• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.687-690
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• 1999

Bacillus stearothermophilus No. 236 isolated from the soil is a strong xylan degrader producing all the xylanolytic enzymes. However, the strain was discovered to be highly intractable to its transformation. In the present study, we have developed a reliable method for transformation of B. stearothermophilus No. 236 by a systematic examination of several factors which might have an influence on the efficiency of electrotransformation. Notably, we found that the most critical factor influencing the transformation efficiency (TE) was the incubation temperature after pulsing, with its optimum incubation of $37^{\circ}C.\; At\; 50^{\circ}C$, the optimum growth temperature of the B. stearothermophilus strain, the transformants could not be obtained at a recognizable level. The combination of field strength of 7.5 kV/cm along with pulse duration of 10 msec (resistance of $400{\Omega}\; and\; capacitance\; of\; 25{\mu}F$) was shown to be the best electrical parameters at the incubation temperature of $37^{\circ}$. A higher TE was obtained when the cells were harvested at an early-exponential phase. Twenty percent of PEG-8000 in a suspension buffer and an addition of 0.1% glycine in the growth medium resulted in about 4-fold and 3-fold increases in TE, respectively. We also found that the plasmid DNA which had been cycled through the host B. stearothermophilus cells enhanced TE by one order of magnitude higher. Under the presently described conditions, $2.5{\times}10^{5} transformants per ${\mu}g$ DNA was attained.

• Korean Journal of Environment and Ecology
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• v.38 no.2
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• pp.143-147
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• 2024

This study aimed to determine the egg-hatching period of boreal digging frogs, Kaloula borealis, and investigate whether the incubation temperature affects the hatching period. In this study, the egg hatching was recorded based on the appearance of the tadpole. The results of this study showed that all the eggs hatched within 48 hours after spawning, with 28.1% (±10.8, n=52) hatching within 24 hours and 99.9% (±0.23, n=49) within 48 hours after spawning. The mean hatching rate of tadpoles showed significant differences depending on the difference in water temperature. The mean hatching rate between 15 and 24 hours after spawning was higher at a water temperature of 21.1 (±0.2) ℃ than at 24.1 (±0.2) ℃. The results suggest rapid hatching occurs at relatively low water temperatures because the spawning habits that spawn eggs in temporary ponds or puddles in the rainy season require rapid hatching before the puddles dry out. The results of this study are helpful for understanding the most suitable temperature conditions for the incubation of eggs of the endangered species, boreal digging frog.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.35 no.5
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• pp.62-71
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• 2003

This study was carried out to develop the bio-chemical pulping method to enhance the energy saving and decrease the capital cost through the soda pulping under atmospheric pressure ($100^{\circ}C$). Nonwood substrates, rice straw, were pretreated by white-rot fungi, Pleurotus cornucopiae. Several basic pieces of data that can be applied in soda pulping were acquired. The results of this study were as follows. Under the conditions without any nutrients or with glucose, N and glucose + N, the weight losses of rice straws inoculated by Pleurotus cornucopiae were 12.1∼32.6 %, 12.0∼26.3 %, 13.0∼25.4 % and 15.3∼24.7 % for 5, 10, 15, 20, 25 and 30 days incubation periods respectively. The more the fungal incubation was extended, the more the weight losses were gained. The yield of untreated rice straw was 54.8 % after pulping. When any nutrients was not added or glucose, N and glucose ＋ N were added for the pretreatment, the total yields were ranged to 57.3∼42.9 %, 51.0∼43.3 %, 51.7∼43.9 % and 52.1∼46.1 % for 5 different incubation periods respectively. The yields were gradually decreased based on the extending of the incubation periods. The physical properties of the rice straw soda pulp without fungal treatment, the density, breaking length, burst index, tear index and folding endurance were 0.24g/㎤, 2.32 Km, 0.91 kPaㆍ$m^2$/g, 46.7 mNㆍ$m^2$/g and 21 times, respectively. In the case of pretreatment without any nutrients or with glucose, N and glucose + N as nutrients, the density was 0.24g/㎤, the breaking length was 3.30∼6.46 Km, the burst index was 1.36∼3.01 kPaㆍ$m^2$/g, the tear index was 33.0∼57.0 mNㆍ$m^2$/g and the folding endurance was 14∼381 times at most incubating periods, when pulping was done. The physical properties were increased as the incubation duration was extended. Especially, when N and glucose ＋ N were added, the physical properties showed superior results during each incubation period.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.97-104
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• 2008

