• Title/Summary/Keyword: Benzoate

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Improved pregnancy rate and sex ratio in fresh/frozen in vivo derived embryo transfer of Hanwoo (Bos taurus coreanae) cows

  • Jihyun Park;Wonyou Lee;Islam M. Saadeldin;Seonggyu Bang;Sanghoon Lee;Junkoo Yi;Jongki Cho
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.779-791
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    • 2023
  • This study aimed to assess the effects of embryonic developmental stage, quality grade, and fresh or frozen/thawed conditions on the pregnancy rate and sex ratio of live offspring in Hanwoo (Bos taurus coreanae) cows. The quality and developmental stage of in vivo-derived (IVD) transferred embryos were evaluated using the standard criteria of the International Embryo Technology Society. The recipient cows were synchronized using conventional (estradiol benzoate and progesterone) protocols before embryo transfer. Embryos were transferred to 297 cows, and pregnancy was monitored for 60-70 days after embryo transfer. The pregnancy rates of fresh and frozen/thawed embryos were 56.90% and 52.49%, respectively. Pregnancy rates varied according to embryo quality (56.18% for grade 1 vs. 36.67% for grade 2). Pregnancy rates also varied by developmental stage and cryopreservation (67.86% vs. 63.49% for stage 4-1, 64.00% vs. 54.72% for 5-1, and 50.00% vs. 47.83% for 6-1, in fresh embryos vs. frozen/thawed embryos, respectively). For stage 7-1, the pregnancy rates were 72.73% for fresh embryos and 20.00% for frozen/thawed embryos. In 66 fresh embryos, the sex ratio of live offspring was 5:5, whereas it was 4(female):6(male) for frozen/thawed embryos among the 95 frozen/thawed embryos. The miscarriage rate was approximately 3% higher for frozen/thawed embryos than for fresh embryos (18.1% for fresh vs. 21.1% for frozen). Seasonal fertility rates were 33.3% in spring, 55.67% in summer, 52.8% in autumn, 60.0% in winter. The following male-to-female ratios were observed in different seasons: 6.7:3.3 in spring, 4.0:6.0 in summer, 5.5:4.5 in autumn, and 3.3:6.7 in winter. The current data revealed no significant differences in pregnancy rates between fresh and frozen/thawed IVD embryos. However, there was a lower pregnancy rate with advanced-stage frozen/thawed embryos (stage 7-1). The current study provides comprehensive results for the better optimization of embryo transfer in Hanwoo cattle to obtain the desired fertility rate, pregnancy rate, and sex ratio of calves. These results provide important insights into the factors that influence the viability and success of IVD embryo transfer in Hanwoo cows and may have practical applications for improving breeding programs and reducing production costs.

Effect of Additives on the Physicochemical Properties of Acetaminophen Liquid Suppository (아세트아미노펜 액상좌제의 물리화학적 특성에 미치는 첨가제의 영향)

