During the rapid phase of gonadal development of the freshwater teleost, the catfish (Silurus asotus), the influence of hCG upon the inducement of final oocyte maturation and spawning was investigated electrophoretically and ultrastructurally. The electrophoretic patterns obtained were different in the presence and absence of some of the major or minor zones, because of the hormone level in catfish. The vitellogenin of hormone-treated fish was stained more intensively than that of sham-treated fish. These proteins showed some minor or main bands of egg extracts which migrated at positions corresponding to molecular weights of approximately 90,000. However, the thickness of electrophoretic band in molecular weight for hCG-treated fish was slightly lower than that for saline control. It seemed the plasma protein with molecular weight of approximately 45,000 in hCG-treated fish disappeared. In contrast to the control fish, the ovaries in the catfish treated with hCG shows a marked ultrastructural change under the electron microscope. No dilated profiles were seen in the granulosa cells of the mature oocyte before ovulation. After germinal vesicle breakdown (GVBD), the zona radiata interna (ZRI) becomes more compact, and there is a loss of all the processes from the pore canals. There is a wide space between the vitelline membrane and zona radiata. Also, during final maturation, the microvillar processes from the oocyte are seen no longer to penetrate deeply into the extracellular spaces of the overlying granulosa cells, and the reticulate patterns of the zona radiata interna becomes occluded, giving the zona radiata a more solid appearance. It has been possible to initiate 100% oocyte maturation in yolk granules and follicles in vivo by treatment with hCG and a high water temperature ($27^{\circ}C$). In hCG-treated fish, the percentages of successful artificial fertilization and hatching were maximal at 15 h after a single injection. It seems clear that a long acting preparation containing hCG can be successfully used in prespawning fish to advance the final events of gonadal maturation and initiate spawning. Further studies are necessary to evaluate the potential of hCG to either stimulate or inhibit the reproductive development of fish at other stages of the seasonal reproductive cycle.
Purpose : This research was conducted to investigate the effect of sweet bee venom pharmacopuncture and low level laser acupuncture on paw edema, pain index, anti-inflammatory factor, AST, ALT and complete blood cell count of a rat model with Complete Freund's Adjuvant-induced arthritis. Methods : Five experimental groups were formed with each consisting of six rats: normal group, control group, sweet bee venom pharmacopuncture group, lower level laser acupuncture group, and sweet bee venom pharmacopuncture, lower level laser acupuncture group. The experimental model of arthritis was induced by two injections of Freund's adjuvant into the left knee joint of Sprague Dawley(SD) rats. The second injection of Freund's adjuvant was given ten days after the first one. Ten days later, sweet bee venom pharmacopuncture and low level laser acupuncture were administered separately or together by assigned groups at $GB_{34}$ and $GB_{39}$ of rats twice a week for a total of six times. Thereafter, edema rate, pain index, tumor necrosis factor-${\alpha}$, interleukin-6, aspartate aminortansferase, alanine aminotransferase and complete blood cell count were measured. Results : We noticed synergic effects of sweet bee venom pharmacopuncture and low level laser acupuncture according to the results of the paw edema and Von Frey pain index. The sweet bee venom pharmacopuncture(BVA) and sweet bee venom pharmacopuncture+ low level laser acupuncture(BVA+LLA) groups experienced a more significant effect when compared with the control group. Conclusions : These results suggest that Sweet Bee Venom Pharmacopuncture and low level laser acupuncture at GB34 and GB39 have a significant anti-inflammatory effect on Freund's adjuvant arthritis in rats.
