Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.
Renal proximal tubular hypertrophy and hyperfunction are known to be early manifestations of experimental and human diabetes. As the hypertrophy and hyperfunction have been suggested to be central components in the progression to renal failure, an understanding of their underlying causes is potentially important for the development of therapy. A primary rabbit kidney proximal tubule cell culture system was utilized to evaluate the possibility that the renal proximal tubular hypertrophy and hyperfunction observed in vivo in diabetes mellitus, can be attributed to effects of elevated glucose levels on membrane transport systems. Primary cultures of rabbit proximal tubules, which achieved confluence at 10 days, exhibited brush-border characteristics typical of proximal tubular cells. Northern analysis indicated $2.2{\sim}2.3$ and 2.0 kb Na/glucose cotransporter RNA species appeared in fresh and cultured proximal tubule cells after confluence, repectively. The cultured cells showed reduced Na/glucose cotransporter activity compared to fresh proximal tubules. Primary cultured proximal tubule cells incubated in medium containing 20 mM glucose have reduced ${\alpha}-MG$ transport compared to cells grown in 5 mM glucose. In the proximal tubule cultures incubated in medium containing 5 mM or 20 mM glucose, phlorizin at 0.5 mM inhibited 0.5 mM ${\alpha}-MG$ uptake by 84.35% or 91.85%, respectively. The uptake of 0.5 mM ${\alpha}-MG$ was similarly inhibited by 0.1 mM ouabain (41.97% or 48.03% inhibition was observed, respectively). In addition, ${\alpha}-MG$ uptake was inhibited to a greater extent when $Na^{+}$ was omitted from the uptake buffer (81.86% or 86.73% inhibition was observed, respectively). In cell homogenates derived from the primary cells grown in 5 mM glucose medium, the specific activity of the Na/K-ATPase $(6.17{\pm}1.27\;{\mu}mole\;Pi/mg\;protein/hr)$ was 1.56 fold lower than the values in cell homogenates treated with 360 mg/dl D-glucose, 20 mM $(9.67{\pm}1.22\;{\mu}mole\;Pi/mg\;protein/hr)$. Total $Rb^{+}$ uptake occurred at a significantly higher rate (1.60 fold increase) in primary cultured rabbit kidney proximal tubule cell monolayers incubated in 20 mM glucose medium $(10.48{\pm}2.45\;nM/mg\;protein/min)$ as compared with parallel cultures in 5 mM glucose medium. $Rb^{+}$ uptake rate in 5 mM glucose medium was reduced by 28% when the cultures were incubated with 1 mM ouabain. The increase of the $Rb^{+}$ uptake by rabbit kidney proximal tubule cells in 20 mM glucose could be attributed primarily to an increase in the rate of ouabain-sensitive $Rb^{+}$ uptake $(5\;mM\;to\;20\;mM;\;4.68{\pm}0.85\;to\;8.38{\pm}1.37\;nM/mg\;protein/min)$. In conclusion, the activity of the renal proximal tubular Na,K-ATPase is elevated in high glucose concentration. In contrast, the activity of the Nafglucose cotransport system is inhibited.
About 500 bacterial and fungal strains from a wide variety of natural habitats were screened for a new type II restriction endonuclease. Among the 500 species, we selected one species that produced a new restriction endonuclease. This strain has an optimum temperature of $30^{circ}C$ for growth. Morphological, cultural, and physiological characteristics were examined for identification of the isolated strain J-482. This strain was found to belong to the genus Alcaligenes. The restriction endonuclease was named as AspJI and partially purified from Alcaligenes sp. J-482 by DEAE-Sephadex A-50 column chromatography and gel filtration. Most of other nucleases were removed by the purification steps. The AspJI has a substrate specificity to ${lambda}$ DNA, pBR322 and Adenovirus-2 DNA. For its maximal activity, the isolated enzyme requires $MgCl_2$, which should be at least 12.5 mM and it does not need any other cofactors. It is maximally active in the absence of NaCl and is completely inactivated at 100 mM NaCl. The pH and temperature optima for activity were pH 7.5 and $37^{circ}C$, respectively. The DNA fragments generated by digesting ${lambda}$ DNA, pBR322, and Adenovirus-2 DNA with AspJI were the same as that produced by AatII. This suggests that AspJI is an isoschizomer of AatII.
