-0 states (A and B) are located within respective free energy wells. The tier-2, tier-1, and tier-0 dynamics, which are defined as conformational interconversion between respective substates/states, involve the side chain rotations on ps timescale, loop motions on ns timescale, and collective motions of the entire structure on timescale of μs to ms, respectively. The tire-0 dynamics are a result of the accumulation of the tier-1 and tier-2 dynamics. By exacerbating the competitive interactions between the protein and solvent and between atoms within the protein, the solvent mobility plays its role in facilitating the cascade amplification of microscopic motions of atoms and atomic groups into the global collective motions of the protein (for details, see the text). Full article ">

1754 KiB

Open AccessArticle

Assessment of Radiation Induced Therapeutic Effect and Cytotoxicity in Cancer Patients Based on Transcriptomic Profiling

by Sajjad Karim, Zeenat Mirza, Adeel G. Chaudhary, Adel M. Abuzenadah, Mamdooh Gari and Mohammed H. Al-Qahtani

Int. J. Mol. Sci. 2016, 17(2), 250; https://doi.org/10.3390/ijms17020250 - 19 Feb 2016

Cited by 15 | Viewed by 7227

Abstract

Toxicity induced by radiation therapy is a curse for cancer patients undergoing treatment. It is imperative to understand and define an ideal condition where the positive effects notably outweigh the negative. We used a microarray meta-analysis approach to measure global gene-expression before and [...] Read more.

Toxicity induced by radiation therapy is a curse for cancer patients undergoing treatment. It is imperative to understand and define an ideal condition where the positive effects notably outweigh the negative. We used a microarray meta-analysis approach to measure global gene-expression before and after radiation exposure. Bioinformatic tools were used for pathways, network, gene ontology and toxicity related studies. We found 429 differentially expressed genes at fold change >2 and p-value <0.05. The most significantly upregulated genes were synuclein alpha (SNCA), carbonic anhydrase I (CA1), X-linked Kx blood group (XK), glycophorin A and B (GYPA and GYPB), and hemogen (HEMGN), while downregulated ones were membrane-spanning 4-domains, subfamily A member 1 (MS4A1), immunoglobulin heavy constant mu (IGHM), chemokine (C-C motif) receptor 7 (CCR7), BTB and CNC homology 1 transcription factor 2 (BACH2), and B-cell CLL/lymphoma 11B (BCL11B). Pathway analysis revealed calcium-induced T lymphocyte apoptosis and the role of nuclear factor of activated T-cells (NFAT) in regulation of the immune response as the most inhibited pathways, while apoptosis signaling was significantly activated. Most of the normal biofunctions were significantly decreased while cell death and survival process were activated. Gene ontology enrichment analysis revealed the immune system process as the most overrepresented group under the biological process category. Toxicity function analysis identified liver, kidney and heart to be the most affected organs during and after radiation therapy. The identified biomarkers and alterations in molecular pathways induced by radiation therapy should be further investigated to reduce the cytotoxicity and development of fatigue. Full article

(This article belongs to the Collection Radiation Toxicity in Cells)

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Agglomerative average-linkage hierarchical clustering for differentially expressed genes between radiation treatment stage and controls. Dendrogram obtained using Partek GS 6.6 software shows the change in expression levels of genes (n = 429, 32-up and 397-downregulated) in RT treated cancer patients compared to untreated controls, Differentially Expressed Genes (DEGs) on X axis and treatment stage on Y axis. The cluster color represents the normalized expression level of a given gene in response to radiation treatment, Purple denotes upregulation and green denotes downregulation according the color scale. Full article ">Figure 2
Inhibition of Calcium-induced T Lymphocyte Apoptosis pathway. Based on overlap of identified DEGs to Calcium-induced T Lymphocyte Apoptosis pathways, IPA had predicted its inhibition. CD3, CAMK4, TRGV9, NFAT, ZAP70, LCK, PRKC, HLA-DOB, ITPR1 genes involved in this pathway were downregulated as shown by the purple circles. XXXX line indicate DNA strand. The white end arrow means “translocation” and the dark end arrow means “acts on”. Full article ">Figure 3
Functional analysis and regulatory effect of differentially expressed genes. Using Ingenuity Pathways Analysis (IPA), Differentially Expressed Genes (DEGs) were overlaid on to the network to find a biological regulatory functional effect based on previously reported interactions in the literature. Function colored with blue denote inhibition and orange denotes activation; development of hematopoietic progenitor cells and development of lymphocytes were inhibited whereas cell death of immune cells as activated function. Full article ">Figure 4
Gene Ontology of differentially expressed genes. Pie Chart obtained using Partek GS 6.6 software shows the change in the biological process of the immune system, biological adhesion, response to stimulus, multi-organism process, biological regulation and developmental process as a significantly affected process. Full article ">

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Open AccessReview

Energy Metabolism Plays a Critical Role in Stem Cell Maintenance and Differentiation

by Chenxia Hu, Linxiao Fan, Panpan Cen, Ermei Chen, Zhengyi Jiang and Lanjuan Li

Int. J. Mol. Sci. 2016, 17(2), 253; https://doi.org/10.3390/ijms17020253 - 18 Feb 2016

Cited by 94 | Viewed by 10912

Abstract

Various stem cells gradually turned to be critical players in tissue engineering and regenerative medicine therapies. Current evidence has demonstrated that in addition to growth factors and the extracellular matrix, multiple metabolic pathways definitively provide important signals for stem cell self-renewal and differentiation. [...] Read more.

Various stem cells gradually turned to be critical players in tissue engineering and regenerative medicine therapies. Current evidence has demonstrated that in addition to growth factors and the extracellular matrix, multiple metabolic pathways definitively provide important signals for stem cell self-renewal and differentiation. In this review, we mainly focus on a detailed overview of stem cell metabolism in vitro. In stem cell metabolic biology, the dynamic balance of each type of stem cell can vary according to the properties of each cell type, and they share some common points. Clearly defining the metabolic flux alterations in stem cells may help to shed light on stemness features and differentiation pathways that control the fate of stem cells. Full article

(This article belongs to the Special Issue Stem Cell Activation in Adult Organism)

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1293 KiB

Open AccessArticle

Metabolic Activity of Radish Sprouts Derived Isothiocyanates in Drosophila melanogaster

by Nieves Baenas, Stefanie Piegholdt, Anke Schloesser, Diego A. Moreno, Cristina García-Viguera, Gerald Rimbach and Anika E. Wagner

Int. J. Mol. Sci. 2016, 17(2), 251; https://doi.org/10.3390/ijms17020251 - 18 Feb 2016

Cited by 48 | Viewed by 9330

Abstract

We used Drosophila melanogaster as a model system to study the absorption, metabolism and potential health benefits of plant bioactives derived from radish sprouts (Raphanus sativus cv. Rambo), a Brassicaceae species rich in glucosinolates and other phytochemicals. Flies were subjected to a [...] Read more.