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin. PA/plasmin system playa role in mammalian fertilization and motility and acrosome reaction of sperm. The present study was undertaken to identify PAs in porcine gametes and investigate a possible role of plasminogen in in vitro fertilization in the pig. When boar spermatozoa were preincubated in a fertilization medium (mTBM) for 0, 2, 4 or 6 h, the activity of tPA-PAI ($110{\sim}117\;kDa$), tPA ($62{\sim}70\;kDa$), and uPA ($34{\sim}38\;kDa$) was observed in the sperm incubation medium and sperm sample. PA activities in the sperm incubation medium significantly (p<0.05) increased according to increasing incubation times, while PA activities in sperm significantly (p<0.05) decreased at the same times. In addition, the rate of acrosome reaction in spermatozoa increased by increasing culture times. When oocytes were separated from porcine cumulus-oocytes complexes at 0, 22 or 44 h of maturation culture, no PA activities were observed in cumulus free-oocyte just after aspiration from follicles. However, the activity of tPA-PAI ($108{\sim}113\;kDa$) and tPA ($75{\sim}83\;kDa$) was observed at 22 h of in vitro culture and significantly (p<0.05) increased as the duration of the culture increased. On the other hand, when porcine oocytes were activated by sperm penetration or calcium ionophore, plasminogen significantly (p<0.05) increased ZP dissolution time (sec) in activated oocytes by sperm penetration. These results suggest that supplementation of plasminogen to fertilization medium may playa positive role in the improvement of in vitro fertilization ability in the pig.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.2
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• pp.240-245
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• 2010

The effects of controlled energy restriction and duration of pre-incubation egg holding on fertility, hatchability and hatch losses were evaluated in aged broiler breeders (64 wk). The energy (ME) required for maintenance, activity, growth and anticipated egg production was calculated and offered to a control group (283-471 kcal/kg) from 21-64 weeks of age. In three other groups, ME was quantitatively reduced either by 20% (SER; severe energy restriction) or 10% (MER; moderate energy restriction) and increased by10% (EEF; excess energy feeding) over the control group (CER; controlled energy restriction). Each diet was offered to 130 pullets in individual cages, and the quantity of ME increased with age. At the end of 64 weeks, fertile eggs were collected from each dietary group for 11 consecutive days and grouped under 4 holding periods based on the length of storage (2, 5, 8 or 11 d). The influence of energy regimes, egg holding intervals and their interaction was evaluated on fertility, hatch losses and hatchability. Broiler breeders maintained on SER regime (231-419 kcal/d) produced maximum number of eggs (993) followed by MER (819), CER (624) and EEF (438) during the 11-day period. The percent fertility and hatchability was significantly (p$\leq$0.05) higher in SER and MER groups compared to CER and EEF. However, energy regimes did not influence the loss in egg weight during pre-incubation storage, shell weight, shell thickness or hatch losses as dead germs and dead in shell. The improvement in hatchability in SER and MER groups appeared to be closely related to higher fertility and lower embryonic mortality. Holding of eggs for 11 days showed a linear loss in egg weight with the length of storage, but did not influence the fertility and hatch losses. The percent hatchability on eggs set was maximum when storage period was restricted to 5 days. The interaction between energy regimes and egg holding periods exhibited better hatchability results with SER regime when eggs were held for 5 days. Response to MER was not different from SER. It was obvious that energy restriction during production period had a positive influence on egg number, fertility and hatchability in aged breeders. At 64 weeks of age, holding of fertile eggs for 5 days prior to incubation was adequate for optimum hatchability in breeders.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.6
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• pp.832-840
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• 2014

This study aimed to investigate the effects of storage duration and temperature on the characteristics of wet brewers grains (WBG) as feeds for ruminant animals. Four storage temperatures ($5^{\circ}C$, $15^{\circ}C$, $25^{\circ}C$, and $35^{\circ}C$) and four durations (0, 1, 2, and 3 d) were arranged in a $4{\times}4$ factorial design. Surface spoilage, chemical composition and microorganism density were analyzed. An in vitro gas test was also conducted to determine the pH, ammonia-nitrogen and volatile fatty acid (VFA) concentrations after 24 h incubation. Surface spoilage was apparent at higher temperatures such as $25^{\circ}C$ and $35^{\circ}C$. Nutrients contents decreased concomitantly with prolonged storage times (p<0.01) and increasing temperatures (p<0.01). The amount of yeast and mold increased (p<0.05) with increasing storage times and temperatures. As storage temperature increased, gas production, in vitro disappearance of organic matter, pH, ammonia nitrogen and total VFA from the WBG in the rumen decreased (p<0.01). Our results indicate that lower storage temperature promotes longer beneficial use period. However, when storage temperature exceeds $35^{\circ}C$, WBG should be used within a day to prevent impairment of rumen fermentation in the subtropics such as Southeast China, where the temperature is typically above $35^{\circ}C$ during summer.

