• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.317-325
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• 2001

This study investigated the effect of ferrous sulfate (Fe$^{2+}$) and/or ascorbic acid (Asc) on fertilizing ability in vitro of frozen-thawed boar spermatozoa. Using chlortetracycline (CTC) fluorescence, the spermatozoa was treated in preincubation medium with control, Fe$^{2+}$(1 mM), Asc (0.5 mM) and Fe$^{2+}$Asc to assessed for acrosome reaction, and the oocyte penetration test to determine whether the Fe$^{2+}$ and/or Asc can promote the penetration ability in vitro. When frozen-thawed spermatozoa was washed with preincubation medium, there were significantly (P ＜ 0.05) more acrosome-reacted in medium with Fe$^{2+}$Asc (38%) than control (27%). The penetration rates were also significantly (P ＜ 0.05) higher in medium with Fe$^{2+}$Asc (76%) than control (55%). Next, the lipid peroxidation of sperm was evaluated on the basis of malondialdehyde production following same treatments. The addition of Fe$^{2+}$Asc to sperm suspension increases the formation of malondialdehyde. However, there were not significantly different under the all conditions. The sperm suspension were also treated with control, Fe$^{2+}$, Asc and Fe$^{2+}$/Asc and assayed for sulfhydry1(-SH) group content. In the Fe$^{2+}$/Asc group, sperm-SH group were higher than another groups. In spermatozoa treated with Fe$^{2+}$ and/or Asc, however, no changes in sperm -SH-groups were detected when compared to controls. In another experiment, the activity of sperm binding to zona pellucida was evaluated through binding to salt-stored porcine oocytes. In control and Asc treatment groups, sperm binding to zona pellucida were significantly (P ＜ 0.05) higher than in medium with Fe$^{2+}$. On the other hand, there is not a significant increase in binding to zona pellucida with spermatozoa treated by Fe$^{2+}$/Asc. In summary, the present study suggests that Fe$^{2+}$/Asc causes an enhancement in fertilizing ability that is associated with penetration rate increased without change of spermatozoa binding capacity to homologous zona pellucida.o homologous zona pellucida.

• The Korean Journal of Nuclear Medicine Technology
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• v.25 no.1
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• pp.29-33
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• 2021

Purpose Plasma renin activity (PRA) test is important for the diagnosis of primary aldosteronism. PRA is an easily deformed substance in vitro and affected by temperature changes. Laboratory of ASAN medical center has consistently found that there was a difference between the initial and re-experimental results. We compared and analyzed the differences in PRA test results according to the sample storage status. Materials and Methods The measurement of PRA was performed by using the radioimmunoassay. From August to September 2020, 43 PRA re-test samples were tested with different sample storage condition. The first group was re-examined by freezing the plasma-separated samples at -18℃, and the second group was re-examined with refrigerated EDTA sample. Also, additional tests were conducted on 13 PRA samples to verify the effect on thawing temperature differences in plasma-separated samples. The same samples were divided into two parts and stored frozen at -18℃, respectively, and thawing samples in room temperature and those in refrigerator were were conducted. Each result was compared and analyzed based on the initial experimental results. Results The results of re-examination after frozen storing plasma separation samples showed a lower correlation than the results of re-examination with EDTA plasma samples in refrigerator. When calculating the percentage based on the initial test results, the average percentage of each was 404.9% and 133.8%. The correlation coefficient was also R=0.8501 and R=0.9966, respectively, showing a higher correlation between plasma in the refrigerated sample EDTA tube. In comparison experiments with differences in thawing temperature, average percentage of the results of initial test and room temperature thawing was 94.3% and the average percentage of the results of refrigerated thawing was 88.0%. After again freezing the sample, the average percentage of the second room temperature thawing result is 107.5%, and the second refrigerated thawing group is 112.7%. Both groups showed an increase from first thawing. Conclusion A comparative analysis of retesting according to differences in sample storage methods in PRA tests showed a higher correlation between the results of retesting of the refrigerated EDTA plasma. And repeated freezing and melting of plasma separation samples, regardless of temperature during defrosting, has been shown to affect results. Therefore, retest of PRA should re-collect plasma from original EDTA plasma to increase reproducibility.

• Applied Microscopy
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• v.16 no.2
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• pp.1-13
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• 1986

The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1273-1273
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• 2001

