This study was carried out to investigate the fine structural changes of rat hepatocytes by repeated treatment of cyclosporin A that has been widely used for immunosuppressive drug in organ transplantation. Sprague-Dawley rats were kept in experimental circumstances for 2 weeks and 50mg/kg B.W of cyclosporin A was injected once a day subcutaneously for 7 days and sacrificed at 1 hour, 1 day, 3 days, 7 days, 14 days, 28 days after the last injection. Fine structural changes were observed by transmission electron microscope (JEM 1200EX II) and the results obtained were as follows. 1. Accumulation of lipid droplets in hepatocytes was prominently increased in 1 hour and 1 day lapse groups, and this finding was slightly reduced in 3 days lapse group and remarkably reduced from 7 days lapse group enough to be recovered completely in 14 days lapse group. 2. Dilatation of rough endoplasmic reticule cisternae, detachment of membrane bound ribosomes, proliferation of smooth endoplasmic reticula were observed in 1 hour and 1 day lapse groups, and these findings were mild in 3 days lapse group and abruptly reduced from 7 days lapse group enough to be recovered completely in 28 days lapse group. 3. Small myelin figures were observed in 3 days lapse group after CsA-treatment. 4. Swelling of mitochondria and destruction of their cristae were observed in 1 hour and 1 day lapse groups, and these findings were recovered from 3 days lapse group. 5. Dilatation of bile canaliculi and remarkable loss of microvilli in the pericanalicular wall were observed in 1 hour lapse group and the most severe change was shown in 1 day lapse group and lasted to 3 days lapse group, and these findings were reduced gradually from 7 days lapse group enough to recovered completely in 28 days lapse group.
In this study, the relationships between sensitivity to oxyfluorfen, absorption of the herbicide, protoporphyrin IX(Proto IX) accumulation and activities of antioxidative enzymes were examined to identify the tolerance mechanism against oxyfluorfen in various rice cultivars having different level of tolerance to this herbicide. Absorption of oxyfluorfen in tolerant rice cultivars was slower than in susceptible cultivars. Proto IX accumulation in various rice cultivars treated with oxyfluorfen was higher in susceptible cultivars than in tolerant ones. In susceptible cultivars especially, Proto IX accumlated rapidly during the herbicide treatment in the dark. Large amounts of Proto IX accumulation were considered to cause membrane lipid peroxidation in the light. However, among the tested rice cultivars, there was little relationship between their tolerance to oxyfluorfen and the activities of antioxidative enzymes. Therefore, it is assumed that differential susceptibility of rice cultivars to oxyfluorfen was due to difference in their capability to absorb the herbicide and to subsequently accumulate Proto IX.
Jha, Pankaj Kumar;Paul, Ashit Kumar;Rahman, M. Bozlur;Tanjim, M.;Bari, Farida Yeasmin;Alam, M. Golam Shahi
Journal of Embryo Transfer
/
v.28
no.1
/
pp.31-39
/
2013
Alpha-tocopherol as an antioxidant acts in preservation of chilled semen by preserving cell membrane damage from lipid peroxidation. Optimum concentrations of ${\alpha}$-tocopherol in egg yolk-citrate (EYC) extender need to be studied in crossbred bull's semen. Different concentrations of ${\alpha}$-tocopherol viz. 0, 1, 2, 4 and 6mg per ml of extender were used. Semen was collected once a week from four bulls used to regular collection, aged 4 to 7 years, weighing 320 to 450 kg, and with body condition score 4 to 4.5 and scrotal circumference 23 to 32 cm. Semen was evaluated routinely and sperm morphology was viewed under light microscope at ${\times}1,000$ magnification after fixing with buffered formal saline. Over 90% had normal head, acrosome, mid-piece and tail. Semen was diluted with egg-yolk-citrate extender to produce $15{\times}10^6$ spermatozoa/ml and 0, 1, 2, 4 and 6 mg/ml ${\alpha}$-tocopherol were added. The semens amples were kept at $8^{\circ}C$. Sperm motility and viability were examined daily up to 5 days under light microscopy at ${\times}200$ magnification. Sperm viability was acceptable (${\geq}40%$) up to the $4^{th}$ day with all concentrations of ${\alpha}$-tocopherol and up to the $5^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. Sperm motility was acceptable (${\geq}40%$) up to the $3^{rd}$ day irrespective of ${\alpha}$-tocopherol concentration, and up to the $4^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. It is suggested that the lifespan of chilled semen may be extended up to 4 days by adding 2mg/ml ${\alpha}$-tocopherol.
