Nitric oxide (NO) is an important regulator of renal blood flow, glomerular hemodynamics, and tubule transport processes in the kidney. There is also evidence that NO is involved in cell cycle regulation and mitotic division. During development the nNOS expression pattern differs from that observed in adult animals. However, little is known about temporal and spatial patterns of nNOS expression in the developing kidney. The purpose of this study was to establish the time of expression and the distribution of nNOS in the developing rat kidney. Kidneys from 14-, 16-, 17-, 18-, and 20-day-old fetuses, 1-, 4-, 7-, 14-, and 21-day-old pups, and adult animals were preserved and processed for immunohistochemistry. In the adult kidney, nNOS was detected in the parietal epithelium of Bowman s capsule, macula densa, descending thin limb and inner medullary collecting duct. nNOS immunoreactivity appeared first in the distal tubule anlage at 15 days of gestation, and in all epithelial cells of developing thick ascending limbs (TAL) as well as macula densa of 17- and 18-day-old fetuses. From 20 days of gestation to 14 days after birth, nNOS was expressed in the newly formed cortical TAL, which are located in the medullary ray, whereas in mature TAL of juxtamedullary nephrons, nNOS immunolabeling gradually decreased in intensity and became restricted to the macula densa. In inner medullary collecting ducts, nNOS immunoreactivity appeared first at 7 days after birth in the papillary tip and gradually ascended to the border between outer and inner medulla. In the descending thin limb and parietal epithelium of Bowman's capsule, weak nNOS immunoreactivity was observed at 14 days after birth and labeling gradually increased to adult levels at 21 days after birth. These results suggest that differential expression of nNOS in the developing kidney is an important physiological regulator of renal function during kidney maturation.
Kim, Hyun-Jung;Kim, Chung-Hyon;Lee, Joong-Yeup;Kwon, Jae-Hee;Hwang, Do-Yeong;Kim, Ki-Chul
Clinical and Experimental Reproductive Medicine
/
v.37
no.1
/
pp.57-64
/
2010
Objectives: Likewise fresh cycle, it is also important to select right blastocysts for transfer in purpose of improving the pregnancy and implantation rates in frozen-thawed embryo transfer (ET) cycles. To investigate the relationship between the developmental velocity at the time of cryopreservation and pregnancy rates, we compared pregnancy rates between the day 5 cryopreservation group and the day 6 cryopreservation group. Methods: Transfers of frozen-thawed blastocysts which had been cryopreserved by vitrification on day 5 or day 6 were performed between January 2006 and June 2007. Ethylene glycol, DMSO, and pull and cut straws were used for vitrification and artificial shrinkage was done in expanded blastocysts. Thawing was performed on the day before transfer and thawed blastocysts were cultured in for 15~18 hrs in Quinn's blastocyct media. Blastocyst survival was assessed before transfer and post-thaw survival was defined as >50% of cells remaining intact and blastocoele re-expansion by the time of transfer. Results: Transfers of thawed blastocyst had been cryopreserved on day 5 were 52 cycles and 41 transfer cycles were cryopreserved on day 6. Patient characteristics, the number of transferred embryos and the survival rate of thawed blastocysts were not different in each cryopreservation day. But the biochemical pregnancy, clinical pregnancy, ongoing pregnancy, and implantation rate were significantly high in transfer of frozen-thawed blastocyst which were cryopreserved on day 5. Conclusions: The clinical pregnancy and implantation rate of day-5 blastocyst showed significantly higher than those of day-6 blastocyst in frozen-ET cycles. This result indicated that developmental rate of blastocyst at cryopreservation time in frozen-thawed cycle is discriminative marker of pregnancy outcome as like in fresh cycle.