To develop multifunctional microbial inoculant, an insluble phosphate-solubilizing bacterium with antifungal activity was isolated from plant rhizospheric soil. On the basis of its morphological, cultural and physiological characteristics and Biolog analysis, this bacterium was identified as Pseudomonas fluorescens RAF15. P. fluorescens RAF15 showed antifungal activities against phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% of glucose, 0.005% of urea, 0.3% $MgCl_2{\cdot}6H_2\;0.01%\;of\;MgSO_4{\cdot}7H_2O\;0.01%,\;of\;CaCl_2{\cdot}2H_2O$, and 0.05% of NaCl along with initial pH 7.0 at $30^{\circ}C$. The soluble phosphate production under optimum condition was 863 mg/L after 5 days of cultivation. The solubilization of insoluble phosphates was associated with a drop in the pH of the culture medium. P. fluorescens RAF15 showed resistance against different environmental stresses like $10-35^{\circ}C$ temperature, 1-4% salt concentration and pH 2-11 range. The strain produced soluble phosphate to the culture broth with the concentrations of 971-1121 mg/L against $CaHPO_4$, 791-908 mg/L against $Ca_3(PO_4){_2}$, and 844 mg/L against hydroxyapatite, respectively. However, the strain produced soluble phosphate to the culture broth with the concentrations of 15 mg/L against $FePO_4$, and 5 mg/L against $AlPO_4$, respectively.
Kim, Man-Chul;Kim, Ju-Sang;Kim, Yun-Beom;Harikrishnan, Ramasamy;Han, Yong-Jae;Heo, Moon-Soo
Korean Journal of Microbiology
/
v.45
no.4
/
pp.397-403
/
2009
For the research of the natural antioxidant from marine sources, an antioxidant-producing marine actinomycetes was isolated from sea water in Jeju coastal area. The strain was identified based on 16S rDNA sequencing, the morphology by a method of scanning electron microscopy, physiological and biochemical characteristics and cellular fatty acid analysis. The isolated strain ACT-1 cell size was $0.5\sim1.0{\mu}m$ and gram positive, aerobic, nonmotile, substrate mycelium are red and gray aerial mycelium. 16S rRNA sequence analysis showed that were Gram-positive bacteria grouped on Streptomyces genus. Results of cellular fatty acid analysis showed that major cellular fatty acids were $C_{15:0}$ anteiso (39.33%), $C_{16:1}$ cis 9 (11.96%), $C_{16:0}$ (13.08%) and $C_{17:0}$ anteiso (10.99%). Finally, strain was identified Streptomyces sp. ACT-1. The antioxidant activity of methanol extract from Streptomyces sp. ACT-1 was evaluated by measuring DPPH, hydroxyl, and alkyl radical scavenging activity using an electron spin resonance (ESR) spectrometer. DPPH radical scavenging activity of SBME-1 (Streptomyces broth methanol extract) was 67% at $1,000{\mu}g$/ml. Hydroxyl radical scavenging activity of SBME-1 was 84% at $500{\mu}g$/ml. Alkyl radical scavenging activity of SBME-1 was 71% at $1,000{\mu}g$/ml.
Purpose: This study was to examine the frequency, distribution and characteristics of researches using biological measurement published from 2000 to 2004 in 10 major nursing journals in Korea, Design: Literature analysis. Method: Journals including papers using biological measurements, publishing year, research design and outcome variables were analyzed. Results: 1. Researches using biological measurement were 318(13.3%). 2. Researches using biological measurement in the Korean Academy of Nursing were highest(97papers, 17.5%) among the nursing journals. The proportion of papers using biological measurement to total number of papers was the highest in the Journal of Korean Biological Nursing Science as 77.3%(51papers). 3. The 233 papers(73.3%) were experimental researches among 318 papers using biological measurement which showed the highest proportion. 4. Patients were highest subjects of researches using biological measurement(197papers, 61.9%). 5. Blood test was most frequently used as physiological variables from 2001 to 2004. Conclusion: Researches using biological measurement of 10 Korean Nursing Journals in year 2000-year 2004 were very low. We need more researches using biological measurement to provide more objective evidence for nursing practice.
