• Korean Journal of Veterinary Research
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• v.56 no.2
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• pp.67-74
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• 2016

Tomato extract has been shown to exert antiplatelet activity in vitro and to change platelet function ex vivo, but with limitations. In this study, antiplatelet activity of water soluble tomato concentrate (Fruitflow I) and dry water soluble tomato concentrate (Fruitflow II) was investigated using rat platelets. Aggregation was induced by collagen and adenosine diphosphate and granule-secretion, $[Ca^{2+}]_i$, thromboxane B2, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels were examined. The activation of integrin ${\alpha}_{IIb}{\beta}_3$ and phosphorylation of signaling molecules, including mitogen-activated protein kinase (MAPK) and PI3K/Akt, were investigated by flow cytometry and immunoblotting, respectively. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were examined. Moreover, in vivo thrombus weight was tested by an arteriovenous shunt model. Fruitflow I and Fruitflow II significantly inhibited agonist induced platelet aggregation, adenosine triphosphate and serotonin release, $[Ca^{2+}]_i$, and thromboxane B2 concentration, while having no effect on cAMP and cGMP levels. Integrin ${\alpha}_{IIb}{\beta}_3$ activation was also significantly decreased. Moreover, both concentrates reduced phosphorylation of MAPK pathway factors such as ERK, JNK, P38, and PI3K/Akt. In vivo thrombus formation was also inhibited. Taken together, these concentrates have the potential for ethnomedicinal applications to prevent cardiovascular ailments and can be used as functional foods.

• Biomolecules & Therapeutics
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• v.7 no.3
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• pp.227-233
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• 1999

The effects of 2-bromo-3-(3,5-tert-butyl-4-hydroxylphenyl)-1,4-naphthalenedione(TPN2), a synthetic vitamin K derivative, on platelet aggregation and its action mechanisms were investigated in rat platelet. TPN2 inhibited the platelet aggregation induced by collagen($10\mu\textrm{g}$/ml), thrombin(0.1 U/ml), A23187($10\mu\textrm{M}$) and arachidonic acid($100\mu\textrm{M}$) in concentration-dependent manner with $IC_{50}$ values of 6.5$\pm$1.3, 59.3$\pm$4.5, 13.0$\pm$2.37 and 2.9$\pm$$1.0\mu\textrm{M}$, respectively. Collagen-induced serotonin release was significantly reduced by TPN2. The elevation of intracellular free $Ca^{2+}$ concentration ([$Ca^{2+}$]i) by collagen stimulation was greatly decreased by the pretreatment of TPN2, which was due to the inhibition of calcium release from intracellular store and influx from outside of the cell. TPN2 also significantly reduced the thromboxane $A_2$($TXA_2$) formation in a concentration-dependent manner. The collagen-induced arachidonic acid (AA) release in [$^3H$]-AA incorporated platelet, an indicative of the phospholipase $A_2$ activity, was decreased by TPN2 pretreatment. TPN2 significantly inhibited the activity of thromboxane synthase, but did not affect the cyclooxygenase activity. From these results. it is suggested that TPN2 exert its antiplatelet activity through the inhibition of the intra-cellular $Ca^{2+}$ mobilization and the decrease of the $TXA_2$ synthesis.

• KSBB Journal
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• v.17 no.3
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• pp.290-294
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• 2002

The present work describes the improvement of an erythritol-producing strain to lower the formation of glycerol, which is a characteristic by-product of the strain and could cause difficulties in the recovery and purification of the final product. The yeast-like fungi Moniliella suaveolens var. nigra, isolated previously in the same laboratory from beehives, was mutated by exposing it to a 4 g/L NTG solution. From a total of 2000 mutated strains, Em6j30-14 was selected as the one having the most desirable properties. Cultivating the strain for seven days in 300 mL flasks containing 30 mL of a 400 g/L glucose medium resulted in an erythritol yield of 43%. The glycerol yield was 5%, which is a value 50% lower as compared with the wild type. However, attempts to reproduce the above results in a 5L-fermenter failed, resulting in a similar erythritol concentration but a much higher formation of glycerol. Possible reasons for such a different behaviour could be oxygen limitation or the aggregation of cells, but the exact mechanism could not yet be identified. Foam formation, which is another major problem in large-scale fermentation, tended to be much lower for the mutant strain.