Song Dong-Keun;Cho Hyun-Mi;Lee Tae-Hee;Suh Hong-Won;Kim Yung-Hi
The Korean Journal of Pharmacology
/
v.31
no.1
s.57
/
pp.11-15
/
1995
Serotonergic neurons in medulla oblongata play an important role in the endogenous descending pain inhibitory system. To illucidate the factors involved in the regulation of medullary serotonergic neurons, we studied the effects of cholecystokinin (CCK) and agents acting on various second messenger systems on 5-hydroxytryptamine (5-HT) release from cultured neurons of rat fetal (gestational age 14th day) medulla oblongata. Cultured cells maintained for 10 days in vitro were stimulated for 48 hours with CCK or other neuropeptides at 10 micromolar concentration. CCK ($10{\mu}M$) and substance P ($10{\mu}M$) significantly increased. 5-HT release. However, somatostatin, proctolin, thyrotropin releasing hormone, and interleukin-6 did not have any effects on 5-HT release. Nimodipine ($1{\mu}M$), a calcium channel blocker, almost completely, and calmidazolium ($1{\mu}M$), a calmodulin antagonist, significantly inhibited the CCK-induced 5-HT release. The total 5-HT content (intracellular 5-HT plus released 5-HT) was significantly increased by CCK. However, the intracellular 5-HT content was not significantly changed by CCK. Forskolin ($5{\mu}M$), an adenylate cyclase activiator, but not $2{\mu}M$ phorbol myristate acetate (PMA), a protein kinase C activator, significantly enhanced 5-HT release. The total 5-HT content (intracellular 5-HT plus released 5-HT) was significantly increased by forskolin. However, the intracellular 5-HT content was not significantly changed by forskolin. PMA had no effect on intracellular 5-HT levels. These results suggest that CCK regulates serotonergic neurons in the medulla oblongata by enhancing 5-HT secretion through calcium influx and caimodulin, and that cyclic AMP system but not protein kinase C system is involved in 5-HT release.
Journal of the Korean Society of Food Science and Nutrition
/
v.33
no.3
/
pp.576-581
/
2004
Foxtail Millet Yakju were treated with heat ($65^{\circ}C$/15 min) or high hydrostatic pressure ($25^{\circ}C$ (RT) and $65^{\circ}C$ (HT)/400 MPa/10 min), and stored for 64 days at l$0^{\circ}C$, $25^{\circ}C$ and 37$^{\circ}C$. Changes in microbial counts, enzyme activity and quality of Yakju during storage were measured. Total viable cells were about 10$^2$ CFU/mL, and remained almost constant during storage at l$0^{\circ}C$ and $25^{\circ}C$, while decreased significantly at 37$^{\circ}C$, and undetected after 55 days of storage in heat- and pressure(RT)-treated, but after 25 days in pressure(HT)-treated Yakju. Lactic acid bacteria and yeast in heat- and pressure-treated Yakju were not detected during storage. The relative activities of a -amylase in heat- and pressure(RT)-treated were more than 100%, while those in pressure(HT)-treated were less than 40% during storage of 64 days at l$0^{\circ}C$. However, at $25^{\circ}C$ and 37$^{\circ}C$ the relative activities in untreated and pressure(RT)-treated were decreased greatly and then reached at the point of the activities of heat- and pressure(HT)-treated. The relative activities of glucoamylase in untreated and pressure(RT)-treated were decreased as the increase of the storage temperature during storage, while those in heat- and pressure(HT)-treated increased slightly as the increase of storage period at 1$0^{\circ}C$ and $25^{\circ}C$, and had no change at 37$^{\circ}C$. pH in heat- and pressure-treated had almost no change. Turbidity and reducing sugar in heat- and pressure-treated increased as the increase of storage temperature during storage.
Tributyltin (TBT) is one of endocrine disrupters which are known as having similar function to sex steroid hormone inducing apoptosis in various tissues of rodents. Recently, it has been reported that TBT induces apoptosis in thymus causing the decreased thymic function, but little is known about the mechanism. To elucidate the mechanism, three-week-old SD female rats were orally administrated with TBT 1, 10, and 25 mg per body weight (kg) and sesame oil as a control for 7 days. On day 8, the thymi were obtained and weighed, and then the number of thymocytes was counted. We also performed H&E staining, TUNEL assay, and Annexin V flow cytometric analysis to examine the apoptosis rates and the structure in the thymus. Next, we investigated the adipogenesis and apoptosis-related mRNA expression levels in the thymi by real-time PCR. The thymic weight and the number of thymocytes were decreased by TBT in a dose-dependent manner. As a result of the H&E staining, the boundary between cortical and medullary area was blurred in the thymi of TBT treated rats compared to those of controls. In the results of TUNEL assay and Annexin V flow cytometric analysis, apoptosis rates in the thymus were increased after TBT treatment. The expression levels of thymic epithelial cell marker genes such as EVA, KGF, AIRE, and IL-7 were significantly decreased in the thymi of TBT treated rats, but $PPAR{\gamma}$, aP2, PEPCK, and CD36 were significantly increased. The expression of $TNF{\alpha}$ and TNFR1 as apoptosis-related genes also was significantly increased after TBT treatment. The present study demonstrates that TBT can increase the expression of adipogenesis and apoptosis-related genes leading to apoptosis in the thymus. These results suggest that the increased adipogenesis of thymus by TBT exposure might induce apoptosis in the thymus resulting in a loss in thymic immune function.