We used Drosophila melanogaster as a model system to study the absorption, metabolism and potential health benefits of plant bioactives derived from radish sprouts (Raphanus sativus cv. Rambo), a Brassicaceae species rich in glucosinolates and other phytochemicals. Flies were subjected to a diet supplemented with lyophilized radish sprouts (10.6 g/L) for 10 days, containing high amounts of glucoraphenin and glucoraphasatin, which can be hydrolyzed by myrosinase to the isothiocyanates sulforaphene and raphasatin, respectively. We demonstrate that Drosophila melanogaster takes up and metabolizes isothiocyanates from radish sprouts through the detection of the metabolite sulforaphane-cysteine in fly homogenates. Moreover, we report a decrease in the glucose content of flies, an upregulation of spargel expression, the Drosophila homolog of the mammalian PPARγ-coactivator 1 α, as well as the inhibition of α-amylase and α-glucosidase in vitro. Overall, we show that the consumption of radish sprouts affects energy metabolism in Drosophila melanogaster which is reflected by lower glucose levels and an increased expression of spargel, a central player in mitochondrial biogenesis. These processes are often affected in chronic diseases associated with aging, including type II diabetes mellitus. Full article

(This article belongs to the Section Biochemistry)

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Glucosinolates in radish sprouts and their corresponding hydrolysis to isothiocyanates. Full article ">Figure 2
Effect of 10-day radish sprout supplementation on male Drosophila melanogaster. (a) relative food intake analyzed by the gustatory assay (n = 3 + SEM; extraction from 3 × 15 flies); (b) relative fitness score detected by the RING assay (n = 3 + SEM). Full article ">Figure 3
(a) Metabolites present in Drosophila melanogaster following the consumption of radish sprouts for 10 days; (b) Representative chromatogram of metabolites found in Drosophila melanogaster following the consumption of radish sprouts for 10 days. F.W. = fresh weight, SFE = sulforaphene, SFN = sulforaphane, I3C = indole-3-carbinole, SFN–CYS = sulforaphane–cysteine; n = 3 + SD. Full article ">Figure 4
Effect of 10-day radish sprout supplementation on male Drosophila melanogaster. (a) relative glucose levels (n = 9 + SEM; extraction from 9 × 5 flies); (b) relative mRNA levels of spargel related to the housekeeping gene RpL32 (n = 3 + SEM; extraction from 3 × 5 flies). * indicates significant differences between control and radish sprout-fed flies (p < 0.05, Student’s t-test). Full article ">

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Open AccessArticle

Antinociceptive and Anti-Inflammatory Effects of Zerumbone against Mono-Iodoacetate-Induced Arthritis

by Ting-Yi Chien, Steven Kuan-Hua Huang, Chia-Jung Lee, Po-Wei Tsai and Ching-Chiung Wang

Int. J. Mol. Sci. 2016, 17(2), 249; https://doi.org/10.3390/ijms17020249 - 18 Feb 2016

Cited by 40 | Viewed by 7242

Abstract

The fresh rhizome of Zingiber zerumbet Smith (Zingiberaceae) is used as a food flavoring and also serves as a folk medicine as an antipyretic and for analgesics in Taiwan. Zerumbone, a monocyclic sesquiterpene was isolated from the rhizome of Z. zerumbet and is [...] Read more.

The fresh rhizome of Zingiber zerumbet Smith (Zingiberaceae) is used as a food flavoring and also serves as a folk medicine as an antipyretic and for analgesics in Taiwan. Zerumbone, a monocyclic sesquiterpene was isolated from the rhizome of Z. zerumbet and is the major active compound. In this study, the anti-inflammatory and antinociceptive effects of zerumbone on arthritis were explored using in vitro and in vivo models. Results showed that zerumbone inhibited inducible nitric oxide (NO) synthase (iNOS), cyclooxygenase (COX)-2 expressions, and NO and prostaglandin E₂ (PGE₂) production, but induced heme oxygenase (HO)-1 expression in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. When zerumbone was co-treated with an HO-1 inhibitor (tin protoporphyrin (SnPP)), the NO inhibitory effects of zerumbone were recovered. The above results suggest that zerumbone inhibited iNOS and COX-2 through induction of the HO-1 pathway. Moreover, matrix metalloproteinase (MMP)-13 and COX-2 expressions of interleukin (IL)-1β-stimulated primary rat chondrocytes were inhibited by zerumbone. In an in vivo assay, an acetic acid-induced writhing response in mice was significantly reduced by treatment with zerumbone. Furthermore, zerumbone reduced paw edema and the pain response in a mono-iodoacetate (MIA)-induced rat osteoarthritis model. Therefore, we suggest that zerumbone possesses anti-inflammatory and antinociceptive effects which indicate zerumbone could be a potential candidate for osteoarthritis treatment. Full article

(This article belongs to the Special Issue Phenolics and Polyphenolics 2015)

