Park, Kee-Jai;Jung, Sung-Won;Kim, Jong-Hoon;Jeong, Jin-Woong
Applied Biological Chemistry
/
v.38
no.2
/
pp.141-146
/
1995
Changes of physicochemical properties of citron juice prepared by two different extraction methods, rotary-crushing and belt-pressing method, were investigated during the storage at $5^{\circ}C$ and $-20^{\circ}C$. Temperature drop of citron juice extracted by belt-pressing method was faster than that of citron juice prepared by rotary-crushing method and its freezing point was $0.8{\sim}0.9^{\circ}C$. During the storage, pH of stored citron juice with rotary-crushing method was increased up to 3.5 after 6 months storage while that of citron juice extracted by belt-pressing method was not changed significantly during the same storage time. Acidity of rotary-crushed citron juice was reduced a little more than that of belt-pressed citron juice during the storage. However, changes of soluble solid content were influenced largely by the storage temperature than by the extraction method. Contents of formol nitrogen and vitamin C were reduced remarkably in all of stored citron juice and $92{\sim}82%$ of farmol nitrogen and $72{\sim}43%$ of vitamin C were remained after 6 months of storage. Among the changes of color value, L values were reduced in the whole stored citron juice and a and b value had a different change pattern respectively according to the extraction and storage temperature. Changes in the content of both amino acid and fatty acid compositions was also observed after same storage period. Especially, in the case of change of fatty acid composition, content of linoleic acid and linolenic acid were reduced after 6 months storage, while those of palmitic acid, stearic acid and oleic acid were increased.
This study was carried out to determine the optimal condition for successful and efficient c cryopreservation of zygotes, 1-cell embryos, using EFS40 which was 40% (v/v) ethylene glycol diluted in DPBS medium containing 30% Fic-oll (w/v) and 0.3 M sucrose. After mouse zygote produced by IVF was vitrified by two freezing methods, the post-warming survival rates of 1-cell zygotes were assessed as cleavage to the 2-cell stage and development into the hatching blastocysts at 5 day. In the one-step method, when embryos were directly exposed to the vitrification solution at 25$^{\circ}C$ for 1 min., survival and development rates of zygotes were 85.5% and 31.9% In the two-step method, embryos were equilibrated with a dilute 20% EG for 1, 3, 5 min. before 1 min. exposure to EFS40, re-spectively. However, the rates of development (17.7, 3.3, 0%) were lower than that of one-step method. The highest survival rate (95.9%) was obtained by one-step method which exposes embryos in EFS40 for 30 sec. In this condition, 63. 8% of cleaved 2-cell developed into hatching blastocysts. In the cell number of Total and ICM using differential labelling technique, there are no significant differences in the cell number of Total and ICM between blastocysts devel oped in vitrified-thawed embryos (63.2${\pm}$16.9, 1 13.5${\pm}$4.0) and control balstocysts (54.0${\pm}$15.2, 1 12.3${\pm}$4.6). Therefore, these results show that mouse zygotes can be successfully cryopreserved by a simple vitrification method although developmental rates of vitrified embryos were reduced. In conclusion, this proposed vitrifi cation procedures can be useful in the cryopreservation of mouse IVF zygotes.
Retrogradation of non-waxy rice (NWR) and waxy rice (WR) cakes (45% moisture) stored at 5$^{\circ}C$, $25^{\circ}C$ and -2$0^{\circ}C$ was studied by differential scanning calorimetry (DSC) and enzymatic ($\beta$-amylase-puuulanase) method. With DSC, endotherms did not appear with rice cakes stored at room ($25^{\circ}C$) and deep freezing (-2$0^{\circ}C$) temperatures but did with samples stored at low temperature (5$^{\circ}C$), showing accelerated retrogradation by low temperature. Onset temperature (To) and peak temperature (Tp) did not change under 14 days at 5$^{\circ}C$ but enthalpy values ($\Delta$H) increased rapidly within one day and increased steadily until 5th day of storage, then equilibrated. Higher $\Delta$H were obtained with WR cakes than NWR cakes. It was suggested that more amylopectin recrystallization occured with WR than NWR. Degrees of gelatinization of rice cakes determined by enzymatic method increased in the following order: 5$^{\circ}C$ < $25^{\circ}C$ < -2$0^{\circ}C$. In contrast with DSC results, dogrees of gelatinization of NWR cakes, were relatively lower than that of WR cakes. However, increased retrogradation extents (melting enthalpies) caused reduced enzyme susceptibilities to $\beta$-amylase-pullulanase system, among NWR or WR cakes stored at 5$^{\circ}C$. The degrees of retrogradation of rice cakes stored at 5$^{\circ}C$ were higher than those stored at $25^{\circ}C$ and -2$0^{\circ}C$ without regard to the kind of rice. The higher sensitivity of the enzymatic method was obtained than that of DSC method when the degrees of retrogradation of rice cakes were determined during storage under this experiment conditions.
