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Frontiers in Cell and Developmental... 2022Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development...
Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development and circulatory failure, suggesting additional roles for TF in embryonic development beyond coagulation. In this study, we characterized the role of one of the TF paralogs () using a zebrafish model. The expression of during embryonic developmental stages was determined by RT-PCR. Spatiotemporal expression pattern of revealed (high expression from 28 to 36 hpf) the role of in the development of the yolk sac, circulation, and fins. Morpholinos (MO), an antisense-based oligonucleotide strategy, was used to knockdown and examined for defects in morphological appearance, bleeding, and vascular patterning. MO-injected embryos showed morphological abnormalities, including shorter body lengths and crooked tails. O-dianisidine staining showed MO-injected embryos exhibited bleeding in the trunk (5.44%) and head (9.52%) regions. Imaging of endothelial-specific transgenic lines () showed a 3-fold decreased caudal vein plexus (CVP) in morphants versus controls at 48 hpf, suggesting a potential role for in angiogenesis. These findings confirm that is essential for angiogenesis, in addition to its involvement in hemostasis.
PubMed: 35386206
DOI: 10.3389/fcell.2022.852989 -
Analytical Sciences : the International... Jan 2022The aim of this work was the development of an automatic sequential injection analysis method to monitor the ozonation process for water disinfection. The determination...
The aim of this work was the development of an automatic sequential injection analysis method to monitor the ozonation process for water disinfection. The determination was based on the reaction between bromate and o-dianisidine in the presence of bromide in acidic medium. The determination parameters were studied and adjusted to enable bromate quantification in the range 0.35-4.0 mg BrO/L with a limit of detection of 20 μg BrO/L. The choice of a sequential injection procedure enabled a minimal consumption of reagents and no need for sample pre-treatment. The developed sequential injection proved to be accurate with < 5% relative deviation when compared to ICP-MS and an average of 101% in recovery percentages studies. It was effectively applied to monitor an ozonation process enabling the follow-up of the process with real-time quantification of the bromate content.
Topics: Bromates; Disinfection; Ozone; Water; Water Supply
PubMed: 35287215
DOI: 10.2116/analsci.21P191 -
Environmental Pollution (Barking, Essex... Nov 2021The hydrothermal preparation of o-dianisidine and triazine interlinked porous organic polymer and its successive derivatisation via metal infusion (Ni, Cu) under...
The hydrothermal preparation of o-dianisidine and triazine interlinked porous organic polymer and its successive derivatisation via metal infusion (Ni, Cu) under hydrothermal and calcination conditions (700 °C) to yield pristine (ANIPOP-700) and Ni/Cu decorated porous carbon are described here (Ni-ANIPOP-700 and Cu-ANIPOP-700). To confirm their chemical and morphological properties, the as-prepared materials were methodically analyzed using solid state C and N NMR, X-ray diffraction, Raman spectroscopy, field emission scanning and high resolution transmission electron microscopic techniques, and x-ray photoelectron spectroscopy. Furthermore, the electrocatalytic activities of these electrocatalysts were thoroughly investigated under standard oxygen evolution (OER) and hydrogen evolution reaction (HER) conditions. The results show that all of the materials demonstrated significant activity in water splitting as well as displayed excellent stability (22 h) in both acidic (HER) and basic conditions (OER). Among the electrocatalysts reported in this study, Ni-ANIPOP-700 exhibited a lower overpotential η of 300 mV in basic medium (OER) and 150 mV in acidic medium (HER), as well as a lower Tafel slope of 69 mV/dec (OER) and 181 mV/dec (HER), indicating 30% lower energy requirement for overall water splitting. Gas chromatography was used to examine the electrolyzed products.
Topics: Carbon; Electrolysis; Polymers; Porosity; Seawater
PubMed: 34343751
DOI: 10.1016/j.envpol.2021.117861 -
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 Colloid and Interface Science Jul 2021The hybridization of enzymes and inorganics in controlled manner is challenging, however, critical for the development of chemo-enzymatic cascade catalyst with high...
The hybridization of enzymes and inorganics in controlled manner is challenging, however, critical for the development of chemo-enzymatic cascade catalyst with high efficiency and selectivity. Here, proteins and metal oxide clusters can be facilely co-assembled on the surface of colloid of poly(4-vinylpyridine) (P4VP) via hydrogen bonding, due to their enriched surface hydrogen bonding donors. The co-assembly method can be generally applied for preparing chemo-enzymatic catalyst within the selected database of various proteins and metal oxide clusters while the assembly units retain their structures and activities. Typically, a 2.5 nm metal oxide cluster {MoFe}, with peroxidase-like activity, are complexed with glucose oxidase (GOX) on P4VP for the catalysis against the oxidization of o-dianisidine (ODA) with the existence of glucose. Due to the synergistic effects of chemical and enzymatic catalysis, the co-assemblies show even higher ODA oxidation activity compared to GOX/catalase bi-enzymatic system, confirming the effectiveness of the co-assembly protocol for cascade catalysis and enabling its applications in rapid glucose detection and biomass conversion.