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Anti-inflammatory action of zerumbone on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells after treatment for 6 h. (A) Inducible nitric oxide (NO) synthase (iNOS), cyclooxygenase (COX)-2, and heme oxygenase (HO)-1 protein expressions; (B) Quantitational and statistical analysis of protein expressions, * p < 0.05 compared to control; (C) prostaglandin E2 (PGE2) production level, * p < 0.05, the zerumbone group compared to the control group; (D) NO production inhibition of zerumbone co-treated with tin protoporphyrin (SnPP), * p < 0.05, the zerumbone group compared to the group co-treated with SnPP. C: control, cells were pretreated with vehicle and LPS (500 ng/mL) B: blank, the cells incubated with vehicle alone. Data are summarized and expressed as Mean ± SEM of three individual experiments (n = 3). Full article ">Figure 2
(A) Effects of zerumbone inhibiting matrix metalloproteinase (MMP)-13 expressions on interleukin (IL)-β-induced chondrocytes; (B) Quantitational and statistical analysis of protein expressions, * p < 0.05 compared to control; Data were used from three separate experiments; a picture of one is shown. C: control, cells were pretreated with vehicle and IL-1β (10 ng/mL) B: blank. Full article ">Figure 3
Abdominal writhing response of acetic-acid induces in mice. There were six mice per group. Results are expressed as Mean ± SEM. The positive control (PC) was morphine (5 mg/kg). * p < 0.05, compared to the control group. Full article ">Figure 4
Change in the swelling volume due to paw edema after a mono-iodoacetate (MIA) injection in the ankle of a rat on days 1 to 4. The positive control (PC) was indomethacin (10 mg/kg). *** p < 0.0005 compared to the control. Full article ">Figure 5
Distribution ratio of right and left hind-limb weight-bearing with mono-iodoacetate (MIA)-induced rat osteoarthritis on day 6. The positive control (PC) was indomethacin (10 mg/kg). * p < 0.05, *** p < 0.0005 compared to the control group. Full article ">Figure 6
Chemical structure of zerumbone isolated from the fresh rhizome of Zingiber zerumbet. Full article ">Figure 7
The experimental schedule of MIA-induced osteoarthritis model. Full article ">

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Open AccessArticle

Hinokitiol Inhibits Melanogenesis via AKT/mTOR Signaling in B16F10 Mouse Melanoma Cells

by Yu-Tzu Tsao, Yu-Fen Huang, Chun-Yu Kuo, Yu-Chiang Lin, Wei-Cheng Chiang, Wei-Kuang Wang, Chia-Wei Hsu and Che-Hsin Lee

Int. J. Mol. Sci. 2016, 17(2), 248; https://doi.org/10.3390/ijms17020248 - 18 Feb 2016

Cited by 28 | Viewed by 9544

Abstract

H inokitiol purified from the heartwood of cupressaceous plants has had various biological functions of cell differentiation and growth. Hinokitiol has been demonstrated as having an important role in anti-inflammation and anti-bacteria effect, suggesting that it is potentially useful in therapies for hyperpigmentation. [...] Read more.

H inokitiol purified from the heartwood of cupressaceous plants has had various biological functions of cell differentiation and growth. Hinokitiol has been demonstrated as having an important role in anti-inflammation and anti-bacteria effect, suggesting that it is potentially useful in therapies for hyperpigmentation. Previously, hinokitiol inhibited the production of melanin by inhibiting tyrosinase activity. The autophagic signaling pathway can induce hypopigmentation. This study is warranted to investigate the mechanism of hinokitiol-induced hypopigmentation through autophagy in B16F10 melanoma cells. The melanin contents and expression of microthphalmia associated transcription factor (MITF) and tyrosinase were inhibited by treatment with hinokitiol. Moreover, the phosphorylation of the protein express levels of phospho-protein kinase B (P-AKT) and phospho-mammalian targets of rapamycin (P-mTOR) were reduced after hinokitiol treatment. In addition, the microtubule associated protein 1 light chain 3 (LC3) -II and beclin 1 (autophagic markers) were increased after the B16F10 cell was treated with hinokitiol. Meanwhile, hinokitiol decreased cellular melanin contents in a dose-dependent manner. These findings establish that hinokitiol inhibited melanogenesis through the AKT/mTOR signaling pathway. Full article

(This article belongs to the Special Issue Biochemistry and Mechanisms of Melanogenesis)

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Chemical structure of hinokitiol. Full article ">Figure 2
Effects of hinokitiol on cell viability and melanin production in B16F10 cell. B16F10 cells were treated with indicated concentrations of hinokitiol for 48 h. (a) Cell viability was measured by WST-1 assay; (b) Effect of hinokitiol on cellular melanin content; (c) Photograph of precipitated B16F10 melanoma cells. Cells were incubated for 48 h with (1.25–125 nM) and without hinokitiol; (d) Tyrosinase activity was measured. * p < 0.05 (mean ± SD, n = 6). Each experiment was repeated three times with similar results. Full article ">Figure 3
The expression levels of tyrosinase and microphthalmia-associated transcription factor (MITF) after hinokitiol treatment. B16F10 cells were treated with hinokitiol at the concentration of 1.25, 12.5 or 125 nM for 48 h. The protein expression was determined by immunoblotting. Inserted values indicated relative proteins expression in comparison with β-actin. Each experiment was repeated three times with similar results. (mean ± SD, n = 3). Full article ">Figure 4
Constitutively active-AKT reduced the expression of tyrosinase and MITF. The B16F10 (105) cells were transfected with constitutively active AKT plasmid (5 μg) for 16 h prior to treated with hinokitiol (125 nM) for 48 h. The expression of P-AKT, AKT, m-TOR, P-mTOR, MITF tyrosinase and LC3 protein in B16F10 cells was determined. The inserted values indicate relative protein expression compared to β-actin. This experiment was repeated with similar results. (mean ± SD, n = 3). Full article ">Figure 5
Hinokitiol reduces melanin content through AKT pathway. The B16F10 (105) cells were transfected with constitutively active AKT plasmid (5 μg) for 16 h prior to treated with hinokitiol (125 nM) for 48 h. The melanin expression were measured. * p < 0.05; *** p < 0.001 (mean ± SD, n = 6). Each experiment was repeated three times with similar results. Full article ">

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Open AccessReview

Systems Pharmacology in Small Molecular Drug Discovery

by Wei Zhou, Yonghua Wang, Aiping Lu and Ge Zhang

Int. J. Mol. Sci. 2016, 17(2), 246; https://doi.org/10.3390/ijms17020246 - 18 Feb 2016

Cited by 107 | Viewed by 15019

Abstract

Drug discovery is a risky, costly and time-consuming process depending on multidisciplinary methods to create safe and effective medicines. Although considerable progress has been made by high-throughput screening methods in drug design, the cost of developing contemporary approved drugs did not match that [...] Read more.

