Assessment of Potential Anticancer Activity of Brown Seaweed Compounds Using Zebrafish Phenotypic Assay

Despite the extensive work on anticancer drug discovery, the number of potent lead compounds that enter the preclinical and clinical trials thus far is still low due to the poor selectivity and understanding in pharmacodynamics. In view of the homology between zebrafish embryogenesis and carcinogenesis in human, zebrafish embryos can be used in the screening platform to elucidate the molecular targets of potential anticancer compounds. In the present study, the possible targets modulating the potential anticancer effects of selected brown seaweed-derived compounds (ie alginate, fucoidan, phloroglucinol, fucosterol, and fucoxanthin) were examined. Teratogenic effects induced by the compounds were observed after 72 hours post-fertilization. Fucoidan, phloroglucinol, and fucosterol were observed to significantly reduce the pigmentation of the zebrafish in a dose-dependent manner at low concentrations (fucoidan, <60 µg/mL; phloroglucinol, <10 µg/mL; fucosterol, <3 µg/mL). On the other hand, embryos treated with fucoxanthin at 200 µg/mL and 300 µg/mL exhibited either phenotypes of curved trunk or bent tail. Further validation work using dual antiplatelet therapy (DAPT) and dorsomorphin as positive controls suggest that fucoxanthin might target the Notch and bone morphogenetic protein (BMP) pathways, respectively. Findings from this exploratory study henceforth have demonstrated the utility of zebrafish embryo to accelerate the discovery of potential compounds for targeted anticancer therapy.

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AV513 Enhances Tissue Factor Activity by Inhibiting TFPI in Hemophilia A Plasma: Potential TFPI Target Sites and Potentiation of Factor VIII Activity.

Blood ◽

10.1182/blood.v110.11.497.497 ◽

2007 ◽

Vol 110 (11) ◽

pp. 497-497 ◽

Cited By ~ 1

Author(s):

Sabine Knappe ◽

Nicholas Kripalani ◽

Susannah Patarroyo-White ◽

Kirk W. Johnson ◽

Srinivasa Prasad

Keyword(s):

Human Plasma ◽

Tissue Factor ◽

Thrombin Generation ◽

Hemophilia A ◽

Brown Seaweed ◽

Full Length ◽

Dependent Manner ◽

Severe Hemophilia ◽

Low Concentrations ◽

Dose Dependent

Abstract AV513, a sulfated polysaccharide prepared from brown seaweed and enriched for fucoidan accelerates clotting in hemophilia A and B plasma. Prior studies have demonstrated that AV513 functions mainly by inhibiting TFPI activity. In this report, we have further studied AV513 mechanism of action using human plasma thromboelastography (TEG) model and a thrombin generation assay in FVIII deficient plasma. In the TEG assay, AV513 enhanced tissue factor (TF) induced clotting in human hemophilia A pooled plasma in a dose-dependent manner. A dose-dependent potentiation effect by AV513 was observed with reduction of TEG R time to normal levels at 0.5 μg/mL of AV513. An AV513 dose dependent increase in thrombin generation was observed in hemophilia A pooled plasma with a 3-fold increase in peak thrombin generation at 2.5 μg/mL AV513 (basal: 40 nM vs. treated: 140 nM). When TFPI was immunodepleted from hemophilia A plasma (FVIII/TFPI deficient), sub-optimal concentrations of TF enhanced thrombin generation which was not further enhanced by addition of AV513. The increase in thrombin generation in FVIII /TFPI deficient plasma was inhibited by 93% after the addition of exogenous full-length TFPI or by 47% after the addition of truncated rTFPI (amino acids 1–161) that lacks the Kunitz 3 domain and the C-terminal tail. AV513 at low concentrations reversed exogenous full length TFPI activity almost completely in FVIII/TFPI-deficient plasma but had only a small effect on truncated TFPI. In a human plasma TEG combination study of AV513 and low concentrations of rFVIII (ReFacto), AV513 at a concentration of 0.25 μg/mL reduced the TEG R time by 35% in severe hemophilia A plasma compared to 10 mU/mL of rFVIII alone. These results indicate that AV513 functions as a pro-coagulant by inhibiting TFPI at least in part via targeting the C-terminal region. Moreover, AV513, in the presence of low rFVIII (ReFacto) levels, improves severe hemophilia A plasma clotting - as measured by TEG - beyond that observed with either agent alone.

