For understanding physiological nature of phototaxis in Synechocystis sp. PCC 6803 PTX(S. 6803 PTX), we examined the effects of some metabolic inhibitors and cation ionophore on the phototactic movement. In the presence of DCMU, which blocks the photosynthetic electron transport just after photosystem II acceptor, there was no inhibitory effect on the phototaxis up to $100\;\mu\textrm{M}$. Instead, the respiratory electron chain inhibitor such as sodium azide dramatically impaired the phototaxis in S. 6803 PTX. These observations indicate that the phototaxis is linked not to photo-phosphorylation, but to respiratory phosphorylation. When the cells were treated with un couplers such as CCCP or DNP, which dissipate the electrochemical gradient of proton($\Delta\mu_{H}+$) across the cytoplasmic membrane, these chemicals did not affect phototaxis. In contrast, when cells were treated with DCCD or NBD which deprive cells of A TP but leave $\Delta\mu_{H}+$ intact across the membrane, the phototactic movement was severly reduced. These results imply that ATP production, not proton motive force, is involved in the phototactic movement in this organism as a driving motive force. The application of specific calcium ionophore A23187 strongly impaired positive phototaxis. Calcium fluxes should be engaged in the sensory trans-duction of phototactic orientation. Finally, when ethionine was supplimented to culture media, the photomovement of this organism was inhibited. This implies that methylation/demethylation mechanism controls the process of phototaxis in S. 6803 PTX like chemotaxis in E. coli and Salmonella typhimurium.murium.
In order to investigate the histopathological, mechanism of Rompun-induced shock, the development of mammary carcinoma, the numerical changes and the morphological findings of the mast cells appeared in the carcinoma were microscopically observed in the rat treated with DMBA and each chemical of compound 48/80 and Rompun. Also mast cell degranulation induced by Rompun was observed with electron microscope. The results observed were summarized as follows: Tumor appeared in 100% of the animals. Tumors grew more rapidly to $10{\times}10mm$ in rats depleted of mast cells ($37.7{\pm}4.2$ days) than was observed in the control group ($42.5{\pm}4.7$ days) (p<0.005). The mean number of tumors per rat was $2.8{\pm}1.3$ in the compound 48/80- treated group in contrast to $3.4{\pm}1.3$ in the control group. No significant difference was apparent in the tumor induction time of Rompun treated group compared with the compound 48/80-treated group, but the tumor measuring at least $10{\times}10mm$ appeared more quickly in the Rompun treated group than in the control group (p<0.005). The numbers of mast cells in the control group were inclined to increase significantly according to the mammary tumor development (p<0.005). In contrast, the mast cells were fewer significantly in the compound 48/80-treated group and Rompun-treated group than in the control group (p<0.005). The numbers of mast cells in the compound 48/80-treated group and Rompun-treated group were inclined to reduce significantly according to the stages of the mammary carcinoma growth in contrast to the control group respectively. The ultrastructural morphologies of mast cells at 30 minutes after Rompun injection were appeared many normal granules in the cytoplasm, but many normal and degranulated granules were scattered along the cell membrane. And at 1 hour after Rompun injection mast cell granules were disappeared nearly or rarely seen. many long cytoplasmic projections were folded back to adhere to their own surface membrane. and mast cells resulted in a reduced size of these cells. Otherwise. compound 48/80 caused extensive degranulation of mast cells by disrupting cell membrane. but mast cell degranulation by Rompun was observed exocytosis of granules through a channel. From the above results. it is concluded that the Rompun may give rise to the dealth of animals as a shock caused by mast cell degranulation.
