• Korean Journal of Medicinal Crop Science
• /
• v.13 no.1
• /
• pp.57-62
• /
• 2005

Salt-tolerant transgenic Panax ginseng plants were produced by introducing the SAL1 geue (3'(2'), 5'-bis-phosphate nucleotidase) that confers tolerance to the salts through Agrobacterium tumefaciens co-cultivation. Cotyledon explants of immature ginseng zygotic embryos cultured on Murashige and Skoog medium lacking growth regulators formed somatic embryos directly with below 10%, but the 74% tranformation rate were observed at the treatment of phytohormone with 1.0 mg/l 2,4-D and 0.5 mg/l kinetin. Somatic embryos were initially cultured on MS medium supplemented with 250 mg/l cefotaxime for 3 weeks and subsequently subcultured five times to a medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime. Upon development into the cotyledonary stage, these somatic embryos were transferred to on the medium containing 50 mg/l kanamycin and 10 mg/l gibberellic acid to induce germination and strong selection. Integration of the transgene into the plants was confirmed by polymerase chain reaction with specific primers. The ginseng transformants with well-developed shoots and roots were successfully acclimatized in a greenhouse when they were planted in soil.

• BMB Reports
• /
• v.37 no.3
• /
• pp.370-375
• /
• 2004

There has been increasing interest in the value of using soybean to delay or reduce the tumor incidence. This study was undertaken to investigate the possible protective effects of soybean against hepatocarcinogenesis induced by DL-ethionine. Accordingly, we measured biochemical changes occurring in serum and liver of rats treated with DL-ethionine in the presence or absence of soybean. Male albino rats were fed a control diet containing the hepatocarcinogen, DL-ethionine, or the control diet plus soybean 30%, or the control diet plus soybean plus DL-ethionine 0.25% for three months and then returned to a control diet for up to nine months. Rats fed a control diet plus DL-ethionine showed a gradual decrease in liver DNA, RNA, total protein, and liver weight and enzyme activites of liver transaminases (GOT and GPT) and alkaline phosphatase over the 7-month study period. This was followed by a large increase in the liver parameters at the end of the $9^{th}$ month, except for 5'-nucleotidase and glucose-6-phosphatase that showed a large decrease. On the other hand, a gradual increase in the serum enzyme activities of GOT, GPT, 5-nucleotidase, alkaline phosphatase, and in the albumin/globulin (A/G) ratio is observed in the group of rats fed a control diet plus DL-ethionine compared to the control group over 8 months, and this was followed by a large increase in all serum parameters studied at nine-months. The administration of 30% soybean to the rat diet in addition to DL-ethionine maintained all parameters studied at near control values until the end of the $9^{th}$ month. This study suggests that soybean has a protective effect against the hepatocarcinogenesis induced by DL-ethionine.

• Clinical and Experimental Reproductive Medicine
• /
• v.48 no.2
• /
• pp.124-131
• /
• 2021

Objective: Approximately 30% of preeclamptic pregnancies exhibit abnormal liver function tests. We assessed liver injury-associated enzyme levels and circulating transforming growth factor beta (TGF-β) levels in an arginine vasopressin (AVP)-induced pregnant Sprague-Dawley rat model. Methods: Pregnant and non-pregnant Sprague-Dawley rats (n=24) received AVP (150 ng/hr) subcutaneously via mini-osmotic pumps for 18 days. Blood pressure was measured, urine samples were collected, and all animals were euthanized via isoflurane. Blood was collected to measure circulating levels of TGF-β1-3 isomers and liver injury enzymes in pregnant AVP (PAVP), pregnant saline (PS), non-pregnant AVP (NAVP), and non-pregnant saline (NS) rats. Results: The PAVP group showed significantly higher systolic and diastolic blood pressure than both saline-treated groups. The weight per pup was significantly lower in the AVP-treated group than in the saline group (p<0.05). Circulating TGF-β1-3 isomer levels were significantly higher in the PAVP rats than in the NS rats. However, similar TGF-β1 and TGF-β3 levels were noted in the PS and PAVP rats, while TGF-β2 levels were significantly higher in the PAVP rats. Circulating liver-type arginase-1 and 5'-nucleotidase levels were higher in the PAVP rats than in the saline group. Conclusion: This is the first study to demonstrate higher levels of TGF-β2, arginase, and 5'-nucleotidase activity in PAVP than in PS rats. AVP may cause vasoconstriction and increase peripheral resistance and blood pressure, thereby elevating TGF-β and inducing the preeclampsia-associated inflammatory response. Future studies should explore the mechanisms through which AVP dysregulates liver injury enzymes and TGF-β in pregnant rats.