Drug discovery is a risky, costly and time-consuming process depending on multidisciplinary methods to create safe and effective medicines. Although considerable progress has been made by high-throughput screening methods in drug design, the cost of developing contemporary approved drugs did not match that in the past decade. The major reason is the late-stage clinical failures in Phases II and III because of the complicated interactions between drug-specific, human body and environmental aspects affecting the safety and efficacy of a drug. There is a growing hope that systems-level consideration may provide a new perspective to overcome such current difficulties of drug discovery and development. The systems pharmacology method emerged as a holistic approach and has attracted more and more attention recently. The applications of systems pharmacology not only provide the pharmacodynamic evaluation and target identification of drug molecules, but also give a systems-level of understanding the interaction mechanism between drugs and complex disease. Therefore, the present review is an attempt to introduce how holistic systems pharmacology that integrated in silico ADME/T (i.e., absorption, distribution, metabolism, excretion and toxicity), target fishing and network pharmacology facilitates the discovery of small molecular drugs at the system level. Full article

(This article belongs to the Section Biochemistry)

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6132 KiB

Open AccessReview

The Potential of Plant Phenolics in Prevention and Therapy of Skin Disorders

by Magdalena Działo, Justyna Mierziak, Urszula Korzun, Marta Preisner, Jan Szopa and Anna Kulma

Int. J. Mol. Sci. 2016, 17(2), 160; https://doi.org/10.3390/ijms17020160 - 18 Feb 2016

Cited by 490 | Viewed by 27092

Abstract

Phenolic compounds constitute a group of secondary metabolites which have important functions in plants. Besides the beneficial effects on the plant host, phenolic metabolites (polyphenols) exhibit a series of biological properties that influence the human in a health-promoting manner. Evidence suggests that people [...] Read more.

Phenolic compounds constitute a group of secondary metabolites which have important functions in plants. Besides the beneficial effects on the plant host, phenolic metabolites (polyphenols) exhibit a series of biological properties that influence the human in a health-promoting manner. Evidence suggests that people can benefit from plant phenolics obtained either by the diet or through skin application, because they can alleviate symptoms and inhibit the development of various skin disorders. Due to their natural origin and low toxicity, phenolic compounds are a promising tool in eliminating the causes and effects of skin aging, skin diseases, and skin damage, including wounds and burns. Polyphenols also act protectively and help prevent or attenuate the progression of certain skin disorders, both embarrassing minor problems (e.g., wrinkles, acne) or serious, potentially life-threatening diseases such as cancer. This paper reviews the latest reports on the potential therapy of skin disorders through treatment with phenolic compounds, considering mostly a single specific compound or a combination of compounds in a plant extract. Full article

(This article belongs to the Special Issue Phenolics and Polyphenolics 2015)

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Open AccessArticle

Investigation of the Antiproliferative Properties of Natural Sesquiterpenes from Artemisia asiatica and Onopordum acanthium on HL-60 Cells in Vitro

by Judit Molnár, Gábor J. Szebeni, Boglárka Csupor-Löffler, Zsuzsanna Hajdú, Thomas Szekeres, Philipp Saiko, Imre Ocsovszki, László G. Puskás, Judit Hohmann and István Zupkó

Int. J. Mol. Sci. 2016, 17(2), 83; https://doi.org/10.3390/ijms17020083 - 17 Feb 2016

Cited by 18 | Viewed by 6504

Abstract

Plants and plant extracts play a crucial role in the research into novel antineoplastic agents. Four sesquiterpene lactones, artecanin (1), 3β-chloro-4α,10α-dihydroxy-1α,2α-epoxy-5α,7αH-guaia-11(13)-en-12,6α-olide (2), iso-seco-tanapartholide 3-O-methyl ether (3) and 4β,15-dihydro-3-dehydrozaluzanin C (4), were isolated from two [...] Read more.

Plants and plant extracts play a crucial role in the research into novel antineoplastic agents. Four sesquiterpene lactones, artecanin (1), 3β-chloro-4α,10α-dihydroxy-1α,2α-epoxy-5α,7αH-guaia-11(13)-en-12,6α-olide (2), iso-seco-tanapartholide 3-O-methyl ether (3) and 4β,15-dihydro-3-dehydrozaluzanin C (4), were isolated from two traditionally used Asteraceae species (Onopordum acanthium and Artemisia asiatica). When tested for antiproliferative action on HL-60 leukemia cells, these compounds exhibited reasonable IC₅₀ values in the range 3.6–13.5 μM. Treatment with the tested compounds resulted in a cell cycle disturbance characterized by increases in the G1 and G2/M populations, while there was a decrease in the S phase. Additionally, 1–3 elicited increases in the hypodiploid (subG1) population. The compounds elicited concentration-dependent chromatin condensation and disruption of the membrane integrity, as revealed by Hoechst 33258–propidium staining. Treatment for 24 h resulted in significant increases in activity of caspases-3 and -9, indicating that the tested sesquiterpenes induced the mitochondrial pathway of apoptosis. The proapoptotic properties of the sesquiterpene lactones were additionally demonstrated withannexin V staining. Compounds 1 and 2 increased the Bax/Bcl-2 expression and decreased the expressions of CDK1 and cyclin B2, as determined at the mRNA level by means of RT-PCR. These experimental results indicate that sesquiterpene lactones may be regarded as potential starting structures for the development of novel anticancer agents. Full article

(This article belongs to the Special Issue Molecular Research in Plant Secondary Metabolism 2015)

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Open AccessEditorial

Toward Precision Medicine: How Far Is the Goal?

by Gloria Ravegnini and Sabrina Angelini

Int. J. Mol. Sci. 2016, 17(2), 245; https://doi.org/10.3390/ijms17020245 - 17 Feb 2016

Cited by 5 | Viewed by 4664

Abstract

The accomplishment of the Human Genome Project, followed by the availability of high-throughput technologies, has led to an impressive change in biomedical research.[...] Full article

(This article belongs to the Special Issue Pharmacogenetics and Personalized Medicine)

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Open AccessArticle

Therapeutic Effects of Erythroid Differentiation Regulator 1 on Imiquimod-Induced Psoriasis-Like Skin Inflammation

by Kyung Eun Kim, Younkyung Houh, Hyun Jeong Park and Daeho Cho

Int. J. Mol. Sci. 2016, 17(2), 244; https://doi.org/10.3390/ijms17020244 - 17 Feb 2016

Cited by 14 | Viewed by 7190

Abstract

Psoriasis is a common skin disease accompanied by chronic inflammation. In previous studies, erythroid differentiation regulator 1 (ERDR1) was shown to have a negative correlation with proinflammatory cytokine IL-18. However, the role of ERDR1 in the inflammatory skin disease psoriasis has not been [...] Read more.

