Metabolomics aims at the comprehensive, qualitative and quantitative analysis of wide arrays of endogenous metabolites in biological samples. It has shown particular promise in the area of toxicology and drug development, functional genomics, system biology and clinical diagnosis. In this study, analytical technique of MS instrument with high resolution mass measurement, such as time-of-flight (TOF) was validated for the purpose of investigation of amino acids, sugars and fatty acids. Rat urine and serum samples were extracted by selected each solvent (50% acetonitrile, 100% acetonitrile, acetone, methanol, water, ether) extraction method. We determined the optimized liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS) system and selected appropriated columns, mobile phases, fragment energy and collision energy, which could search 17 metabolites. The spectral data collected from LC/TOFMS were tested by ANOVA. Obtained with the use of LC/TOFMS technique, our results indicated that (1) MS and MS/MS parameters were optimized and most abundant product ion of each metabolite were selected to be monitorized; (2) with design of experiment analysis, methanol yielded the optimal extraction efficiency. Therefore, the results of this study are expected to be useful in the endogenous metabolite fields according to validated SOP for endogenous amino acids, sugars and fatty acids.
This study aimed to develop polyclonal antibodies to regional inedible adipocytes of Korean native cattle (Hanwoo) and investigate cross-reactivity of the antibodies. Patterns in plasma membrane proteins (PMPs) from abdominal and subcutaneous adipocytes of Hanwoo isolated by collagenase digestion were investigated using SDS-PAGE. As antigens, abdominal and subcutaneous adipocyte PMPs of Hanwoo were injected to sheep 3 times at 3 wk intervals for passive immunization, and non-immunized serum and antisera were collected before and after the injections. Titers of the antisera obtained from sheep and their cross-reactivities with heart, kidney, liver, lung, muscle, and spleen of Hanwoo were determined by ELISA. Isolation and culture of abdominal and subcutaneous adipocytes of Hanwoo were performed for analysing LDH concentration. Based on the SDS-PAGE analysis, specific proteins of PMPs in abdominal and subcutaneous adipocytes appeared despite rather similar patterns between both adipocytes. At the level of 1:1,000 dilution, little antibody reactivity appeared in non-immunized serum whereas the antisera had relatively strong reactivity up to the level of 1:128,000 and 1:64,000 dilution. These findings may indicate that strong antibodies against adipocyte PMPs can be developed using an immunological approach. Extremely low reactivities of abdominal and subcutaneous adipocyte antisera were detected with PMPs of the organs. Both antisera strongly reacted with each adipocyte PMPs and showed statistically (p<0.01) higher cross-reactivities compared with non-immunized serum. In conclusion, these results may indicate that the present polyclonal antibodies against regional inedible adipocyte PMPs are well developed and have safety in cross-reactivities with body organs. Further studies on in vivo cross-reactivity and fat reduction of the antibodies against abdominal and subcutaneous adipocytes PMPs of Hanwoo should be required for inedible fat-reduced high quality beef production.
Protective effects of natural components including genistein (4',5,7-trihydroxyisoflavone) from Glycine max MERRILL on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. Genistein $(10{\sim}100\;{\mu}m)$ suppressed photohemolysis in a concentration-dependent manner, and was more effective than the lipid peroxidation chain blocker, ${\alpha}$-tocopherol (Vit. E). Glycoside of genistein, genistin, the water-soluble antioxidant, L-ascorbate, and the iron chelator, myo-inositol hexaphosphoric acid dodecasodium salt (sodium phytate) did not exhibit protective effect against photohemolysis. L-Ascorbate and sodium phytate stimulated photohemolysis at high concentration $(500\;{\mu}m)$. ${\alpha}$-Carotene 3,3'-diol (lutein), a singlet oxygen $(^1O_2)$ quencher, exhibited pronounced protective effect, an indication that $^1O_2$ is important in photohemolysis sensitized by rose-bengal. Reactive oxygen scavenging activities $(OSC_{50})$ of natural antioxidants including genistein on reactive oxygen species (ROS) generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay were in the order of sodium phytate > L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. $OSC_{50}$ value of genistein, genistin, ${\alpha}$-tocopherol, L-ascorbate, and sodium phytate were 41.0, 109.0, 9.0, 5.2, and $0.56{\mu}m$ respectively. The order of free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ was L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. These results indicate that genistein can function as an antioxidant in biological systems, particularly skin exposed to solar UV radiation by scavenging $^1O_2$ and other ROS, and to protect cellular membranes against ROS.