• Applied Biological Chemistry
• /
• v.27 no.2
• /
• pp.100-106
• /
• 1984

Five subcellular fractions were obtained by successive centrifugation from the liver of rats within 6 hours of life and characterized by comparing marker compound or marker enzyms. After incubating $3{\beta}$-hydroxy-$5{\alpha}$-pregnan-20-one with the each fraction, the steroids were analyzed by TLC, GLC and GC-MS. A $6{\alpha}$-hydroxylase which hydroxylizes the tetra-hydrogenated compound of progesterone, $3{\beta}$-hydroxy-$5{\alpha}$-pregnan-20-one, was localized in the crude plasma membrane fraction, but not in the microsome fraction. The maximum 6α-hydroxylation was observed at pH 7.0. While this 6α-steroid hydroxylase was not able to hydroxlyze the progesterone, the $3{\alpha}$-isomer was hydroxylized at the $6{\alpha}$-position.

• Journal of Applied Biological Chemistry
• /
• v.48 no.4
• /
• pp.183-186
• /
• 2005

Arabidopsis AHL gene encodes a 3'(2')-phosphoadenosine 5'-phosphate (PAP)-specific phosphatase that plays a role in the sulfate activation pathway. We complemented E. coli cysQ mutant defective in cysteine biosynthesis with the AHL gene. AHL cDNA was cloned into the prokaryotic expression vector pKK388-1 and transformed into the bacterial mutant. Since cysQ mutant is a leaky cysteine auxotroph only under aerobic conditions, the bacteria were grown in liquid media with vigorous shaking to provide more aeration. In cysteine-free medium, cysQ mutant and the mutant harboring empty vector did not grow well, whereas cells harboring AHL cDNA exhibited significantly improved growth with doubling time of approximately 3 h. cysQ is known to encode a 3'(2'),5'-diphosphonucleoside 3'(2')-phosphohydrolase (DPNPase). However, our data suggest that cysQ protein has PAP-specific phosphatase activity in addition to DPNPase activity. Microbial complementation procedure described in this paper is useful for structure-activity studies of PAP-specific phosphatases identified from microbes and plants.

• Archives of Pharmacal Research
• /
• v.13 no.4
• /
• pp.325-329
• /
• 1990

Synaptosomal plasma membrane vesicles (SPMV) were isolated from fresh bovine cerebral cortex by several well-known procedures, and the best procedures was selected by enzymatic and morphological standards. The SPMV isolated by the modified procedure of Smith and Loh showed the highest purity. The specific activities of Na, K-ATPase, acetylcholinesterase and 5'-nucleotidase were about 5, 6-fold, 2, 5-fold and 3, 3-fold, respectively, enriched in the plasma membrane fraction as compared to those of the crude homogenate. Electron microscpic examination also showed that the membranes were in vesicular form.

• Archives of Pharmacal Research
• /
• v.12 no.2
• /
• pp.128-134
• /
• 1989

$Na^+-Ca^{2+}$ exchange process in sarcolemmal vesicles isolated from mesenteric arteries of Wistar-Kyoto normotensive(WKY) and spontaneously hypertensive rats(SHR) was investigated. The sarcolemmal fractions isolated after homogenization and sucrose density gradient centrifugation were enriched with 5'-nucleotidase and ouabain sensitive, $K^+-dependent$ phosphatase activities. When the vesicles were loaded with $Na^+$, a time dependent $Ca^{2+}$ uptake was observed. However, very little $Ca^{2+}$ uptake was observed when the vesicles were loaded with $K^+$, or $Ca^{2+}$ uptake of the $Na^+-loaded$ vesicles was carried out in high sodium medium so that there was no sodium gradient. When the vesicles loaded with $Ca^{2+}$ by $Na^+-Ca^{2+}$ exchange were diluted into potassium medium containing EGTA, $Ca^{2+}$ was rapidly released from the vesicles. $Na^+-dependent\;Ca^{2+}$ uptake was increased in SHR compared to WKY, but passive efflux of preaccumulated $Ca^{2+}$ from the vesicles was decreased in SHR. The data indicate that the membrane vesicles of rat mesenteric arteries exhibit $Na^+-Ca^{2+}$ exchange activity. It is also suggested that changes of this process in vascular smooth muscle cell membrane of SHR may be involved in higher intracellular $Ca^{2+}$ concentration and higher basal tone in SHR.

