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Journal of Cancer Research and Clinical... Dec 2017Our previous experiments show that the main constituent of green-tea catechins, (-)-epigallocatechin gallate (EGCG), completely prevents tumor promotion on mouse skin... (Review)
Review
PURPOSE
Our previous experiments show that the main constituent of green-tea catechins, (-)-epigallocatechin gallate (EGCG), completely prevents tumor promotion on mouse skin initiated with 7,12-dimethylbenz(a)anthracene followed by okadaic acid and that EGCG and green tea extract prevent cancer development in a wide range of target organs in rodents. Therefore, we focused our attention on human cancer stem cells (CSCs) as targets of cancer prevention and treatment with EGCG.
METHODS
The numerous reports concerning anticancer activity of EGCG against human CSCs enriched from cancer cell lines were gathered from a search of PubMed, and we hope our review of the literatures will provide a broad selection for the effects of EGCG on various human CSCs.
RESULTS
Based on our theoretical study, we discuss the findings as follows: (1) Compared with the parental cells, human CSCs express increased levels of the stemness markers Nanog, Oct4, Sox2, CD44, CD133, as well as the EMT markers, Twist, Snail, vimentin, and also aldehyde dehydrogenase. They showed decreased levels of E-cadherin and cyclin D1. (2) EGCG inhibits the transcription and translation of genes encoding stemness markers, indicating that EGCG generally inhibits the self-renewal of CSCs. (3) EGCG inhibits the expression of the epithelial-mesenchymal transition phenotypes of human CSCs. (4) The inhibition of EGCG of the stemness of CSCs was weaker compared with parental cells. (5) The weak inhibitory activity of EGCG increased synergistically in combination with anticancer drugs.
CONCLUSIONS
Green tea prevents human cancer, and the combination of EGCG and anticancer drugs confers cancer treatment with tissue-agnostic efficacy.
Topics: Animals; Biomarkers, Tumor; Catechin; Humans; Neoplasms; Neoplastic Stem Cells
PubMed: 28942499
DOI: 10.1007/s00432-017-2515-2 -
Molecules (Basel, Switzerland) Nov 2020Alzheimer's disease (AD), which is among the most prevalent neurodegenerative diseases, manifests as increasing memory loss and cognitive decline. Tau phosphorylation...
Alzheimer's disease (AD), which is among the most prevalent neurodegenerative diseases, manifests as increasing memory loss and cognitive decline. Tau phosphorylation and aggregation are strongly linked to neurodegeneration, as well as associated with chronic neuroinflammatory processes. The anti-inflammation effects of natural products have led to wide recognition of their potential for use in treating and preventing AD. This study investigated whether eupatin, a polymethoxyflavonoid found in species, has inhibitory effects on neuroinflammation and tau phosphorylation. We treated mouse macrophages and microglia cells with lipopolysaccharides (LPSs) to activate inflammatory signals, and we treated neuronal cells with a protein phosphatase 2A inhibitor, okadaic acid (OA), or transfection with pRK5-EGFP-Tau P301L plasmid to induce tau phosphorylation. The results indicated that eupatin significantly reduced the LPS-induced protein expression and phosphorylation of p65 and inducible nitric oxide synthase as well as downstream products interleukin 6 and nitrite, respectively. Furthermore, eupatin markedly inhibited the expression of phospho-tau in response to OA treatment and plasmid transfection. We discovered that this inhibition was achieved through the inhibition of glycogen synthase kinase 3β (GSK3β), and molecular docking results suggested that eupatin can sufficiently bind to the GSK3β active site. Our results demonstrate that eupatin has neuroprotective effects, making it suitable for AD treatment.
Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Flavonoids; Macrophages; Mice; Microglia; Molecular Docking Simulation; Neuroprotective Agents; Phosphorylation; Phytotherapy; Plant Extracts; Zingiberaceae; tau Proteins
PubMed: 33266202
DOI: 10.3390/molecules25235652 -
Toxins Nov 2018Several species of the dinoflagellate genus produce toxins that accumulate in bivalves when they feed on populations of these organisms. The accumulated toxins can lead... (Review)
Review
Several species of the dinoflagellate genus produce toxins that accumulate in bivalves when they feed on populations of these organisms. The accumulated toxins can lead to intoxication in consumers of the affected bivalves. The risk of intoxication depends on the amount and toxic power of accumulated toxins. In this review, current knowledge on the main processes involved in toxin accumulation were compiled, including the mechanisms and regulation of toxin acquisition, digestion, biotransformation, compartmentalization, and toxin depuration. Finally, accumulation kinetics, some models to describe it, and some implications were also considered.
Topics: Alveolata; Animals; Bivalvia; Cell Compartmentation; Digestion; Marine Toxins
PubMed: 30400229
DOI: 10.3390/toxins10110453 -
ACS Chemical Neuroscience Oct 2022Alzheimer's disease (AD) is a progressive, fatal, neurodegenerative disorder for which only treatments of limited efficacy are available. Despite early mentions of...