Psoriasis is a common skin disease accompanied by chronic inflammation. In previous studies, erythroid differentiation regulator 1 (ERDR1) was shown to have a negative correlation with proinflammatory cytokine IL-18. However, the role of ERDR1 in the inflammatory skin disease psoriasis has not been evaluated. In this study, to investigate the role of ERDR1 in psoriasis, recombinant ERDR1 was injected intraperitoneally into a psoriasis mouse model. Recombinant ERDR1 (rERDR1) significantly alleviated the symptoms of psoriasis-like skin inflammation and reduced the mRNA of various psoriasis-related markers, including keratin 14, S100A8, and Th17-related cytokines IL-17 and IL-22, suggesting that rERDR1 exerts therapeutic effects on psoriasis via the regulation of Th17 functions. Additionally, the expression of CCL20, a well-known Th17 attracting chemokine, was determined. CCL20 expression significantly decreased in the rERDR1-injected group compared with the vehicle (PBS)-injected group. CCR6 expression in the psoriatic lesional skin was also decreased by rERDR1 administration, implying the inhibition of CCR6-expressing Th17 cell chemotaxis via the downregulation of CCL20. Taken together, this study provides the first evidence that ERDR1 may be a potential therapeutic target for psoriasis. Full article

(This article belongs to the Special Issue Inflammatory Skin Conditions)

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Recombinant erythroid differentiation regulator 1 (ERDR1) exerts therapeutic effects on psoriasis-like skin inflammation. (a) Photos showing redness and scales of psoriatic lesional skin induced by topical application of Aldara cream (5% imiquimod) on the shaved back skin of female 6-week-old C57BL/6J mice treated by 10 or 100 μg/kg intraperitoneally injected recombinant ERDR1 (rERDR1) versus PBS vehicle control. Five mice were used for each group, experiments were repeated independently three times, and data shown represent one mouse from each group; (b) H&E staining of the lesional skin shows decreased inflammatory infiltration, acanthosis, parakeratosis, and severe desquamation in rERDR1-injected mice compared to controls (×100 magnification); (c) Severities of redness, thickness, and scales of psoriasis-like skin lesions were evaluated from 0 to 4 (0, none; 1, mild; 2, moderate; 3, severe; and 4, very severe). Scoring was based on the severity of redness, thickness, and scales. Data are reported as mean ± standard deviation (SD). All values were analyzed using an unpaired Student’s t-test. * p < 0.05, ** p < 0.01, *** p < 0.0005; (d) The overall local clinical score was totaled from the sum of each category, including redness, thickness, and scales. Data are reported as mean ± SD. All values were analyzed using an unpaired Student’s t-test. ** p < 0.01; (e) Back skin thickness of the rERDR1-injected group vs. vehicle (PBS)-injected group, as measured using a thickness gauge. Data are reported as mean ± SD. All values were analyzed using an unpaired Student’s t-test. * p < 0.05. Data shown represent one of three independent experiments. Full article ">Figure 2
Expression of various biomarkers for psoriasis is regulated by rERDR1 administration. (a) Expression of hyper-proliferating marker, keratin 6, keratin 14, keratin 16, and antimicrobial peptide, S100A8, was detected by RT-PCR. The Krt1, Krt14, Krt16 and S100a8 mRNAs increased by imiquimod treatment were significantly decreased by rERDR1 administration; (b) Transcription of effector cytokines IL-17 and IL-22 that are produced by Th17 cells and act in psoriasis pathogenesis, as well as the inflammatory cytokine TNF-α, were detected in psoriatic lesional skin tissues. rERDR1 reduced mRNA expression of IL-17, IL-22, and TNF-α; (c) Erdr1 mRNA expression was detected in psoriatic lesional skin tissues. Erdr1 mRNA was significantly decreased following imiquimod treatment, and levels increased following rERDR1 administration. Data shown represent one of three independent experiments. Full article ">Figure 3
Recombinant ERDR1 inhibits CCL20 expression, resulting in decreased CCR6+ cells in psoriatic lesional skin. (a) Expression of CCL20, a chemokine for the trafficking of Th17 cells, was detected using RT-PCR. The increased CCL20 mRNA expression by imiquimod treatment was markedly reduced by rERDR1 administration; (b) To detect CCR6 expression in the skin tissues, immunohistochemistry was performed with a polyclonal rat anti-mouse CCR6 antibody. Staining results were visualized under a microscope (original magnification; ×100). Data shows one of three independent experiments. Full article ">

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Open AccessArticle

Integrative Transcriptome, Genome and Quantitative Trait Loci Resources Identify Single Nucleotide Polymorphisms in Candidate Genes for Growth Traits in Turbot

by Diego Robledo, Carlos Fernández, Miguel Hermida, Andrés Sciara, José Antonio Álvarez-Dios, Santiago Cabaleiro, Rubén Caamaño, Paulino Martínez and Carmen Bouza

Int. J. Mol. Sci. 2016, 17(2), 243; https://doi.org/10.3390/ijms17020243 - 17 Feb 2016

Cited by 27 | Viewed by 7209

Abstract

Growth traits represent a main goal in aquaculture breeding programs and may be related to adaptive variation in wild fisheries. Integrating quantitative trait loci (QTL) mapping and next generation sequencing can greatly help to identify variation in candidate genes, which can result in [...] Read more.

Growth traits represent a main goal in aquaculture breeding programs and may be related to adaptive variation in wild fisheries. Integrating quantitative trait loci (QTL) mapping and next generation sequencing can greatly help to identify variation in candidate genes, which can result in marker-assisted selection and better genetic structure information. Turbot is a commercially important flatfish in Europe and China, with available genomic information on QTLs and genome mapping. Muscle and liver RNA-seq from 18 individuals was carried out to obtain gene sequences and markers functionally related to growth, resulting in a total of 20,447 genes and 85,344 single nucleotide polymorphisms (SNPs). Many growth-related genes and SNPs were identified and placed in the turbot genome and genetic map to explore their co-localization with growth-QTL markers. Forty-five SNPs on growth-related genes were selected based on QTL co-localization and relevant function for growth traits. Forty-three SNPs were technically feasible and validated in a wild Atlantic population, where 91% were polymorphic. The integration of functional and structural genomic resources in turbot provides a practical approach for QTL mining in this species. Validated SNPs represent a useful set of growth-related gene markers for future association, functional and population studies in this flatfish species. Full article

(This article belongs to the Special Issue Fish Molecular Biology)

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Open AccessArticle

Delivery of Flavonoids and Saponins from Black Bean (Phaseolus vulgaris) Seed Coats Incorporated into Whole Wheat Bread

by Rocio A. Chávez-Santoscoy, Marco A. Lazo-Vélez, Sergio O. Serna-Sáldivar and Janet A. Gutiérrez-Uribe

Int. J. Mol. Sci. 2016, 17(2), 222; https://doi.org/10.3390/ijms17020222 - 17 Feb 2016

Cited by 23 | Viewed by 8206

Abstract

Cereal-based products can be used as vehicles for the delivery of relevant bioactive compounds since they are staple foods for most cultures throughout the world. The health promoting benefits of flavonoids and saponins contained in black bean seed coats have been previously described. [...] Read more.

