• Title/Summary/Keyword: Protein A

Search Result 30,038, Processing Time 0.059 seconds

Studies on the Maturation of the Follicular Oocytes by Xenoplastical Transplantation in the Anterior Chamber of the Eye (異種動物의 眼前房에 이식된 濾胞卵子의 成熟에 관한 연구)

  • Cho, Wan Kyoo
    • The Korean Journal of Zoology
    • /
    • v.13 no.3
    • /
    • pp.68-74
    • /
    • 1970
  • In the previous studies, the present author found that high proportion of the follicular oocytes from mouse and rabbit ovaries are able to resume their maturation division in the anterior chamber of the eye in which they have been incubated by auto- or homoplastic transplantation. Especially in the case of the homoplastic transplantation, it was known that no trouble has been detected in the process of resumption of the oval maturation in particular connection with the antigen-antibody reaction between donor and recipient. These findings provide a possibility that the follicular oocytes from various animals would be matured in the eye even after the xenoplastic transplantation. Under such an assumption, the present studies were performed to examine the behavior of the follicular oocytes in the eye chamber of the animals of different species. For the donor of the follicular oocytes, domestic rabbits, albino rats of Sprague-Dowley strain, and albino mice of A-strain bred in our laboratory were used. The oocytes obtained from the ovarian follicules were introduced to the anterior chamber of the eye of different species of animals, with an exception of rabbit in which only the female animals were used as a recipient. The procedures of collection of ova, introduction to the eye, harvest from the eye ball, fixation, and staining were the same as mentioned in the previous reports (Cho, 1967b; Cho and Kim, 1968). The conclusions obtained are summarized as below. 1. The rabbit follicular oocytes are able to mature in the eye chambers of both male mouse and rat, although the proportion of the maturation is lower than when they are incubated autoplastically in the eye. When the ova were incubated in the male mouse eye for 24 hours, 21 per cent of them showed chromosomes at metaphase I and II, whereas the rate was 32 per cent when they were incubated in the eye of the male rat. These are apparently low comparing to the rate of 52 per cent of autoplastic transplantation. 2. When rat follicular oocytes were transferred into the mouse eye chamber and recovered after 24 hours, 43 per cent of them produced the mataphase I and II chromosomes. This proportion was higher than the result of the homoplastic transplantation which yielded 23 per cent of the ova on maturation. 3. The most striking result was found in the experiment with mouse follicular oocytes. Seventy-six per cent of the oocytes resumed their maturation division within 24 hours after they were transferred into the male rat eye chamber, and this figure was significantly high compared to the result o 55 per cent obtained by the homoplastic transplantation. In the rat eye, the induction of the degenerative ova also was low (19%). On the other hand, the proportion of the oval maturation decreased to 45 per cent, while that of degeneration increased 33 per cent when they were incubated in the eye of the female rabbit. 4. It was apparent from the present experiments that the follicular oocytes can reveal their activation to maturation in the eye chamber which contains aqueous humor which is known to be composed of low protein content and of very little gamma-globulin which acts as an antibody(Oser, 1965), and that it shows higher osmolarity than blood serum(Levene, 1958). Taking these properities into consideration the humor may provide unfavourable environment to the cells and tissues incubated in. However, it could be noteworthy finding that only the follicular oocytes in the eye of the different species can grow in healthy condition although the maturation rates are varied with the animal species. The fact that the rabbit follicular oocytes show the lower proportion in maturation may be due to the greater amount of the yolk granules in the egg cytoplasm than those in the mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat resumed their maturation process in greater proportion would e explained by the fact that the rat eye chamber particularly provides the better environment to the mouse oocytes than the eye chamber of mouse does.

  • PDF

A Wide Region of Tropical Asia Adaptable Japonica Rice 'Asemi' (아시아 광지역 적응성 자포니카 벼 '아세미')

  • Jeong, Eung-Gi;kang, Kyeong-Ho;Hong, Ha-Cheol;Cho, Young-Chan;Jung, O-Young;Jeon, Yong-Hee;Chang, Jae-Ki;Lee, Jeom-Ho;Won, Yong-Jae;Yang, Un-Ho;Jung, Kuk-Hyun;Yeo, Un-Sang;Kim, Bo-Kyeong
    • Journal of the Korean Society of International Agriculture
    • /
    • v.31 no.1
    • /
    • pp.76-81
    • /
    • 2019
  • 'Asemi' is a rice variety derived from a cross between 'Jinmibyeo' which has translucent milled rice and medium maturity and 'Cheolwon46', an elite line with high yield and early maturity by the rice breeding team at NICS, RDA in 2013. The heading date of 'Asemi' is August 1, six days earlier than the check variety 'Hwaseong'. It has 82 cm culm length and 109 spikelets per panicle. 'Asemi' is resistant to blast disease, stripe virus and tungro virus, but susceptible to other viruses and planthoppers. The milled rice of this variety exhibits translucent, clear non-glutinous endosperm and short grain shape. It has protein content (6.7%) higher than 'Hwaseong', and amylose content (19.5%) similar to 'Hwaseong'. The milled rice recovery rate of 'Asemi' is similar to that of 'Hwaseong'. However, the head rice rate of 'Asemi' is higher than that of 'Hwaseong'. Milled rice yield of 'Asemi' is 5.23 MT/ha in ordinary cultivation. ' Asemi' could be adaptable to the wide region of tropical Asia (Registration No. 5639).