This study analyzed the antibacterial activity of the ethanol extract of Schizandra chinensis Baillon against food pathogenic microorganisms to determine its capabilities as a natural antimicrobial agent. A paper disc diffusion test, minimum inhibitory concentration (MIC) determination, and time-kill assay showed that the ethanol extract strongly inhibits the growth of Listeria monocytogenes, Bacillus cereus, Escherichia coli O157:H7, and Pseudomonas aeruginosa. Release of cytoplasmic ${\beta}$-galactosidase was detected in E. coli, E. coli O157:H7, S. aureus, and P. aeruginosa treated with the ethanol extract. An increase of outer membrane permeability caused by the ethanol extract was also observed. An outward flow of cell constituents was detected in the Gram negative strains treated with the ethanol extract. These results imply that the inner and outer membranes of cells were partially destroyed and cell constituents were released by the treatment of the S. chinensis Baillon ethanol extract. The results of this study indicate that ethanol extract of S. chinensis Baillon evidences a fairly good antibacterial effect.
G protein coupled receptors (GPCRs) transmit various extracellular signals into the cells. Upon binding of the ligands, conformational changes in the extracellular and/or transmembrane (TM) domains of CPCRs were propagated into the cytoplasmic (CP) domain of the molecule leading to the activation of their cognate heterotrimeric C proteins and kinases. Constitutively active GPCR mutants causing the activation of C Protein signaling even in the absence of ligand binding are of interest for the study of activation mechanism of GPCRs. Two classes of constitutively active mutations, categorized by their effects on the salt bridge between Ell3 and K296, were found in the TM domain of rhodopsin. Opsin mutants containing combinations of the mutations were constructed to study the conformational changes required for the activation of rhodopsin. Rhodopsin chromophore regenerated with 11-cis-retinal showed a thermal stability inversely correlated with its constitutive activity. In contrast, rhodopsin mutants exhibited a binding affinity to an agonist, all-trans-retinal, in a constitutive activity-dependent manner. In order to test whether the conformational changes responsible for the activation of trans-ducin (Gt) are the same as the conformation required for the recognition of rhodopsin kinase, analysis of the mutants were carried out with phosphorylation by rhodopsin kinase. Rhodopsin mutants containing combinations of different classes of the mutations showed a strong synergistic effect on the phosphorylation of the mutants in the dark as similar to that of Gt activation. The results suggest that at least two or three kinds of segmental and independent conformational changes are required for the activation of rhodopsin and the conformational changes responsible for activating rhodopsin kinase and Gt are similar to each other.
Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.
Park, Hyo-Jin;Kim, Jin-Woo;Park, Jae-Young;Yang, Seul-Gi;Jung, Jae-Min;Kim, Min-Ji;Koo, Deog-Bon
Journal of Embryo Transfer
/
v.32
no.1
/
pp.17-24
/
2017
Ganglioside GD1a is specifically formed by the addition of sialic acid to ganglioside GM1a by ST3 ${\beta}$-galactoside ${\alpha}$-2,3-sialyltransferase 2 (ST3GAL2). Above all, GD1a are known to be related with the functional regulation of several growth factor receptors, including activation and dimerization of epidermal growth factor receptor (EGFR) in tumor cells. The activity of EGF and EGFR is known to be a very important factor for meiotic and cytoplasmic maturation during in vitro maturation (IVM) of mammalian oocytes. However, the role of gangliosides GD1a for EGFR-related signaling pathways in porcine oocyte is not yet clearly understood. Here, we investigated that the effect of ST3GAL2 as synthesizing enzyme GD1a for EGFR activation and phosphorylation during meiotic maturation. To investigate the expression of ST3GAL2 according to the EGF treatment (0, 10 and 50 ng/ml), we observed the patterns of ST3GAL2 genes expression by immunofluorescence staining in denuded oocyte (DO) and cumulus cell-oocyte-complex (COC) during IVM process (22 and 44 h), respectively. Expression levels of ST3GAL2 significantly decreased (p<0.01) in an EGF concentration (10 and 50 ng/ml) dependent manner. And fluorescence expression of ST3GAL2 increased (p<0.01) in the matured COCs for 44 h. Under high EGF concentration (50 ng/ml), ST3GAL2 protein levels was decreased (p<0.01), and their shown opposite expression pattern of phosphorylation-EGFR in COCs of 44 h. Phosphorylation of EGFR significantly increased (p<0.01) in matured COCs treated with GD1a for 44 h. In addition, ST3GAL2 protein levels significantly decreased (p<0.01) in GD1a ($10{\mu}M$) treated COCs without reference to EGF pre-treatment. These results suggest that treatment of exogenous ganglioside GD1a may play an important role such as EGF in EGFR-related activation and phosphorylation in porcine oocyte maturation of in vitro.