• Microbiology and Biotechnology Letters
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• v.43 no.2
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• pp.91-96
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• 2015

Prions are proteinaceous infectious particles that cause neurodegenerative diseases, such as scrapie in sheep, bovine spongiform encephalopathy in cattle and Creutzfeldt-Jakob disease (CJD) in humans. Although the detailed process, regarding the abnormal conversion of prion proteins (PrP), remains to be fully elucidated, a number of environmental factors appear to affect the formation of misfolded PrP, termed PrP^Sc. Because oceanic algae contain mycosporine-like amino acids (MAAs), which exhibit cellular defensive activities under a variety of stress conditions, we investigated the level of PrP^Sc in prion-infected neuroblastoma cells using mycosporine-glycine, porphyra-334 and shinorine. When judged by the level of protease-resistant PrP^Sc in western blots, porphyra-334 and shinorine increased the level of PrP^Sc in cells, but mycosporine-glycine did not. The current results indicate that the MAAs tested in this study enhance the formation of PrP^Sc.

• Macromolecular Research
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• v.11 no.4
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• pp.260-266
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• 2003

Ethylene vinyl acetate (EVA)/clay nanocomposites were synthesized by blending a solution of ethylene vinyl acetate copolymer containing 12% vinyl acetate abbreviated as EVA-12 in toluene and dispersion of dodecyl ammonium ion intercalated montmorillonite (l2Me-MMT) in N,N-dimethyl acetamide (DMAc). X-ray patterns of sodium montmorillonite ($Na^+$-MMT) and 12Me-MMT exhibited $d_{001}$ peak at $2{\theta}=7.4^{\circ}$ and $2{\theta}=5.6^{\circ}$ respectively; that is, the interlayer spacing of MMT increased by about 0.39 nm due to intercalation of dodecyl ammonium ions. The XRD trace of EVA showed no peak in the angular range of $3-10^{\circ}(2{\theta})$. In the XRD patterns of EVA/clay hybrids with clay content up to 6 wt% the basal reflection peak of 12Me-MMT was absent. leading to the formation of delaminated configuration of the composites. When the 12Me-MMT content was 8 wt% in the EVA-12 matrix, the hybrid revealed a peak at about $2{\theta}=5.6^{\circ}$, owing to the aggregation of aluminosilicate layers. Transmission electron microscopic photograph exhibited that an average size of 12-15 nm clay layers were randomly and homogeneously dispersed in the polymer matrix, which led to the formation of nanocomposite with delaminated configuration. The formation of delaminated nanocomposites was manifested through the enhancement of mechanical properties and thermal stability, e.g. tensile strength of an hybrid containing only 2 wt% 12Me-MMT was enhanced by about 36% as compared with neat EVA-12.

• Preventive Nutrition and Food Science
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• v.1 no.1
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• pp.134-142
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• 1996

Diabetes carries an increased risk of atherosclerotic disease that is not fully explained by known car-diovascular risk factors. There is accumulating evidence that advanced glycation of structural proteins, and oxidation and glycation of circulating lipoproteins, are implicated in the pathogenesis of diabetic ather-osclerosis. Reactions involving glycation and oxidation of proteins and lipids are believed to contribute to atherogenesis. Glycation, the nonenzymatic binding of glucose to protein molecules, can increase the ather-ogenic potential of certain plasma constituents, including low density lipoptotein(LDL). Glycation of LDL is significant increased in diabetic patients compared with normal subjects, even in the presence of good glycemic control. Metabolic abnormalities associated with glycation of LDL include diminished recognition of LDL by the classic LDL receptor; increased covalent binding of LDL in vessel walls ; enhanced uptake of LDL by the macrophages, thus stimulating foam cell formation ; increased platelet aggregation; formation of LDL-immune complexes ; and generation of oxygen free radicals, resulting on oxidative damage to both the lipid and protein components of LDL and to any nearby macromolecules. Oxidized lipoproteins are characterzied by cytotoxicity, potent stimulation of foam cell formation by macrophages, and procoagulant effects. Combined glycation and oxidation, "glycoxidation" occurs when oxidative reactions affect the initial products of glycation, and results in irreversible structural alterations of proteins. Glycoxidation is of greatest significance in long lived proteins such as collagen. In these proteins, glycoxidation products, believed to be atherogenic, accumulate with advancing age : in diabetes, their rate of accumulate is accelerated. Inhibition of glycation, oxidation and glycoxidation may form the basis of future antiaterogenic strategies in both diabetic and nondiabetic individuals.dividuals.