Kim, Kyeongnam;Lee, Byung-Ho;Park, Jeong Sun;Yang, Jeong Oh;Lee, Sung-Eun
Journal of Applied Biological Chemistry
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v.60
no.3
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pp.271-277
/
2017
Ethyl formate has been used for the control of insect pests by fumigation. However, there were not many reports to show its target site of fumigant toxicity on insect pests since its first use in the agricultural industry. In the present study, we showed the presumable target sites of ethyl formate fumigation in insect pests using Myzus persicae nymphs. After ethyl formate fumigation, the nymphs of this species were collected and the changes at the biochemical and molecular level were determined. The activity of cytochrome c oxidase (COX) was approximately two-fold higher after ethyl formate fumigation. In addition, the expression levels of acetylcholinesterase (AChE) decreased gradually with increasing ethyl formate concentration. These two findings suggested that COX and AChE might be the major target sites of ethyl formate fumigation. In addition to these results, the analysis of lipid content using MALDI-TOF MS/MS identified 9 phospholipids differently generated 2-fold higher in the ethyl formate-treated nymphs than that in the control nymphs, thereby leading to changes in cell membrane composition in M. persicae nymphs. Therefore, the ethyl formate fumigation caused lethal effects on M. persicae nymphs by changing COX activity, AChE gene expression, and phospholipid production.
Journal of Fisheries and Marine Sciences Education
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v.27
no.4
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pp.1031-1040
/
2015
Ultrstructural studies of germ cell differentiation and vitellogenesis in the oocytes of the female Rapana venosa in the brackish water area of Seomjin River were investigated by transmission electron microscope observations. In the early vitellogenic oocytes, the Golgi complex and mitochondria were involved in the formation of glycogen particle, lipid droplets, and yolk granules. In the late vitellogenic oocytes, the rough endoplasmic reticulum and multivesicular bodies were involved in the formation of proteid yolk granules in the cytoplasm. However, heterosynthetic vitellogenesis in this species were not observed in vitellogenic oocytes during oogenesis. A mature yolk granule was composed of three components: crystalline core, electron lucent cortex and the limiting membrane. As shown in some large gastropods, vitellogenesis in R. venosa occurred by way of endogenous autosynthesis without heterosythetic vitellogenesis (exogeneous endocytosis), which are found in the oocytes in bivalves. The mating period and spawning activity were related with the increases of seawater temperatures and salinities.
