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Theranostics 2021Tanshinone, a type of diterpenes derived from , is a particularly promising herbal medicine compound for the treatment of cancers including acute myeloid leukemia...
Tanshinone, a type of diterpenes derived from , is a particularly promising herbal medicine compound for the treatment of cancers including acute myeloid leukemia (AML). However, the therapeutic function and the underlying mechanism of Tanshinone in AML are not clear, and the toxic effect of Tanshinone limits its clinical application. Our work utilizes human leukemia cell lines, zebrafish transgenics and xenograft models to study the cellular and molecular mechanisms of how Tanshinone affects normal and abnormal hematopoiesis. WISH, Sudan Black and O-Dianisidine Staining were used to determine the expression of hematopoietic genes on zebrafish embryos. RNA-seq analysis showed that differential expression genes and enrichment gene signature with Tan I treatment. The surface plasmon resonance (SPR) method was used with a BIAcore T200 (GE Healthcare) to measure the binding affinities of Tan I. methyltransferase assay was performed to verify Tan I inhibits the histone enzymatic activity of the PRC2 complex. ChIP-qPCR assay was used to determine the H3K27me3 level of EZH2 target genes. We found that Tanshinone I (Tan I), one of the Tanshinones, can inhibit the proliferation of human leukemia cells and in the xenograft zebrafish model, as well as the normal and malignant definitive hematopoiesis in zebrafish. Mechanistic studies illustrate that Tan I regulates normal and malignant hematopoiesis through direct binding to EZH2, a well-known histone H3K27 methyltransferase, and inhibiting PRC2 enzymatic activity. Furthermore, we identified and as two possible downstream genes of Tan I's effects on EZH2. Together, this study confirmed that Tan I is a novel EZH2 inhibitor and suggested and as two potential therapeutic targets for myeloid malignant diseases.
Topics: ATP Binding Cassette Transporter, Subfamily G, Member 2; Abietanes; Animals; Animals, Genetically Modified; Antineoplastic Agents, Phytogenic; Cell Line, Tumor; Cell Proliferation; Chromatin Immunoprecipitation; Enhancer of Zeste Homolog 2 Protein; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Hematopoiesis; Histones; Humans; Leukemia; Matrix Metalloproteinase 9; Neoplasm Proteins; Polycomb Repressive Complex 2; Protein Binding; RNA-Seq; Salvia miltiorrhiza; Surface Plasmon Resonance; Transcriptome; Xenograft Model Antitumor Assays; Zebrafish
PubMed: 34093860
DOI: 10.7150/thno.53170 -
Journal of Hazardous Materials Oct 2023In this study, we developed a colorimetric ozone passive sampler (OPS) incorporating o-dianisidine, a redox dye, into a polydimethylsiloxane sheet. The reaction between...
In this study, we developed a colorimetric ozone passive sampler (OPS) incorporating o-dianisidine, a redox dye, into a polydimethylsiloxane sheet. The reaction between ozone (O) and o-dianisidine result in a visible yellowish color change. Unlike previous passive methods that rely on nitrate extraction or the color disappearance of indigotrisulfonate, the OPS offered improved recognition of average O exposure. To optimize OPS based on time-weighted average (TWA), we extracted and quantified the amount of reacted o-dianisidine after exposing OPS to O by varying concentrations (0-200 ppb) within 8 h. Colorimetric changes of OPS were further analyzed by capturing images, and the effective absorbance of blue scale showed the best fit (EA, R =0.997). OPS validation on visual detection assessed by six parameters: limit of detection, limit of quantification, reproducibility, sampling rate, selectivity to interfering gases, and sensitivity to environmental factors. To enhance visibility, the OPS was assembled with coloration exposure guidelines, and a smartphone app was developed to quantify average O exposures. We further conducted field tests that showed the significant disparity between O concentrations and personal O exposures, which is considered more crucial for assessing health risks. The OPS was optimized to monitor O exposure levels and raise awareness among workers and occupants regarding invisible indoor hazards.