  • Choi, Han-Gon;Jung, Jae-Hee;Ryu, Jei-Man;Lee, Mi-Kyung;Kim, In-Sook;Lee, Beom-Jin;Kim, Chong-Kook
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.290-295
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    • 1998
  • To optimize the formulation of acetaminophen liquid suppository, the effect of additives on the physicochemical properties of liquid suppository base was investigated. The physi cochemical properties of P 407/P 188 (15/15%) (abbreviated in 15/15) and P 407/P l88 (15/20%) (abbreviated in 15/20) were measured after the addition of following additives; 2.5% acetaminophen as an active ingredient, vehicle components (5% ethanol, 5% propylene glycol, 5% glycerin), preservatives (0.1% sodium benzoate, 0,1% methylparahydroxybenzoate, 0.1% propylparahydroxybenzoate) and 1% of sodium chloride as an ionic strength controlling agent. Poloxamer gel was prepared with three different buffer solutions (pH 1.2, 4.0 and 6.8) and the physicochemical properties, gelation temperature, gel strength and bioadhesive force, were determined. In the results, the effect of additives on the physicochemical properties was dependent on their bonding capacities including hydrogen bonding and cross-linking bonding. Because the hydrogen-bonding capacities of acetaminophen, ethanol and propylene glycol were smaller than that of poloxamer, the binding force of poloxamer gel became weak by their putting in between poloxamer gel. Therefore, the gelation temperature (15/15, $35.7^{\circ}C$ vs 37.0, 39.4 $38.2^{\circ}C$; 15/20, $29.2^{\circ}C$ vs 31.2, 32.0, $30.3^{\circ}C$) increased, and gel strength (15/15, 4.03 see vs 2.72, 2.08, 3.12sec; 15/20, 300g vs 50, 50, 200g) and bioadhesive force (15/15, $6.8{\times}10^2\;dyne/cm^2$ vs 3.2, 6.0, $6.0{\times}10^2\;dyne/cm^2$; 15/20, $97.3{\times}10^2\;dyne/cm^2$ vs 11.1, 89.5, $92.0{\times}10^2\;dyne/cm^2$) decreased. Furthermore, the binding force of poloxamer gel became strong due to the hydrogen-bonding capacities of glycerin and the cross-liking bonding of sodium salt. Then, the gelation temperature (15/15, 35.0, $32.1^{\circ}C$; 15/20, 26.0, $21.0^{\circ}C$) decreased, and gel strength (15/15, 6.51 see, 300g; 15/20, 500, 650g) and bioadhesive force (15/15, 7.2, $81.6{\times}10^2\;dyne/cm^2$; 15/20, 112.3, $309.2{\times}10^2\;dyne/cm^2$) increased. The effect of pH on the physicochemical properties of poloxamer gel was dependent on the ingredients with which the buffer solutions were prepared. Poloxamer gels prepared with pH 1.2 and 4.0 buffer solutions had the increasing gelation temperature (15/15, 37.5, $38.1^{\circ}C$; 15/20, 33.1, $34.0^{\circ}C$) and the decreasing gel strength (15/15, 2.98, 3.81sec; 15/20, 200, 200g) and bioadhesive force (15/15, $7.0{\times}10^2dyne/cm^2$; 15/20, $74.0{\sim}88.1{\times}10^2dyne/cm^2$) owing to HCl. Poloxamer gel prepared with pH 6.8 buffer solutions had the decreasing gelation temperature (15/15, $27.2^{\circ}C$; 15/20, $22.3^{\circ}C$) and the increasing gel strength (15/15, 400g; 15/20, 550g) and bioadhesive force (15/15, $207.0{\times}10^2dyne/cm^2$; 15/20, $215.0{\times}10^2dyne/cm^2$) due to the cross-linking bonding of $NaH_2PO_4\;and\;K_2HPO_4$.

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A Survey on the Application of Preservatives to Processed Food Types (보존료의 가공식품 유형별 사용 현황 연구)

  • Jeong, Eun-Jeong;Jin, Kyoung Nam;Choi, Hyeonjeong;Jeong, Yusang;Kim, Yong-Suk
    • Journal of Food Hygiene and Safety
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    • v.35 no.3
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    • pp.261-270
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    • 2020
  • The application of color retention agents (3 items), preservatives (17 items), and bleaching agents (6 items) as food additives in processed foods were investigated by food type. Among color retention agents, sodium nitrite was used the most with 257 cases, mainly in seasoned jeoktal (71.21%), ready-to-eat foods (7.78%), and breads (4.87%). Of the benzoates (1,236 cases) used as a preservative, sodium benzoate showed up most, in 1,215 cases, while 81.16% of these were in beverages such as beverage base (39.51%), mixed beverages (22.47%), and ginseng/red ginseng beverages (8.89%). Grapefruit seed extracts (3,291 cases) were applied to 44 types of processed foods such as sauces (54.65%), liquid tea (10.46%), and other products (5.15%). Ethyl p-hydroxybenzoate (2,957 cases) was applied to products (total 96.44%) such as sauces (92.15%), blended soy sauce (2.77%), and pickled foods (1.52%). Potassium sorbate was applied to a total of 789 cases, mainly pickled foods (40.43%) and processed fishery products (47.15%). All 27 cases of sorbic acid were applied to fish paste (100%). Of the bleaching agents, sodium bisulfite and sodium hydrosulfite were mainly used in confectioneries, breads or rice cakes, and potassium metabisulfite, sodium metabisulfite, and sulfur dioxide were mainly found in alcoholic beverages including fruit wine, while sodium sulfite was mostly used in pickled foods. These results are deemed useful in applying food additives to processed foods.