The efficiency of the luminal fermentation process influences overall efficiency of luminal production, animal health and reproduction. Ruminant production systems have a significant impact on the global environment, as well. Animal wastes contribute to pollution of the environment as ammonia volatilized to the air and nitrate leached to ground water. Microbial protein synthesis in the rumen satisfies a large proportion of the protein requirements of animals. Quantifying the microbial synthesis is possible by using markers for lumen bacteria and protozoa such as nucleic acids, purine bases, some specific amino acids, or by isotopic $^{15}N,^{32}P,\;and\;^{35}S$ labelled feeds. All those methods require cannulated animals, they are time-consuming and some methods are very expensive as well. Many attempts have been made to find an alternative method for indirect measurement of microbial synthesis in intact animals. The present investigations aimed to assess possibilities of NIRS for prediction of purine nitrogen excretion and ruminal microbial nitrogen synthesis by NIR spectra of urine. Urine samples were collected from 12 growing sheep,6 of them male, and 6- female. The sheep were included in feeding experiment. The ration consisted of sorghum silage and protein supplements -70:30 on dry matter basis. The protein supplements were chosen to differ in protein degradability. The urine samples were collected daily in a vessel containing $60m{\ell}$ 10% sulphuric acid to reduce pH below 3 and diluted with tap water to 4 liters. Samples were stored in plastic bottles and frozen at $-20^{\circ}C$ until chemical and NIRS analysis. The urine samples were analyzed for purine derivates - allantoin, uric acid, xantine and hypoxantine content. Microbial nitrogen synthesis in the lumen was calculated according to Chen and Gomes, 1995. Transmittance urine spectra with sample thickness 1mm were obtained by NIR System 6500 spectrophotometer in the spectral range 1100-2500nm. The calibration was performed using ISI software and PLS regression, respectively. The following statistical results of NIRS calibration for prediction of purine derivatives and microbial protein synthesis were obtained.(Table Omitted). The result of estimation of purine nitrogen excretion and microbial protein synthesis by NIR spectra of urine showed accuracy, adequate for rapid evaluation of microbial protein synthesis for a large number of animals and different diets. The results indicate that the advantages of the NIRS technology can be extended into animal physiological studies. The fast and low cost NIRS analyses could be used with no significant loss of accuracy when microbial protein synthesis in the lumen and the microbial protein flow in the duodenum are to be assessed by NIRS.

• Korean Journal of Food Science and Technology
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• v.10 no.2
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• pp.203-208
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• 1978

A study was made on the changes in Acid Value (AV) and Peroxide Value (POV) of the Mackerel ordinary muscle during low temperature storage. Three different samples were used-natural (control), ascorbic acid (As. A)-added and NaCl-added. The samples were stored at the temperature of $-15^{\circ}C$ during 75days immediately after quick freezing at $-23^{\circ}C$ The change in the amount of fatty acids was also measured by means of GLC method. 1. In comparison with the control, the As. A -added showed lower AV and POV, whereas the Nacl-added gave higher values. 2. Among the various fatty acids, which are constituents of the fish lipids, the amount of palmitic acid (16:0), stearic acid (18:0), and oleic acid (18:1) had been increased during the storage, while there had been steady decrease in the amounts of unsaturated fatty acid such as linolenic acid (18:3), arachidonic acid (20:4), clupanodonic acid (22:5), and docosahexaenoic acid (22:6). 3. For the first 30 days after freezing, the decreasing rates of the amounts of docosahexaenoic acid in three samples of the As. A-added, the control, and the NaCl-added-were 7%, 14% and 36% respectively. However, after 45 days these samples showed the same decreasing rates of more than 57 percent.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1649-1656
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• 2015

Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.13-13
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• 2001

This study was undertaken to evaluate the effects of catalase using xanthine (X) - xanthine oxidase (XO) system on in vitro maturation and fertilization in pig. When follicular oocytes were cultured in maturation medium with X and/or XO, the maturation rates were not significantly different between in medium with and without catalase despite of different culture periods. However, significantly (P<0.05) higher maturation rates were obrained in culture with X-XO system. The rates of degenerated oocytes were increased with culture periods prolonged, and were significantly (P<0.05) higher in medium without than with catalase at 120 h of culture. On the other hand, the parthenogenetic oocytes were observed with high proportions at 72 h of culture, hut were not different in medium with and without catalase at various times of culture. In another experiment, the frozen-thawed boar spermatozoa treated with X-XO system for in vitro fertilization. The penetration rates were higher in medium with that than without catalase during the in vitro fertilization with, none (P<0.05), XO and X＋XO. On the other hand, when sperm were treated with none, X, XO and X＋XO, lipid peroxidation were higher in medium without that than with catalase. However, the changes in sperm penetration and lipid peroxidation showed opposite patterns. The sperm suspensions were also treated with X and/or XO for assay of sulfhydryl (-SH) group content. Under the above all conditions, sperm-SH group were higher detected In medium with that than without catalase. The activity of sperm binding to zona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were higher than in medium with X, XO and X＋XO groups. No significant differences, however, were observed between medium with and without catalase. In conclusion, the exposure of follicular oocytes and spermatozoa to X-XO system may be caused stimulating in vitro maturation and fertilization in pig. This work was supported by grant No. 2000-1-22200-001-3 from the Basic Research Program of the Korea Science & Engineering Foundation.