Topics: Humans; Colorimetry; Dianisidine; Reproducibility of Results; Levonorgestrel; Ozone
PubMed: 37703734
DOI: 10.1016/j.jhazmat.2023.132510 -
International Journal of Nanomedicine 2023Curcumin (CUR) and piperine (PP) are bioactive compounds with prominent pharmacological activities that have been investigated for the treatment of various diseases. The...
Development of Curcumin and Piperine-Loaded Bio-Active Self-Nanoemulsifying Drugs and Investigation of Their Bioactivity in Zebrafish Embryos and Human Hematological Cancer Cell Lines.
PURPOSE
Curcumin (CUR) and piperine (PP) are bioactive compounds with prominent pharmacological activities that have been investigated for the treatment of various diseases. The aim of the present study is to develop Bio-SNEDDS for CUR and PP as a combined delivery system for cancer therapy.
METHODS
CUR and PP loaded Bio-SNEDDSs with varying compositions of bioactive lipid oils, surfactants, and cosolvents were prepared at room temperature. Bio-SNEDDSs were characterized using a Zetasizer Nano particle size analyzer and further examined by transmission electron microscopy (TEM) for morphology. The in vivo toxicity of the preparations of Bio-SNEDDS was investigated in wild-type zebrafish embryos and cytotoxicity in THP-1 (human leukemia monocytic cells), Jurkat (human T lymphocyte cells) and HUVEC (non-cancerous normal) cells.
RESULTS
Bio-SNEDDSs were successfully developed with black seed oil, Imwitor 988, Transcutol P and Cremophor RH40 at a ratio of 20/20/10/50 (%w/w). The droplet size, polydispersity index and zeta potential of the optimized Bio-SNEDDS were found to be 42.13 nm, 0.59, and -19.30 mV, respectively. Bio-SNEDDS showed a spherical structure evident by TEM analysis. The results showed that Bio-SNEDDS did not induce toxicity in zebrafish embryos at concentrations between 0.40 and 30.00 μg/mL. In TG (fli1: EGFP) embryos treated with Bio-SNEDDS, there was no change in the blood vessel structure. The O-dianisidine staining of Bio-SNEDDS treated embryos at 48 h post-fertilization also showed a significant reduction in the number of blood cells compared to mock (DMSO 0.1% V/V) treated embryos. Bio-SNEDDS induced significant levels of cytotoxicity in the hematological cell lines THP-1 and Jurkat, while low toxicity in normal HUVEC cell lines was observed with IC50 values of 18.63±0.23 μg/mL, 26.03 ± 1.5 μg/mL and 17.52 ± 0.22 μg/mL, respectively.
CONCLUSION
Bio-SNEDDS exhibited enhanced anticancer activity and could thus be an important new pharmaceutical formulation to treat leukemia.
Topics: Animals; Humans; Pharmaceutical Preparations; Curcumin; Zebrafish; Drug Delivery Systems; Solubility; Administration, Oral; Surface-Active Agents; Hematologic Neoplasms; Leukemia; Emulsions; Nanoparticles; Biological Availability
PubMed: 37051315
DOI: 10.2147/IJN.S400330 -
Heliyon Jan 2021Various aromatic compounds that are structurally analogous to lignin were tested as possible/preferred substrates for purified laccase from newly isolated white rote...