• Journal of Ginseng Research
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• v.41 no.4
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• pp.548-555
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• 2017

Background: Korean Red Ginseng has been used for several decades to treat many diseases, enhancing both immunity and physical strength. Previous studies have documented the therapeutic effects of ginseng, including its anticancer, antiaging, and anti-inflammatory activities. These activities are mediated by ginsenosides present in the ginseng plant. Ginsenoside Rg3, an effective compound from red ginseng, has been shown to have antiplatelet activity in addition to its anticancer and anti-inflammatory activities. Platelets are important for both primary hemostasis and the repair of the vessels after injury; however, they also play a crucial role in the development of acute coronary diseases. We prepared ginsenoside Rg3-enriched red ginseng extract (Rg3-RGE) to examine its role in platelet physiology. Methods: To examine the effect of Rg3-RGE on platelet activation in vitro, platelet aggregation, granule secretion, intracellular calcium ($[Ca^{2+}]_i$) mobilization, flow cytometry, and immunoblot analysis were carried out using rat platelets. To examine the effect of Rg3-RGE on platelet activation in vivo, a collagen plus epinephrine-induced acute pulmonary thromboembolism mouse model was used. Results: We found that Rg3-RGE significantly inhibited collagen-induced platelet aggregation and $[Ca^{2+}]_i$ mobilization in a dose-dependent manner in addition to reducing ATP release from collagen-stimulated platelets. Furthermore, using immunoblot analysis, we found that Rg3-RGE markedly suppressed mitogen-activated protein kinase phosphorylation (i.e., extracellular stimuli-responsive kinase, Jun N-terminal kinase, p38) as well as the PI3K (phosphatidylinositol 3 kinase)/Akt pathway. Moreover, Rg3-RGE effectively reduced collagen plus epinephrine-induced mortality in mice. Conclusion: These data suggest that ginsenoside Rg3-RGE could be potentially be used as an antiplatelet therapeutic agent against platelet-mediated cardiovascular disorders.

• Bulletin of the Korean Chemical Society
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• v.35 no.4
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• pp.1169-1176
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• 2014

In this study, water-dispersible ZnS:Mn nanocrystals were synthesized by capping the surface with conventional and simple structured amino acid ligands: $\small{L}$-Glycine and $\small{L}$-Alanine. The ZnS:Mn-Gly and ZnS:Mn-Ala nanocrystal powders were characterized by XRD, HR-TEM, EDXS, ICP-AES, and FT-IR spectroscopy. The optical properties were measured by UV-Visible and photoluminescence (PL) spectroscopy. The PL spectra for the ZnS:Mn-Gly and ZnS:Mn-Ala showed broad emission peaks at 599 nm and 607 nm with PL efficiencies of 6.5% and 7.8%, respectively. The measured average particle size from the HR-TEM images were $6.4{\pm}0.8$ nm (ZnS:Mn-Gly) and $4.1{\pm}0.5$ nm (ZnS:Mn-Ala), which were also supported by Debye-Scherrer calculations. In addition, the degree of aggregation of the nanocrystals in aqueous solutions were measured by a hydrodynamic light scattering method, which showed formation of sub-micrometer size aggregates for both ZnS:Mn-Gly ($273{\pm}94$ nm) and ZnS:Mn-Ala ($233{\pm}34$ nm) in water due to the intermolecular attraction between the capping amino acids molecules. Finally, the cytotoxic effects of ZnS:Mn-Gly and ZnS:Mn-Ala nanocrsystals over the growth of wild type E. coli were investigated. As a result, no toxicity was shown for the ZnS:Mn-Gly nanocrystal in the colloidal concentration region from 1 ${\mu}g/mL$ to 1000 ${\mu}g/mL$, while ZnS:Mn-Ala showed significant toxicity at 100 ${\mu}g/mL$.

