A hyperthermophilic bacteria (strain HJ6) was isolated from a hot springs located in the Arima-cho, Hyogo, Japan. The cells were long-rod type ($2-4{\mu}m$), about $0.4{\mu}m$ in diameter. The pH and temperature for optimal growth were 6.5 and $80^{\circ}C$, respectively. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that HJ6 belonged to the genus Thermus thermophilus (Tt). The gene encoding the Trehalose synthase (TS) was cloned and sequenced. The open reading frame (ORF) of the TtTS gene was composed of 2,898 nucleotides and encoded a protein (975 amino acids) with a predicted molecular weight of 110.56 kDa. The deduced amino acid sequence of TtTS showed 99% and 83% identities to the Thermus caldophilus TS and Meiothermus ruber TS, respectively. TtTS gene was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for Trehalose synthase activity were found to be $80^{\circ}C$ and 7.5, respectively. The half-life of heat inactivation was about 40 min at $90^{\circ}C$. The maximum trehalose conversion rate of maltose into trehalose by the enzyme increased as the substrate concentration increased, and reached 55.7% at the maltose concentration of 500 mM, implying that the enzyme conversion was dependent of the substrate concentration.
This study is to diagnose the drought tolerance of twenty broad-leaved tree species by the pressure-volume(P-V) curves. As for the diagnosis of drought tolerance, the valuable water relations parameters obtained from P-V curves are the osmotic potential at full turgor, ${\Psi}_0{^{sat}}$, osmotic potential at incipient plasmolysis, ${\Psi}_0{^{tlp}}$, maximum bulk modulus of elasticity, $E_{max}$, and relative water content at incipient plasmolysis, $RWC^{tlp}$. Also, the figures related to the diagnosis of drought tolerance are the free water content (FWC) versus leaf water potential(${\Psi}_L$), volume-averaged turgor pressure ($P_{vat}$) versus leaf water potential (${\Psi}_L$), and H$\ddot{o}$fler diagram. In this study, the relatively high drought tolerant species are Fraxinus rhynchophylla, Quercus acutissima, Quercus serrata, Quercus aliena, and Populus alba${\times}$glandulosa ; the relatively low drought tolerant species are Fraxinus mandshurica, Betula platyphylla var. japonica, Populus euramericana, Kalopanax pictum, Carpinus loxiflora, Carpinus cordata, Prunus sargentii, Prunus leveilleana, and Cornus controversa ; medium species are Quercus mongolica, Acer mono, Acer triflorum, Acer pseudo-sieboldianum, Ulmus davidiana, and Zelkova serrata.
Lee Chang-Hee;Bahn Kyeong-Nyeo;Cho Tae-Yong;Lee Ju-Yeon;Lee Young-Ja;Chae Gae Yong
Journal of Food Hygiene and Safety
/
v.20
no.4
/
pp.267-271
/
2005
Lecithin is a naturally occurring group of phospholipids found in nearly every living cell and has been widely used as the ingredient of functional foods. Lecithin has high content of phosphatidylcholine(PC), pharmaceutical material which promotes metabolism through the cell membrane. This study was carried out to improve the present inconvenient analytical method of PC in law for health & functional foods. The commodities used in this experiment, were two kinds of egg yolk and eight kinds of soybean lecithin functional foods. PC was separated with isocratic elution with hexane : isopropanol : D.W (30:60:8) through silica column (2.1$\times$150 mm) by HPLC with Evaporative Light-Scattering Detector (ELSD). The flow rate of the eluent was 0.5 ml/mim and infect volume was 10ul. The neubilizer temperature of detector was $60^{\circ}C$, drift tube temperature of that was $75^{\circ}C$ and gas flow was 30 psi. Quantification was carried out by external standardization. Limit of quantification was 0.15ppm. Lecithin contents of egg yolk and soybean Products were > $66\%$ and > $81\%$), respectively. Phosphatidylcholine contents of egg yolk and soybean products were > $74\%$ and > $18\%$, respectively.
The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.