Various aromatic compounds that are structurally analogous to lignin were tested as possible/preferred substrates for purified laccase from newly isolated white rote fungi, WRF03. The pH optima were tested using different substrates and kinetic studies were conducted at these pH optima. The pH optima in the presence of ABTS, α-naphthol, o-dianisidine, and catechol were 4.5 but 5.0 and 5.5 in the presence of guaiacol and pyrogallol, respectively. The initial velocities obtained from the kinetic study were analyzed using Graph Pad Prism 7 and Lineweaver-Burk plot to obtain kinetic constants ( and ) which were used to calculate substrate specificity. Amongst all the substrates tested, ABTS had the highest specificity-constant (181.51 Ms), and therefore, the most preferred substrate was followed by α-naphthol, -dianisidine, guaiacol, pyrogallol, and catechol. Resorcinol, orcinol, and veratryl alcohol did not display any considerable chemical shift in the presence of WRF03 laccase. Also, oxidation of phenolic substrates appeared to be dependent on the nature of the substituent groups and their relative position on the aromatic nucleus. Since most of these substrates are structural analogs of lignin and many recalcitrant environmental pollutants, the enzyme may find application in delignification, treatment of wastewater containing dyes, and polycyclic aromatic hydrocarbons (PAHs).
PubMed: 33537494
DOI: 10.1016/j.heliyon.2021.e06080 -
Polymers Dec 2020The pronouncedly low thermal conductivity of polymers in the range of 0.1-0.2 W m K is a limiting factor for their application as an insulating layer in microelectronics...
The pronouncedly low thermal conductivity of polymers in the range of 0.1-0.2 W m K is a limiting factor for their application as an insulating layer in microelectronics that exhibit continuously higher power-to-volume ratios. Two strategies can be applied to increase the thermal conductivity of polymers; that is, compounding with thermally conductive inorganic materials as well as blending with aromatic units arranged by the principle of π-π stacking. In this study, both strategies were investigated and compared on the example of epoxy-amine resins of bisphenol A diglycidyl ether (BADGE) and 1,2,7,8-diepoxyoctane (DEO), respectively. These two diepoxy compounds were cured with mixtures of the diamines isophorone diamine (IPDA) and -dianisidine (DAN). The epoxy-amine resins were cured without filler and with 5 wt.-% of SiO nanoparticles. Enhanced thermal conductivity in the range of 0.4 W·m·K was observed exclusively in DEO-based polymer networks that were cured with DAN (and do not contain SiO fillers). This observation is argued to originate from π-π stacking of the aromatic units of DAN enabled by the higher flexibility of the aliphatic carbon chain of DEO compared with that of BADGE. The enhanced thermal conductivity occurs only at temperatures above the glass-transition point and only if no inorganic fillers, which disrupt the π-π stacking of the aromatic groups, are present. In summary, it can be argued that the bisphenol-free epoxy-amine resin with an epoxy compound derivable from natural resources shows favorably higher thermal conductivity in comparison with the petrol-based bisphenol-based epoxy/amine resins.
PubMed: 33375238
DOI: 10.3390/polym13010065 -
The Journal of Biological Chemistry May 1979Extracts of aerobically grown Escherichia coli B exhibit both catalase and dianisidine peroxidase activities. Polyacrylamide gel electrophoresis demonstrates two...
Extracts of aerobically grown Escherichia coli B exhibit both catalase and dianisidine peroxidase activities. Polyacrylamide gel electrophoresis demonstrates two distinct catalases which have been designated hydroperoxidases I and II (HP-I and HP-II) in order of increasing anodic mobility. HP-I has been purified to essential homogeneity and found to be composed of four subunits of equal size. Its molecular weight is 337,000, and it contains two molecules of protoheme IX per tetramer. Its amino acid composition is unusual, for so large a protein, in lacking half-cystine. HP-I is a very efficient catalase with an activity optimum at pH 7.5, a Km for H2O2 of 3.9 mM, and a turnover number of 9.8 x 10(5) per min. It is also a broad specificity peroxidase capable of acting upon dianisidine, guaiacol, p-phenylenediamine, and pyrogallol. Dianisidine acted as a powerful reversible inhibitor of the catalatic activity of HP-I and as a suicide substrate when HP-I functioned in its peroxidatic mode.