The effect of Antiseptics on the Galactolipid Metabolism of Chlorella ellipsoidea Chloroplast and Thylakoid Envelope (Chlorella ellipsoidea 엽록체막과 틸라코이드막의당지질 대사에 미치는 식품보존제의 효과)

  • 최은아;장재선;이종삼
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.221-231
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    • 1998
  • The biosynthesis of galactolipid and the composition of fatty acid in chloroplast and thylakoid envelope isolated from C. ellipsoidea treated with antiseptics (potassium sorbate: PS, sodium benzoate:SB, calcium propionate:CP) were analyzed. The contents of monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and total lipid in treatment with antiseptics were lower to compared with the control. The major fatty acid utilized for biosynthesis of MGDG in chloroplast envelope were palmitoleic acid (ave. 15.55%), oleic acid (ave. 15.09%) in control. Otherwise, the major fatty acids in P.S treatment were utilized for oleic acid (ave. 13.71%), linolenic acid (ave. 14.36%), palmitoleic acid (ave. 18.26%), oleic acid (ave. 17.26%) in S.B treatment, and oleic acid (ave. 16.88%), palmitoleic acid (ave. 16.31%) in CP treatment. It was showed that the major fatty acids in chloroplast envelope DGDG were oleic acid (ave. 15.75%), linolenic acid (ave. 17.74%) in control, oleic acid (ave. 14.90%), palmitoleic acid (ave. 15.97%) in P.S treatment, palmitoleic acid (ave. 13.29%), oleic acid (ave. 15.74%) in S.B treatment, and oleic acid (ave. 14.52%), palmitoleic acid (ave. 14.03%) in C.P treatment. The major fatty acid utilized for biosynthesis of MGDG in thylakoid envelope were linolenic acid (ave. 14.78%), oleic acid (ave. 12.90%) in control. Otherwise, the major fatty acids were utilized for palmitoleic acid (ave. 13.00%), palmitic acid (ave. 13.00%) in P.S treatment, palmitoleic acid (ave. 12.94%), oleic acid (ave. 12.43%) in S.B treatment, and oleic acid (ave. 12.43%), palmitoleic acid (ave. 12.43%) in C.P treatment. It was showed that the major fatty acids in thylakoid envelope DGDG were linolenic acid (ave. 18.01 %), oleic acid (ave. 15.53%) in control, linolenic acid (ave. 19.20%), linoleic acid (ave. 14.14%) in P.S treatment, palmitoleic acid (ave. 9.03%), oleic acid (ave. 14.85%) in S.B treatment, oleic acid (ave. 13.90%), linolneic acid(ave. 12.66%) in C.P treatment.

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Effects of PEG on Embryo Production in Superovulated Hostein Cows (젖소 과배란 처리시 PEG(Polyethylene Glycol) 처리가 수정란 생산에 미치는 영향)

  • Choi S. H.;Ryu I. S.;Han M. H.;Cho S. R.;Choe C. Y.;Kim H. J.;Son D. S.;Kim Y. K.;Lee J. W.
    • Journal of Embryo Transfer
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    • v.20 no.3
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    • pp.317-322
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    • 2005
  • This study was conducted to improve the efficiency of embryo recovery and to establish the protocols of superovulation in Holstein cows. Sixteen Holstein cows were used the test the efficacy of three superovulation regimens using Folltropin. In the case of regimen 1, CIDR plus with E2 capsule was inserted in cows at the random stage of estrous cycle and the total of 400 mg Folltropin V was adminstered twice a day for 4 days(Folltropin V group). In regimen 2, CIDR was inserted and 3.0 mg estradiol benzoate was administered i.m. next day and the total of 400 mg Folltropin was adminstered twice a day for 4 days(Folltropin V+EB group). For regimen 3, CIDR insertion was same as in the regimen 2 and the total of 400 mg Folltropin diluted with $10\%$ PEG 8,000 was administered once(Folttropin V+PEG 8,000 group). In all the regimens, CIDR were removed on 12th day and 45 mg dinoprost was administered i.m. simultaneously. The heat detected donors were administered 200 ug LH-RH and inseminated twice with 2 straws of frozen semen 12 hours apart. Embryo were collected using Foley catherter in each uterine homs on 6${\~}$8 days after inseminations. The evaluation of collected embryos were according to the IETS manual. The CL responses according to the superovulation treatments were 5.8, 20.6, 24.0 in the Folltropin V, Folltropin+EB and Folltropin V+PE 8,000 groups, respectively and there were significant different among the treatments(p<0.01). Transferable embyos collected were 3.6$\pm$2.4, 3.3$\pm$l.8 and 2.8$\pm$2.3, in the Folltropin V, Folltropin+EB and Folltropin V+PE 8,000 groups, respectively. Degenerated and unfertilized embryos in regimen 2 and 3 than regimen 1. These results indicates that superovulation treatments with both multiple injections and a single injection using PEG of Folltropin combined with CIDR insertion at the random stage of estrus cycle can be used to produce Holstein embryos.