Chon, Soon-Ho;Paik, Doo-Jin;Lee, Chul Burm;Kim, Hyuck;Chung, Won Sang;Kim, Young Hak;Kang, Jung Ho;Jee, Heng Ok
Tuberculosis and Respiratory Diseases
/
v.59
no.4
/
pp.397-405
/
2005
Background : Laminin-1 is known to have regular functions in the development and course of differentiation of the lungs. The morphogenesis and distribution of laminin-1 still remains as a mystery and its distribution and changes in the molecular structure of laminin-1 in the pathogenesis of the lung still is a subject of great controversy. In this study, experiments were done to delineate the distribution and changes in the amount of laminin-1 after inducing inflammation of the lungs by exposing experimental animals to CS gas and especially, to find compositions of laminin-1 within type II pneumocytes. Materials and Methods : The experimental subjects of study were newborn rats and the extracted tissue from the experimental rats were viewed under light microscope and electron microscope after the sections were treated with immunohistochemical methods and immunogold reaction methods using bounded gold particles. Results : 1) Lymphocytes and mononuclear phagocytes invaded the alveolar septa in the 2 day group rats after CS gas exposure and intense interstitial inflammation was seen in the 3 day group. 2) Laminin immunoreactions decreased to a moderate degree in the 2 and 3 day group rats after CS gas exposure and strong laminin immunoreactions were seen again in the 5 and 7 day group rats. 3) Gold particles in basal lamina of the lung blood-air barrier decreased and in the type I pneumocytes decreased in the 2 and 3 day group rats after CS gas exposure. 4) Gold particles were seen only on the surface of the cell membranes of type II pneumocytes of the 1 and 2 day group rats after CS gas exposure. 5) Few gold particles around the lamellar bodies and cytoplasm of type II pneumocytes in the control rat group and at 12 hours after CS gas exposure. Gold particles are seen only on the surface of type II pneumocytes of the 1 and 2 day group rats after CS gas exposure and are evenly distributed in small amounts in the cells of the 3 day group after CS gas exposure. Conclusion : CS gas exposure in the rats caused inflammation of lung alveolar septa and also induced a decrease in laminin-1 in basal lamina and loss of laminin-1 in the cytoplasm of type II pneumonocytes. As the inflammatory cells disappeared, an increase in the distribution of laminin-1 occurred. This reflects tissue regeneration functions of laminin-1 in the pneumocytes of rats and the distribution of laminin-1 in type II pneumocytes can be seen through the electron microscope using immunogold methods.
An experiment was conducted to investigate the effects of four water temperatures (10, 15, 20, and $25^{\circ}C$) in combination with three photoperiods (24L:0D, 12L: 12D, and OL:24D) on the oxygen consumption rate of juvenile dark-banded rockfish, Sebastes inermis (mean body weight $20.5{\pm}0.7g$). The oxygen consumption rates of S. inermis were measured in triplicate for 24 hours using a continuous flow-through respirometer. Different combinations of water temperatures and photoperiods resulted in significant differences in the mean oxygen consumption rate of S. inermis (P<0.001). The oxygen consumption increased with increasing water temperatures for all photoperiod treatments (P<0.01). Mean oxygen consumption rates at 10, 15,20 and $25^{\circ}C$ ranged $178.3\sim283.5,\;386.7\sim530.7,\;529.2\sim754.3$ and $590.0\sim785.5mg\;O_2kg^{-1}h^{-1}$, respectively. $Q_{10}$ values ranged $3.17\sim5.51$ between 10 and $15^{\circ}C,\;1.87\sim2.10$ between 15 and $20^{\circ}C$ and $1.08\siml.24$ between 20 and $25^{\circ}C$, respectively. Fish held in continuous darkness (OL:24D) used consistently less okygen than fish exposed to continuous light (P<0.05). The mean oxygen consumption offish in a 12L:12D photoperiod was higher than that offish in 24L:0D and 0L:24D photoperiods under all temperature treatments except $10^{\circ}C$. The oxygen consumption of fish exposed to the 12L:12D photoperiod was significantly higher during the light phase than during the dark phase under all temperature treatments except $10^{\circ}C\;(P<0.05)$. This study provides empirical data for estimating oxygen consumption of S. inermis under given condition. This result has application for culture management and bioenergetic model for growth of this species.