Topics: Amino Acids; Dianisidine; Escherichia coli; Kinetics; Macromolecular Substances; Molecular Weight; Peroxidases; Substrate Specificity
PubMed: 374409
DOI: No ID Found -
Indian Journal of Clinical Biochemistry... Jan 2009Serum ceruloplasmin is one of the most commonly used screening tests for Wilson's disease. However immunological assays for ceruloplasmin are not recommended for...
Serum ceruloplasmin is one of the most commonly used screening tests for Wilson's disease. However immunological assays for ceruloplasmin are not recommended for diagnosis and management of Wilson's disease through calculation of free copper index. Enzymatic methods using non-physiological substrates have toxicity and stability problems, making them difficult to automate. Ferroxidase assays may be a satisfactory alternative for measuring serum ceruloplasmin. The o-dianisidine hydrochloride manual method for estimation of serum ceruloplasmin enzyme activity was compared with an automated method using the ferroxidase activity of ceruloplasmin in measurement in a double blind study in 91 consecutive patients screened for Wilson's disease. The o-dianisidine and ferroxidase methods both successfully identified 7 patients with Wilson's disease. Values for these 7 patients in the o-dianisidine and ferroxidase methods were median 5.0 (range 0-16.0 U/L) and median 45.0 (range 4-166 U/L) respectively. There were 7 other positive values (<62 U/L) with the o-diansidine method and 2 (<200 U/L) with the ferroxidase method, where WD was not confirmed. ROC curves for both methods showed area under the curve of 0.998 for o-dianisidine and 0.997 for ferroxidase. Using literature cut off values of 62 U/L and 200 U/L respectively both methods had 100% sensitivity and specificity was 91.7% (o-dianisidine) and 97.6% (ferroxidase). For the o-dianisidine assay, specificity was improved to 98.8% using a cut off of 22.5 U/L. In the 84 persons (46 adults and 38 children) in whom the diagnosis of Wilson's disease was not established, the mean value for ceruloplasmin activity by the o-dianisidine and ferroxidase methods was 124.7 ± 48.7 U/L and 571.4 ± 168.1 U/L respectively. There were no significant differences between sex or age of patients (p > 0.29). In a subsequent evaluation with 372 specimens, the Pearson correlation coefficient between the assays was 0.908, p < 0.01, slope 4.06, intercept 265.8, with the manual assay as the x-axis. The ferroxidase assay is a suitable replacement for the o-dianisidine assay in detecting patients with Wilson's disease.
PubMed: 23105801
DOI: 10.1007/s12291-009-0003-4 -
Ecotoxicology and Environmental Safety Jan 2021The environmental effects of additives have attracted increasing attention. Sodium dehydroacetate (DHA-S), as an approved preservative, is widely added in processed...
The environmental effects of additives have attracted increasing attention. Sodium dehydroacetate (DHA-S), as an approved preservative, is widely added in processed foods, cosmetics and personal care products. However, DHA-S has been recently reported to induce hemorrhage and coagulation aberration in rats. Yet little is known about the ecotoxicological effect and underlying mechanisms of DHA-S. Here, we utilized the advantage of zebrafish model to evaluate such effects. DHA-S induced cerebral hemorrhage, mandibular dysplasia and pericardial edema in zebrafish after 24 h exposure (48-72 hpf) at 50 mg/L. We also observed the defective heart looping and apoptosis in DHA-S-treated zebrafish through o-dianisidine and acridine orange staining. Meanwhile, DHA-S induced the deficiency of Ca and vitamin D3 in zebrafish. We further demonstrated that DHA-S stimulated Ca influx resulting in Ca-dependent mitochondrial damage in cardiomyocytes. Additionally, DHA-S inhibited glucose uptake and repressed the biosynthesis of amino acids. Finally, we identified that sodium bicarbonate could rescue zebrafish from DHA-S induced cardiovascular toxicity. Altogether, our results suggest that DHA-S is a potential risk for cardiovascular system.