Effect of Multiple Superovulation and Parity on Embryo Production in Hanwoo (한우의 반복 과배란 및 산차가 수정란 생산에 미치는 영향)

  • Choi S. H.;Ryu I. S.;Son D. S.;Cho S. R.;Han M. H.;Kim H. J.;Choe C. Y.;Kim Y. K.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.185-190
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    • 2005
  • This study was performed to investigate the effects of multiple superovulation and parity on embryo production in Hanwoo cows. Donors were superovulate 4 times $1\~2$ months interval and inserted CIDR plus (with the capsule of estradiol benzoate 10mg) on Day 10 from standing heat for 9 days and injected 2.5ml FSH (Antorin R-10) 2 times in a day on 6th day to 10th day from insertion of CIDR and the doses of FSH were decreasing 0.5ml on every 2 times. On 3th day of FSH injection, 25ml $PGF_2{\alpha}$ were injected i.m. and on 4th day, CIDR was removed. After 2 days from removing CIDR, AI was performed 2 times 12 hour apart with 2 straws of Korean Proved frozen Semen and simultaneously 200ug/ml GnRH was injected and embryos were recovered on 7th day from Al. The response rates of superovulated donors were $85.7\%,\;90.5\%,\;62.5\%,\;100\%$ from 1 to 4 times of superovulation, respectively. There were significant differences among No. of superovulation times (P<0.05). The results of transferable embryos were 3.7, 3.4, 3.4, 5.7 from 1 to 4 times of superovualtion, respectively. There were no differences among No. of superovulation times. The results of transferable embryos were 2.5, 3.0, 5.3, 3.0, 3.4 form heifer, first born to 4 the born, respectively. There were significant differences among the parities of donors (P<0.05). These results suggested that even 4 times of superovulations of Hanwoo donors could be able to recover transferable embryos, it might be used the donors maximally and improved the adaptation of embryo transfer to farms safely.

Risk Factors for the Probability of Pregnancy Following Synchronization Protocols in Dairy Cows (젖소에서 배란동기화 프로그램 적용 후 임신율에 영향을 미치는 요인 분석 연구)

  • Jeong, Jae-Kwan;Kang, Hyun-Gu;Jung, Young-Hun;Hur, Tai-Young;Kim, III-Hwa
    • Journal of Veterinary Clinics
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    • v.31 no.5
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    • pp.382-388
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    • 2014
  • The objective of this study was to determine the risk factors associated with pregnancy following 3 synchronization protocols in dairy cows. Data were collected on 1,952 cows from 22 dairy farms, including synchronization protocols ($PGF_{2{\alpha}}$ + estradiol benzoate [PG+EB], Ovsynch, and CIDR-ovsynch), cow parity, body condition score (BCS), and dates of previous calving, insemination and conception. The odds ratio (OR) for pregnancy were analyzed by logistic regression using the LOGISTIC procedure in SAS. The analysis revealed that farm (p = 0.005), cow parity (p = 0.0001), BCS (p < 0.005), and AI season (p < 0.05) significantly affected and calving to AI interval tended to affect (p < 0.1) the probability for pregnancy. Although synchronization protocols did not affect the probability for pregnancy (p > 0.05), cow parity and synchronization protocols showed a significant interaction (p < 0.005); the OR (0.60) was significantly lower (p < 0.0001) for multiparous cows compared to primiparous cows using PG+EB, whereas the OR (1.44) tended to be higher (p < 0.1) for multiparous cows compared to primiparous cows using the Ovsynch, and the probability for pregnancy did not differ between multiparous and primiparous cows using the CIDR-ovsynch (p > 0.05). Cows with BCS ${\geq}$ 3.00 were more likely pregnant (OR: 1.41) compared with cows having BCS ${\leq}$ 2.75, whereas cows inseminated during summer had a lower OR (0.73) compared with those inseminated during spring. Cows with a calving to AI interval > 150 days were more likely to be pregnant (OR: 1.20) compared with cows with a calving to AI interval ${\leq}$ 150 days. In conclusion, the OR for pregnancy following synchronization protocols in dairy cows was affected by farm, parity, BCS, calving to AI interval of the cow, and AI season, and there was a significant interaction between cow parity and synchronization protocols; the OR for pregnancy was lower for multiparous cows compared with primiparous cows using the PG+EB protocol.