This study was conducted to screen for plant food materials that improve human intestinal microflora, especially microorganisms associated with obesity. Among 30 tested plant food materials, the extract of Schizandra chinensis inactivated Eubacterium limosum, Bacteroides fragilis and Clostridium spp. Additionally, S. chinensis extract was also found to have a growth-promoting effect on Bifidobacterium spp.. The antimicrobial activity and antioxidant activity of the water extract did not decrease in respond to heating. Additionally, the water extract of S. chinensis did not show a toxic effect on the growth of Caco-2 cells. In vivo feeding tests were performed to investigate the influence of extract on the intestinal microflora in rats. Although the extract did not reduce obesity induced by a high fat diet, it led to significant increase in the population of Bifidobacterium spp. and a decrease in the population of Clostridium spp. in rats. Taken together, these results indicate that S. chinensis could be useful as a functional food component to control intestinal microbial flora.
Kim, Chong-Ju;Yeon, Kyu-Min;Kim, Shin-Tae;Wang, Joon-Ho;Yoo, Kwang-Ha
Tuberculosis and Respiratory Diseases
/
v.52
no.6
/
pp.608-615
/
2002
Background : ADA is an enzyme found in most cells, and is involved in purine metabolism, but its chief role concerns the proliferation and differentiation of lymphocytes, especially T-lymphocytes. For that reason ADA has been looked on as a marker of cell-mediate immunity, which is the key mechanism of the tuberculous pleural effusion. Thus, the pleural fluid ADA activity is increased in the tuberculous pleural effusion. Age associated immune decline is characterized by decreases in both B and T-lymphocyte function and the former may be largely a result of the latter. Therefore, the pleural fluid ADA activity would be lower in old rather than in young, patients with tuberculous pleural effusion. We studied the relationship between age, and pleural fluid ADA activity, in patients with tuberculous pleural effusion. Materials and Methods : In the 46 patients with tuberculous pleural effusion enroll in this study, the pleural fluid ADA activities were measured by means of an automated kinetic method. Results : The mean age of the patients was $53.0{\pm}22.0$ years, with a male to female ratio of 30:16. The patients were divided into two groups, young patients, regarded as < 65 and old regarded as ${\geq}65$ years with 28 and 18 patients, respectively. The pleural fluid ADA activity in both groups show significant differences : $99.4{\pm}22.6$ IU/L(young patients) Vs. $75.8{\pm}30.9$ IU/L(old patients)(p<0.05), but a negative correlation with age (r=-0.311, p<0.05). Conclusion : Although pleural fluid ADA activity was not adequately increased, tuberculous pleural effusion, in older patients, would have to be considered clinically suspicious tuberculous pleural effusion.
Jang, Yoonah;Lee, Hye Jin;Choi, Chang Sun;Um, Yeongcheol;Lee, Sang Gyu
Journal of Bio-Environment Control
/
v.23
no.4
/
pp.383-390
/
2014
This study was conducted to investigate the growth characteristics of cucumber scion and pumpkin rootstock under different levels of light intensity (photosynthetic photon flux, PPF) and plug cell size in a closed transplant production system with artificial lighting. Cucumber scion and pumpkin rootstock seedlings were grown under the combinations of three levels of PPF (PPF 165, 248, and $313{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) and five types of plug tray (50, 72, 105, 128, and 200 cells in the tray) for nine days. The shoot dry weight and relative growth rate increased with increasing PPF and plug cell size. As PPF increased, cucumber scion and pumpkin rootstock seedlings had higher dry matter, lower specific leaf area, and lower hypocotyl length. The first true leaf of cucumber scion and pumpkin rootstock unfolded at eight and seven days after sowing, respectively, except the treatment using 200-cell plug tray. The unfolding of first true leaf of seedlings grown in 200-cell plug tray was delayed by one day. Accordingly, it was considered that the use of small cell size such as 200-cell plug tray would require more time for the production of scion and rootstock. Based on the results, we suggest that cucumber scion and pumpkin rootstock be grown in 105-cell to 128-cell plug tray for eight days and 72-cell to 105-cell plug tray for seven days, respectively, when using splice grafting method with root-removed rootstock. Additionally, higher PPF is suggested to improve the growth and quality of scion and rootstock.
