• Journal of Life Science
• /
• v.21 no.2
• /
• pp.183-193
• /
• 2011

Migration and differentiation of mesenchymal stem cells are crucial for tissue regeneration in response to injury. Sphingosine-1-phosphate (S1P) is a bioactive lipid that regulates a variety of biological processes, including proliferation, survival, differentiation and motility. In the present study, we determined the role of S1P in migration and differentiation of human bone marrow-derived mesenchymal stem cells (BMSCs). S1P stimulated migration of BMSCs in a dose- and time-dependent manner, and pre-incubation of the cells with pertussis toxin completely abrogated S1P-induced migration, suggesting involvement of Gi-coupled receptors in S1P-induced cell migration. S1P elicited elevation of intracellular concentration of $Ca^{2+}$ ($[Ca^{2+}]_i$) and pretreatment with VPC23019, an antagonist of $S1P_1/S1P_3$, blocked S1P-induced migration and increase of $[Ca^{2+}]_i$. Small interfering RNA-mediated knockdown of endogenous $S1P_1$ attenuated S1P-induced migration of BMSCs. Furthermore, S1P treatment induced expression of $\alpha$-smooth muscle actin ($\alpha$-SMA), a smooth muscle marker, and pretreatment with VPC23019 abrogated S1P-induced $\alpha$-SMA expression. S1P induced phosphorylation of p38 mitogen-activated protein kinase (MAPK), and pretreatment of cells with SB202190, an inhibitor of p38 MAPK, or adenoviral overexpression of a dominant-negative mutant of the p38 MAPK blocked S1P-induced cell migration and $\alpha$-SMA expression. Taken together, these results suggest that S1P stimulates migration and smooth muscle differentiation of BMSCs through an $S1P_1$-p38 MAPK-dependent mechanism.

• Journal of Life Science
• /
• v.23 no.11
• /
• pp.1351-1359
• /
• 2013

The anti-proliferative activity of ${\beta}$-ionone was investigated on human non-small lung cancer A-549 cells (designated A-549 cells). A-549 cells were treated with various concentrations of ${\beta}$-ionone (1, 5, 10, and 15 ${\mu}M$) for two, four, and six days. Biochemical markers related to the growth inhibition of A-549 cells by ${\beta}$-ionone were measured at the second day of incubation. ${\beta}$-Ionone inhibited the growth of A-549 cells by dose-and time-dependent manners, resulting in an $IC_{50}$ of 5.0 ${\mu}g/ml$ at the second day of incubation. ${\beta}$-Ionone induced apoptosis by a dose-dependent manner. ${\beta}$-Ionone increased levels of p53, p21, and Bax proteins, but suppressed expression of the Bcl-2 protein. Similarly, ${\beta}$-ionone enhanced cytochrome c release from the mitochondria to the cytosol, and induced activation of caspase-9 and -3. Additionally, ${\beta}$-ion-one reduced $cPLA_2$ and COX-2 protein levels. These results suggest that the ${\beta}$-ionone inhibits the proliferation of A-549 cells through reciprocal regulation of Bax and Bcl-2 gene expression and suppression of $cPLA_2$ and COX-2 protein expressions.

