A 990 bp full-length gene (xynAHJ2) encoding a 329-residue polypeptide (XynAHJ2) with a calculated mass of 38.4 kDa was cloned from Bacillus sp. HJ2 harbored in a saline soil. XynAHJ2 showed no signal peptide, distinct amino acid stretches of glycoside hydrolase (GH) family 10 intracellular endoxylanases, and the highest amino acid sequence identity of 65.3% with the identified GH 10 intracellular mesophilic endoxylanase iM-KRICT PX1-Ps from Paenibacillus sp. HPL-001 (ACJ06666). The recombinant enzyme (rXynAHJ2) was expressed in Escherichia coli and displayed the typical characteristics of low-temperature-active enzyme (exhibiting optimum activity at $35^{\circ}C$, 62% at $20^{\circ}C$, and 38% at $10^{\circ}C$; thermolability at ${\geq}45^{\circ}C$). Compared with the reported GH 10 low-temperature-active endoxylanases, which are all extracellular, rXynAHJ2 showed low amino acid sequence identities (<45%), low homology (different phylogenetic cluster), and difference of structure (decreased amount of total accessible surface area and exposed nonpolar accessible surface area). Compared with the reported GH 10 intracellular endoxylanases, which are all mesophilic and thermophilic, rXynAHJ2 has decreased numbers of arginine residues and salt bridges, and showed resistance to $Ni^{2+}$, $Ca^{2+}$, or EDTA at 10 mM final concentration. The above mechanism of structural adaptation for low-temperature activity of intracellular endoxylanase rXynAHJ2 is different from that of GH 10 extracellular low-temperature-active endoxylanases. This is the first report of the molecular and biochemical characterizations of a novel intracellular low-temperature-active xylanase.
Background & Objective : Naeso-san(NSS) has been used for the treatment of functional dyspepsia, regarded as a gastric dysmotility disease. A main cause of gastric dysmotility is antral dilatation or antroduodenal uncoordination. Therefore, we investigated the effect of NSS on gastric motility and its mechanism of action, as well as the morphologic changes in antral dilatated rats. Methods : Antral dilatated rats were induced by wrapping a nonabsorbable rubber ring(D:6mm, W:4mm, T:1mm) around the 1st portion of the duodenum for 8 weeks. Then morphologic changes were investigated and compared with normal intact rats before and after 8 weeks. Gastric emptying was measured by administration of normal saline(NS) or NSS in normal intact and antral dilatated rats. In another series of experiments to evaluate the mechanism of NSS under delayed conditions, normal intact rats were treated with atropine sulfate(1mg/kg, s.c.), quinpirole HCl(0.3mg/kg, i.p.), $NAME(N^{G}-nitro-L-arginine$ methyl ester, 75mg/kg, s.c.) and cisplatin(10mg/kg, i.p.), respectively. The myoelectrical activity of the gastric smooth muscle was recorded in normal intact and antral dilatated rats. The contractile waves were measured for 30 minutes before and after administration of each solution(NS, NSS). Results : Body weight gain of antral dilatated rats was significantly lower than that of the controls. Futhermore, we found the thickness of the mucosal and muscular layers and surface area of the stomach increased significantly compared with controls. NSS 278㎎/㎏ improved gastric emptying more than normal saline or NSS 93mg/kg in normal intact(p=0.026) and antral dilatated rats(p=0.03). NSS enhanced gastric emptying significantly in the NAME treated group(p=0.002). NSS 278mg/kg increased the significant postprandial dominant power than that of NS in normal intact rats, whereas there was no statistical significance in antral dilatated rats. Conclusions : NSS stimulates gastric motility through the cholinergic pathway. We expect that pathologic model with antral dilatation can be used as an exprimental tool which is similar to dyspepsia and NSS would be effective especially in dysmotility-like functional dyspepsia with antral dilatation or impaired reservoir functions such as gastric adaptive relaxation.