Topics: Catalysis; Glucose Oxidase; Oxidation-Reduction; Oxides; Polymers
PubMed: 33794409
DOI: 10.1016/j.jcis.2021.03.090 -
Aquatic Toxicology (Amsterdam,... May 2021Exposure to oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) at critical developmental time-points in fish models impairs red blood cell concentrations in a...
Exposure to oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) at critical developmental time-points in fish models impairs red blood cell concentrations in a regioselective manner, with 2-hydroxychrysene being more potent than 6-hydroxychrysene. To better characterize this phenomenon, embryos of Japanese medaka (Oryzias latipes) were exposed to 2- or 6-hydroxychrysene (0.5, 2, or 5 μM) from 4 h-post-fertilization (hpf) to 7 d-post-fertilization. Following exposure, hemoglobin concentrations were quantified by staining fixed embryos with o-dianisidine (a hemoglobin-specific dye) and stained embryos were imaged using brightfield microscopy. Exposure to 2-hydroxychrysene resulted in a concentration-dependent decrease in hemoglobin relative to vehicle-exposed embryos, while only the highest concentration of 6-hydroxychrysene resulted in a significant decrease in hemoglobin. All tested concentrations of 2-hydroxychrysene also caused significant mortality (12.2 % ± 2.94, 38.9 % ± 14.4, 85.6 % ± 11.3), whereas mortality was not observed following exposure to 6-hydroxychrysene. Therefore, treatment of embryos with 2-hydroxychrysene at various developmental stages and durations was subsequently conducted to identify key developmental landmarks that may be targeted by 2-hydroxychrysene. A sensitive window of developmental toxicity to 2-hydroxychrysene was found between 52-100 hpf, with a 24 h exposure to 10 μM 2-hydroxychrysene resulting in significant anemia and mortality. Since exposure to 2-hydroxychrysene from 52 to 100 hpf, a window that includes liver morphogenesis in medaka, resulted in the highest magnitude of toxicity, liver development and function may have a role in 2-hydroxychrysene developmental toxicity.
Topics: Animals; Chrysenes; Embryo, Nonmammalian; Embryonic Development; Hemoglobins; Oryzias; Stereoisomerism; Water Pollutants, Chemical
PubMed: 33714762
DOI: 10.1016/j.aquatox.2021.105791 -
Comparative Biochemistry and... Jun 2021We investigated whether ferulic acid (FA), a nutraceutical could mitigate the arsenic (As) induced cardiotoxicity. Zebrafish larvae (60 and 72 h post-fertilization...
We investigated whether ferulic acid (FA), a nutraceutical could mitigate the arsenic (As) induced cardiotoxicity. Zebrafish larvae (60 and 72 h post-fertilization [hpf]) were used to study the effect of FA on As at different time points (24 and 48 h after exposure). The FA exposure was given as pre-treatment (60 hpf) and simultaneous treatment (72 hpf) to translate the results for As contaminated areas. To accomplish this, the lethality assay was done, and based on the results, the dosage for As (1 mM) and FA (30 μM) was fixed. The FA intervention (30 μM) as 12 h pre-treatment (60 hpf) and simultaneous treatment along with As (72 hpf) decreased the As content in zebrafish larvae as evidenced by inductively coupled plasma-mass spectrometry. As exposure showed congenital deformities especially cardiac malformations in zebrafish larvae after 24 and 48 h. These teratogenic effects induced by As were reduced by FA supplementation in both groups. Also, o-dianisidine staining demonstrated that As treated larvae encountered abnormal cardiac function with reduced blood circulation, while FA supplementation reversed these effects. The acetylcholinesterase activity, a biomarker of As-induced cardiotoxicity was also found to be decreased in As group, which was rescued by FA. The modulation in the expression of the genes involved in cardiogenesis (nkx2.5, bmp2b, gata4, gata5, myh6, myl7, and tnnt2) further confirmed the ameliorative effect of FA on As induced malformations.
Topics: Animals; Arsenic; Cardiotoxicity; Coumaric Acids; Embryo, Nonmammalian; Embryonic Development; Gene Expression Regulation, Developmental; Water Pollutants, Chemical; Zebrafish; Zebrafish Proteins
PubMed: 33631344
DOI: 10.1016/j.cbpc.2021.109021 -
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 -
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 -
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