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Effects of ethanol on gastric epithelial cell phospholipid dynamics and cellular function

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1987.252.2.g237 ◽

1987 ◽

Vol 252 (2) ◽

pp. G237-G243

Author(s):

R. E. Bailey ◽

R. A. Levine ◽

J. Nandi ◽

E. H. Schwartzel ◽

D. H. Beach ◽

...

Keyword(s):

Membrane Structure ◽

Lipid Membrane ◽

Proton Nuclear Magnetic Resonance ◽

Resonance Spectroscopy ◽

Dependent Manner ◽

Chloride Uptake ◽

Peak Intensity ◽

Low Concentrations ◽

Resonance Band ◽

Dose Dependent

The lipid profile of isolated gastric superficial epithelial cells (SEC) was evaluated by proton nuclear magnetic resonance spectroscopy (1H-NMR). The most conspicuous resonance band in SEC spectra was due to the protons of +N(CH3)3 groups of phosphatidylcholine and, to a lesser degree, other phospholipid derivatives, on the basis of their chemical shift and addition of purified phospholipids. NMR of cell lysates and phospholipid extracts of SEC in deutero-chloroform provided further spectral resolution of these components. Phospholipase or ethanol treatments of SEC produced membrane disorganization reflected as increased peak intensity of the phospholipid signals. In addition, ethanol, in a dose-dependent manner, attenuated paranitrophenyl phosphatase activity, which correlated with inhibition of total and ouabain-sensitive 86Rubidium chloride uptake by SEC. This study suggests that NMR used in conjunction with other biochemical techniques can monitor SEC membrane structure-function relationships. NMR is a potentially powerful noninvasive probe to show changes in lipid membrane organization induced by low concentrations of ethanol (1%) and may indicate an early sign of "cytotoxicity" in intact SEC.

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Mode of Action of the Hypertrehalosaemic Peptides from the American Cockroach

Zeitschrift für Naturforschung C ◽

10.1515/znc-1985-9-1015 ◽

1985 ◽

Vol 40 (9-10) ◽

pp. 670-676 ◽

Cited By ~ 16

Author(s):

Gerd Gäde

Keyword(s):

Cyclic Amp ◽

Adenylate Cyclase ◽

Mode Of Action ◽

Glycogen Phosphorylase ◽

Fat Body ◽

American Cockroach ◽

Dependent Manner ◽

Low Concentrations ◽

First Time ◽

Dose Dependent

Abstract Although crude extracts of cockroach (Periplaneta amencana) corpora cardiaca have been shown previously to affect the activity of adenylate cyclase and phosphorylase, we demonstrate in the present study for the first time that low concentrations (0.5 to 5 pmol) of the synthetic myoactive peptides. M I and M II, also affect these systems; these myoactive peptides are identical to the hypertrehalosaemic hormones I and II, and cause an increase in the concentration of the second messenger cyclic AMP in the fat body.In addition, both octapeptides activate fat body glycogen phosphorylase and promote breakdown of fat body glycogen. Both peptides increase the levels to haemolymph carbohydrate in a dose-dependent manner.

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Biphasic Dose–Response Induced by PCB150 and PCB180 in HeLa Cells and Potential Molecular Mechanisms

Dose-Response ◽

10.1177/1559325820910040 ◽

2020 ◽

Vol 18 (1) ◽

pp. 155932582091004

Author(s):

Ainy Zehra ◽

Muhammad Zaffar Hashmi ◽

Abdul Majid Khan ◽

Tariq Malik ◽

Zaigham Abbas

Keyword(s):