Cereal-based products can be used as vehicles for the delivery of relevant bioactive compounds since they are staple foods for most cultures throughout the world. The health promoting benefits of flavonoids and saponins contained in black bean seed coats have been previously described. In the present work, the effect of adding flavonoids and saponins from black bean seed coat to the typical yeast-leavened whole wheat bread formulation in terms of bread features, organoleptic properties and phytochemical profile was studied. The retention of bioactive compounds was determined and the inhibitory effects of in vitro enzyme digested samples on two colon cancer cell lines (Caco-2 and HT29) was evaluated. The addition of bioactive compounds did not significantly affect baking properties or texture parameters. Among organoleptic properties of enriched breads, only crumb color was affected by the addition of bioactive compounds. However, the use of whole wheat flour partially masked the effect on color. More than 90% of added flavonoids and saponins and 80% of anthocyanins were retained in bread after baking. However, saponins were reduced more than 50% after the in vitro enzyme digestion. The black bean seed coat phytochemicals recovered after in vitro enzyme digestion of enriched breads significantly reduced by 20% the viability of colon cancer cells without affecting standard fibroblast cells (p < 0.05). Full article

(This article belongs to the Special Issue Advances in Molecular Research of Functional and Nutraceutical Food)

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1600 KiB

Open AccessArticle

Deep Sequencing and Screening of Differentially Expressed MicroRNAs Related to Milk Fat Metabolism in Bovine Primary Mammary Epithelial Cells

by Binglei Shen, Liying Zhang, Chuanjiang Lian, Chunyan Lu, Yonghong Zhang, Qiqi Pan, Runjun Yang and Zhihui Zhao

Int. J. Mol. Sci. 2016, 17(2), 200; https://doi.org/10.3390/ijms17020200 - 17 Feb 2016

Cited by 36 | Viewed by 6884

Abstract

Milk fat is a key factor affecting milk quality and is also a major trait targeted in dairy cow breeding. To determine how the synthesis and the metabolism of lipids in bovine milk is regulated at the miRNA level, primary mammary epithelial cells [...] Read more.

Milk fat is a key factor affecting milk quality and is also a major trait targeted in dairy cow breeding. To determine how the synthesis and the metabolism of lipids in bovine milk is regulated at the miRNA level, primary mammary epithelial cells (pMEC) derived from two Chinese Holstein dairy cows that produced extreme differences in milk fat percentage were cultured by the method of tissue nubbles culture. Small RNA libraries were constructed from each of the two pMEC groups, and Solexa sequencing and bioinformatics analysis were then used to determine the abundance of miRNAs and their differential expression pattern between pMECs. Target genes and functional prediction of differentially expressed miRNAs by Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes analysis illustrated their roles in milk fat metabolism. Results show that a total of 292 known miRNAs and 116 novel miRNAs were detected in both pMECs. Identification of known and novel miRNA candidates demonstrated the feasibility and sensitivity of sequencing at the cellular level. Additionally, 97 miRNAs were significantly differentially expressed between the pMECs. Finally, three miRNAs including bta-miR-33a, bta-miR-152 and bta-miR-224 whose predicted target genes were annotated to the pathway of lipid metabolism were screened and verified by real-time qPCR and Western-blotting experiments. This study is the first comparative profiling of the miRNA transcriptome in pMECs that produce different milk fat content. Full article

(This article belongs to the Special Issue MicroRNA Regulation)

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Sequencing reads distribution of known miRNAs in pMECs and mammary gland tissues. The distribution sequencing reads in the bovine lactating mammary gland was reported by Zhen et al. [<a href="#B15-ijms-17-00200" class="html-bibr">15</a>]. Full article ">Figure 2
Validation of known miRNAs using stem-loop qPCR. (a) 13 known miRNAs were detected by stem-loop qPCR, using three replicates. The relative expression was calculated using the 2−ΔΔCt method after the threshold cycle (Ct) and was normalized using the Ct of U6. The relative expression levels were presented as the 2−ΔΔCt means ± standard error. The error bars indicate the standard error of the 2−ΔΔCt mean values. *: 0.01 < p < 0.05, **: p < 0.01; (b) Comparison of expression patterns for known miRNAs (identified by Solexa sequencing) between pMECs and mammary gland (MG) tissues. X-axis: the miRNA types; Y-axis: expression differences of miRNAs in high and low fat samples. Because the methods used to calculate differential expression by stem-loop qPCR differs from that used by Solexa sequencing, significance analysis was not performed, and the figure only shows the general trend of miRNA expression. Full article ">Figure 2 Cont.
Validation of known miRNAs using stem-loop qPCR. (a) 13 known miRNAs were detected by stem-loop qPCR, using three replicates. The relative expression was calculated using the 2−ΔΔCt method after the threshold cycle (Ct) and was normalized using the Ct of U6. The relative expression levels were presented as the 2−ΔΔCt means ± standard error. The error bars indicate the standard error of the 2−ΔΔCt mean values. *: 0.01 < p < 0.05, **: p < 0.01; (b) Comparison of expression patterns for known miRNAs (identified by Solexa sequencing) between pMECs and mammary gland (MG) tissues. X-axis: the miRNA types; Y-axis: expression differences of miRNAs in high and low fat samples. Because the methods used to calculate differential expression by stem-loop qPCR differs from that used by Solexa sequencing, significance analysis was not performed, and the figure only shows the general trend of miRNA expression. Full article ">Figure 3
The number of miRNA predicted target genes mapped by pathway. Blue column: the significantly enriched pathway of target genes. Pink column: the annotated pathway related to the lipid metabolism. Full article ">Figure 4
Six miRNAs and their target genes that are involved in milk fat metabolism in bovine mammary gland. The oval color represents the pathway that the target genes involved. Blue oval: Unsaturated fatty acid synthesis. Green oval: fatty acid biosynthesis. Red oval: fatty acid metabolism. Black oval: the targets involved in more than one metabolic pathway. The colored boxes represent the expression trend. Black rectangle: down-regulated miRNAs in pMEC-LL. Blue rectangle: up-regulated miRNAs in pMEC-LL. Full article ">Figure 5
Relative expression analysis of miRNAs and their predicted target genes. (a) Bta-miR-33a was down-regulated, which was opposite to the expression level of ELOVL5, ELOVL6 and SC4MOL in MG-LL; (b) Bta-miR-152 was down-regulated in MG-LL and the significantly up-regulated genes were PTGS2, PRKAA1 and UCP3; (c) Bta-miR-224 was up-regulated in MG-LL and the corresponding down-regulated genes were ALOX15, PTGS1, LPL and GST; (d) Western-blotting results of five target genes. All of the results are expressed as the mean ± S.D. of three replicates. * p < 0.05; GAPDH is internal control. Full article ">