• Food Science of Animal Resources
• /
• v.18 no.2
• /
• pp.185-191
• /
• 1998

Human milk and bovine milk in normal stage were fractionated four parts : whey, skimmilk membrane, and casein pellet. The specific activity (nmole / mim / mg protein) and distribution ratio(%) of suborganella marker enzymes in each separated milk fraction were determined. Especially, neutral $Ca^{2+}$-ATPase, acid $Ca^{2+}$-ATPase, NADH-cytochrome C reductase, and acid phosphatase were higher in human milk. However, both $Ca^{2+}$-ATPases were not detected in all fractions of bovine milk. On the other hand, 5'-nucleotidase, phosphodiesterase I, alkaline phosphatase, and $\gamma$-glutamyl transpeptidase activities in bovine milk were higher than in human milk. Most of the marker enzymes were highly distributed in cream fraction of either human milk or bovine milk, and their specific activities were high to 24 fold from 3 fold when compared with that of whole milk. These results suggest that marker enzymes in mammary epitherial cell are transfered into cream fraction by the membrane rearrangement, and different biochemical reaction between human and bovine exists for milk secretion in mammary gland.

• Toxicological Research
• /
• v.12 no.2
• /
• pp.251-258
• /
• 1996

Panax ginseng has been used for various diseases including hepatic disorders. The aim of the present study was to investigate the hepatoprotective effects of ethanol extract and saponin of Panax ginseng in thioacetamide-intoxicated rats and to compare with silymarin, a known hepatoprotective agent. Male Sprague-Dawley rats were given single intragastric administration of thioacetamide. Aqueous solutions of ethanol extract and saponin of Panax ginseng with or without silymarin were administered intragastrically daily for six days from four days before until one day after thioacetamide administration. At the end of the treatment, the rats were fasted overnight and sacrificed. As a result, thioacetamide caused significant increase in serum levels of AST, ALT, 5'-nucleotidase and bilirubin. Thioacetamide increased $Ca^++$ content but decreased protein content in liver tissue. These thioacetamide-induced biochemical changes were prevented both by ethanol extract of ginseng and silymarin, but not by ginseng saponin. Silymarin did not potentlate the effect of either ethanol extract or saponin of ginseng on these parameters. Thioacetamide-induced confluent necrosis was not protected by the test drugs. In conclusion, ethanol extract of ginseng protects the liver possibly by stabilizing the cell membrane and by inhibiting thioacetamide-induced $Ca^++$ increase in the hepatocytes, which was comparable to that of silymarin.

• Journal of Chest Surgery
• /
• v.34 no.6
• /
• pp.454-464
• /
• 2001

Background: It has been recognized that systemic inflammatory reaction and oxygen free radical formed by activated leukocyte in the procedure of cardiopulmonary bypass(CPB) frequently produce postoperative cardiac and pulmonary dysfunction. The purpose of this study was to evaluate the efficacy of leukocyte-depleting filters in the cardiopulmonary bypass circuit for patients undergoing open heart surgery(OHS). Material and method: The study involved 15 patients who underwent OHS with a Leukoguard-6 leukocyte filter placed in the arterial limbs of the bypass circuit(filter group, n=15) and 15 patients who did not have the filter(control group, n=15). We analyzed the differences between the groups in intraoperative changes of peripheral blood leukocyte and platelet counts, pre- and postbypass changes of malondialdehyde(MDA), troponin-T(TnT), 5'-nucleotidase(5'-NT) in coronary sinus blood, spontaneous recovery rate of heart beat after CPB, pre-and postoperative cardiac index(Cl) and pulmonary vascular resistance(PVR), and the amounts of postoperative bleeding and sternal wound complication. Result: During CPB, total leukocyte count of the filter group(9,567$\pm$ 842/㎣) was significantly less than that of the control group(13,573+1,167/㎣) (p<0.01), but there was no significant difference in platelet count between the groups. Postoperative levels of MDA(3.78+0.32 $\mu$mol/L vs 5.86+0.65 $\mu$mo1/L, p<0.01), TnT(0.40$\pm$0.04 ng/mL vs 0.59$\pm$0.08 ng/mL, p<0.05) and 5'-NT(3.88$\pm$0.61 U/L vs 5.80$\pm$0.90 U/L, p<0.05) were all significantly lower in the filter group than the control group. Postoperative Cl was higher in the filter group than the control group(3.26$\pm$0.18 L/$m^2$min vs 2.75$\pm$0.17 L/$m^2$/min, p=0.05). PVR of the filter group was lower than that of the control group(65.87$\pm$7.59 dyne/sec/cm$^{5}$ vs 110.80+12.22 dyne/sec/cm$^{5}$ , p<0.01). Spontaneous recovery rate of heart beat in the filter group was higher than that in the control group(12 patients vs 8 patients, p<0.05). Postoperative wound infection occurred in one case in the filter group and 4 case in the control group(p<0.05). Postoperative 24 hour blood loss of the filter group was more than that of the control group (614$\pm$107 mL vs 380+71 mL, p=0.05).