This study was conducted to investigate the developmental ability of interspecies cloned embryos after nuclear transfer of goat fetal fibroblast cells into porcien oocytes. Recipient porcine and goat oocytes were obtained from slaughterhouse and matured in vitro according to established protocols. Enucleation was accomplished by aspirating the first polar body and cytoplasm and a single donor cell was individually microinjected into vitelline space of the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol fusion medium. After electro-fusion, interspecies reconstituted embryos were cultured in PZM-3 for 7 days. In porcine interspecies nuclear transfer with goat fetal fibroblast cells, the cleavage rate of reconstituted embryos were 58.9% which was no significant different from that in porcine nuclear transfer embryos (67.4%). However, the developmental rate into blastocyst stage was 5.4% in interspecies nuclear transfer which was significantly lower than that in porcine intraspecies nuclear transfer (13.6%). When the developmental ability of porcine interspecies nuclear transfer with goat cells was compared with goat intraspecies nuclear transfer, the cleavage rate of embryos were 59.2% and the developmental rate into morular and blastocyst stage was 13.6% in interspecies nuclear transfer which were significantly lower than those in intraspecies nuclear transfer embryos. This result indicated that porcine interspecies nuclear transfer with goat fetal fibroblast cells showed the developmental potential in vitro with lower cleavage and developmental rate compared with intraspecies nuclear transfer.
Yu, Hyo Jung;Park, Eun Ae;Kim, Ji Young;Cho, Soo Jin;Kim, Young Ju;Park, Hye Sook;Ha, Eun Hee
Clinical and Experimental Pediatrics
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v.51
no.7
/
pp.754-759
/
2008
Purpose : Abnormal activation patterns of Th1/Th2-cells have been suggested to increase the prevalence of allergic diseases. Prevention is regarded as an important corner stone in the management of allergic diseases. In this study, we have investigated the relationship between cord blood levels of IL-4, IL-10, and IL-12 in preterm newborns and the development of allergic respiratory diseases in infancy Methods : Forty-six preterm newborns born at the Ewha Womans University Mokdong Hospital between January 2003 and July 2005, were enrolled for this study, and consent was obtained to test their cord blood samples. Clinical history was obtained from the hospital records. Cord blood was obtained at birth and kept frozen until it was tested. The levels of IL-4, IL-10, and IL-12 were determined by enzyme-linked immunosorbent assay (ELISA). Results : All infants were followed-up for a median of $16.0months{\pm}13.2d$ (range, 12.0 to 36.0 months). Eighteen infants who developed wheezing showed lower cord blood levels of IL-12 ($366.60{\pm}140.40$ vs $435.09{\pm}91.20pg/mL$, P=0.009). Cord blood levels of IL-4 and IL-10 showed no significant difference between the two groups. Four newborns who later developed asthma, and infants with asthma showed lower IL-12 level in the cord blood than other groups. Conclusion : Lower concentration of cord blood levels of IL-12 in newborns who later developed wheezing and asthma suggested that they had abnormal activation patterns of Th1/Th2-cells at the time of birth, and cord blood IL-12 level can be used as a predictor of allergic respiratory diseases.
The early life history of Korean bullhead, Pseudobagrus fulvidraco was studied to obtain some information required in aquaculture and reinforcement of natural population. The fertilized eggs were almost spherical in shape and demersal. The egg membranes were transparent with minute folds on the surface, causing them to stick to other substrates. Yolk is yellowish without oil droplets. The eggs just after fertilization were measuring $1.4{\pm}0.03mm$(1.3~1.5mm, n=10) and expanded to $1.7{\pm}0.08mm$(1.6~1.8mm, n=10) in diameter in 1.5 hr. The blastodisc was formed in 30 min and cleavage started in 1 hr after fertilization, and the intervals of each stage of cleavage was about 30 min at $25.0^{\circ}C$. The yolk from 32-cell stage to gastrula stage partly depressed and the depressed part moved clockwise. Hatching occurred in 53 hr after fertilization and hatched embryos had 18~19+20~21(38~40) myomeres measuring 4.2~4.3mm in total length. At the age 7 d after hatching, 4 pairs of barbels were already formed ; 1 pair on the posterior part of the nostril, 1 pair on the upper jaw, and 2 pairs on the lower jaw. And the posterior margin of caudal fin changed into two folds. The lateral band and the form of all fins were completed in 3 weeks.