Hela Cells ◽

Molecular Mechanisms ◽

Dependent Manner ◽

Genotoxic Effects ◽

Species Formation ◽

Low Concentrations ◽

Extracellular Signal ◽

High Concentrations ◽

High Doses ◽

Dose Dependent

The polychlorinated biphenyls (PCBs) are persistent and their dose-dependent toxicities studies are not well-established. In this study, cytotoxic and genotoxic effects of PCB150 and PCB180 in HeLa cells were studied. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay indicated that the cell proliferation was stimulated at low doses (10−3 and 10−2 µg/mL for 12, 24, 48, and 72 hours) and inhibited at high doses (10 and 15 µg/mL for 24, 48, and 72 hours) for both PCBs. Increase in reactive oxygen species formation was observed in the HeLa cells in a time- and dose-dependent manner. Malondialdehyde and superoxide dismutase showed increased levels at high concentrations of PCBs over the time. Glutathione peroxidase expression was downregulated after PCBs exposure, suggested that both PCB congeners may attributable to cytotoxicity. Comet assay elicited a significant increase in genotoxicity at high concentrations of PCBs as compared to low concentrations indicating genotoxic effects. PCB150 and PCB180 showed decrease in the activity of extracellular signal–regulated kinase 1/2 and c-Jun N-terminal kinase at high concentrations after 12 and 48 hours. These findings may contribute to understanding the mechanism of PCBs-induced toxicity, thereby improving the risk assessment of toxic compounds in humans.

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Purification and some properties of the 45 kDa diphenylene iodonium-binding flavoprotein of neutrophil NADPH oxidase

Biochemical Journal ◽

10.1042/bj2650095 ◽

1990 ◽

Vol 265 (1) ◽

pp. 95-100 ◽

Cited By ~ 46

Author(s):

C M Yea ◽

A R Cross ◽

O T G Jones

Keyword(s):

Polyclonal Antibodies ◽

Specific Inhibitor ◽

Small Subunit ◽

Dependent Manner ◽

Superoxide Generation ◽

Diaphorase Activity ◽

Low Concentrations ◽

Diphenylene Iodonium ◽

Neutrophil Superoxide ◽

Dose Dependent

The 45 kDa diphenylene iodonium-binding flavoprotein of the human neutrophil superoxide-generating oxidase has been purified by affinity chromatography. The polypeptide was eluted from Blue Memsep or 2′,5′-ADP-agarose columns with either NADP or low concentrations of the specific inhibitor diphenylene iodonium. The purified protein was shown to bind FAD at a ratio of 1.09 mol of FAD/mol of protein. The reconstituted flavoprotein had a fluorescence spectrum similar, but not identical, to that of free FAD. It had an isoelectric point of approx. 4.0. The reconstituted flavoprotein displayed no diaphorase activity towards a range of artificial electron acceptors. Polyclonal antibodies raised against the pure protein inhibited superoxide generation by solubilized oxidase in a dose-dependent manner, and inhibited superoxide generation when incubated with either cytosol or membrane fractions in a reconstituted system. These antibodies precipitated the 45 kDa polypeptide together with a haem-containing 23 kDa protein thought to be the small subunit of cytochrome b-245. Antibodies raised against cytochrome P-450 reductase also precipitated these two polypeptides. These results are consistent with the 45 kDa polypeptide being the flavoprotein of the neutrophil superoxide-generating oxidase.

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Inhibition of Zika Virus by Marine Algae

10.20944/preprints201703.0087.v1 ◽

2017 ◽

Cited By ~ 2

Author(s):

Claudio Cirne-Santos ◽

Caroline De Souza Barros ◽

Caio Cesar Richter Nogueira ◽

Renata Mendonça Campos ◽

Valéria Teixeira ◽

...

Keyword(s):

Zika Virus ◽

Marine Algae ◽

Kappaphycus Alvarezii ◽

Strong Association ◽

Public Health Problem ◽

Brazilian Coast ◽

Dependent Manner ◽

Sulphated Polysaccharides ◽

Low Concentrations ◽

Dose Dependent

For many years marine algae has been subject of numerous researches and as a source of natural products with antiviral activity, such as terpenes, alkaloids and sulphated polysaccharides. However, the anti-Zika virus (ZIKV) potential of algae has not been studied. In this study we evaluated extracts seven species of the three major classes of seaweeds (Phaeophyceae, Chlorophyceae and Rhodophyceae) against ZIKV. All seaweeds tested are native of the Brazilian coast, except for Kappaphycus alvarezii that can be cultivated. ZIKV a mosquito-borne flavivirus, has become a public health problem. In recent years there has been an increase in the number of cases and a strong association between ZIKV outbreak and the spread of cases of Guillain-Barré Syndrome and microcephaly. All seaweed extracts tested in this work inhibits ZIKV replication in a dose-dependent manner. Caulerpa racemosa, Kappaphycus alvarezii and Osmundaria obtusiloba extracts were able to inhibit viral replication at low concentrations with EC50 values ranging from 1.38 to 1.98 µg/mL. We observed that O. obtusiloba presented a significant virucidal effect. Our results suggest that extracts of C. racemosa, K. alvarezii and O. obtusiloba presented very promising results, being excellent candidates for further studies, demonstrating that marine algae are an interesting source for the development of novel anti-ZIKV agents.