In order to prevent the pine mushrooms, Tricholoma matsutake, from being damaged by the pine needle gall midges, Thecodiplosis japonensis, and thereby, to increase their production and improve their quality, a sprinkler system was installed on the mushroom field. A low-concentration insecticide (deltamethrin 1% EC, x2,000) was sprayed once at insects' most active time every day during the period of insects' adult occurrence and thereafter, the irrigation by ground water spraying was periodically enforced. Such a test was conducted at Yangyang-Gun, Kwangwon-do, Korea for 2 years from 2000 through 2001. The pine needle gall midges generally emerged for about 40 days from late May to early July. 50% emergence of them was about June 6, and peak emergence (more than 80%) was early or mid-June. Gall formation rate was 3.5% on average with this ground insecticide spraying, while 51.3% when not treated. Control effectiveness of this insecticide spraying was 92.3%, which was higher than 82.5% by the conventional injection of insecticide into tree stems. Pine mushrooms emerged for about 35 days from mid-September through earlier October, and around 80% of them did for about 15 days from late September through early October. As a result of the periodic ground water-spraying (30 mm per week) for 2 months (from August to October), the production of mushrooms increased by 74.3% (110% in terms of weight), with their quality improvement. The mushrooms produced from the treated stand by the spraying system were priced 8,670,000 wons per hectare, and thus, the net income deducting the facility and management cost was 4,310,000 wons, about 5% higher than value from the control stand. It was analyzed that this treatment was significantly cost effective when the facilities are used more than 5 years.
A rapid, selective and sensitive reversed-phase HPLC method for the determination of promethazine in human serum was developed, validated, and applied to the pharmacokinetic study of promethazine. Promethazine and internal standard, chlorpromazine, were extracted from human serum by liquid-liquid extraction with n-hexane containing 0.8% isopropanol and analyzed on a Capcell Pak CN column with the mobile phase of acetonitrile-0.2 M potassium dihydrogen phosphate (42:58, v/v, adjusted to pH 6.0 with 1 M NaOH). Detection wavelength of 251 nm and flow rate of 0.9 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed promethazine concentration (10 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 1-40 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was 1 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 96.15 to 105.40% for promethazine with overall precision (% C.V.) being 6.70-11.22%. The relative mean recovery of promethazine for human serum was 63.54%. Stability (freeze-thaw and short-term) studies showed that promethazine was stable during storage, or during the assay procedure in human serum. However, the storage at $-80^{\circ}C$ for 4 weeks showed that promethazine was not stable. Extracted serum sample and stock solution were not allowed to stand at ambient temperature for 12 hr prior to injection. The peak area and retention time of promethazine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of promethazine in human serum samples for the pharmacokinetic studies of orally administered Himazin tablet (25 mg as promethazine hydrochloride) at three different laboratories, demonstrating the suitability of the method.
Osteoarthritis is a disease that affects the articular cartilage and osseous tissue, and can be worsened by aging, overweight status, and post-traumatic arthritis. The present study aimed to evaluate the effect of ID-CBT5101 (tyndallized Clostridium butyricum) on bone metabolism and the inflammatory response in a monosodium iodoacetate-induced rat model of osteoarthritis. ID-CBT5101 was administered orally at doses of $10^8$ or $10^{10}CFU/day$ for 2 weeks before direct injection of monosodium iodoacetate ($3mg/50{\mu}l$ of 0.9% saline) into the intra-articular space of the rats' right knees. The rats subsequently received the same doses of oral ID-CBT5101 for another 4 weeks. We evaluated the treatment effects based on serum biomarkers, mRNA expression, morphological and histopathological analyses of the knee joints, and weight-bearing distribution analysis. Compared with those in control rats, the ID-CBT5101 treatments significantly reduced the serum concentration of inflammation and bone metabolism markers (i.e., COX-2, IL-6, $LTB_4$, and COMP), and significantly increased the concentration of $IFN-{\gamma}$ and glycosaminoglycans. In addition, the ID-CBT5101 treatments inhibited the mRNA expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases (i.e., MMP-2, MMP-3, MMP-9, MMP-13, TIMP-1, and TIMP-2). Furthermore, the ID-CBT5101 treatments effectively preserved the knee cartilage and synovial membrane, and significantly decreased the amount of fibrous tissue. Moreover, compared with that of the negative control group, the ID-CBT5101 treatments increased the weight-bearing distribution by ${\geq}20%$. The results indicate that ID-CBT5101 prevented and alleviated osteoarthritis symptoms. Thus, ID-CBT5101 may be a novel therapeutic option for the management of osteoarthritis.