For veterinary imaging diagnosis, we obtained MR images of the canine brain, spine, kidney and pelvis from 3T MRI system which was equipped with the world first 3T active shield magnet. Spin echo (SE) and fast Spin Echo (FSE) images were obtained from the canine brain, spine, kidney and pelvis of normal and sick dogs using a homemade birdcage and transverse electromagnetic (TEM) resonators operating in quadrature and tuned to 128 MHz. In addition, we employed a homemade saddle shaped RF coil. Typical common acquisition parameters were as follows: matrix=512$\times$512, field of view (FOV)=20cm, slice thickness=3 w, number of excitations (NEX)=1. For T1-weighted MR images, we used TR=500 ms, TE=10 or 17.4 ms. For T2-weighted MR images, we used TR=4000 ms, TE=108 ms. Signal to noise ratio (SNR) of 3T system was measured 2.7 times greater than that of prevalent 1.57 system. The high resolution images acquired in this study represent more than a 4-fold increase in in-plane resolution relative to conventional images obtained with a 20 cm field of view and a 5 mm slice thickness. MR images obtained from 3T system revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter The present results demonstrate that the MR images from 3T system could provide better diagnostic quality of resolution and sensitivity than those of 1.5T system. The elevated SNR observed in the 3T high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the microscopic structural level under in vivo conditions. These images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods. Moreover, MRI technique could begin to apply for veterinary medicine in Korea.
Park, Seong-sik;Choi, Jae-young;Chung, Min-suk;Kim, Yi-suk;Lee, Je-man;Lee, Kyung-ae;Cho, Gyu-seon;Lee, Ji-young;Park, Eun-kyung
Journal of Sasang Constitutional Medicine
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v.10
no.1
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pp.81-99
/
1998
1. Background : Sasang Constitutional Medicine divides mankind into Taeyangin(太陽人), Taeumin(太陰人), Soyangin(少陽人) and Soeumin(少陰人) according to appearance, moral nature and characteristic of constitutional symptoms, and it is a medical science to study about different treatment for each constitution. In Sasang Constitutional Medicine, diagnosis of constitution was so important that we had the necessity of studying constitutional diagnostic method which had presented wholly and intuitively in Dongyi Soose Bowon(東醫壽世保元) with objective and analytic thinking. There were several attempts to classify Sasang Constitutional Types through morphological investigation of each parts of body. The one of them was to use measuring the body. In this study, we purposed to make clear whether the analysis of fingerprints and palm, one of the physical anthropologic methods would be helpful to classify Sasang Constitution. 2. Method : After practicing Questionnaire I and QSCC(Questionnaire of Sasang Constitution) II from 347 healthy Korean adults(242 males and 106 females), we analyzed fingerprints and palm prints of 208 people(142 males and 66 females) who were classified same constitution from two questionnaires. We analyzed the types of fingerprints according to the number of triradius and counted the number of dermal ridges from fingerprint center to triradius. We found triradius in the lower part of each finger and analyzed the courses of dermal ridges and counted the number of dermal ridges between each triradius. We mesured angle atd, and then researched relationship between fingerprints, palm prints and Sasang Constitution. 3. Result and Conclusion : The results of Sasang Constitution analysis of 208 Korean adults showed 76 Taeumin(36.5%), 81 Soeumin(39.0%) and 51 Soyangin(24.5%). The analytic results of fingerprints and palm prints are as follows : The types of fingerprints were helpful to sort Taeumin and the types of palm prints were helpful to sort Soyangin and Soeumin especially. In addition, the number of dermal ridges in fingerprints and palm prints were helpful to sort Soyangin. Fingerprints and palm prints have characteristic in each constitutions, so the investigation of fingerprints and palm prints seems to helpful to classify Sasang Constitution. We are planning to report fingerprints and palm prints of more subjects.