Liposome as a carrier of topotecan (TPT), a promising anticancer drug, has been reported in attempt to improve the stability and antitumor activity of TPT. However, the biodistr ibution pattern of TPT liposome in vivo and PEG-modified liposome containing TPT have not been studied systemically. In this paper, the in vitro stability and in vivo biodistribution behavior of several liposomes containing TPT with different lipid compositions and PEG-modification were studied. Compared with the 'fluid' liposome (S-Lip) composed of soybean phosphatidylcholine (SPC), the 'solid' liposome (H-Lip) composed of hydrogenated soybean phosphatidylcholine HSPC decreased the leaking efficiency of TPT from liposome and enhanced the stability of liposome in fetal bovine serum (FBS) or human blood plasma (HBP). The results of biodistribution studies in S$_{180}$ tumor-bearing mice showed that liposomal encapsulation increased the concentrations of total TPT and the ratio of lactone form in plasma. Compared with free TPT, S-Lip and H-Lip resulted in 5- and 19- fold increase in the area under the curve (AUC$_{0\rightarrow\propto}$), respectively. PEG- modified H-Lip (H-PEG) showed 3.7-fold increase in AUC$_{0\rightarrow\propto}$ compared with H-Lip, but there was no significant increase in t$_{1/2}$ and AUC$_{0\rightarrow\propto}$ for PEG-modified S-Lip (S-PEG) compared with S-Lip. Moreover, the liposomal encapsulation changed the biodistribution behavior, and H-Lip and H-PEG dramatically increased the accumulation of TPT in tumor, and the relative tumor uptake ratios were 3.4 and 4.3 compared with free drug, respectively. There was also a marked increase in the distribution of TPT in lung when the drug was encapsulated into H-Lip and H-PEG. Moreover, H-PEG decreased the accumulation of TPT in bore marrow compared with unmodified H-Lip. All these results indicated that the membrane fluidity of liposome has an important effect on in vitro stability and in vivo biodistribution pattern of liposomes containing TPT, and PEG-modified 'solid' liposome may be an efficient carrier of TPT.
Ginseng has been used as a general tonic agent to invigorate human body. In the present study, we isolated novel glycolipoproteins from ginseng that activate $Ca^{2+}$-activated $Cl^-$ channel (CaCC) in Xenopus oocytes and transiently increase intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) in mouse Ehrlich ascites tumor cells. We named the active ingredients as gintonin. Gintonin exists in at least six different forms. The native molecular weight of gintonin is about 67 kDa but its apparent molecular weight is about 13 kDa, indicating that gintonin might be a pentamer. Gintonin is rich in hydrophobic amino acids. Its main carbohydrates are glucose and glucosamine. Its lipid components are linoleic, palmitic, oleic, and stearic acids. Gintonin actions were blocked by U73122, a phospholipase C inhibitor, 2-aminoethxydiphenyl borate, an inositol 1,4,5-trisphosphate receptor antagonist, or bis (o-aminophenoxy) ethane-N,N,N0,N0-tetracetic acid acetoxymethyl ester, a membrane permeable $Ca^{2+}$ chelator. In the present study, we for the first time isolated novel gintonin and showed the signaling pathways on gintonin-mediated CaCC activations and transient increase of $[Ca^{2+}]_i$. Since $[Ca^{2+}]_i$ as a second messenger plays a pivotal role in the regulation of diverse $Ca^{2+}$-dependent intracellular signal pathways, gintonin-mediated regulations of $[Ca^{2+}]_i$ might contribute to biological actions of ginseng.
The present study was undertaken to demonstrate the surface structure of Paragonimus westermani metacercaria in Korea with special reference to the distribution of sensory papillae. Metacercariae were isolated from crayfish, one of the second intermediate tost of P. westermani in Bogil island, Chollanam-do (Province), Korea, where has been known as an endemic area of human paragonimiasis. Isolated metacercariae were encysted and examined with light. scanning and transmission electron microscopes for morphological features. On the surface of iBetacercariae, three types of sensory FaFillae were identified. Large domed papillae ($3~5{\mu\textrm{m}}$), which were covered with wrinkled plasma n!embrane of the worm, were distributed on the oral and ventral suckers only. On the oral sucker, these large