• Journal of Plant Biology
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• v.38 no.1
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• pp.95-105
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• 1995

Complete microsporogenesis of Hibiscus syriacus L. were carried out employing LM, TEM, and SEM to investigate the pollen ontogeny that undergoes considerable structural differentiation. The process first began with several cell diYisions in the anther primordium that produces 3 different tissues of epidennal, archesporial, and connective tissues. Only archesporial tissue involved further differentiation into the tapetum and formation of reproductive cells, pollen mother cells (PMC). The tapetum and PMC were closely associated with each other structurally and metabolically by exhibiting numerous plasmodesmata, mitochondria, and many small vacuoles in their dense cytoplasm. A callosic wall began to surround the PMC while meiosis took place in the PMC to produce 4 microspores. When thick callose encircled each microspore as a frame, the sporodenn development initiated from the plasma membrane of a pollen grain in a tetrad. The first fonned sporoderm layer was bacules and tectum of sexine that originated from the plasma membrane. After the dissolution of a callose, further development Qf sporoderm continued in the order of nexine 1, nexine 2, and intine layer. The nexine layer was thicker (ca. $2-3.5\;\mu\textrm{m}$) than the intine layer whose thickness was about $0.9-1.5\;\mu\textrm{m}$. Upon completion of the sporoderm development, that is after intine formation, spines and apertures of pollen surface ornamentation initiated from the tectum. Spines were dimorphic, about $4-9\;\mu\textrm{m}\;an;15-20\;\mu\textrm{m}$ in length, and no basal cushion was detected. The mature pollen grains ranged $100-200\;\mu\textrm{m}$ in diameter, but their average was about $170\;\mu\textrm{m}$. About 120 spines were observed over the spheroidal pollen surface. Apertures were simple punctures of $2-3\;\mu\textrm{m}$ in diameter and about 50 apertures were arranged somewhat helically over the surface. Comparing such features of form and size of the pollen, sporodenn sculpture and structure, and aperture and spine conditions with known evolutionary trends in the genus Hibiscus, Hibiscus syriacus seemed to possess many advanced features in the sporodenn differentiation.iation.

• Korean Journal of Food Science and Technology
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• v.33 no.6
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• pp.753-759
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• 2001

To evaluate antiinflammation of Aloe vera peel, antiimflammation substances were extracted from Aloe vera peel and identified, and we investigated the effect of the its substance the inhibitory effect on the activity of hyaluoronidase, elastase, collagenase and prostaglandin endoperoxide synthase. The water extract from Aloe vera peel were successfully purified with solvent fractionation, silica gel column chromatography, preparative thin layer chromatography and UV spectrometer. Two purified active substances were identified as aloe-emodin and barbaloin by Mass Spectrometer, $^1H-NMR$ and FT-IR. Aloe-emodin and barbaloin. $IC_{50}$ values of aloe-emodin and barbaloin against hyaluronidase activity were 40 and $70\;{\mu}g/mL$, respectively. Leuckocyte elastase, which is related to the destruction of various tissue, $IC_{50}$ values of them were 50 and $60\;{\mu}g/mL$, respectively. $IC_{50}$ values of aloe-emodin and barbaloin against collagenase activity were 40 and $60\;{\mu}g/mL$, respectively. and $IC_{50}$ values of aloe-emodin and barbaloin aganist the prostaglandin endoperoxide synthase, which play an important role in inflammatory reactions, were 40 and $70\;{\mu}g/mL$, respectively. Inhibitory effects of aloe-emodin, barbaloin and aspirin against carrageenan paw edema were 74.9, 52.9 and 51.9% as inhibiton percentage, respectively, at dose of 100 mg/kg and that of indomethancin was 49.7 at dose of 10 mg/kg. Cell cytotoxicity of barbaloin against human gingival cells was lower than that of aloe-emodin. Aloe-emodin and barbaloin did not show cytotoxicity against human gingival cells at concentration of 1.0 and $5.0\;{\mu}g/mL$, However, aloe-emodin and barbaloin showed less cytotoxicity than chlorhexidine, which usually have been used as the agent of anticaries and antiinflammation. These results suggested that aloe-emodin and barbaloin from Aloe vera peel have the effect of anticaries and antiinflammation.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.238-245
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• 1999