This study was carried out to obtain the fundamental data for the mass seedling production of grunt, Hapalogenys nitens in terms of the natural spawning and some characteristics of the eggs spawned. The wild grunt were reared at indoor tanks for three years. The adults spawners were 34.0∼44.0 cm (38.6$\pm$4.0 cm, n=7) in total length, 1.00∼2.23 kg (1.62$\pm$0.50 kg, n=7) in body weight. Spawning were observed 9 times from September 22 to October 1, 2000 and 37 times from August 22 to October 3, 2001, with a water temperature range of 19.8$\pm$28.5$^{\circ}C$. The total number of eggs collected was 2.29${\times}$10$^{7}$ (1.7${\times}$10$^{3}$/ml). The relative proportion of floating eggs to total eggs was 41.7%. The fertilization rate of floating eggs was ranged between 85.0 and 99.9% and the hatching rate was ranged between 2.9 and 93.0%. Fertilized eggs were buoyant and spherical in shape, and were 0.85∼0.98 mm in diameter. Each egg contained 1-5 oil globules which were, 0.18∼0.25 mm in diameter. The incubation time from fertilization to blastodisc formation was 10 minutes, to blastula was 3 hours, and to the hatched larvae at 26$^{\circ}C$ was 20 hours 30 minutes. The newly hatched larvae attained total length of 1.81$\pm$0.18 mm. The time required from fertilization to hatching was 31∼34 hours at 23$^{\circ}C$ and 17∼20 hours at 29$^{\circ}C$.
In this study the extracellular production of 5-aminolevulinic aicd (ALA) by recombinant E. coli BL2l (DE3) pLysS harboring the plasmid pFLS45 are investigated. Optimum concentrations of succinic acid and glycine for cell growth and ALA production were found to be 30 mM and 15 mM, respectively. Levulinic acid (LA) as an inhibitor of ALAD was added to the culture medium in the end of exponential cell growth phase and its optimum concentration was 30 mM. Growth of recombinant E. coli BL2l (DE3) pLysS (pFLS45) was largely dependent upon the pH value of culture medium. When the pH of culture medium was in the range of 6.0 and 6.5, high cell mass and ALA production were obtained. IPTG induction for the expression of the fusion gene did not enhance the production of ALA. Recombinant cell grew at 30't faster than at 37$^{\circ}C$, but ALA productivity was lower than at 37$^{\circ}C$. Repeated addition of glycine, succinic acid, and LA increased the production of ALA and the inhibition of intracellular ALA dehydratase activity, with up to 1.3 g/L ALA having been produced in the cultivation.
Predictive growth model of putrefactive bacteria of surimi-based imitation crab in the modified surimi-based imitation crab (MIC) broth was investigated. The growth curves of putrefactive bacteria were obtained by measuring cell number in MIC broth under different conditions (Initial cell number, $1.0{\times}10^2,\;1.0{\times}10^3$ and $1.0{\times}10^4$ colony forming unit (CFU)/mL; temperature, $15^{\circ}C,\;20^{\circ}C\;and\;25^{\circ}C$) and applied them to Gompertz model. The microbial growth indicators, maximum specific growth rate constant (k), lag time (LT) and generation time (GT), were calculated from Gompertz model. Maximum specific growth rate (k) of putrefactive bacteria was become fast with rising temperature and fastest at $25^{\circ}C$. LT and GT were become short with rising temperature and shortest at $25^{\circ}C$. There were not significant differences in k, LT and GT by initial cell number (p>0.05). Polynomial model, $k=-0.2160+0.0241T-0.0199A_0$, and square root model, $\sqrt{k}=0.02669$ (T-3.5689), were developed to express the combination effects of temperature and initial cell number, The relative coefficient of experimental k and predicted k of polynomial model was 0.87 from response surface model. The relative coefficient of experimental k and predicted k of square root model was 0.88. From above results, we found that the growth of putrefactive bacteria was mainly affected by temperature and the square root model was more credible than the polynomial model for the prediction of the growth of putrefactive bacteria.
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