This study was to test whether in vitro matured Hanwoo oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5~10 min, exposed in EG30 for 30 sec, each oocyte was individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures 〔1.0 M sucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS〕 at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were pregnant and three of them were ongoing-pregnant by manual palpation at 250 days after transfer. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.
Disappearance rate of injected $D_2O$ from the arterial blood as well as the effect of histamine on the rate were studied in rabbits. The concentrations of $D_2O$ in the serial arterial samples obtained through a Polyethylene tubing inserted into the carotid artery were assayed by the freezing point elevation method of Reaser. At zero time 3 ml of isotonic $D_2O$ in normal saline was injected into the jugular vein and at the same time serial sampling of arterial blood started. The serial sampling interval was either 7.7 sec or 12.3 sec. In the histamine treated animals histamine diphosphate (0,5 mg of histamine base) was injected intravenously 30 minutes prior to the zero time. The following results were obtained. 1. $D_2O$ concentration in arterial plasma water, x, was empirically obtained by the sum of 2 exponential terms of time, $x=Ae^{-k1t}+Be^{-k2t},$ and its theoretical basis was sought. The first term of the right member of the equation was regarded to be attributable to the compartment P which possessed instantaneous exchange of water with plasma. The second term was postulated to represent the poorly exchangeable compartment. 2. The constant A of the equation was evaluated as 4,37% and 14.3% in the control and histamine treated groups, respectively. B was 1.19% in the control and 0.849% in histamine treated animals. 3. The disappearance rates determined were; $k_1=0.0519{\pm}0.0221\;sec^{-1}\;K_2=0.00454{\pm}0.00247\;sec^{-1}$ in the control group. $k_1=0.1137{\pm}0.0290\;sec^{-1}\;K_2=0.00499{\pm}0.00204\;sec^{-1}$ in the histamine group. 4. In the histamine treated animals the disappearance rate of the first term was larger than that of the control animals, suggesting an enlarged size of the rapidly exchangeable compartment with regard to the plasma water. On the other hand the constant B was decreased by histamine administration, suggesting a distribution of $D_2O$ in an enlarged volume. This view was also made clear by comparing the apparent asymptotes to which the concentration curves of $D_2O$ approached in respective groups. The asymptotes in the histamine treated group showed lower values.
The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{\circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{\circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{\pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{\pm}2.7%$ and Water: $27.5{\pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{\pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{\pm}3.2%$ and Water: $67.6{\pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{\pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{\pm}0.2%$ and Water: $0.5{\pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{\pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{\pm}1.8%$ and Water: $36.2{\pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{\pm}0.4%$) and all ($61.9{\pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{\pm}3.3%$, Water: $87.8{\pm}2.9%$ within live sperm and Tissue: $49.28{\pm}3.7%$, Water: $42.0{\pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.
Bearing capacity of pile foundations in cold region is dominated by adfreeze bond strength between surrounding soil and pile perimeter. It denotes that adfreeze bond strength is the most important design parameter for foundations in cold region. Adfreeze bond strength is affected by various factors like 'soil type', 'frozen temperature', 'normal stress acting on soil/pile interface', 'loading rate', 'roughness of pile surface', etc. Several methods have already been proposed to estimate adfreeze bond strength during past 50 years. However, most methods have not considered the effect of normal stress for adfreeze bond strength. In this study, both freezing temperature and normal stress have been controlled as primary factors affecting adfreeze bond strength. A direct shear box was used to measure adfreeze bond strength between sand and aluminum under different temperature conditions. Based on the test results, the relation between shear strength of frozen sand and adfreeze bond strength have been investigated. The test results showed that both of shear strength and adfreeze bond strength tend to increase with decreasing frozen temperature or increasing confining pressure. The ratio of shear strength and adfreeze bond strength, expressed as $r_s$, decreased initially frozen section but increased at much lower frozen temperature and there were uniform intervals under the different normal stress conditions. A method for predicting adfreeze bond strength using $r_s$ has finally been proposed in this study.