Pathophysiological mechanism of hemorrhagic fever with renal syndrome (HFRS) is not fully understood. Major clinical findings of HFRS patients are widespread hemorrhage, acute renal failure and shock. Basic lesion is vascular injury with microvascular hemorrhage and relatively little inflammation. According to autopsy findings, renal medulla shows focal hemorrhage, tubular necrosis and interstitial mononuclear infiltrates. The predominant cell type in the renal and pulmonary interstitium is a fibroblast and it participates in the healing process at the injury site by secreting a large amount of extracellular matrix proteins. Cultured human lung fibroblasts and Mongolian gerbil fibroblasts were known to be good host cells for the hantaan virus. It is possible that not only the endothelial cell but also the fibroblast is a target of Hantaan virus and the fibroblast might be involved in the pathogenesis and the healing process in HFRS. Integrins are adhesion molecules, and act as receptors for many extracellular matrix proteins. Recently, there are many reports that cell surface integrins influence on some viral infections or reversely viruses influence on the expression of integrins. The ${\alpha}_5{\beta}_1$ integrin is a major receptor for the fibronectin which is an important extracellular matrix protein secreted by fibroblasts. In this study, the role of ${\alpha}_5{\beta}_1$ integrin in the infection of Hantaan virus was examined by using anti-${\alpha}_5{\beta}_1$, integrin, anti-${\alpha}_5$ integrin and anti-${\beta}_1$, integrin antibodies in chicken embryo fibroblasts (CEF) and Mongolian gerbil fibroblasts(MGF). The treatment of anti-${\alpha}_5{\beta}_1$, integrin antibody in CEF reduced the virion titers 26.8% and the amount of nucleocapsid N protein 32.6% when compared with control CEF. When MGF were treated with anti-${\alpha}_5$, anti-${\beta}_1$ and anti-${\alpha}_5{\beta}_1$ integrin antibodies, virion titers were reduced by 26.5%, 29.4% and 28.7% and the amount of nucleocapsid N protein were reduced by 65.2%, 59.7% and 72.6%. These results suggested that ${\alpha}_5{\beta}_1$ integrin might act as a receptor for the Hantaan virus or blocking of ${\alpha}_5{\beta}_1$ integrin influences on the viral replication in CEF and MGF. It is also possible that the blocking of only one subunit of integrin represents similar results in that of whole molecule.
The principal objective of the current study was to isolate a purpurogallin derivative as an oxidation product from gallic acid, in an effort to assess the anti-inflammatory effects of this compound. Purpurogallin derivative is known to be the one of the oxidation products of gallic acid. This compound has been identified as a minor chemical component in fermented tea products. It has been previously demonstrated that theaflavins, the oxidation products of catechins found in fermented tea products, exert profound antioxidant and anti-inflammatory effects. However, the biological activities of a minor chemical component in fermented teas have yet to be evaluated. Purpurogallin carboxylic acid (PCA) was identified as a major oxidation product of gallic acid from a peroxidase/hydrogen peroxide oxidation model system. The identity of the PCA was verified by $^{1}H$ NMR, $^{13}C$ NMR and MS techniques. PCA treatment significantly suppressed the generation of pro-inflammatory mediators including nitric oxide and IL-6 in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. According to the nitrite assay, PCA 100, 75, and $50{\mu}g/mL$ treatment dose-dependently inhibited NO production by 57.6, 41.5, and 21.8%, respectively, in LPS-stimulated RAW264.7 murine macrophage cells. Moreover, IL-6 production was inhibited to a significant degree with PCA treatment of 100 and $75{\mu}g/mL$ at 43.1 and 23.9%, respectively. PCA treatment also significantly suppressed $PGE_2$ production at levels of 100 and $75{\mu}g/mL$. These results showed that PCA exerts inhibitory effects on the production of inflammatory mediators.
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