Antioxidative effects of the water or ethanol extracts from Rhus verniciflua Stokes (RVS) were measured by protection against hydroxyl radicals in mouse brain tissue culture. In the water extracts from RVS, cell viabilities were estimated 60.0, 66.0, 72.0, 84.0 and 90.0% at addition of 1, 2, 4, 7 and $10{\mu}L$, respectively, compared with GO (20 mU/mL) alone. The cell viability in the ethanol extracts was similarly with water extracts. In the antitumor effects, the results showed that percentages of the HeLa cell death were approximately 24% for 12 hrs, 57% for 48 hrs at addition of 10%/well ethanol extracts respectively. To know inhibition of tumor growth, in vivo, mice (BALB/c) were inoculated with 0.25 mL CT-26 $(1{\times}10^6\;cells/mL)$ subcutaneously. After the generation of tumor, the results of RVS extracts (ethanol, water) injection showed generally that the tumor size in BALB/c was reduced. For physicochemical characterization of the RVS extracts, purified substances of water or ethanol extracts were analized with SDS-PAGE and ICP spectrometer. In electrophoresis, gel showed 2 bands (210, 230 KDa). The results of ICP verified that RVS extracts contain $Cu^{2+}$ in both samples. Conclusively, this substance might be a laccase which has a biological effective function, as a natural bioactive substance.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.13-22
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• 2005

By extracting the sample of Ulmus davidiana Planch(Ulmaceae), which was known to have the protection of damaged organ and the anti-inflammation action, it was experimented whether it is available for the application of treatment of osteoporosis. In the previous experiment, the extracts from Ulmus davidiana Planch(Ulmaceae) were confirmed to inhibit Cathepsin K through treating the cell of long bone, which contains osteoclast. Through this, it is suggested that Ulmus davidiana Planch(Ulmaceae) can play a role of prodrug as an inhibitor of absorbing bone ash in the treatment of osteoporosis. In the present experiment, a research in vitro Ulmus davidiana Planch(Ulmaceae) on the growth and sensibilization of osteoblast in a state that induced osteosis by using the cell tissue of MC3T3-El pre-osteoblastic was conducted. As a result, it could be confirmed that Ulmus davidiana Planch(Ulmaceae) has the strengthening function by enhancing the dosage and the activity of ALP depending on the time. The dosage was observed at the minimum of $50{\mu}g/m{\ell}$ and the maximum of $150{\mu}g/m{\ell}$. The enhancement in bone morphogenetic protein-2 at $100{\mu}g/m{\ell}$ UD could be observed, and it also increased the concentration of ALP mRNA within the cell of MC3T3-El. At $60{\mu}g/m{\ell}$ UD which indicated a little increase in Type I collagen mRNA for a long time of culture. However, it was shown to sharply inhibit the expression of gene in the culture between 15-20 days. These results suggest that Ulmus davidiana Planch(Ulmaceae) has an influence upon bone metabolism through thje sensibilization of osteoblast. Therefore, it could be known that utilized Ulmus davidiana Planch(Ulmaceae) can be positively applied for the general disease of bone metabolism through future studies.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.2
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• pp.290-303
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• 2006

Tooth eruption is a complex and tightly regulated process that involves cells of the tooth organ and the surrounding alveolus. Osteoclast precursors must be recruited into the dental follicle prior to the onset of eruption. This function of dental follicle may be regarded as the ability of bone remodeling characterized by the interaction of osteoclasts and osteoblasts. This is because tooth eruption is a localized event in which many of the genes required for eruption are expressed in the dental follicle. RANKL is a membrane-bound protein that is a member of the TNF ligand family. which is present on bone marrow stromal cells and osteoblasts, and induces osteoclast formation and activation from precursor cell. The biologic effect of RANKL is inhibited by OPG and, in bone, the relative ratio of RANKL and OPG modulates osteoclastogenesis. To evaluate the roles of RANKL and OPG in tooth eruption and the relations with the expression pattern of Runx2, in situ hybridization was performed with mandibles of mice at postnatal stage 1, 3, 5, 7, 9 and 11. mRNA of RANKL, OPG, and Runx2 are expressed in dental follicle and surrounding tissue from P1 to 11. To determine the sites of osteoclastic activity during tooth eruption, mandibles were dissected. Peak osteoclastic activity in alveolar bone along the occlusal and basal regions was observed from P5 to 9, with osteoclasts in these regions being large and strongly TRAP-positive The specific spatio-temporal expression patterns of RANKL, OPG, and Runx2 in our study suggest that tooth eruption could be progressed through the interactions of molecular signaling among dental follicle, dental organ and alveolar bone, furthermore it means that dental follicle is quite important in tooth eruption In addition, it indicates that these genes (RANKL, OPG, and Runx2) play critical roles in tooth eruption.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.2
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• pp.181-191
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• 2006