Kim, Sung Eun;Kim, Myoung-Hwan;Woo, Joohan;Kim, Sung Joon
The Korean Journal of Physiology and Pharmacology
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v.24
no.6
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pp.555-561
/
2020
TWIK-related two-pore domain K+ channel-2 (TREK-2) has voltage-independent activity and shows additional activation by acidic intracellular pH (pHi) via neutralizing the E332 in the cytoplasmic C terminal (Ct). We reported opposite regulations of TREK-2 by phosphatidylinositol 4,5-bisphosphate (PIP2) via the alkaline K330 and triple Arg residues (R355-357); inhibition and activation, respectively. The G334 between them appeared critical because its mutation (G334A) endowed hTREK-2 with tonic activity, similar to the mutation of the inhibitory K330 (K330A). To further elucidate the role of putative bent conformation at G334, we compared the dual mutation forms, K330A/G334A and G334A/R355-7A, showing higher and lower basal activity, respectively. The results suggested that the tonic activity of G334A owes to a dominant influence from R355-7. Since there are additional triple Arg residues (R377-9) distal to R355-7, we also examined the triple mutant (G334A/R355-7A/R377-9A) that showed tonic inhibition same with G334A/R355-7A. Despite the state of tonic inhibition, the activation by acidic pHi was preserved in both G334A/R355-7A and G334A/R355-7A/R377-9A, similar to the R355-7A. Also, the inhibitory effect of ATP could be commonly demonstrated under the activation by acidic pHi in R355-7A, G334A/R355-7A, and G334A/R355-7A/R377-9A. These results suggest that the putative bent conformation at G334 is important to set the tug-of-war between K330 and R355-7 in the PIP2-dependent regulation of TREK-2.
The bacteriocin produced by Lactobacillus sp. GM7311 showed strong inhibitory activity against the growth of three Gram positive bacteria, Listeria monocytogenes, Bacillus subtilis, and Staphylococcus aureus. When the bacteriocin was added to the culture at different phases, viable cells of all of the tested strains were decreased, although the most inhibited phase was different. Thereby, when the bacteriocin $(100\;Bu/m{\ell})$ was added to exponential and stationary phase of L. monocytogenes, the rapid reduction of viable cell counts occured. And, in the case of B. subtilis, the highest inhibitory effect occured at lag phase and mid-exponential phase by the addition of the bacteriocin under same condition as mentioned above. Also, we can observe the accelerated reduction of survivors counts for the all of the phase except stationary phase in the S. auresus. Transmission electron microscopic observation of L. monocrogenes and B. subtilis treated with bacteriocin revealed apparent Iysis of the cell wall and excretion of the cell contents, indicating bacteriolysis. Also, the amino acids and fatty acids compositions were different from controls. However, the Iysis of cell wall didn't occur in S. aureus, though the cytoplasmic materials were reduced. This result indicates that the bacteriocin inhibits the synthesis of nuclear materials such as DNA, RNA and proteins.