• Journal of the Korean Professional Engineers Association
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• v.32 no.4
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• pp.104-118
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• 1999

Alaska Pollack (Theragramma), Mackerel (Socomber japonicus), yellow corvenia (Pseudosc iance manchurica) were dressed, and then meat was separated from the other parts through a fish meat separator. After dehydration, the meat was ground with a silent cutter, packaged in polyethylene bags, and stored at -15。C for days. Samples were taken at regular intervals throughout the storage period and investigated for changes in puality characteristics such as the amounts of nitrogenous compounds, degree of lipid oxidation, fatty acid composition, and organoleptic factors. The resrlts obtained in this study were as follows; 1. The volatile basic nitrogen[VBN] contents of Alaska pollack, and yellow corvenia meat pastes increased from 14.4, 11.2 and 10.8mg% to 41.6, 38.3 and 40.6mg%, respectively during a 120 day storage period, whereas the trimethylamine oxide nitrogen [TMAO-N] contents decreased from 117.2, 12.8 and 17.2mg% to 40.3, 2.6 and 7.1mg% during the same period. 2. The TBA value of the mackerel meat paste showed a maximum peak after 60 days, and then decreased gradually, whereas the TBAvalues of the alaska pollack and yellow corvenia meat pastes increased steadily during the same period. The acid values of the meat pastes increased during the storage period, while the iodine values decreased. 3. The fatty acid composition of the total lipid of the meat pastes changed considerably during ghe 120 days storage period : saturated fatty acids in the total lipid such as myristic, palmitic, and stearic acid increased, while unsaturated fatty acids such as linoleic, gadoleic, eicosapentaenoic, erucic and docosahexaenoic acid decreased steadily. The initial percentage contents of the unsaturated fatty acids in the total lipid of the Alaska pollack, mackerel, and yellow corvenia meat pastes were 87.2%, 63.9%, and 75.9% respectively. However, the contents decreased to 46.0%, 42.5% and 51.3% after the 120day storage period. 4. The color of the meat pastes changde gradually into dark brown. L values of the meat paste measured with a thistimulus colorimeter decreased steadily during the storage period, while a and b values increased during same period. 5. Judging from the results of organoleptic evaluation on the fish odor, color and overall acceptability, significant difference were found between the odor and color of the mackerel and those of the yellow corvenia meat pastes. Overall acceptability score of yellow corvenia was higher than that of Alaska pollack or mackerel meat pastes.

• Food Science and Preservation
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• v.23 no.2
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• pp.204-210
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• 2016

This study aimed to find an effective preservation method of lotus leaves for the preparation of lotus-leaf Yakju throughout the year. The characteristics of Yakju containing lotus leaves pretreated by blanching, steaming, roasting, and drying were investigated in this study. The chemical properties of Yakju (pH, amino acidity, alcohol content, and volatile acid content) were significantly different for all treatments, except for the total acid content. The polyphenol content ranging from 315.89 to 462.63 ppm (p<0.05) was significantly different depending on pretreatment method L color value for Yakju was not significantly different all treatments, while a value was significantly different in Yakju containing frozen stored leaves after pretreatment. The b value was lowest for the blanching treatment, and was significantly different for all treatments. Among the organic acids present in Yakju, the oxalic, citric, and malic acid contents were significantly different for all treatments. In sensory evaluation, lotus-leaf Yakjus containing fresh leaves and dried leaves were preferred to the others in color and flavor among Yakjus prepared with lotus leaves before cold storage. However, among the Yakjus prepared with lotus leaves after cold storage, that containing blanched leaves was preferred to the others in terms of flavor, taste, and overall acceptability.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.1
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• pp.37-45
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• 1975

The present study was ,conducted to evaluate the effects of condensed phosphates on the refeezing damage of Alaska pollack muscle. The fillets were dipped in such solution as 5 and $10\%$ sodium polyphosphate, 1 and $5\%$ mixture of sodium polyphosphate and sodium pyrophosphate (1:1, w/w) for 1 and 5 minutes, respectively, before refreezing. And fillets were frozen at $27^{\circ}C\~28^{\circ}C$ and stored for 15 days at $-18^{\circ}\~-20^{\circ}$. The degree of denaturation was estimated by determining amounts of drip relased, content of total solids, nitrogen, and DNA in the drip an cooking-weight-loss. Phosphorus absorbed in the muscle was also determined. Phosphorus absorbed in the fillets treated with loft solution of sodium polyphosphate for 5 minutes amounted to 101 mg/100g muscle as $P_2O_5$. The absorption was dependent on tile concentration of treating solution rather than on the dipping time. The increase of phosphorus absorption seemed to affect to reduction of drip. Among the treating conditions, $10\%$, 5 minutes and $10\%$ 1 minute with sodium polyphosphate appeared most effective ones on drip reduction. The effect of $5\%$, minutes with the mixture of sodium polyphosphate and sodium pyrophosphate did not show so benefitable effect in refrozen fillets. As a tendency total solids, nitrogen, and DNA in tile drip varied proportionally to the amount of drip released. And the content of DNA was lower than the amount. Treatment, at higher the concentration and longer the dipping time, resulted in the lower cooking-weight-loss and the better quality on organoleptic test of thawed fillets.