The purpose of this study was to investigate the anti-obesity effects of crude saponin (CS) in Korean red ginseng (KRG) on rats fed a high fat (HF) diet. Methods: Male Sprague-Dawley (SD) rats were divided into a normal diet group and a high-fat (HF) diet group. The HF diet group became obese from being fed a HF diet over five weeks. The normal diet group were fed a normal diet. Each CS group of the normal diet group and HF diet group was administered CS (200 mg/kg, i.p.) for three weeks, the control group of both types was administered normal saline (1ml/kg, i.p.) instead of CS. Each group had 5 rats. Results: After administration of CS, the body weight, food consumption, adipose tissues, and expression of appetite peptides such as leptin and neuropeptide-Y (NPY) were investigated in the HF diet group as well as the normal diet group. Administration of CS reduced body weight, food intake, and fat content in the HF and normal diet groups. After CS administration, NPY expression and leptin were lower in the HF diet group. Conclusions: Our results suggest that CS may be useful in the treatment of obesity, especially of type qixu (氣虛).
Air pressure decay(APD) rate and ultrafiltration rate(UFR) tests were performed on new and saline rinsed dialyzers as well as those roused in patients several times. C-DAK 4000 (Cordis Dow) and CF IS-11 (Baxter Travenol) reused dialyzers obtained from the dialysis clinic were used in the present study. The new dialyzers exhibited a relatively flat APD, whereas saline rinsed and reused dialyzers showed considerable amount of decay. C-DAH dialyzers had a larger APD(11.70$\pm$1.32mmHg/min)compared to CF dialyzers(4.32$\pm$0.55mmHg/min)(p<0.05). However, there was no observable difference in the UFR between the two dialyzers. Neither APD nor UFR showed any significant increase with an increasing number of reuses for up to more than 20reuses. A substantial number of failures observed in APD(larger than 20mmHe/min)on the reused dialyzers(2 out of 40 CP and S out 26 C-DAK) were attributed to the Possible damage on the fibers. The CF 15-11 HFDs which failed APD test did not show changes in the UFR compared to normal dialyzers indicating that APD is a more sensitive test than UFR test to evaluate the integrity of the fibers.
Three hundred gram of Korean ginseng root was extracted with 95% ethanol on a boiling water bath for about 300 hr. Evaporation of alcohol yieled 50.2g of dark brown residue which was used by dissolving 4 mg of the residue in 1 ml of physiological saline. The ginseng group and the saline group received each day 0.5 ml per 100 g body weight of ginseng extract and physiological saline, respectively. Both the ginseng and saline group with stress were exposed to positive radial acceleration (1∼29g), cold (5$^{\circ}C$, 0$^{\circ}C$ &-10$^{\circ}C$) and heat (35$^{\circ}C$) environment, and surgical stress. After termination of the last stress, the tolerance, body weight, visceral organ weight, basal metabolism rate, rectal temperature, the number of erythrocyte and leucocyte, hemoglobin level, hematocrit ratio, total serum protein content and it's fraction and the content of adrenal ascorbic acid in the experimental animal exposed to stress were measured and at the corresponding periods, the same measurements were also carried out with the ginseng and the saline groups without stress exposure (serving as control). Results obtained were as follows. 1. Administration of ginseng does depressed the decrease of the tolerance, body weight, visceral organ weight, basal metabolism rate, the number of erythrocyte, hemoglobin value, hematocrit ratio and the A/G ratio in the mice and rats exposed to various stress. 2. The change of the rectal temperature, eosinophile counts, total serum protein content and the content of adrenal ascorbic acid of ginseng group that exposured to various stress facilitates the reaction to, and accelerates the recovery from the stress. 3. Even after hypophysectomy which served the link between the central and the peripheral portion of the stress mechanism, the adrenal ascorbic acid content of ginseng group decreased significantly more than that of the saline group 30 min. after administration of ACTH, while the value approached the normal level significantly closer in the ginseng group than in the saline group 1 and 2 hr after ACTH administration. Judging from the above results, it is concluded that administration of ginseng extract tolerated the experimental animals under the environment of stressfu1 stmuli, although the ginseng has no significant influence upon the stress mechanism in the absence of stressful stimuli. The site of action of the ginseng appears to be in the peripheral portion of the stress mechanism.