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Open AccessBrief Report

Atypical Antipsychotics in the Treatment of Acute Bipolar Depression with Mixed Features: A Systematic Review and Exploratory Meta-Analysis of Placebo-Controlled Clinical Trials

by Michele Fornaro, Brendon Stubbs, Domenico De Berardis, Giampaolo Perna, Alessandro Valchera, Nicola Veronese, Marco Solmi and Licínia Ganança

Int. J. Mol. Sci. 2016, 17(2), 241; https://doi.org/10.3390/ijms17020241 - 16 Feb 2016

Cited by 42 | Viewed by 9875

Abstract

Evidence supporting the use of second generation antipsychotics (SGAs) in the treatment of acute depression with mixed features (MFs) associated with bipolar disorder (BD) is scarce and equivocal. Therefore, we conducted a systematic review and preliminary meta-analysis investigating SGAs in the treatment of [...] Read more.

Evidence supporting the use of second generation antipsychotics (SGAs) in the treatment of acute depression with mixed features (MFs) associated with bipolar disorder (BD) is scarce and equivocal. Therefore, we conducted a systematic review and preliminary meta-analysis investigating SGAs in the treatment of acute BD depression with MFs. Two authors independently searched major electronic databases from 1990 until September 2015 for randomized (placebo-) controlled trials (RCTs) or open-label clinical trials investigating the efficacy of SGAs in the treatment of acute bipolar depression with MFs. A random-effect meta-analysis calculating the standardized mean difference (SMD) between SGA and placebo for the mean baseline to endpoint change in depression as well as manic symptoms score was computed based on 95% confidence intervals (CI). Six RCTs and one open-label placebo-controlled studies (including post-hoc reports) representing 1023 patients were included. Participants received either ziprasidone, olanzapine, lurasidone, quetiapine or asenapine for an average of 6.5 weeks across the included studies. Meta-analysis with Duval and Tweedie adjustment for publication bias demonstrated that SGA resulted in significant improvements of (hypo-)manic symptoms of bipolar mixed depression as assessed by the means of the total scores of the Young Mania Rating Scale (YMRS) (SMD −0.74, 95% CI −1.20 to −0.28, n SGA = 907, control = 652). Meta-analysis demonstrated that participants in receipt of SGA (n = 979) experienced a large improvement in the Montgomery–Åsberg Depression Rating Scale (MADRS) scores (SMD −1.08, 95% CI −1.35 to −0.81, p < 0.001) vs. placebo (n = 678). Publication and measurement biases and relative paucity of studies. Overall, SGAs appear to offer favorable improvements in MADRS and YMRS scores vs. placebo. Nevertheless, given the preliminary nature of the present report, additional original studies are required to allow more reliable and clinically definitive conclusions. Full article

(This article belongs to the Special Issue Antipsychotics)

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3591 KiB

Open AccessArticle

Role of miR-222-3p in c-Src-Mediated Regulation of Osteoclastogenesis

by Shinya Takigawa, Andy Chen, Qiaoqiao Wan, Sungsoo Na, Akihiro Sudo, Hiroki Yokota and Kazunori Hamamura

Int. J. Mol. Sci. 2016, 17(2), 240; https://doi.org/10.3390/ijms17020240 - 16 Feb 2016

Cited by 25 | Viewed by 6566

Abstract

MicroRNAs (miRNAs) are small non-coding RNAs that play a mostly post-transcriptional regulatory role in gene expression. Using RAW264.7 pre-osteoclast cells and genome-wide expression analysis, we identified a set of miRNAs that are involved in osteoclastogenesis. Based on in silico analysis, we specifically focused [...] Read more.

MicroRNAs (miRNAs) are small non-coding RNAs that play a mostly post-transcriptional regulatory role in gene expression. Using RAW264.7 pre-osteoclast cells and genome-wide expression analysis, we identified a set of miRNAs that are involved in osteoclastogenesis. Based on in silico analysis, we specifically focused on miR-222-3p and evaluated its role in osteoclastogenesis. The results show that the inhibitor of miR-222-3p upregulated the mRNA levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) and tartrate-resistant acid phosphatase (TRAP), while its mimicking agent downregulated their mRNA levels. Western blot analysis showed that its inhibitor increased the protein levels of TRAP and cathepsin K, while its mimicking agent decreased their levels. Genome-wide mRNA expression analysis in the presence and absence of receptor activator of nuclear factor κ-B ligand (RANKL) predicted c-Src as a potential regulatory target of miR-222-3p. Live cell imaging using a fluorescence resonance energy transfer (FRET) technique revealed that miR-222-3p acted as an inhibitor of c-Src activity, and a partial silencing of c-Src suppressed RANKL-induced expression of TRAP and cathepsin K, as well as the number of multi-nucleated osteoclasts and their pit formation. Collectively, the study herein demonstrates that miR-222-3p serves as an inhibitor of osteoclastogenesis and c-Src mediates its inhibition of cathepsin K and TRAP. Full article

(This article belongs to the Special Issue MicroRNA Regulation)

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Food-Derived Bioactives Can Protect the Anti-Inflammatory Activity of Cortisol with Antioxidant-Dependent and -Independent Mechanisms

by Erik J. B. Ruijters, Guido R. M. M. Haenen, Mathijs Willemsen, Antje R. Weseler and Aalt Bast

Int. J. Mol. Sci. 2016, 17(2), 239; https://doi.org/10.3390/ijms17020239 - 15 Feb 2016

Cited by 12 | Viewed by 6524

Abstract

In chronic inflammatory diseases the anti-inflammatory effect of glucocorticoids (GCs) is often decreased, leading to GC resistance. Inflammation is related with increased levels of reactive oxygen species (ROS), leading to oxidative stress which is thought to contribute to the development of GC resistance. [...] Read more.