Comparison of Spodoptera frugiperda Control Effects for Corn According to the Control Thresholds and Chemical Spraying Methods (열대거세미나방에 대한 옥수수의 요방제 수준 및 약제 살포방법에 따른 방제 효과 비교)

  • You Kyoung Lee;Hyun Ju Kim;Nak Jung Choi;Bo Yoon Seo;June Yeol Choi
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.25 no.3
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    • pp.142-150
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    • 2023
  • As global warming continues, the time of invasion of Spodoptera frugiperda has been advanced and the inflow rate has been increasing, leading to great increases in damage to crops. In this study, in order to minimize crop damage caused by S. frugiperda, the control period was set for corn fields through control thresholds, and the control effects according to the chemical spraying methods were investigated in forage corn filed. Even under the condition of 4% injury level during the corn silking stage, the damage rate of ear was 70%, showing an aspect of extensive damage. The economic injury level of S. frugiperda second instar larvae was shown to be 0.7 larvae per stalk, and the control threshold level was shown to be 0.6 larvae. The income was calculated by applying the corn wholesale unit price, and according to the result, even under the condition of injury level of 4%, there was a loss of KRW 895,221/10a, and the higher the injury level, the greater the decrease in income. To control S. frugiperda, the insecticidal effects of 10 single formulations registered for S. frugiperda were tested, and according to the results, four types(emamectin benzoate, chlorantraniliprole, indoxacarb, and spinetoram) showed high insecticidal activity not lower than 93.3%, and three types (chloran- traniliprole, spinetoram, and indoxacarb) were considered to be effective in controlling S. frugiperda as they showed high residual effects through insecticidal effect persistence tests. Therefore, conventional control and aerial control were conducted twice at 7-day intervals with indoxacarb SC and chlorantraniliprol WP, which show high activity against S. frugiperda, respectively, prior to the silking of forage corn. As a result, conventional control showed higher control values, 46.3%p in the case of indoxacarb SC and 21.7%p in the case of chlorantraniliprol WP, than aerial control through the primary control. In the secondary control too, higher control values of 26.7%p in the case of indoxacarb SC and 40.4%p in the case of chlorantraniliprol WP were found in conventional control than in aerial control. Therefore, it is considered necessary to prepare measures to improve the control effects in the recent situation where alternative methods for manpower control are widely used.

An Ultrastructural Study on the Development of the Knee Joint in the Human Fetus (인태아 슬관절 발육에 관한 전자현미경적 연구)

  • Kim, Baik-Yoon;Joo, Ki-Jung;Nam, Kwang-Il;Yoon, Jae-Rhyong
    • Applied Microscopy
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    • v.30 no.2
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    • pp.213-232
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    • 2000
  • The development of the knee joint was studied by electron microscopy in human fetuses ranging from 20 mm to 260 mm crown-rump length ($7\sim30$ weeks of gestational age). The appearance of the primordium of the meniscus and cruciate ligament was conspicuous as the mesenchymal cells , preceeding that of joint space at 30 mm fetus. The primitive joint cavity was first seen in the interzone from the 40 mm fetus and its intermediate layer proceeded developing as a narrow cleft which was closely incorporated with two chondrogenic layers. Poorly differentiated mesenchymal cells of the meniscus at 40 mm fetus containing predominantly free ribosomes differentiated into fibroblasts at 60 mm fetus. By 100 mm fetus, the fibroblast in inner zone of the meniscus presented as oval profiles with a short cell processes, whereas middle and peripheral zones presented as elongated cells. Differentiation of the synovial membrane coincided with clarification of the joint cavity When dilatation of the synovial cavity occurred, the two types of synovial cells were identified at 60 mm fetus. By 100 mm fetus a majority of the intimal cells were B-type. B-type cells were clearly distinguishable from A-type cells by their content of extensive rough endoplasmic reticula and well developed Golgi complexes. In contrast, A-type cells had numerous filopodia, pinocytotic vesicles, lysosomes and large vacuoles. At 260 mm fetus the B-type cells were also a majority of intimal cells. At 260 mm fetus the inner zone of the meniscus was filled with parallel oriented fascicles of collagenous fibers and oval fibroblasts. The middle zone was constituted of parallel and radially arranged fibers and fibroblasts. The outer zone was populated by elongated fibroblasts encircled by crossed collagenous fibers with the blood vessels. At 30 mm fetus the fibroblasts of the cruciate ligament contained rough endoplasmic reticula and mitochondria. Collagen fibrils were noted within narrow cytoplasmic processes which were continued with the extracellular space. Collagen fibrils of ligament were filled in the bulk of extracellular space at 100 mm fetus. By $150\sim260mm$ fetus, the cruciate ligaments were constituted of longitudinally oriented bundle of collagen fibrils with irregular rows of round cells between.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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