This is the study of the effects of Aqua-acupuncture with Gardeniae Fructus on thc recovery of rat's liver which was damaged by 0.3ml/ea of $CCI_4$. Rats were divided into 4 groups; Normal-group(None treated group), Control-group(Not treated after $CCI_4$-intoxicated), Exp. I(Treated with Saline Aqua-acupuncture after $CCI_4$-intoxicated) and Exp. ll(Treated with Gardeniae Fructus Aqua-acupuncture after $CCI_4$-intoxicated). Biochemical assays for each serum enzyme activities of AST, ALT, Albumin, LDH, ${\gamma}$-GT, TG and Total cholesterol were performed. The results were summarized as follows: 1. AST activities in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In companson with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua-acupuncture treated group *The professor of Dept. of Acupuncture & Moxibustion, 2. At T activities in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In com pan son with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated worked effectively to rat's damaged liver. 3. Albumin in serum increased in the Gardeniae Fructus Aqua- acupurkture treated group after $CCI_4$-intoxicated. 4. LDH in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In comparison with Saline-treated group after CClcintox icated, the Gardeniae Fructus Aqua acupuncture treated group after $CCI_4$-intoxicated worked highly effectively to rat's damaged liver. 5. ${\gamma}$-GT In serum significantly decreased In the Gardeniae Fructus Aqua-acupuncture trea ted group after $CCI_4$-intoxicated. In compan son with Saline-treated group after $CCI_4$-intoxicated, the Crardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated was not recognized significantly. 6. TG in serum significantly decreased in the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In comparison with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua -acupuncture treated group after $CCI_4$-intoxicated worked highly effectively to rat's damaged liver. 7. Total cholesterol in serum decreased in the Gardenias Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated. In comparison with Saline-treated group after $CCI_4$-intoxicated, the Gardeniae Fructus Aqua-acupuncture treated group after $CCI_4$-intoxicated worked highly effectively to rat's damaged liver. The results from ahove show that the Gardeniae Fructus Aqua- acupuncture has highly effects on the damaged liver caused by $CCI_4$. Therefore it is expected that the Gardeniae Fructus Aqua- acupuncture could be used to cure the damaged liver.
A study was conducted to monitor the changes in serum protein profile, cholesterol and blood glucose during endotoxic shock in buffalo calves and also to assess the role of prophylactic supplementation of vitamin E and selenium in alleviating the endotoxic effects. Fifteen male buffalo calves (6-8 months of age) were divided into three groups: Group I (control)-infused with 0.9% saline solution; Group II-infused with E. coli endotoxin at 5${\mu}g/kg$ body weight in normal saline solution; Group III- supplemented prophylactically with 250 mg vitamin E and 7.5 mg selenium by i/m injections at weekly intervals for one month prior to the induction of endotoxic shock. The blood samples were collected at 0, 1, 3, 6, 9, 12, 24, 48 and 72 h after the induction of shock. Endotoxin caused a significant (p<0.05) hypoproteinemia from 3-12 h post infusion in group II but this hypoproteinemia was less pronounced and only from 3-9 h post infusion in vitamin E and selenium supplemented calves. Hypoglycemia was observed in group II from 3-24 h and blood glucose level returned to normal at 72 h. However hypoglycemia was mild in group III and blood glucose returned to normal at 48h. Hypocholesterolaemia and hypoalbuminemia were found in both groups II and III but these changes were less pronounced in group III i.e. vitamin E and Se supplemented calves. Serum electrophoretic protein patterns of group III were quite similar to those of control group but animals of group II had different electrophoretic pattern. It was concluded that the antioxidant effects of vitamin E and Se prevent the liver against oxidative stress during endotoxic shock.
Objectives : The objective of this study was to evaluate the effect of Dokhwaljihwang-tang (DJT) intravenous pharmacopuncture on cisplatin-induced emesis and gastric mobility disorder in Wistar rats. Methods : Thirty rats were randomly divided into six groups and cisplatin was administered to all groups except the normal group. The cisplatin group (n=5) received a cisplatin injection only. The saline group (n=5) was injected with cisplatin followed by 0.4 mL of saline. Groups DJT-1, DJT-2, and DJT-3 were injected with cisplatin, followed by 0.315 g/kg, 0.104 g/kg, and 0.034 g/kg of DJT, respectively. Body weight, food intake, and kaolin intake of rats were measured 12 h, 24 h, and 36 h after cisplatin injection. Residual food in the stomach was measured 48 h after cisplatin injection. Results : There was no significant difference in weight. The food intake was not significantly different 12 h after cisplatin administration. All groups except the normal group showed significantly decreased food intake after 24 h. After 36 h, food intake was not significantly different between groups DJT-1, DJT-2, and DJT-3 and the normal group. The kaolin intake of groups DJT-1 and DJT-2 was significantly decreased at 12 h and 24 h after cisplatin injection. Kaolin intake and residual food in the stomach were significantly decreased in groups DJT-1, DJT-2, and DJT-3. Conclusion : In a Wistar rat model, DJT intravenous pharmacopuncture is suggested to be effective for cisplatin-induced emesis and gastric motility disorder. In the future, it is necessary to study the mechanism and chemical composition of each individual constitutive drug.