The intrauterine inseminator (IUI) was developed to provide the method of depositing dog semen into the uterine body instead of the vagina. The IUI consists of a vaginal endoscope, a balloon sheath, and injection catheter. When the endoscope is inserted into the vagina and the balloon expanded with air, the cervical os becomes visible so a injection catheter can be inserted through the cervix for deposition of the frozen-thawed semen. The efficacy of the IUI device was compared to intra-vaginal artificial insemination using semen that had been collected and frozen from pooled sperm-rich fraction of ejaculates collected from two Jindo dog donors. Aliquots of semen were extended with a Tris-egg yolk diluent, centrifuged, the seminal plasma removed, the pellet resuspended with the same diluent, and cooled to $5^{\circ}C$ over a 2 h period. A Tris-egg yolk-glycerol extender was added at $5^{\circ}C$; after 1 h, semen was loaded into 0.5 ml straws, and straws were frozen in LN vapor for 5 min, and immersed in LN for storage. The final sperm concentration for freezing was approximately $100{\times}10^{6}cells/ml$. The straws were thawed at $70^{\circ}C$ for precisely 6 sec, 1.5 ml Tris-egg yolk buffer at $38^{\circ}C$ added, and the 2 ml of thawed semen was used for a single insemination using the IUI device. Each bitch was inseminated at optimal insemination point, which was estimated by vaginal epithelial cells staining and progesterone concentration analysis. Use of the IUI device resulted in 21 of 26 females giving birth to 89 pups ($4.2{\pm}1.6$ pups per litter), while intra-vaginal AI resulted in 6 of 15 females whelping a total of 17 pups ($2.8{\pm}1.2$ pups per litter). We believe the IUI device is easier to use than previously described devices used for intrauterine insemination. In our experience the expansion of the balloon has a calming effect on the bitch that aids the inseminator. These results indicate that the IUI device was able to provide high fertility with 50 million frozen sperm per insemination and two inseminations.
Sixty one commercial seed samples of three cruciferous vegetable crops were examined for seed-borne Alternaria spp. by deep freezing blotter method. Alternaria raphani, A. brassicicola and A. brassicae were detected from 7, 6 and 1 seed samples of chinese cabbage out of 20 with average infection ratio of 3.8, 1.8 and 0.5, respectively. A. raphani and A. brassicae were detected from 11 and 2 seed samples of radish out of 38 with average infection ratio of 7.2 and 1.0, respectively. A. brassicicola was detected from 3 samples of cabbage seed with average infection ratio of 21.8. Most of the Alternaria spp. detected were recovered from the seed coat and caused seed rot and seedling infections. A. brassicicola was recovered from the embryos of radish, chinese cabbage and cabbage in low infections and A. raphani was recovered from the embryos of radish. In inoculation experiments, A. brassicicola, A. brassicae and A. raphani produced black leaf spots on old leaves of three creciferous vegetable crops and the spots gradually increased in size appearing gray to light brown lesions which later coalesce and caused defoliation.
This study was carried out to investigate properties of the bread prepared by applying emulsifiers to the frozen dough, Doughs made by the sponge and dough method with the sweet dough formula were quickly frozen at $-40^{\circ}C$ and stored for 6 weeks at $-20^{\circ}C$. The effects of emulsifiers on the number of yeast cells, the volume of the bread, the hardness and the quality evaluation were investigated after frozen doughs were thawed, fermented and baked every week. In the effect of the number of yeast cells, SSL 0.3% and DATEM 0,2% produced a more effective result than others during the freezing storage, The highest loaf volume was formed in bread supplemented with SSL 0,3% and DATEM 0,2%, In the moisture content, bread supplemented with SSL 0,5% showed the highest value, Bread supplemented with SSL 0,3% and DATEM 0,2% produced the lowest value of bread hardness and received the highest score in quality evaluation, In this study, the addition of SSL 0,3% and DATEM 0,2% in making frozen dough led to better bread quality as compared to others.
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