A Study on Rumen Fermentation Characteristics and Greenhouse Gas Emission of Forages in South Korea (국내 조사료별 반추위 내 발효특성과 온실가스 발생량에 관한 연구)

  • Young Ho Joo;Ji Yoon Kim;Myeong Ji Seo;Chang Hyun Baeg;Seung Min Jeong;Sam Churl Kim
    • Journal of The Korean Society of Grassland and Forage Science
    • /
    • v.43 no.4
    • /
    • pp.268-273
    • /
    • 2023
  • The present study estimated rumen fermentation characteristics and greenhouse gas emissions of different forages. Alfalfa, timothy, tall fescue, Italian ryegrass, and rice straw as the main forage sources for Hanwoo were used in the present study. Crude protein was highest in alfalfa but lowest in rice straw (p<0.05). Ether extract was higher in alfalfa and Italian ryegrass than in the other forages (p<0.05). Crude ash was highest in rice straw but lowest in tall fescue (p<0.05). Neutral detergent fiber was highest in tall fescue but lowest in alfalfa (p<0.05). Acid detergent fiber was highest in Italian ryegrass and rice straw but lowest in alfalfa (p<0.05). In vitro digestibilities of dry matter (DMD) and neutral detergent fiber (NDFD) were highest in timothy but lowest in rice straw (p<0.05). Rumen pH was highest (p<0.05) in alfalfa, while ammonia-N was higher (p<0.05) in alfalfa and Italian ryegrass than in the other forages. Total volatile fatty acid was highest (p<0.05) in timothy, while acetate and propionate were highest (p<0.05) in alfalfa and rice straw, respectively. Acetate to propionate ratio was higher (p<0.05) in alfalfa, timothy, and Italian ryegrass than in rice straw. Rice straw had lowest total gas (mL) (p<0.05) but highest its per DMD and NDFD. Rice straw had higher (p<0.05) CO2 (per DMD and NDFD) compared to alfalfa (per DMD and NDFD), timothy (per DMD and NDFD), tall fescue (per NDFD), and Italian ryegrass (per DMD). Again, rice straw had higher (p<0.05) CH4 (per DMD and NDFD) compared to timothy (per DMD and NDFD) and tall fescue (per NDFD). Therefore, this study indicates that timothy has a higher nutrient digestibility and volatile fatty acid in the rumen leading to a reduction of greenhouse gas emission.

Effects of the Processing Methods of Forage Rye (Secale cereale) on Rumen Fermentation Characteristics and Greenhouse Gas Emissions In Vitro of Hanwoo (호밀 조사료 가공방법이 한우의 반추위 내 발효특성과 온실가스 발생량에 미치는 영향)

  • Ji Yoon Kim;Seung Min Jeong;Young Ho Joo;Chang Hyun Baeg;Bu Gil Choi;Arrynda Rachma Dyasti Wardani;Sam Churl Kim
    • Journal of The Korean Society of Grassland and Forage Science
    • /
    • v.44 no.2
    • /
    • pp.99-105
    • /
    • 2024
  • This study was conducted to estimate the effects of the forage process on rumen fermentation characteristics and greenhouse gas emissions of rye. Rye was grown at the Taeyoung Livestock farm and harvested at the heading stage. The harvested rye (5 kg) was sub-sampled for fresh forage, hay, and silage in triplicates. The sub-sampled rye was freeze-dried or air-dried for fresh forage or rye hay, respectively. For rye silage, the sub-sampled rye forage was ensiled into a 10 L mini bucket silo and stored for 90 days. For 72 h rumen incubation, each forage (0.3 g) was placed into the incubation bottle with the rumen mixture (30 mL) in quadruplicates. After the incubation, total gas was measured and sub-sampled for CO2 and CH4 analyses, and the bottle content was centrifuged for in vitro digestibilities of dry matter (IVDMD) and neutral detergent fiber (IVNDFD), and rumen fermentation characteristics. Silage had higher crude protein, crude ash, and acid detergent fiber concentrations than fresh forage and hay but lower non-fiber carbohydrates and relative feed value (p<0.05). And, silage had higher lactic acid bacteria than the other forages but lower pH (p<0.05). After 72 h incubation in the rumen, fresh forage had higher IVDMD and butyrate content than the other forages (p<0.05). However, silage had higher rumen pH and propionate content than the other forages but lower A:P ratio (p<0.05). Regarding greenhouse gases, silage had lowest total gas (mL/g DMD and NDFD) and CH4 (mL/g DMD and NDFD) emissions, while fresh forage had lowest CO2 (mL/g DMD) emission (p<0.05). Therefore, this study concluded that the ensiling process of rye can effectively mitigate greenhouse gas emissions of Hanwoo.