• Horticultural Science & Technology
• /
• v.31 no.1
• /
• pp.80-88
• /
• 2013

Carrot (Daucus carota L. var. sativa) is one of the most extensively used vegetable crops in the world and a significant source of nutrient because of its high content of ${\beta}$-carotene, well known as the precursor of vitamin A carotenoid. However, seed-hairs generated and elongated from the epidermal cell of seeds inhibit absorption and germination by various factors such as carotol and so on. Accordingly, mechanical hair removal process is essential before commercialization of carrot seeds. Because of this process, producers will have additional losses such as time consuming, manpower, capital and so on. Furthermore, physical damage of seeds causes irregular germination rate. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed and studies for molecular markers related to seed-hair characteristic is needed for a new breeding program. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-SMR 616 OP 659-1 line, hairy-seed phenotype CT-SMR 616 OP 677-14 line and short-hair seed phenotype CT-ATR 615 OP 666-13 line, hairy-seed phenotype CT-ATR 615 OP 671-9 were constructed, respectively. Furthermore, 1,248 ESTs in each line, total 4,992 ESTs were sequenced. As a result, 19 SNP sites and 14 SNP sites in each of 2 combinations were confirmed by analyzing these EST sequences from short-hair and hairy-seed lines. Then we designed SNP primer sets from EST sequences of SNP sites for high resolution melting (HRM) analysis. Designed HRM primers were analyzed using hairy seed phenotype CT-SMR 616 OP 1040 line and short-hair seed phenotype CT-SMR 616 OP 1024, 1025, 1026 lines. One set of HRM primers showed specific difference between the melting curves of hairy and short-hair seed phenotype lines. Based on this result, allele-specific (AS) PCR primers were designed for easier selection between hairy-seed carrot and hairless seed carrot. These results of HRM and AS-PCR are expected to be useful in breeding of hairless seed carrot cultivar as a molecular marker.

• Horticultural Science & Technology
• /
• v.33 no.1
• /
• pp.93-105
• /
• 2015

This study was performed to analyze genetic relationships of the four major cucurbitaceous crops including watermelon, melon, cucumber, and squash/pumpkin. Among 120 EST-SSR primer sets selected from the International Cucurbit Genomics Initiative (ICuGI) database, PCR was successful for 51 (49.17%) primer sets and 49 (40.8%) primer sets showed polymorphisms among eight Cucurbitaceae accessions. A total of 382 allele-specific PCR bands were produced by 49 EST-SSR primers from 24 Cucurbitaceae accessions and used for analysis of pairwise similarity and dendrogram construction. Assessment of the genetic relationships resulted in similarity indexes ranging from 0.01 to 0.85. In the dendrogram, 24 Cucurbitaceae accessions were classified into two major groups (Clade I and II) and 8 subgroups. Clade I comprised two subgroups, Clade I-1 for watermelon accessions [I-1a and I-1b-2: three wild-type watermelons (Citrullus lanatus var. citroides Mats. & Nakai), I-1b-1: six watermelon cultivars (Citrullus lanatus var. vulgaris S chrad.)] a nd C lade I -2 for melon and cucumber accessions [I-2a-1 : 4 melon cultivars(Cucumis melo var. cantalupensis Naudin.), I-2a-2: oriental melon cultivars (Cucumis melo var. conomon Makino.), and I-2b: five cucumber cultivars (Cucumis sativus L.)]. Squash and pumpkin accessions composed Clade II {II-1: two squash/ pumpkin cultivars [Cucurbita moschata (Duch. ex Lam.)/Duch. & Poir. and Cucurbita maxima Duch.] and II-2: two squash/pumpkin cultivars, Cucurbita pepo L./Cucurbita ficifolia Bouche.}. These results were in accordance with previously reported classification of Cucurbitaceae species, indicating that watermelon EST-SSRs show a high level of marker transferability and should be useful for genetic study in other cucurbit crops.

• Journal of Nutrition and Health
• /
• v.44 no.1
• /
• pp.5-15
• /
• 2011

Although iron is an essential mineral, excess iron intake during pregnancy may increase oxidative stress in tissues. This study was conducted to investigate the effects of iron overload during pregnancy on iron status and oxidative stress in maternal rats. Ten week-old female Sprague-Dawley rats were mated with male rats. Non-pregnant (control) and pregnant rats were fed diets containing normal Fe (35 mg/kg diet), high Fe (350 mg/kg diet), or excess Fe (1,050 mg/kg diet) during pregnancy. Rats were sacrificed on pregnancy day 19. No significant difference in weight gain, diet intake, or litter size was observed according to iron intake levels. Furthermore, serum iron, hemoglobin, and hematocrit were not different among the rats administered the three levels of Fe both in the control and pregnant groups. However, the iron levels were lower in pregnant rats than those in the control. The liver and spleen iron contents increased significantly in the excess Fe group. An increase in liver ferritin levels with increasing iron intake was observed. Protein carbonyl content, as a marker of oxidative stress, increased significantly in liver with increasing iron intake but not malondialdehyde. Glutathione peroxidase activity in the liver of pregnant rats fed excess iron decreased significantly. Bcl-2 protein expression in the liver declined remarkably with increasing maternal iron intake in pregnant rats. Taken together, iron overload during pregnancy had little effect on hematology. However, the deposits of iron in the liver and the decline in antioxidant enzyme activity implied increased oxidative stress in tissues of the excess Fe group. These results suggest that excess iron intake during pregnancy increases oxidative stress in maternal tissues and may also affect fetal tissues.