domed papillae were 12~13 in number. On the other hand large domed papillae on the ventral sucker were constantly 6 in number and hexagonal in distribution. Small domed papillae ($2~3{\mu\textrm{m}}$), of which surface was more smooth than those of large ones, were distributed symmetrically on the ventral (30~32 pairs) and dorsal surfaces (40~42 Pairs). Ciliated Papillae ($0.8~1.5{\mu\textrm{m}}$) were observed about 5~6 in number around the oral sucker and 3~5 pairs each on the ventral and dorsal surface of the body. Single Fcinted spines covered the entire surface of the body except around the excretory pore. Spines on the anterior fart of the body were 0.9~2.0${\mu\textrm{m}}$ in length and $45~55/100{\mu\textrm{m}}$2 in number, and were gradually reduced in length ($0.4~1.4{\mu\textrm{m}}$) and in nuns.her ($12~27/100{\mu\textrm{m}}$2) toward the posterior part. The body wall of p. westermoni metacercariae was consisted with anucleated syncytium layer, fibrous interstitial layer and musclar layer. In the anucleated syncytium, biconcave ($0.15~0.55{\mu\textrm{m}}$) and spherical ($0.08~0.16{\mu\textrm{m}}$) secretory granules, which were transferred from epidermal cells via protoplasmic tubules, mitochondria and rihoEorses, T-ere observed. Spines originated around the basement membrane protruded externally. Epidermal cells were consisted with a nucleus and a cytoplasm, and connected to syncytium with protoplasmic tubules. In the cytoplasm many secretory granules, mitochondria, Golgi complex, endoplasmic reticula, ribosomes and lipid droplets were observed.
Kim, Hye Jin;Yoon, Hae Min;Kim, Tae Young;Lee, Won Jun
Journal of Life Science
/
v.26
no.7
/
pp.758-763
/
2016
Fatty acid transporters are key mediators of skeletal muscle lipid metabolism. Several protein groups have been implicated in cellular long-chain fatty acid uptake or oxidation, including fatty acid transporter proteins (FATPs), the plasma membrane fatty acid-binding protein (FABPpm), and the fatty acid translocase (FAT/CD36). FAT/CD36 is highly expressed in skeletal muscle and known to be regulated by various factors such as exercise and hormones. Insulin-like growth factor-I (IGF-I) is a well-known regulator of skeletal muscle cells. However, it has not been studied whether there is any interaction between IGF-I and FAT/CD36 in skeletal muscle cells. In this study, the effects of IGF-I treatment on FAT/CD36 induction were examined. Differentiated C2C12 cells were treated with 20 ng/ml of IGF-I at different time points. Treatment of C2C12 cells with IGF-I resulted in increased FAT/CD36 mRNA and protein expression. After 24 and 48 hr of IGF-I treatment, FAT/CD36 mRNA increased 89% and 24% respectively. The increase of both proteins returned to the control level after 72 hr of IGF-I treatment, suggesting that the FAT/CD36 gene is regulated pretranslationally by IGF-I in skeletal muscle cells. These results suggest that IGF-I can regulate the expression of FAT/CD36 in skeletal muscle cells. In conclusion, IGF-I induces a rapid transcriptional modification of the FAT/CD36 gene in C2C12 skeletal muscle cells and has modulating effects on fatty acid uptake proteins as well as oxidative proteins.
An Actinomycetes strain, MSA-1, containing antimicrobial component was isolated from the black sponge, Halichondzia okadai, and was identified to a genus level by morphological and chemotaxonornic methods. The gray colored spores were oval type with smooth surface and formed flexibilis spore chains. The cell wall of this strain was type I containing D-aminopimellic acid (D-DAP) and no specific sugar was detected. Phospholipid of the cell membrane was PII type including phophoethanolamine and the major fatty acids of total lipid were branched anteiso-15 : 0, iso-16 : 0, 16 : 0 and iso-17 ; 0. From these results and other characteristics described in the Bergey's Manual, this strain was identificated as a Streptomyces sp. Meanwhile, 10mg of pale yellow colored antimicreobial component was isolated by HPLC method from the cultured Streptomyces sp. (70g of cryophillized mycellis). By crystallographyc analysis, HIRESMS and NMR assignment, the antimicrobial component produced from the strain MSA-1 was elucidated as the staurosporine (indolo[2,3-a]carbazole alkaloid).
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