Experimental Studies on Heat Conductivity of Human Bone and Torsional Strength of Pasteurized Porcine Tibia (생체골의 열전도성 및 열처리된 골의 염전력 변화에 대한 실험적 연구)

  • Park, Il-Hyung;Kim, Sin-Gun;Shin, Dong-Kyu;Ihn, Joo-Chul
    • The Journal of the Korean bone and joint tumor society
    • /
    • v.1 no.1
    • /
    • pp.7-16
    • /
    • 1995
  • In countries where confucianism is popular, it is extremely hard to get fresh cadaver bone for allograft. Therefore in Korea, the reimplantation of resected autoclaved autogenous bone segments has been increasingly performed for limb reconstruction of extremities with malignancies. To preserve the bone morphogenetic protein and mechanical strength of heated bone, many studies have reported that pasteurization of bone is far better than autoclaving over $100^{\circ}C$. Based on this assumption, replacement with a pasteurized autogenous bone graft after resection of a malignant bone tumor was deemed feasible for reconstruction. However, little is known about how high a temperature and how much time for pasteurization is needed to make tumors completely necrotic and to maintain the mechanical strength of bone. Consequantly, experimental studies were carried out to test heat conductivity of human bone and torsional strength of porcine tibia after pasteurization. First, two pairs of human proximal tibia and distal femur were used. We used T-type thermocoples to check core temperature of the bone and a computerized data acquisition system to record results. Without reaming of the medullary cavity, in a $60^{\circ}C$-thermostatic saline tub, it took 32 minutes and 50 seconds to raise the core temperature of human proximal tibia from $20^{\circ}C$ to $58^{\circ}C$, and 30 minutes for distal femur. In a $80^{\circ}C$ saline tub, it took 12 minutes and 50 seconds for proximal tibia, and 11 minutes and 10 seconds for distal femur. In contrast, using porcine tibia whose cortical thickness is similar to that of human tibia, after reaming of the medullary canal, it took less than 3 minutes and 30 seconds in a $60^{\circ}C$ saline tub, less than 1 minute and 45 seconds in a $70^{\circ}C$ tub, and less than 1 minute in a $80^{\circ}C$ tub to elevate core temperature from $20^{\circ}C$ to $58^{\circ}C$. Second, based on data of the heat conductivity test, we compared the torsional strength before and after pasteurization. Twenty matched pairs of porcine tibia were used, The left one was used as a non-heated control group and the right one as a pasteurized experimental group. Wighout reaming of the medullary cavity, there was no statistical difference in torsional strength between the pasteurization of the $60^{\circ}C$-35minute and of $80^{\circ}C$-15minute. With reaming, we also found no statistical difference among pasteurization of $60^{\circ}C$-15 minute, of $70^{\circ}C$-15 minute, and of $80^{\circ}C$-15 minute groups. In conclusion, reaming of the medullary canal is very helpful in saving pasteurization time. And, in a $60^{\circ}C$ saline tub, no significant weakness in torsional strength occurs with pasteurization of the bone for up to 35 minutes. Also no significant weakness in torsional strength occurs with an exposure of 15 minutes to the $80^{\circ}C$ saline tub.

  • PDF

Physicochemical Characteristics of Silky Fowl(Gallus domesticus var. silkies) (백봉오골계육의 이화학적 특성)