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Whole Peptidoglycan Extracts from theLactobacillus paracaseisubsp.paracaseiM5 Strain Exert Anticancer ActivityIn Vitro

BioMed Research International ◽

10.1155/2018/2871710 ◽

2018 ◽

Vol 2018 ◽

pp. 1-11 ◽

Cited By ~ 2

Author(s):

Shumei Wang ◽

Xue Han ◽

Lanwei Zhang ◽

Yingchun Zhang ◽

Hongbo Li ◽

...

Keyword(s):

Lactobacillus Paracasei ◽

Molar Ratio ◽

Dependent Manner ◽

Apoptotic Pathway ◽

Trypan Blue Exclusion ◽

Anticancer Effects ◽

Transmission Electron ◽

Sds Page ◽

Dose Dependent ◽

Ht 29

TheLactobacillus paracaseisubsp. paracaseiM5 strain exerted potential anticancer activity through the cell wall. In this study, whole peptidoglycan (WPG) was extracted from theLactobacillus paracaseisubsp. paracaseiM5 strain and was evaluated for anticancer effects as well as its properties. SDS-PAGE analysis confirmed the presence of WPG with dominant bands of approximately 14.4 kDa. Further analysis revealed that the amino acids present in the WPG consisted of alanine, glycine, glutamic acid, and lysine in a molar ratio of approximately 8 : 5 : 3 : 3.5. In addition, the cell viability of HT-29 cells with WPG addition was investigated with methyl thiazolyl tetrazolium (MTT) and trypan blue exclusion (TBE) assays, and cell apoptosis was analyzed with a transmission electron microscope, flow cytometry, and semiquantitative RT-PCR. These results showed that WPG exerted cytotoxic effects on colon cancer HT-29 cells in a dose-dependent manner and upregulated proapoptotic genes, while downregulating antiapoptotic genes. The gene expression study definitively revealed that WPG induced a mitochondria-mediated apoptotic pathway.

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B-Cell Translocation Gene 2 (Btg2) Regulates Vertebral Patterning by Modulating Bone Morphogenetic Protein/Smad Signaling

Molecular and Cellular Biology ◽

10.1128/mcb.24.23.10256-10262.2004 ◽

2004 ◽

Vol 24 (23) ◽

pp. 10256-10262 ◽

Cited By ~ 46

Author(s):

Sean Park ◽

Young Jae Lee ◽

Ho-Jae Lee ◽

Tsugio Seki ◽

Kwon-Ho Hong ◽

...

Keyword(s):

Bone Morphogenetic Protein ◽

Target Gene ◽

Bmp Signaling ◽

Dependent Manner ◽

Tail Bud ◽

Adaptor Molecule ◽

Morphogenetic Protein ◽

Dose Dependent ◽

Transcriptional Target

ABSTRACT Btg2 is a primary p53 transcriptional target gene which may function as a coactivator-corepressor and/or an adaptor molecule that modulates the activities of its interacting proteins. We have generated Btg2-null mice to elucidate the in vivo function of Btg2. Btg2-null mice are viable and fertile but exhibit posterior homeotic transformations of the axial vertebrae in a dose-dependent manner. Consistent with its role in vertebral patterning, Btg2 is expressed in the presomitic mesoderm, tail bud, and somites during somitogenesis. We further provide biochemical evidence that Btg2 interacts with bone morphogenetic protein (BMP)-activated Smads and enhances the transcriptional activity of BMP signaling. In view of the genetic evidence that reduced BMP signaling causes posteriorization of the vertebral pattern, we propose that the observed vertebral phenotype in Btg2-null mice is due to attenuated BMP signaling.

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Lasius fuliginosus Nest Carton as a Source of New Promising Bioactive Extracts with Chemopreventive Potential

International Journal of Molecular Sciences ◽

10.3390/ijms22094392 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4392

Author(s):

Grzegorz Karol Wagner ◽

Magdalena Jaszek ◽

Bernard Staniec ◽

Monika Prendecka ◽

Dominika Pigoń ◽

...