NFI-C null mice demonstrated aberrant odontoblast differentiation and thus abnormal dentin formation while other tissues/organs in the body, including ameloblasts, appear to be unaffected and normal. However little is known about the mechanism of NFI-C function in odontoblast differentiation and dentin formation. Odontoblasts are tall, highly polarized cells that are responsible for formation and maintenance of the predentin and dentin. An indication of their polarity is the acquisition of specialized intercellular junctions. As preodontoblasts differentiate into odontoblasts, they are Joined and attached at the apical end by well developed terminal webs of cytoskeletal actins, and associated tight as well as adherent njunctions. In this study, in order to investigate if disruption of the NFI-C gene interferes with formation of a specific or other structural proteins of the intercellular junctions, we examined morphological characteristic of the aberrant odontoblast in NFI-C null mice using light and electron microscope. In addition, we determined the expression of major structural proteins of intercellular junctions, ZO-1 and occludin, during the differentiation of odontoblasts using immunohitochemistry. The results were as follows : 1. In light microscopy, abnormal odontoblasts of incisors of the NFI-C null mice were round in shape, lost their polarity, and trapped in osteodentin-like mineralized tissue. Mutant molars have relatively normal crowns, but short and abnormal differentiating adontoblasts in root formation area. 2. Electron microscopy of abnormal odontoblasts revealed the dissociation of the round osteoblast-like cells, the loss of their cellular polarity, and the absence of an intercellular junctional complex known as the tight junctions. 3. A mutant incisor showed labeling for ZO-1 at the proximal and distal ends of secreting ameloblasts, while staining for ZO-1 was not observed in the abnormal odontoblasts. 4. A normal incisor showed immunoreactivity for occludin in the differentiating odontoblasts. However, staining for occludin was not observed in the abnormal odontoblasts of mutant incisor. These results suggest that NFI-C gene causes dissociation of odontoblast and thus abberant odontoblast differentiation and abnormal dentin formation by interfering with the formation of intercellular junctions.

• Tuberculosis and Respiratory Diseases
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• v.47 no.6
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• pp.797-806
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• 1999

Background: About 20% of small cell lung cancer(SCLC) patients have bone marrow(EM) metastasis at the time of diagnosis and the remaining patients are also considered with micrometastasis. In an attempt to detect EM micrometastasis, we used cytokeratin(CK)-20 as a molecular marker, which is specific for epithelial cells. Method: A sensitive RT-PCR assay was used to compare CK-20 expression both in SCLC cell line H209 and normal leukocyte and to evaluate EM aspirates of 28 SCLC patients. Result: H209 cell line showed CK-20 expression but normal leukocyte did not, suggesting CK-20 expression is lung tissue-specific. Of 28 patients(11 limited disease, 17 extensive disease), only 2(1/11, 1/17) samples tested revealed positive signal for CK-20. Two patients with CK-20 expression had EM metastasis or multiple bone involvement during follow-up. Conclusion: Although circulating tumor cells were detected in EM of small portion of patients with bone metastasis, CK-20 doesn't seem to be a reliable marker for the detection of micrometastasis in SCLC. This study emphasizes that identification of more specific marker for micrometastsis is mandatory prior to clinical application.

• Tuberculosis and Respiratory Diseases
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• v.39 no.6
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• pp.502-510
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• 1992

Background: Neuron specific enolase (NSE) is a neuronal form of the glycolytic enzyme enolase which was first found in extracts of brain tissue, and later in a variety of APUD cells and neurons of the diffuse endocrine system. SCLC shares many APUD properties with normal neuroendocrine cells. NSE immunostaining and serum NSE measurement may be a useful marker of neuroendocrine differentiation in lung tumors and diagnosis of small cell carcinoma. Methods: NSE immunohistochemical staining was done and at the same time serum NSE levels were measured in 22 small cell lung cancer and 21 non small cell lung cancer which were confirmed histologically. Results: 1) NSE immunoreactivity was detected in 9 of the 18 (50%) small cell lung cancer, in 5 of the 16 non small cell lung cancer. 2) Whereas the mean value in non-small cell lung cancer group was $11.79{\pm}4.47\;ng/ml$, the mean level of serum NSE in small cell lung cancer increased up to $59.3{\pm}77.8\;ng/ml$. In small cell lung cancer patients, mean value of limited disease group was $20.19{\pm}12.91\;ng/ml$, while mean value of extended disease group was $91.9{\pm}94.2\;ng/ml$ showing statistically significant difference. If serum levels above 20 ng/ml were tentatively defined as positive, 16 of 22 (73%) patients with SCLC had positive serum NSE level, but only one patient with NSCLC did. There was no correlation between serum NSE level and immunoreactivity of NSE. Conclusion: These studies indicate that serum NSE measurement may be a useful marker for the diagnosis and disease extent and NSE immunostaining can be used to demonstrate the neuroendocrine components of lung tumor.