• Analytical Science and Technology
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• v.28 no.5
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• pp.331-340
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• 2015

Mutations in Fused in Sarcoma (FUS) have been identified in patients with amyotrophic lateral sclerosis (ALS) or Frontotemporal Dementia (FTD). Pathological FUS is mis-localized to cytosol and forms aggregates associated with stress granules (SG), while FUS is normally localized to nucleus. However, it is largely unknown how pathological FUS forms SG-aggregates and which domains are responsible for this process. In this study, we examined cellular localization and aggregation of ALS-linked FUS missense mutants (P525L, R521C, R521H, R521G), analyzed the domains responsible for cytosolic FUS aggregation in HEK293T cells, and confirmed this in cultured mouse neurons. To do this, we firstly generated missense mutants of FUS and then examined their cellular localization. We found that P525L was mostly mis-localized to cytosol and formed FUS-positive SG aggregates while R521C, R521H, or R521G was localized to both nucleus and cytosol. To further characterize the domains required for aggregate formation of cytosolic FUS, we generated different domain-deletion mutants using FUS-∆17 which has a deletion of nuclear localization signal. Interestingly, cytosolic FUS without SYGQ and RGG1 domain or cytosolic FUS without RGG2-ZnF-RGG3 domain did not form FUS-positive SG aggregates, while cytosolic FUS without RRM domain generated more aggregates compared to FUS-∆17. Taken together, these data suggest that SYGQ-RGG1 or RGG2-ZnF-RGG3 domain contributes to formation of cytosolic aggregate, while RRM domain might interfere with FUS aggregation. Therefore, our studies will provide important insight for understanding cellular pathogenesis of neurodegeneration associated with FUS aggregate as well as finding therapeutic targets for ALS or FTD.

• The Journal of Korean Medicine
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• v.29 no.2
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• pp.151-164
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• 2008

Objective: To investigate the effects of Yeongdamsagantang (YDGT) on apoptosis of neuronal cells that can result in dementia. Method: The water extract of the YDGT was tested in vitro for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with $A{\beta}$ oligomer-related dementias. $A{\beta}$ oligomers derived from proteolytic processing of the ${\beta}-amyloid$ precursor protein (APP), including the $amyloid-{\beta}$ peptide $(A{\beta})$, play a critical role in the pathogenesis of Alzheimer's disease. A neuroblastoma cell line stably expressing an $A{\beta}$ oligomerassociated neuronal degeneration was used to investigate if YDGT inhibits formation of $A{\beta}$ oligomer. To measure the ATP generating level in mitochondrial membrane, luciferin/luciferase luminescence kit (Promega) and luminator was used, and to survey the protein's apparition, confocal microscopy was used. Result: $A{\beta}oligomer$ had a profound attenuation in the increase in CT105 expressing neuro2A cells from YDGT. Experimental evidence indicates that YDGT protected against neuronal damage from cells, but its cellular and molecular mechanisms remain unknown. We demonstrated that YDGT inhibited formation of $amyloid-{\beta}$ $(A{\beta})$ oligomers, which were the behavior, and possibly causative, features of AD. The decreased $A{\beta}$ oligomer in the presence of YDGT was observed in the conditioned medium of this $A{\beta}oligomer-secreting$ cell line under in vitro. In the cells, YDGT significantly attenuated mitochondrion-initiated apoptosis. Conclusion: (i) a direct $A{\beta}$ oligomer toxicity and the apoptosis initiated by the mitochondria; and (ii) multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer aggregation, underlie the neuroprotective effects of YDGT.