Kim, Jung-Eun;Kim, Woo-Yeon;Kim, Ji-Wook;Park, Hyun-Soo;Lee, Seung-Hoon;Lee, Soon-Young;Kim, Min-Ji;Kim, A-Reum;Park, Soo-Nam
Journal of the Society of Cosmetic Scientists of Korea
/
v.36
no.4
/
pp.303-314
/
2010
In this study, the antibacterial, antioxidative effect and component analysis of Pinus koraiensis leaf extracts were investigated. MIC values of the ethyl acetate fraction from P. koraiensis leaf extracts on P. acnes, S. aureus, P. ovale, and E. coli were 0.06 %, 0.25 %, 0.13 % and 0.50 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction on P. acnes, P. ovale. and S. aureus was more prominent. The aglycone fraction of P. koraiensis leaf extracts ($22.93\;{\mu}g/mL$) showed more higher free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. koraiensis leaf extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The 50 % ethanol extract ($0.70\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. Also the ethyl acetate ($1.04\;{\mu}g/mL$) and the aglycone fraction ($1.43\;{\mu}g/mL$) showed very high antoxidant activity. The protective effects of extract/fractions of P. koraiensis leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. koraiensis leaf extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). TLC and HPLC chromatogram of the ethyl acetate fraction obtained from hydrolysis of P. koraiensis leaf extracts revealed 2 main bands (PK-4, PK-6) and peaks (peak 1, peak 2), which were identified as kaempferol-3-O-glucoside (PK-6, peak 1) and kaempferol-3-O-arabinoside (PK-4, peak 2) by LC/ESI-MS/MS, respectively. These results indicate that extract/fractions of P. koraiensis can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membrane against ROS. Extract/fractions of P. koraiensis can be applicable to new cosmeceuticals for antioxidant, antiaging, and antibacterial activity.
This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.
SMZ is one of the most widely used antibacterial agents in veterinary medicine. It is also used as a growth promotant in many species of domestic animals There are marked species differences in its metabolism and pharmacokinetics. However, its pharmacokinetic and metabolism in rabbits. which are ragarded not only as good laboratorty animals hut also as good economical animals in its own, are lacking. Sex-differences in drug metabolism are well recognized in wide range of animal species including rats. Males are known to he more active than females. It is also know that there are Significant differences in the direction of metabolic pathways. But recently, female goats are reported to be more active in the metabolie capacity of SMZ than the other sex by Dutch researchers at Utrecht. Therefore, it is not easy to make general conclusicn of having higher SMZ metal-die capacity in the male compared to the opposite sex in every animal species. In this regard, the study on metabolic pattern of SMZ in rabbits, which are regarded as hervivorous, is of interest because the dietary habbits of rabbit are comparable to thai of goal, NEW Zealand White rabbits of each sex were given SMZ(35mg/kg) as a bolus injection into the marginalean, vein in order to study its pharmacokinetic profiles(using plasma) anc metabolic pattem(24h urine) as specified in the methods anc materials. 1. In the rabbit, the major metabolic pathway of SMZ was the acetylation(the formation of $N_4AcSMZ$). There were hydroxylation pathways(50HSMZ, $6CH_2OHSMZ$) as well, in the metabolism of SMZ in the rabbit, but minor pathways. 2. No sex differences in the metabolic direction of SMZ and its metabolites formation were found from the urinary excreted metabolites of SMZ out of 24h collected urine. 3. The concentration-time curves of SMZ(35mg/kg, iv) in the plasma compartment were fitted to a one-compartment open model when using a computer program(NONLIN). There was also no difference in the pharmacokinetic pattem of SMZ between two sexes. 4. The emergence of $N_4AcSMZ$ metabolized from SMZ was very fast in the plasma of the rabbit The elimination of $N_4AcSMZ$ was prolonged as compared to that of the parent drug Vie found no sex difference in the elimination pattern of $N_4AcSMZ$ in the rabbit.
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