• Tuberculosis and Respiratory Diseases
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• v.44 no.6
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• pp.1332-1342
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• 1997

Background : Acute pulmonary injury by paraquat are caused by multiple mechanisms including direct injury with oxygen free radicals and several mediators released from inflammatory cells. In order to clarify whether vitamin E could reduce tissue damages induced by intraperitoneal administration of paraquat and to investigate the pathogenetic mechanisms of paraquat-induced pulmonary injury, vitamin E as a free radical scavenger was administered. Method : Rats were divided into three groups (group 1 : control, group 2 : paraquat treated group, group 3 : paraquat and vitamin E treated group). Animals were sacrificed on day 1, day 2, day 3, and day 8 after the administration of saline, paraquat, or paraquat/vitamin E. Results : Treatment with vitamin E decreased the death rate of rats treated with paraquat. Comparing with control group ($1.37{\times}10^6/ml$), mean total cell counts recovered from the lavage fluid from animals treated with paraquat($1.65{\times}10^6/ml$) were increased(p=0.06). Magnitudes of increment of the total cell counts on the Day 8 in the vitamin E treated group were smaller than those of the animals treated with paraquat alone. The neutrophils began to appear in significant amounts in the lavage fluid on Day 8 after the administration of paraquat(37.0+12.7%). A significant decreasing neutrophil concentration at Day 8 was observed in the paraquat/vitamin E treated group(20.6+13.4%). Histologically the degree of pulmonary fibrosis was most prominent in the paraquat treated group while diffuse alveolar damage was continuously observed in the paraquat/vitamin E treated group and extensive interstitial lymphocytic infiltration was seen in the paraquat/vitamin E treated group. The paraquat/vitamin E treated group showed the less histologic changes. Conclusion : In this study vitamin E acting as a scavenger of neutrophil-derived free radicals and suppressant of lipid peroxidation, seemed to be the effective antioxidant in the inhibition of paraquat-induced pulmonary injury.

• Journal of Life Science
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• v.16 no.4
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• pp.612-617
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• 2006

Phosphoinositide-specific phospholipase $C-{\gamma}\;1\; (PLC-{\gamma}\;1)$ has pivotal roles in cellular signaling by producing second messengers, inositol 1,4,5-trisphosphate $(IP_3)$ and diacylglycerol (DG). Tubulin is a main component of microtubules and mitotic spindle fibers, which are composed of ${\alpha}-$ and ${\beta}-tubulin$ heterodimers in all eukaryotic cells. In humans, six ${\beta}-tubulin$ isotypes have been identified which display a distinct pattern of tissue expression. Previously we found that $PLC-{\gamma}\;1$ and one of four ${\beta}-tubulin$ isotypes including ${\beta}1$, ${\beta}2$, ${\beta}3$ and ${\beta}6$, colocalized in COS-7 cells and cotranslocated to the plasma membrane to activate $PLC-{\gamma}\;1$ upon agonist stimulation. In the present study, we demonstrate that the remaining two, tubulin ${\beta}4$ and ${\beta}5$, also showed a potential to activate $PLC-{\gamma}\;1$. The phosphatidylinositol 4,5-bisphosphate $(PIP_2)$ hydrolyzing activity of $PLC-{\gamma}\;1$ was substantially increased in the presence of purified ${\beta}4$ and ${\beta}5$ tubulin in vitro, whereas the activity was not promoted by bovine serum albumin, suggesting that tubulin ${\beta}4$ and ${\beta}5$ also activate $PLC-{\gamma}\;1$. Taken together, our results suggest that all the ${\beta}-tubulin$ isotype activates $PLC-{\gamma}\;1$ activity to regulate cellular signaling.