Alzheimer's disease (AD) is a progressive, fatal, neurodegenerative disorder for which only treatments of limited efficacy are available. Despite early mentions of dementia in the ancient literature and the first patient diagnosed in 1906, the underlying causes of AD are not well understood. This study examined the possible role of dopamine, a neurotransmitter that is involved in cognitive and motor function, in AD. We treated adult zebrafish () with okadaic acid (OKA) to model AD and assessed the resulting behavioral and neurochemical changes. We then employed a latent learning paradigm to assess cognitive and motor function followed by neurochemical analysis with fast-scan cyclic voltammetry (FSCV) at carbon fiber microelectrodes to measure the electrically stimulated dopamine release. The behavioral assay showed that OKA treatment caused fish to have lower motivation to reach the goal chamber, resulting in impeded learning and decreased locomotor activity compared to controls. Our voltammetric measurements revealed that the peak dopamine overflow in OKA-treated fish was about one-third of that measured in controls. These findings highlight the profound neurochemical changes that may occur in AD. Furthermore, they demonstrate that applying the latent learning paradigm and FSCV to zebrafish is a promising tool for future neurochemical studies and may be useful for screening drugs for the treatment of AD.
Topics: Alzheimer Disease; Animals; Carbon Fiber; Dopamine; Microelectrodes; Neurotransmitter Agents; Okadaic Acid; Zebrafish
PubMed: 36113115
DOI: 10.1021/acschemneuro.2c00484 -
Harmful Algae May 2024This article presents the first results on shellfish toxicity in the Slovenian sea (Gulf of Trieste, Adriatic Sea) since the analytical methods for the detection of...
Okadaic acid as a major problem for the seafood safety (Mytilus galloprovincialis) and the dynamics of toxic phytoplankton in the Slovenian coastal sea (Gulf of Trieste, Adriatic Sea).
This article presents the first results on shellfish toxicity in the Slovenian sea (Gulf of Trieste, Adriatic Sea) since the analytical methods for the detection of biotoxins (PSP, ASP, DSP and other lipophilic toxins) in bivalve molluscs were included in the national monitoring program in 2013. In addition to toxins, the composition and abundance of toxic phytoplankton and general environmental characteristics of the seawater (surface temperature and salinity) were also monitored. During the 2014-2019 study period, only lipophilic toxins were detected (78 positive tests out of 446 runs), of which okadaic acid (OA) predominated in 97 % of cases, while dinophysistoxin-2 and yessotoxins only gave a positive result in one sampling event each. The number of samples that did not comply with the EC Regulation for the OA group was 17 or 3.8 % of all tests performed, all of which took place from September to November, while a few positive OA tests were also recorded in December, April, and May. This toxicity pattern was consistent with the occurrence pattern of the five most common DSP-producing dinoflagellates, which was supported by the development of warm and thermohaline stratified waters: Dinophysis caudata, D. fortii, D. sacculus, D. tripos and Phalacroma rotundatum. The strong correlation (r = 0.611, p < 0.001) between D. fortii, reaching abundances of up to 950 cells L, and OA suggests that D. fortii is the main cause of OA production in Slovenian waters. Strong interannual variations in OA and phytoplankton dynamics, exacerbated by the effects of anthropogenic impacts in this coastal ecosystem, reduce the predictability of toxicity events and require continuous and efficient monitoring. Our results also show that the introduction of the LC-MS/MS method for lipophilic toxins has improved the management of aquaculture activities, which was not as accurate based on mouse bioassays.
Topics: Okadaic Acid; Animals; Phytoplankton; Marine Toxins; Slovenia; Mytilus; Seafood; Seawater; Dinoflagellida
PubMed: 38830710
DOI: 10.1016/j.hal.2024.102632 -
Molecular & Cellular Proteomics : MCP Aug 2023Protein phosphorylation is an essential regulatory mechanism that controls most cellular processes, including cell cycle progression, cell division, and response to...