Kang, Shin Hyuk;Shin, Kee Cheol;Kim, Woo Seob;Bae, Tae Hui;Kim, Han Koo;Kim, Mi Kyung
Archives of Plastic Surgery
/
v.42
no.2
/
pp.179-185
/
2015
Background Capsular contracture is the most troublesome complication in breast implant surgery. Although capsule formation can be seen as a normal reaction to a foreign body, it can induce pain, hardness, deformity, and other pathologic problems. Surgical intervention is required in severe cases, but even surgery cannot guarantee a successful outcome without recurrence. This experimental study confirms that single topical administration of leukotriene antagonist zafirlukast (Accolate, Astrazeneca) reduces peri-implant capsule formation and prevents capsular contracture. Methods Twelve smooth-surfaced cohesive gel implants were implanted in New Zealand White rabbits. These miniature implants were designed to be identical to currently used products for breast augmentation. The rabbits were divided into 2 groups. In the experimental group (n=6), the implant and normal saline with zafirlukast were inserted in the submuscular pocket. In the control group (n=6), the implant and normal saline alone were used. Two months later, the implants with peri-implant capsule were excised. We evaluated capsule thickness and collagen pattern and performed immunohistochemical staining of myofibroblasts, transforming growth factor $(TGF)-{\beta}1$, 2. Results The thickness of the capsules in the experimental group was reduced in both dorsal and ventral directions. The collagen pattern showed parallel alignment with low density, and the number of myofibroblasts as well as the amounts of $TGF-{\beta}1$ and $TGF-{\beta}2$ were reduced in the experimental group. Conclusions We suggest that single topical administration of leukotriene antagonist zafirlukast can be helpful in reducing capsule formation and preventing capsular contracture via myofibroblast suppression, modulation of fibroblastic cytokines, and anti-inflammatory effect.
Kim, Min-Kyung;Choi, Yoon;Kong, Hyun-Seok;Leem, Joong-Woo;Leem, Hang-Soo;Chung, Soo-Jin;Lee, Cheong
The Korean Journal of Pain
/
v.12
no.2
/
pp.171-176
/
1999
Background: Effect of nitric oxide on the hyperalgesia induced by inflammation is controversial. From our previous experiment, NOS inhibitor, L-NAME given during the induction period decrease mechanical hyperalgesia occured by Freund's complete adjuvant induced inflammation. In addition, we attempted to analyze the effects of L-NAME on mechanical hyperalgesia after the development of inflammation by Freund's complete adjuvant in rat paw. Methods: Male Sprague Dawley rats were divided into four groups; control (normal saline), and three different doses of L-NAME (0.1 mg, 1 mg, 10 mg). Inflammation was induced in rats by injecting 0.15 ml of Freund's complete adjuvant (FCA) intraplantarly. Rats showed typical hyperalgesia within twelve hours after injection and maintained this for about one week. Tests were done 2 days after injection of FCA. After the baseline test either L-NAME or saline was injected under light halothane anesthesia. Effect of L-NAME on hyperalgesia was assessed by measuring mechanical hyperalgesia at 15, 30, 60, 90, 120 minutes. Same experients were repeated on normal rats. Results: When injected at the site of inflammation, L-NAME caused dose dependent decrease in mechanical hyperalgesia. However, normal rats also showed increased mechanical threshold after L- NAME injection. Conclusions: Although L-NAME reduces FCA induced mechanical hyperalgesia, this result may solely be due to inhibition of nitric oxide production and need to be further determined.
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