• Journal of Photoscience
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• v.4 no.2
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• pp.41-48
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• 1997

The plasma membrane and microsomes, isolated from the cells treated with hematoporphyrm derivative (HpD) for 1 and 24 h, accumulated the aggregated porphyrin. The quantity of aggregated porphyrin was same in the plasma membrane and microsomes after isolating them from cells treated with HpD for 1 h whereas the microsomes accumulated higher quantity of aggregated porphyrin when cells were treated with HpD for 24 h. Photodynamic action of aggregated porphyrin on plasma membrane and microsomes was investigated using lipid specific fluorescent probes: 1,6-diphenyl-1,3,5-hexatrine (DPH) and 1-(4-trimethylammonium), 6-diphenyl-1,3,5-hexatrine(TMA-DPH). The time dependent anisotropy of these probes in the membranes was measured and the decay of anisotropy was analyzed using wobbling in cone model. Upon irradiation both the plasma membrane and the microsomes showed an increase in the limiting anis~)tropy and order parameter and a decrease in the cone angle of the lipid probes. The increase in the limiting anisotropy was pronounced in membranes isolated from the cells treated with HpD for 24 h. Photoinduced change in the limiting anisotropy was dependent on the duration of incubation of cells with HpD before isolating the membranes. In both the membranes. the membrane core was affected more as compared to the outer leaflet. In addition to the structural changes, a decrease in Na$^+$-K$^+$-ATPase and NADPH cyt c reductase activity was also observed upon irradiation of HpD treated cells. Inhibition in NADPH cyt c reductase was more when cells were treated with HpD for 24 h, however, Na$^+$-K$^+$-ATPase activity did not depend on the duration of the treatment of cells with HpD before irradiation. Our results suggest that the extent of photoinduced perturbations in the membranes varies as a function of duration of the treatment of cells with HpD and the membrane core is more susceptible to the photodynamic action of aggregated porphyrin.

• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.578-582
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• 1998

In greenhouse studies, control efficacy of water chestnut (Eleocharis Kuroguwai) by Epicoccosorus nematosporus was affected by temperature and dew condition. The appressoria were formed abundantly in the range of 20~28$^{\circ}C$. When stem segments o(30 cm long) of the water chestnut were inoculated with the conidial suspension of E. nematorporus, the mean conidial number attacted amounted to 2,545 conidia. Out of 2545 conidia attacted to the stem pieces, 1,733 (68%) conidia formed appressoria. When these stem pieces were treated for 24 hr at 28$^{\circ}C$ under dew condition, 183,1 (7.2%) lesions were formed 10 days after incubation. The time necessary for the death of the plants was about 24 days. Appressoria were formed at 15~35$^{\circ}C$, but decreased rapidly in their numbers at the temperature lower than 1$0^{\circ}C$ or at 35$^{\circ}C$. The appressoria formation seemed to be depended on the dew duration, which was effective to the lesion formation and plant mortality. Under dew duration of 16~24 hr with temperature range of $25^{\circ}C$ to 3$0^{\circ}C$, the weed control was increased up to 93.9%. There were no differences between the first and second or third dew treatments. A delay of 2 or 3 days in dew treatment did not increase the mortality of plants. For the use of E. nematosporus as a mycoherbicide of water chestnut, a conidial suspension should be applied when dew conditions are kept for 12 hr after inoculation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.6
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• pp.461-467
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• 2005

Background. 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed (CFSE) is the fluorescent labelling agent of living cells and used to trace the cells in vivo after transplatnation of various cells. The CFSE labelled cells can maintain fluorescence for up to 7 days after labelling. The MC3T3-E1 cell line (MC3T3) has been used for many studies about osteoblast, which is well known as a mouse preosteoblast. So the CFSE would be used to trace the transplanted MC3T3. However there are few reports about CFSE labelling of MC3T3. This study is aimed to know about adequate concenturation and incubation time of CFSE to MC3T3. Materials and methods. The MC3T3 was incubated in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ using ${\alpha}$-minimal essential medium (${alpha}$-MEM) containing10% FBS and gentamycin. Ten mM CFSE solution in dimethylsulphoxide (DMSO: 1%) was diluted with phosphate buffered saline (PBS) and final concentration of culture medium was, respectively, 5, 10, 15, 20, 25 and 30 ${{\mu}M$. Then the MC3T3 was incubated with CFSE in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 and 45 minutes in each concentration. The fluorescence of CFSE labelled cells was analysed with a inverted fluorescence microscope. The duration of cell labelling was also studied. Trypan blue dye exclusion test was done for cell viability. Results. For concentration between 5 and 10 ${\mu}M$, CFSE did not significantly label the MC3T3 in vitro. The destruction of MC3T3 was observed at the concentration of 20 ${\mu}M$. In the concentration of 15 ${\mu}M$, the best labelling was obtained at an incubation period between 15 and 30 minutes. The MC3T3 labelled with an incubation period of 15 minutes at 15 ${\mu}M$ was still fluorescent 7 days after CFSE labelling. The mean cell viability was 95.93%. Conclusion. These results suggests an incubation period of 15 minutes at 15 ${\mu}M$ of CFSE provides best labelling of MC3T3 in vitro.

• Journal of Food Hygiene and Safety
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• v.6 no.2
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• pp.79-81
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• 1991

On May 23, 1990, an acute self-limited gastrointestinal illness was reported by twenty-seven persons. They were some of the employees and family members who attended a company open house picnic in Bergen County, New Jersey. Food questionnaires implicated that ziti was the vehicle of transmission (chi square 9.05). Median incubation period was 9.0 hours, and the median duration of illness 24 hours Clostridium perfringens organisms and enterotoxin were isolated from stool samples.