Protein phosphorylation is an essential regulatory mechanism that controls most cellular processes, including cell cycle progression, cell division, and response to extracellular stimuli, among many others, and is deregulated in many diseases. Protein phosphorylation is coordinated by the opposing activities of protein kinases and protein phosphatases. In eukaryotic cells, most serine/threonine phosphorylation sites are dephosphorylated by members of the Phosphoprotein Phosphatase (PPP) family. However, we only know for a few phosphorylation sites which specific PPP dephosphorylates them. Although natural compounds such as calyculin A and okadaic acid inhibit PPPs at low nanomolar concentrations, no selective chemical PPP inhibitors exist. Here, we demonstrate the utility of endogenous tagging of genomic loci with an auxin-inducible degron (AID) as a strategy to investigate specific PPP signaling. Using Protein Phosphatase 6 (PP6) as an example, we demonstrate how rapidly inducible protein degradation can be employed to identify dephosphorylation sites and elucidate PP6 biology. Using genome editing, we introduce AID-tags into each allele of the PP6 catalytic subunit (PP6c) in DLD-1 cells expressing the auxin receptor Tir1. Upon rapid auxin-induced degradation of PP6c, we perform quantitative mass spectrometry-based proteomics and phosphoproteomics to identify PP6 substrates in mitosis. PP6 is an essential enzyme with conserved roles in mitosis and growth signaling. Consistently, we identify candidate PP6c-dependent dephosphorylation sites on proteins implicated in coordinating the mitotic cell cycle, cytoskeleton, gene expression, and mitogen-activated protein kinase (MAPK) and Hippo signaling. Finally, we demonstrate that PP6c opposes the activation of large tumor suppressor 1 (LATS1) by dephosphorylating Threonine 35 (T35) on Mps One Binder (MOB1), thereby blocking the interaction of MOB1 and LATS1. Our analyses highlight the utility of combining genome engineering, inducible degradation, and multiplexed phosphoproteomics to investigate signaling by individual PPPs on a global level, which is currently limited by the lack of tools for specific interrogation.
Topics: Humans; Proteolysis; Protein Serine-Threonine Kinases; Phosphoprotein Phosphatases; Phosphorylation; Threonine; Colorectal Neoplasms; Protein Phosphatase 2
PubMed: 37392812
DOI: 10.1016/j.mcpro.2023.100614 -
Cellular Physiology and Biochemistry :... 2018Okadaic acid (OA) and the structurally related compounds dinophysistoxin-1 (DTX1) and dinophysistoxin-2 (DTX2) are marine phycotoxins that cause diarrheic shellfish...
BACKGROUND/AIMS
Okadaic acid (OA) and the structurally related compounds dinophysistoxin-1 (DTX1) and dinophysistoxin-2 (DTX2) are marine phycotoxins that cause diarrheic shellfish poisoning (DSP) in humans due to ingestion of contaminated shellfish. In order to guarantee consumer protection, the regulatory authorities have defined the maximum level of DSP toxins as 160 µg OA equivalent kg-1 shellfish meat. For risk assessment and overall toxicity determination, knowledge of the relative toxicities of each analogue is required. In absence of enough information from human intoxications, oral toxicity in mice is the most reliable data for establishing Toxicity Equivalence Factors (TEFs).
METHODS
Toxins were administered to mice by gavage, after that the symptomatology and mice mortality was registered over a period of 24 h. Organ damage data were collected at necropsy and transmission electron microscopy (TEM) was used for ultrastructural studies. Toxins in urine, feces and blood were analyzed by HPLC-MS/MS. The evaluation of in vitro potencies of OA, DTX1 and DTX2 was performed by the protein phosphatase 2A (PP2A) inhibition assay.
RESULTS
Mice that received DSP toxins by gavage showed diarrhea as the main symptom. Those toxins caused similar gastrointestinal alterations as well as intestine ultrastructural changes. However, DSP toxins did not modify tight junctions to trigger diarrhea. They had different toxicokinetics and toxic potency. The lethal dose 50 (LD50) was 487 µg kg-1 bw for DTX1, 760 µg kg-1 bw for OA and 2262 µg kg-1 bw for DTX2. Therefore, the oral TEF values are: OA = 1, DTX1 = 1.5 and DTX2 = 0.3.
CONCLUSION
This is the first comparative study of DSP toxins performed with accurate well-characterized standards and based on acute toxicity data. Results confirmed that DTX1 is more toxic than OA by oral route while DTX2 is less toxic. Hence, the current TEFs based on intraperitoneal toxicity should be modified. Also, the generally accepted toxic mode of action of this group of toxins needs to be reevaluated.