Keyword(s):

Antioxidant Properties ◽

Vero Cells ◽

Dependent Manner ◽

Impedance Sensing ◽

Lasius Fuliginosus ◽

Anticancer Effects ◽

Bioactive Properties ◽

Cell Substrate ◽

Chemopreventive Activity ◽

Dose Dependent

Six new water extracts (E1–E6) were obtained from nest carton produced by jet black ants Lasius fuliginosus and tested for their biochemical and bioactive properties, including antioxidative and anticancer effects. The present study demonstrated significant qualitative and quantitative differences in the content of individual biochemical constituents, as well as bioactive properties between the investigated samples. All tested extracts demonstrated antioxidant properties (determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) methods), and the highest antioxidative potential was recorded in extracts E1 and E2 (188.96 and 313.67 μg/mL of ascorbic acid equivalent for ABTS and 176.42 and 202.66 μg/mL for DPPH reagent). Furthermore the six extracts exhibited strong inhibitory activity towards human melanoma cells of the A-375 CRL-1619 line in a dose-dependent manner. The most interesting chemopreventive activity was exhibited by extract E2, which inhibited the proliferation of A-375 cells to the greatest extent, while having a minimal effect on Vero cells. The effect on cancer cells has been confirmed using the Electric Cell-substrate Impedance Sensing (ECIS) technique. Significant impedance changes have been detected in A-375 and Vero cells following the administration of extract E2. The obtained results are really promising and constitute the basis for further research on the nest carton of jet black ant.

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Dual Effects of Silibinin on Human Pancreatic Cancer Cells

10.21203/rs.3.rs-130714/v1 ◽

2020 ◽

Author(s):

Su-Mi Lee ◽

Gil-Woo Lee ◽

Seon-Young Park ◽

Hosouk Joung ◽

Chang-Hwan Park ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cancer Cells ◽

Human Pancreatic Cancer ◽

Dependent Manner ◽

Protein Levels ◽

Ductal Cells ◽

Pancreatic Cancer Cells ◽

Anticancer Effects ◽

Pancreatic Cells ◽

Dose Dependent

Abstract Background Silibinin is a flavonoid with antihepatotoxic properties, and exhibits pleiotropic anticancer effects. However, the molecular mechanisms responsible for its anticancer actions in pancreatic cancer cells, and the effects on such cells and normal pancreatic cells, remain unclear. The objective of this study was to determine the effect of silibinin on human pancreatic cancer cells and normal ductal cells.Methods Human pancreatic cancer cells (MIA PaCa-2 and PANC-1) and normal ductal cells (hTERT-HPNE) were cultured with 0–400 μM silibinin for 48 h. Thereafter, the proliferation, invasion, apoptosis, and signaling pathways of the pancreatic cells were evaluated.Results Silibinin significantly inhibited the proliferation, invasion, and spheroid formation of human pancreatic cancer cells in vitro in a dose-dependent manner (p < 0.05). It also induced apoptosis in a dose-dependent manner. Western blot analysis showed that silibinin downregulated extracellular signaling-regulated kinase (ERK) and serine/threonine protein kinase (AKT) in human pancreatic cancer cells. It also upregulated microtubule associated protein 1 light chain 3 β (LC3B) and cleaved caspase-3 via c-Jun N-terminal kinases (JNK) signaling. On the other hand, silibinin increased the mRNA and protein levels of c-Jun, Twist-related protein 1, and Snail. It also decreased exogenous p53 levels, but increased endogenous c-Jun protein levels in human pancreatic cancer cells. However, silibinin did not affect cell viability and endogenous c-Jun levels in pancreatic normal ductal cells. It increased exogenous p53 levels, but decreased stemness-related gene expression in pancreatic normal ductal cells. Silibinin increased Ki-67 levels in pancreatic cancer cells, but decreased them in pancreatic normal ductal cells.Conclusion Silibinin not only exerted anticancer effects by inhibiting AKT–ERK and JNK signaling, but also upregulated cancer stemness–related genes in human pancreatic cancer cells. These results suggest that silibinin should be used as a therapeutic agent for human pancreatic cancer with caution.

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