In chronic inflammatory diseases the anti-inflammatory effect of glucocorticoids (GCs) is often decreased, leading to GC resistance. Inflammation is related with increased levels of reactive oxygen species (ROS), leading to oxidative stress which is thought to contribute to the development of GC resistance. Plant-derived compounds such as flavonoids are known for their ability to protect against ROS. In this exploratory study we screened a broad range of food-derived bioactives for their antioxidant and anti-inflammatory effects in order to investigate whether their antioxidant effects are associated with the ability to preserve the anti-inflammatory effects of cortisol. The anti-inflammatory potency of the tested compounds was assessed by measuring the oxidative stress–induced GC resistance in human macrophage-like cells. Cells were pre-treated with H₂O₂ (800 µM) with and without bioactives and then exposed to lipopolysaccharides (LPS) (10 ng/mL) and cortisol (100 nM). The level of inflammation was deducted from the concentration of interleukin-8 (IL-8) in the medium. Intracellular oxidative stress was measured using the fluorescent probe 2′,7′-dichlorofluorescein (DCFH). We found that most of the dietary bioactives display antioxidant and anti-inflammatory action through the protection of the cortisol response. All compounds, except for quercetin, revealing antioxidant activity also protect the cortisol response. This indicates that the antioxidant activity of compounds plays an important role in the protection of the GC response. However, next to the antioxidant activity of the bioactives, other mechanisms also seem to be involved in this protective, anti-inflammatory effect. Full article

(This article belongs to the Special Issue Antioxidant 2.0——Redox Modulation by Food and Drugs)

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Full article ">Figure 1
Effect of bioactives on intracellular oxidative stress and glucocorticoid resistance arranged in accordance to their ability to restore the anti-inflammatory effect of cortisol. (A) IL-8 levels (%) in medium after cells were pre-incubated with H2O2 (800 µM) ± bioactives (10 µM) for 1 h. and subsequently exposed to LPS (10 ng/mL) and cortisol (100 nM) for 16 h (N = 6, mean ± SEM); (B) Intracellular oxidative stress levels. Differentiated monocytes were incubated with DCFH for 45 min and then exposed to H2O2 (800 µM) ± bioactives (10 µM) and fluorescence was recorded for 1 h (N = 4, mean ± SD, * p < 0.05 Dunnett’s); (C) Correlation of the ability to reduce intracellular oxidative stress and to protect the cortisol response. Dotted lines represent maximal IL-8 production or fluorescence because no bioactives were added during H2O2 incubation. Full article ">Figure 2
Effect of the flavanol EC and several metabolites on intracellular oxidative stress and glucocorticoid resistance. (A) Cells were pre-incubated with H2O2 (800 µM) ± flavanols (10 µM) for 1 h and subsequently exposed to LPS (10 ng/mL) and cortisol (100 nM) for 16 h. IL-8 levels (%) were measured in the cell medium (N = 6, mean ± SEM); (B) Intracellular oxidative stress. Differentiated monocytes were incubated with DCFH for 45 min, exposed to H2O2 (800 µM) ± flavanols and fluorescence was measured for 1 h (N = 4, mean ± SD, * p < 0.05 Dunnett’s). The dotted line represents IL-8 production or fluorescence of cells pre-incubated with H2O2 (800 µM) without the bioactives, and subsequently exposed to LPS (10 ng/mL) and cortisol (100 nM). Full article ">Figure 3
Effect of quercetin (Q) metabolites on cortisol response. Cells were pre-incubated with H2O2 (800 µM) ± Q metabolites (10 µM) for 1 h and subsequently exposed to LPS (10 ng/mL) and cortisol (100 nM) for 16 h and IL-8 levels (%) measured in the cell medium (N = 6, mean ± SD, * p < 0.05 Mann-Whitney U test). The dotted line represents IL-8 production of cells pre-incubated with H2O2 (800 µM) without the bioactives, and subsequently exposed to LPS (10 ng/mL) and cortisol (100 nM). Full article ">

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Open AccessArticle

The Chinese Herbal Medicine Formula mKG Suppresses Pulmonary Fibrosis of Mice Induced by Bleomycin

by Ying Gao, Li-Fu Yao, Yang Zhao, Li-Man Wei, Peng Guo, Meng Yu, Bo Cao, Tan Li, Hong Chen and Zhong-Mei Zou

Int. J. Mol. Sci. 2016, 17(2), 238; https://doi.org/10.3390/ijms17020238 - 15 Feb 2016

Cited by 24 | Viewed by 6809

Abstract

Pulmonary fibrosis (PF) is a serious progressive lung disease and it originates from inflammation-induced parenchymal injury with excessive extracellular matrix deposition to result in the destruction of the normal lung architecture. Modified Kushen Gancao Formula (mKG), derived from traditional Chinese herbal [...] Read more.

Pulmonary fibrosis (PF) is a serious progressive lung disease and it originates from inflammation-induced parenchymal injury with excessive extracellular matrix deposition to result in the destruction of the normal lung architecture. Modified Kushen Gancao Formula (mKG), derived from traditional Chinese herbal medicine, has a prominent anti-inflammatory effect. The present study is to explore the inhibitory effects of mKG on bleomycin (BLM)-induced pulmonary fibrosis in mice. mKG significantly decreased pulmonary alveolitis, fibrosis scores, and interleukin-6 (IL-6), interleukin-17 (IL-17), transforming growth factor-β (TGF-β) and hydroxyproline (HYP) levels in lung tissue of mice compared with BLM treatment. It markedly alleviated the increase of HYP content in the lung tissues and pathologic changes of pulmonary fibrosis caused by BLM instillation. In conclusion, mKG has an anti-fibrotic effect and might be employed as a therapeutic candidate agent for attenuating pulmonary fibrosis. Full article

(This article belongs to the Section Biochemistry)

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Int. J. Mol. Sci., Volume 17, Issue 2 (February 2016) – 125 articles

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