Hyun Hwangbo;Cheol Park;EunJin Bang;Hyuk Soon Kim;Sung-Jin Bae;Eunjeong Kim;Youngmi Jung;Sun-Hee Leem;Young Rok Seo;Su Hyun Hong;Gi-Young Kim;Jin Won Hyun;Yung Hyun Choi
Biomolecules & Therapeutics
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v.32
no.3
/
pp.349-360
/
2024
Oxidative stress contributes to the onset of chronic diseases in various organs, including muscles. Morroniside, a type of iridoid glycoside contained in Cornus officinalis, is reported to have advantages as a natural compound that prevents various diseases. However, the question of whether this phytochemical exerts any inhibitory effect against oxidative stress in muscle cells has not been well reported. Therefore, the current study aimed to evaluate whether morroniside can protect against oxidative damage induced by hydrogen peroxide (H2O2) in murine C2C12 myoblasts. Our results demonstrate that morroniside pretreatment was able to inhibit cytotoxicity while suppressing H2O2-induced DNA damage and apoptosis. Morroniside also significantly improved the antioxidant capacity in H2O2-challenged C2C12 cells by blocking the production of cellular reactive oxygen species and mitochondrial superoxide and increasing glutathione production. In addition, H2O2-induced mitochondrial damage and endoplasmic reticulum (ER) stress were effectively attenuated by morroniside pretreatment, inhibiting cytoplasmic leakage of cytochrome c and expression of ER stress-related proteins. Furthermore, morroniside neutralized H2O2-mediated calcium (Ca2+) overload in mitochondria and mitigated the expression of calpains, cytosolic Ca2+-dependent proteases. Collectively, these findings demonstrate that morroniside protected against mitochondrial impairment and Ca2+-mediated ER stress by minimizing oxidative stress, thereby inhibiting H2O2-induced cytotoxicity in C2C12 myoblasts.
The study was conducted to obtain the genetic information on heterosis and combining ability of the quantitative characters for F1 hybrid breeding in silkworms. Six parental varieties and each set of 30 diallel crosses in F1's were used as materials, and bred on the randomized complete block design with three replications. Fourteen characters were observed with the twenty samples in each tray. The data were analyzed for (1) heterosis and combining ability in F1 hybrid. The heterosis in the weight and the length of cocoon showed positively high at 24.51%, and 23.4%, respectively and the weight of the whole cocoon as well as the weight of the whole cocoon layer showed a siginificant heterosis ranging from 15.56% to 15.71% and from 17.14% to 19.01%, but the fifth and the total instar period showed negative heterosis. It was found that the combination between, C70XRomogua and N9 X Romogua showed highly a negative heterosis on the fifth instar period and for the cocoon weight. The female of N9+Sansuian and the male of Romogua X Sansurian have a high heterosis effect, on the cocoon shell weight, and Sansurian X Romogua(reciprocal) on the length and the weight of cocoon filament with no regard to sexuality. The significant maternal and cytoplasmic effect on heterosis of the cocoon weight and the cocoon shell weight were observed with the combinations, N9 X C5, N63 X C70 and on the length of the cocoon filament with the combinations, Sansurian X N63, Sansurian X C5, Sansurian X C70 and N9 X C70, N63 X C70 on the weight of cocoon filament. As mean squared of GCA, SCA and RCA were significant with these combining ability for all characters resulted from additive and non-additive altogether and there is a significant difference between reciprocals. Sansurian showed a negative GCA effect on the fifth and total larval duration, but the higher positive GCA effects took places with varieties N9 and C5 on the length, width, weight of cocoon, cocoon shell weight, percentage of cocoon shell weight, length and weight of cocoon filament, percentage of raw-silk with no regard to both generations and silkworm sexuality. The values of SCA between the cross combinations varied generation-wise and sex-wise. It was shown that SCA value for the fifth instar period was highly negative for Sansurian X C70, Romogua X C70, Sansurian X C5, Romogua X C5, but it was positive effect on the cocoon weight, cocoon shell weight with N9 X C5, and C70 X Sansurian, on the length of cocoon filament with N9 X C5, Romogua X Sansurian on the weight of cocoon filament between Romogua and N63 and on the percentage of raw-silk between the combination of Sansurian X Romoga.
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