Journal of Physiology & Pathology in Korean Medicine
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v.23
no.6
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pp.1234-1240
/
2009
This study was to compare the effectiveness and validity of various data-mining algorithm for Sasang type diagnostic test. We compared the sensitivity and specificity index of nine attribute selection and eleven class classification algorithms with 31 data-set characterizing Sasang typology and 10-fold validation methods installed in Waikato Environment Knowledge Analysis (WEKA). The highest classification validity score can be acquired as follows; 69.9 as Percentage Correctly Predicted index with Naive Bayes Classifier, 80 as sensitivity index with LWL/Tae-Eum type, 93.5 as specificity index with Naive Bayes Classifier/So-Eum type. The classification algorithm with highest PCP index of 69.62 after attribute selection was Naive Bayes Classifier. In this study we can find that the best-fit algorithm for traditional medicine is case sensitive and that characteristics of clinical circumstances, and data-mining algorithms and study purpose should be considered to get the highest validity even with the well defined data sets. It is also confirmed that we can't find one-fits-all algorithm and there should be many studies with trials and errors. This study will serve as a pivotal foundation for the development of medical instruments for Pattern Identification and Sasang type diagnosis on the basis of traditional Korean Medicine.
The estrogen-mediated effect of mesenchymal stem cells (MSCs) is a highly critical factor for the clinical application of MSCs. However, the present study is conducted on MSCs derived from adult donors, which have different physiological status with steroid hormonal changes. Therefore, we explores the important role of $17{\beta}$-estradiol (E2) in MSCs derived from female and male newborn piglets (NF- and NM-pBMSCs), which are non-sexually matured donors with steroid hormones. The results revealed that in vitro treatment of MSCs with E2 improved cell proliferation, but the rates varied according to the gender of the newborn donors. Following in vitro treatment of newborn MSCs with E2, mRNA levels of Oct3/4 and Sox2 increased in both genders of MSCs and they may be correlated with both estrogen receptor ${\alpha}$ ($ER{\alpha}$) and $ER{\beta}$ in NF-pBMSCs, but NM-pBMSCs were only correlated with $ER{\alpha}$. Moreover, E2-treated NF-pBMSCs decreased in ${\beta}$-galactosidase activity but no influence on NM-pBMSCs. In E2-mediated differentiation capacity, E2 induced an increase in the osteogenic and chondrogenic abilities of both pBMSCs, but adipogenic ability may increased only in NF-pBMSCs. These results demonstrate that E2 could affect both genders of newborn donor-derived MSCs, but the regulatory role of E2 varies depending on gender-dependent characteristics even though the original newborn donors had not been affected by functional steroid hormones.
This experiment conducted to know physiological characteristics and stress effect on different growth stage of soybean by night illumination. Soybean variety, Shinpaldalkong 2, Keumjungkong and Muhankong were treated by night illumination with 20~30 Lux (0.05~0.08W m$^{-2}$ , 0.24~0.36 $\mu$㏖ S$^{-1}$ m$^{-2}$ ) for 15 days at six different growth stage, seedling, pre-floral initiation, post-floral initiation, pod filling and seed ripening stage. Night illumination delayed flowering to 2~8 days compared to control. Delay of flowering by night illumination severely effected at the pre-floral initiation stage. Stem length was increased all the night illumination treatments except the pod filling stage. Number of nodes in Shinpaldalkong 2 and Keumjungkong 1 were increased until before post-floral initiation stage but in Muhankong were increased until after post-floral initiation stage by night illumination treatments. Number of pods were decreased all the night illumination treatments except seedling stage compared with control. Yield decreased all the treatments and severe the loss rate degree showed the order of prefloral initiation, post-floral initiation, seedling and flowering stage.
This study was conducted to screen an alginate-degrading microorganism and to investigate the characteristics of the alginate-degrading activity of its crude enzyme. A marine bacterium which produces extracellular alginate-degrading enzymes was isolated from the brown alga Sargassum thunbergii. 16S rRNA sequence analysis and physiological profiling resulted in the bacterium's identification as a Vibrio crassostreae strain, named Vibrio crassostreae PKA 1002. Its optimal culture conditions for growth were pH 9, 2% NaCl, $30^{\circ}C$ and a 24 hr incubation time. The optimal conditions for the alginate degrading ability of the crude enzyme produced by V. crassostreae PKA 1002 were pH 9, $30^{\circ}C$, a 48 hr incubation time and 8% alginic acid. The alginate degrading crude enzyme produced 3.035 g of reducing sugar per liter in 4% (w/v) alginate over 1 hr.
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