Kim, Soon-Sun;Rhee, Gyu-Seek;Lee, Rhee-Da;Kwack, Seung-Jun;Lim, Kwon-Jo;Yhun, Hyo-Jung;Park, Kui-Lea
Proceedings of the Korea Society of Environmental Toocicology Conference
/
2003.10a
/
pp.57-69
/
2003
It is well known that many pesticides possess hormonal activity, and affect the developments of wildlife and mammals including human. Currently, pyrethroid insecticides are in worldwide use to control in and outdoor pests, providing potential far environmental exposure. Hormonal activities of these pyrethroid insecticides, however, have been little studied, and the developmental effects of them were no reported. Therefore, we firstly examined the potential estrogenic activities of some pyrethroid insecticides (permethrin, cypermethrin, tetramethrin, deltamethrin, sumithrin, fenvalerate and bioallethrin) by immature rat uterotrophic assay, luciferase reporter gene assay and Calbindin-D$\sub$9k/ (CaBP-9k) gene expression assay. Uterine wet weights were increased by permethrin and the permethrin-induced weights were inhibited by ICI 182780 in the uterolrophic assay. On the other hand tetramethrin significantly reduced uterine and vaginal wet weights, and also inhibited the E2-induced weight increases at all doses tested. Cypermethrin and sumithrin had a tendency to increase uterine weights, although not statistically significant. Permethrin and cypermethrin dose-dependently increased the luciferase activity in reporter gene assay. Northern blot analysis showed that permethrin induced CaBP-9k mRNA expression whereas tetramethrin inhibted. Subsequent studies were conducted to investigate the possible developmental effects of four pyrethroid insecricides (permethrin, cypermethrin, sumithrin and teramethrin). Either diethlbestrol (DES) or 17${\beta}$ -estradiol (E2) was used as a reference control in this study. Pyrethroid insecticides were administered to Sprague Dawley rats via subcutaneous injection at 6 to 18 days of gestation or 1 to 5 days after birth. In utero treatment of permethrin (10mg/kg/day) in female rat resulted in significant increases in uterine and ovarian weights while significant decreases in serum E2 concentration, uterine and ovarian ER${\alpha}$ mRNA levels. Sumithrin and permethrin led to acceleration in vaginal opening of female rat, while delay in preputial separation of male after neonatal treatment. Anogenital distances of PND 18 were significantly reduced in sumthrin-treated, and permerhrin-treated male rats after neonatal treatment. All the pyrethroid insecticides tested caused significant increases in uterine weights on PND 18, while significant reductions in the first diestrus phase when neonataly treated. In addition, exposure to pyrethroids in neonatal period led to significant reduction in relative brain weight in female rat on PND 18, but its weight was recovered in diestrus phase. In summary, Our experimental data demonstrate the possibilities of developmental effects of pyrethroid insecticides via estrogenic or antiestrogenic activity.
The aim of this study was to test if 3,5,3'-triiodothyronine (T3) are involved in the osmoregulatory actions in euryhaline starry flounder Platichthys stellatus. We investigated osmoregulatory parameters ($Na^+,\;Cl^-$ and osmolality), blood cortisol and glucose in starry flounder acclimated to seawater (SW, 33 psu) and that were transferred and allowed to acclimate to freshwater (FW, 0 psu). Fish in SW were injected with T3 (5, 10, and $15{\mu}g$/g body weight) or vehicle (0.9% NaCl), and then transferred to FW. They were sampled 3 days after the transfer. With T3 at $10{\mu}g$/g, levels of plasma $Na^+$ and $Cl^-$ were significantly higher than in sham (only saline) and control fish (without hormone and saline). Osmolality was significantly higher after injection with T3 at 10 and $15{\mu}g$/g than in the control. However, T3 at $5{\mu}g$/g had no effect on hyper-osmoregulation. In this study, all dose of T3 induced a significant increases in plasma cortisol without glucose. These results suggest a positive hyper-osmoregulatory role of T3 in starry flounder to hypoosmotic environment, maybe a positive interaction of T3 with cortisol for maintenance of hyper-osmoregulatory ability.