• Journal of Nutrition and Health
• /
• v.50 no.3
• /
• pp.246-256
• /
• 2017

Purpose: This study aimed to investigate the association of total dietary antioxidant capacity (TAC) with oxidative stress and metabolic markers among patients with metabolic syndrome according to gender. Methods: A total of 346 subjects aged 30~59 years with two or more risk factors of metabolic syndrome were recruited from a general hospital near Seoul in South Korea between 2010 and 2012 based on data from the medical checkup. Biochemical indices for oxidative stress and metabolic markers were measured. Food consumption data from 3-day food records were linked with the antioxidant capacity database for commonly consumed Korean foods to estimate individual's TAC. Results: Average dietary TAC of the study subjects was 132.0 mg VCE/d/1,000 kcal in men and 196.4 mg VCE/d/1,000 kcal in women. Levels of ${\gamma}$-glutamyltransferase (GGT), systolic blood pressure, diastolic blood pressure, and blood triglycerides were reduced significantly according to increasing TAC in men, but there was no significant trend in women. Intakes of total flavonoids and carotenoids were significantly negatively correlated with GGT (p < 0.05) and d-ROMs (p < 0.01) in men, whereas those of ${\alpha}$-tocopherol (p < 0.05) and ${\gamma}$-tocopherol (p < 0.05) were positively correlated with biological antioxidant potential (BAP) in women. The odds ratio of high oxidative stress indices and abnormal metabolic markers according to TAC level were not significant in either men or women. Conclusion: The results show that dietary TAC was partially associated with oxidative stress and metabolic markers among patients with metabolic syndrome. Further research is required for elucidating the association between dietary TAC and incidence of metabolic syndrome and chronic diseases within a large population in prospective studies.

• Current Research on Agriculture and Life Sciences
• /
• v.5
• /
• pp.119-126
• /
• 1987

This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.

• Journal of Korean Society of Forest Science
• /
• v.90 no.1
• /
• pp.155-160
• /
• 2001

Changes in genetic diversity after seed-tree practice were simulated in test plantation of tulip tree at Sokdal-ri, Banwol-myon, Hwasung-gun, Kyungki Provence. I-SSR markers were analyzed in a total of 305 tulip trees. A total of 89 amplicon variants were observed by PCR with 9 I-SSR primers. Genetic diversity for 305 trees was relatively high (S. I. =0.4532). Individuals originated from the seed orchards in U.S.A. showed the highest level of genetic diversity (0.4530), those from Anyang showed the medium level (0.4152), and those from Cheonbuk showed the lowest (0.3929). Simulation of seed-tree practice accompanied by 2 consecutive thinnings was performed on the basis of morphological characteristics and planted distances of the individual trees, which left 37 trees as candidates for seed-trees. Decreasing rate of genetic diversity within seed sources was greatest for individuals from Cheonbuk (28.3%), moderate for those from Anyang (16.3%), and smallest for those from U.S.A. (8.0%). In spite of little difference in decreasing rate of individuals for the 3 seed sources (87.5~88.2%), large difference in decreasing rate of genetic diversity within seed sources might be due to difference in number of mother trees for the 3 seed sources. For example, whereas individuals originated from the seed orchard in U.S.A., which showed the smallest decreasing rate of genetic diversity, might be originated from relatively large number of mother trees, those from Anyang and Cheonbuk might be originated from relatively small number of mother trees. Although mean of 17.5% of the genetic diversity within seed sources was decreased through 2 consecutive thinnings, a decrease in genetic diversity for the whole individuals (37 trees) was only 6.1%. This observation suggests that the seed-tree practice on the basis of the criteria established in the present study may not result in great reduction in overall genetic diversity of the progenies.