Topics: Animals; Apoptosis; Calcium; Cardiotoxicity; Cell Line; Cerebral Hemorrhage; Dose-Response Relationship, Drug; Edema, Cardiac; Embryo, Nonmammalian; Embryonic Development; Heart; Myocardium; Pericardium; Pyrones; Rats; Zebrafish
PubMed: 33396133
DOI: 10.1016/j.ecoenv.2020.111613 -
Nanotoxicology Jun 2018Nowadays, nanotechnology environmental health and safety (nanoEHS) is gaining attention. We previously found that silica nanoparticles (SiNPs) could induce vascular...
Nowadays, nanotechnology environmental health and safety (nanoEHS) is gaining attention. We previously found that silica nanoparticles (SiNPs) could induce vascular endothelial damage. However, the subsequent toxicologic response to SiNPs-induced endothelial damage was still largely unknown. In this study, we explored the inflammation-coagulation response and thrombotic effects of SiNPs in endothelial cells and zebrafish embryos. For in vitro study, swollen mitochondria and autophagosome were observed in ultrastructural analysis. The cytoskeleton organization was disrupted by SiNPs in vascular endothelial cells. The release of proinflammatory and procoagulant cytokines including IL-6, IL-8, MCP-1, PECAM-1, TF and vWF, were markedly elevated in a dose-dependent manner. For in vivo study, based on the NOAEL for dosimetry selection, and using two transgenic zebrafish, Tg(mpo:GFP) and Tg(fli-1:EGFP), SiNPs-induced neutrophil-mediated inflammation and impaired vascular endothelial cells. With the dosage higher than NOAEL, SiNPs significantly decreased blood flow and velocity, exhibiting a blood hypercoagulable state in zebrafish embryos. The thrombotic effect was assessed by o-dianisidine staining, showed that an increasing of erythrocyte aggregation occurred in SiNPs-treated zebrafish. Microarray analysis was used to screen the possible genes for inflammation-coagulation response to SiNPs in zebrafish, and the JAK1/TF signaling pathway was further verified by qRT-PCR and Western blot assays. For in-deepth study, il6st was knocked down with specific morpholinos. The whole-mount in situ hybridization and qRT-PCR analysis showed that the expression jak1 and f3b were attenuated in il6st knockdown groups. In summary, our data demonstrated that SiNPs could induce inflammation-coagulation response and thrombotic effects via JAK1/TF signaling pathway.
Topics: Animals; Blood Coagulation; Cells, Cultured; Embryo, Nonmammalian; Endothelium, Vascular; Humans; Inflammation; Janus Kinase 1; Nanoparticles; Signal Transduction; Silicon Dioxide; Thrombosis; Zebrafish
PubMed: 29658397
DOI: 10.1080/17435390.2018.1461267 -
Heliyon Jan 2021Kinetic and physicochemical properties of peroxidase purified using a novel and cost efficient protocol was investigated with a view to providing information on its...
Kinetic and physicochemical properties of peroxidase purified using a novel and cost efficient protocol was investigated with a view to providing information on its possible biotechnological potentials. peroxidase was purified to homogeneity in two steps, involving ATPS and size exclusion chromatography on Sephadex G-100 with a yield of 84.12 %. In-gel activity staining revealed the presence of one isoform of peroxidase. The purified peroxidase is monomeric with native and subunits molecular weight of 38.9 and 43.5 kDa respectively. Kinetic parameters - , , of the purified enzyme were 2.5 units/mg protein, 0.020 ± 0.04 mM and 1.37 ± 0.18 mM respectively. Its optimum pH and temperature were 5 and 30 °C respectively. The purified enzyme cross-linked BSA into an insoluble matrix with the aid of caffeic acid. The study concluded that the purification scheme adopted is rapid and efficient, the purified enzyme exhibited some physiochemical properties that make it suitable for various biotechnological applications.
PubMed: 33521366
DOI: 10.1016/j.heliyon.2021.e06032