Lee Jae yeong;Kim Joong-hyun;Lee Won guk;Kang Seong soo;Bae Chun sik;Choi Seok hwa
Journal of Veterinary Clinics
/
v.22
no.1
/
pp.21-25
/
2005
This study was performed to assess therapeutic effect of the tibial plateau leveling osteotomy (TPLO) in dogs with experimentally transected cranial cruciate ligaments (CrCL). Nine healthy adult Beagle dogs were transected left CrCL under general anesthesia. The dogs were assigned to TPLO and non-TPLO control groups. The TPLO procedures for correcting the CrCL rupture in the left stifle of dogs were performed under sterile conditions. Before TPLO procedures, all dogs were screened by orthopedic and radiographic examinations. Dogs were lameness free for the previous three months, and when examined at the walk and trot on a hard surface, in a straight line and on a circle. Lateral and craniocaudal radiographs were done to confirm the soundness of the both knee joint in dogs and not detectable lesions were diagnosed. The dogs were intravenously injected with a 10 mci/kg of 99mTechnetium-methylene diphosphonate (99mTc-MDP) under general anesthesia. Scintigraphs were obtained using a large field of view gamma camera equipped a parallel-hole, low-energy about 3 hours after intravenous injection of 99mTc-MDP. Before CrCL transection and 4, 8, and 12 weeks after the procedures, scintigraphy were conducted. Bone uptake of the left stifle joint increased after the procedures in all dogs. When the bone uptake from the TPLO procedure was compared with that of the control, there was a significant difference (p < 0.05). At 12 weeks after the TPLO procedure, the dogs showed normal anatomical posture and gait. It is concluded that TPLO procedure was effective in reconstruct of the stifle joint in dogs with CrCL rupture.
Ga-67 citrate scan has been requested for detection or follow-up of inflammatory or neoplastic disease. Visualization of Ga-67 citrate in the kidneys at 48 and 72 hr post injection is usually interpreted as evidence of renal pathology. But precise mechanisms of abnormal Ga-67 uptake in kidneys were unknown. We undertook a study to determine the clinical value of Ga-67 citrate imaging of the kidneys in 68 patients with primary or secondary nephropathy confirmed by renal biopsy and 66 control patients without renal disease. Renal uptake in 48 to 72 hr images was graded as follows: Grade 0=back-ground activity:1=faint uptake greater than background;2=definite uptake, but less than lumbar vertebrae; 3=same uptake as lumbar vertebrae, but less than liver; 4=same or higher uptake than liver. The results were as follows. 1) 42 of 65 (62%) patients with noninfectious nephritis showed grade 2 or higher Ga-67 renal uptake but only 10 percent of control patients showed similar uptake. 2) In 14 patients with systemic lupus erythematosus, 8 of 9 (89%) patients with lupus nephritis exhibited marked renal uptake 3) 36 of 41 patients (88%) with combined nephrotic syndrome showed Grade 2 or higher renal uptake. 4) Renal Ga-67 uptake was correlated with clinical severity of nephrotic syndrome determined by serum albumin level, 24 hr urine protein excretion and serum lipid levels. 5) After complete remission of nephrotic syndrome, renal uptake in all 8 patients who were initially Grade 3 or 4, decreased to Grade 1 or 0. In conclusion, we think that the mechanism of renal Ga-67 uptake in nephrotic syndrome might be related to the pathogenesis of nephrotic syndrome. In systemic lupus erythematosus, Ga-67 citrate scan is useful in predicting renal involvement.
For the treatments and protection of bacterial fish disease, many requirements are needed for aquatic probiotics so that they are effective in aquaculture animals but are also harmless to humans. In the present study, among 17 candidate probiotic bacteria (CPB) obtained from the edible part of the shellfish, Bacillus sp. CPB-St (CPB-St) were selected and in vitro evaluated for the possibility as a probiotic strain for the control of fish streptococcosis which frequently occurs in the olive flounder farms. CPB-St showed inhibitory effects on the growth of various fish pathogenic bacteria, Streptococcus sp., S. parauberis, S. iniae, Lactococcus garvieae and L. piscium by the double layer test ranging about 18~26 mm of clear zone. Inhibitory activity of CPB-St to Streptococcus sp. was observed 6 hours after and the growth of Streptococcus sp. was decreased to 8~55 folds in the co-culture of CPB-St with Streptococcus sp.. The safety of CPB-St to fish and survival of CPB-St in the intestine were assessed in the olive flounder, Paralichthys olivaceus. Fish mortality was not observed in artificial infection with CPB-St for 2 weeks. CPB-St was entirely excreted from the stomach and intestine 24 hours after oral injection. This results indicate that CPB-St has potential applications as a probiotic for the control of fish streptococcosis in aquaculture.
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