Topics: Administration, Oral; Animals; Body Weight; Chromatography, High Pressure Liquid; Female; Heart; Intestine, Small; Liver; Mice; Myocardium; Okadaic Acid; Protein Phosphatase 2; Pyrans; Stomach; Tandem Mass Spectrometry; Toxicity Tests
PubMed: 30176657
DOI: 10.1159/000493039 -
Ecotoxicology and Environmental Safety Jan 2023Phycotoxins are a class of multiple natural metabolites produced by microalgae in marine and freshwater ecosystems that bioaccumulate in food webs, particularly in... (Review)
Review
Phycotoxins are a class of multiple natural metabolites produced by microalgae in marine and freshwater ecosystems that bioaccumulate in food webs, particularly in shellfish, having a great impact on human health. Phycotoxins are mainly leached and absorbed in the small intestine when human consumers accidentally ingest toxic aquatic products contaminated by them. To assess the intestinal uptake and damage of phycotoxins, a typical in vitro model was developed and widely applied using the human colorectal adenocarcinoma Caco-2 cell line. In this review, the application cases were summarized for multiple phycotoxins, including microcystins (MCs), cylindrospermopsins (CYNs), domoic acids (DAs), saxitoxins (STXs), palytoxins (PLTXs), okadaic acids (OAs), pectenotoxins (PTXs) and azaspiracids (AZAs). The results of the previous studies showed that each group of phycotoxins presented different cytotoxicity and mechanisms to Caco-2 cells, and significant discrepancies in the transport of phycotoxin across the Caco-2 cell monolayers. Therefore, this review describes the evaluation assays of the Caco-2 cell monolayer model, illustrates the principles of several primary cytotoxicity evaluation assays, and summarizes the cytotoxicity of each group of phycotoxins to Caco-2 cells line and their cellular transport, and finally proposes the development of multicellular intestinal models for future comprehensive studies on the toxicity and absorption of phycotoxins in the intestine. It will improve the understanding of Caco-2 cell monolayer models in the toxicology studies on phycotoxins and the potentially detrimental effects of microalgal toxins on the human intestine.
Topics: Humans; Caco-2 Cells; Ecosystem; Intestinal Barrier Function; Marine Toxins; Okadaic Acid; Microalgae
PubMed: 38321666
DOI: 10.1016/j.ecoenv.2022.114447 -
Environmental Science and Pollution... Jan 2024The okadaic acid (OA)-group toxins, including OA, dinophysistoxin-1 (DTX1), dinophysistoxin-2 (DTX2), and dinophysistoxin-3 (DTX3), cause diarrheic shellfish poisoning...
The okadaic acid (OA)-group toxins, including OA, dinophysistoxin-1 (DTX1), dinophysistoxin-2 (DTX2), and dinophysistoxin-3 (DTX3), cause diarrheic shellfish poisoning in humans. To manage OA-group toxins more strictly, Korean regulations were recently revised to consider OA, DTX1, DTX2, and DTX3 combined. Thus, our study characterized the occurrence of OA, DTX1, DTX2, and DTX3 in seafood distributed across South Korea, and a risk assessment of seafood consumption was conducted. Two hundred and seventeen samples from 16 bivalve and 7 non-bivalve species collected from three representative coastal areas in 2021 were analyzed via liquid chromatography-tandem mass spectrometry. OA, DTX1, and DTX3 were detected in 2.3%, 4.1%, and 9.2% of the examined samples, with positive mean levels of 11.3, 16.4, and 40.9 µg/kg, respectively. DTX2 was not detected in any of the samples. At least one OA-group toxin was detected in the bivalve samples, including blood clams, pan shells, hard clams, mussels, and scallops, whereas none were detected in non-bivalves. The estimated acute exposure to OA-group toxins through the intake of seafood in the Korean population and consumer groups was low, ranging from 24.7 to 74.5% of the recommended acute reference dose (ARfD) of 0.33 μg OA equivalents/kg body weight. However, for the scallop consumers aged 7-12 years, acute exposure to OA-group toxins exceeded the ARfD, indicating a possible health risk. These results suggest that including DTX3 in the new regulatory limits is appropriate to protect Korean seafood consumers from exposure to OA-group toxins.
Topics: Animals; Humans; Okadaic Acid; Marine Toxins; Bivalvia; Seafood; Risk Assessment; Pyrans
PubMed: 38147248
DOI: 10.1007/s11356-023-31568-4 -
Foods (Basel, Switzerland) Jun 2022In this investigation, a new approach for developing a sensitive lateral flow immunoassay (LFIA) was proposed for the detection of the hazardous marine toxin okadaic...
In this investigation, a new approach for developing a sensitive lateral flow immunoassay (LFIA) was proposed for the detection of the hazardous marine toxin okadaic acid (OA). It is based on the indirect format with anti-species antibodies labeled by gold nanoparticles (AuNPs) and cascade signal amplification. The latter is performed by first passing a mixture of anti-OA antibodies and a tested sample along the immunochromatographic test strip and then performing several cycles of the interaction of anti-species antibodies conjugated with AuNPs with free antibodies, which bind to anti-species antibodies but are not specific to the target analyte. As a result, branched aggregates are formed, due to which the colorimetric signal intensification occurs. The developed test system enabled the detection of OA with an instrumental detection limit of 30 pg/mL and a cutoff of 1 ng/mL, which exceeds these characteristics in the LFIA without amplification by 7 and 2 times, respectively. The OA recoveries from seawater, fish, and seafood varied from 76.9% to 126%. The test system may be required for point-of-care monitoring of samples for phycotoxin contamination; the developed principle of signal amplification can be used in cases where highly sensitive detection of trace amounts of a contaminant is required.
PubMed: 35741890
DOI: 10.3390/foods11121691