• Journal of Yeungnam Medical Science
• /
• v.6 no.1
• /
• pp.59-68
• /
• 1989

Ceftizoxime sodium is a new synthetic ${\beta}$-lactam antibiotic combining potent antibacterial activity with high stability to a wide range of bacterial ${\beta}$-lactamase. This experiment was achieved to evaluate the antibacterial activities of ceftizoxime sodium againist Gram negative enteric bacteria isolated from in outpatient visiting Yeungnam university hospital and to study the emergence of drug induced bacterial varients which resist to ceftizoxime in vitro. The antibacterial activity of the ceftizoxime was compared with that of antibiotics and its effect on population of normal intestinal flora in mice was observed. The results are summarized as follows : 1. Highly effective antibacterial activity of ceftizoxime against Gram negative enteric bacilli was demonstrated and this antibacterial activity was superior to that of ampicillin. 2. Several test strains shows multiple antibiotic resistence. Among 15 strains of Escherichia coli, 1 strain was resistent to ampicillin, cefadroxyl, gentamicin, tetracycline, and 2 strains were resistent to ampicillin, cefadroxyl, tetracycline, five strains of Escherichia coli and Enterobacter cloacae was resistent to amplicillin, tetracycline and Shigella dysenteria was resistent to ampicillin, gentamicin, tetracycline. 3. The frequency of in vitro emergence of resistent varients among ceftizoxime sensitive bacteria in the presence of increasing concentrations of the compound was found to be low. 4. Plasmid was isolated in 6 of 9 strains (6 strains of Escherichia coli, Shigella dysenteriae, Enterobacter cloaceae and Salmonella typhi) That showed different antibiotic resistance. They were 5 strains of Escherichia coli and 1 strain of Shigella dysenteriae. However, plasmid could not be considered as a hallmark for antibiotic resistance by this. Further studies with curing experiment are to be accomplished for this purpose. 5. Changes in the bacterial count of normal intestinal flora following 25mg/kg/day administration of ceftizoxime over S consecutive days were not significant. In conclusion, ceftizoxime appeared to be a drug of choice in the treatment of Gram negative enteric bacilli infection.

• Laboratory Medicine Online
• /
• v.6 no.1
• /
• pp.25-30
• /
• 2016

Background: The cell cycle-dependent enzyme thymidine kinase 1 (TK1) is known to increase during cancer cell proliferation and has been reported as a prognostic marker for various hematologic malignancies and solid tumors. This study aimed to determine the reference interval in Korean healthy controls and to evaluate the usefulness of TK1 as a biomarker for aggressive clinical behavior in B-cell lymphoma patients. Methods: We enrolled 72 previously untreated patients with B-cell lymphoma and 143 healthy controls. Serum TK1 levels were measured by chemiluminescence immunoassay ($Liaison^{(R)}$, DiaSorin, USA). We established the reference intervals in healthy controls. The diagnostic performance of serum TK1 was studied using receiver operating characteristic (ROC) analysis, and the correlation between the cutoff level for serum TK1 and clinical characteristics of B-cell lymphoma was evaluated. Results: The reference range (95th percentile) of serum TK1 in healthy controls was 5.4-21.8 U/L. There was a clear difference in TK1 levels between patients with B-cell lymphoma and healthy controls ($40.6{\pm}68.5$ vs. $11.8{\pm}4.4U/L$, P <0.001). The area under the curve of serum TK1 for the diagnosis of B-cell lymphoma was 0.73 (cutoff, 15.2 U/L; sensitivity, 59.7%; specificity, 83.2%). An increased TK1 level (${\geq}15.2U/L$) correlated with the advanced clinical stage (P <0.001), bone marrow involvement (P =0.013), international prognostic index score (P =0.001), lactate dehydrogenase level (P =0.001), low Hb level (<12 g/dL) (P =0.028), and lymphocyte count (P =0.023). Conclusions: The serum TK1 level could serve as a useful biomarker for aggressive clinical behavior in B-cell lymphoma patients.