  • Cho, Chae-Min;Park, Chung-Kil;Lee, Min-Young;Lew, In-Deok
    • Food Science of Animal Resources
    • /
    • v.26 no.3
    • /
    • pp.306-314
    • /
    • 2006
  • In this study, the anatomic characteristics and the contents of nutritional ingredients and minerals of silky fowl and yeonsan ogolgye was investigated. Silky fowl is covered with white silky feather. There is a pinch of vertical white tassel on peak of the head, especially a silky fowl cock. The crown of a silky fowl has a nickname of phoenix crown, the crown of a cock is mostly like a rose, while that of a hen like strawberry or mulberry. The ear of silky fowl is mostly peacock green(Light blue turquoise), bronze for a small part. Peacock green is most common for Silky Fowl with a age of $60{\sim}150$ days, over the age of 150 days, the peacock green will be gradually replaced by purplish red. Beaks are leaden blue, short and stout while the face is smooth and fine. The lower jaw of silky fowl has comparatively long tiny hair, similar to beard. The two legs of silky fowl are covered with a handful of feather, or known as 'Putting on trousers'. Each leg of silky fowl has five talons. The whole skin, eyes, mouth, talons of silky fowl are grey black The bone and marrow of silky fowl are light black the periosteum is black. The whole meat, internal organs and abdominal fat of sillry fowl aye black the heart and leg meat are light black. Silky fowl meat had lower moisture and lipids content, but higher Ash and protein content than meats of yeosan ogolgye and general chickens. For mineral contents of leg muscle and breast muscle, silky fowl had higher contents of phosphorus (P), iron (Fe), potassium (K), zinc (Zn) than those of yeonsan ogolgye, while yeonsan ogolgye had higher contents of calcium (Ca) than that of silky fowl. The contents of iron (Fe) and zinc (Zn) is higher in leg muscle than in breast muscle for the silky fowl and yeonsan ogolgye. Leg muscle of silky fowl contains a lot of iron (Fe), about 4 times as much as that of leg muscle of yeonsan ogolgye. For the silky fowl and the yeonsan ogolgye, leg muscles contains a lot of zinc (Zn), about 5 times, 4 times respectively as much as that of breast muscle.

Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis (폐(肺)디스토마(Paragonimus westermani) 감염(感染) 고양이 혈청(血淸)에 대(對)한 ELISA 항체가(抗體價)의 의의(意義))

  • Choi, Won-Young;Yoo, Jae-Eul;Nam, Ho-Woo;Choi, Hyung-Rak
    • Parasites, Hosts and Diseases
    • /
    • v.24 no.2
    • /
    • pp.177-186
    • /
    • 1986
  • This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of $2{\mu}g/ml$ antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: 1. Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at $51{\sim}65%$ precipitate (PA2), followed by $0{\sim}50%$ precipitate (PAl), $66{\sim}80%$ precipitate (PA3), and $81{\sim}90%$ precipitate (PA4). Unprecipitated protein over 90% ammonium sulfate (PA5) showed the lowest antigenicity. 2. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. 3. By passing through the Sephadex G-200 column, PA1 and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2-I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. 4. SDS-polyacrylamide gel electrophoresis of PA1-I, P A1-II, PA2-I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons(KD) to 196 KD; of them 220KD protein was major band. Fraction PA2-I was composed of $255{\sim}225\;KD$, and PA3-I, $255{\sim}240\;KD$, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. 5. Absorbance by ELISA began to increase within $10{\sim}20$ days after the infection and reached the highest on $140{\sim}180$ days, then made plateau thereafter. 6. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. 7. Fraction PA1-II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I, fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at $20{\sim}30$ days after infection.

  • PDF

Studies on the chemical composition of citrus fruits in Korea(I) -The chemical composition of main varieties- (한국산(韓國産) 감귤류(柑橘類)의 화학성분(化學成分)에 관(關)한 연구(硏究)(I) -주요(主要) 감귤품종별(柑橘品種別) 화학성분함량(化學成分含量)에 관(關)하여-)

  • Yang, C.B.;Park, H.;Kim, Z.U.
    • Applied Biological Chemistry
    • /
    • v.8
    • /
    • pp.29-37
    • /
    • 1967
  • Citrus fruits of ten varieties grown in Chaeju island and a few other fruits for the comparison were analyzed to determined the contents of crude fat, crude fiber, total carbohydrates, macroelements and ash. Acids and sugars in fruit juices were also determined and the characteristic of neutralization of fruit juices were investigated. 1. The per cent of edible part of citrus fruits having the range of 48.5(Daingwoochi) to 72(Onju) were lower than that of other fruits. It was lower by about 8 per cent than that of the same variety produced in U.S.A. It was shown that the amount of rind per fruit might be increased in the citrus fruit grown in the low annual temperature. 2. The content of crude protein were around 1% and higher than other's. The contents of crude fat were below 0.1% in three varieties and over 0.1 in others. The contents of crude fiber were between 0.3 to 0.8% and the fruits with the high content of crude protein were inclined to have the high content of crude fiber and it was also shown that the low annual temperature was inclined to increase the amount of crude fiber per fruit. 3. The amount of total acid were from 19.5 m.e per 100 g of fresh fruit in Byongkyul to 44.2 m.e in Daingwoochi. The high percentage of titrable acid was over 90 in two sour varieties, Daingwoochi and Hakyul having pH below 3 and the high content of total acid. These two varieties were above 10 in the ratio of total acid in the edible part to the total acid in the rind(total acid in edible part/total acid in the rind). The content of combined acid was lower than that of titrable acid in the edible part and vice versa in the rind. 4. Navel was highest as 12.82% in the total sugar content and the lowest content was 4.9% of Hakyul. The contents of reducing sugar in the citrus fruits were about half of that in other fruits. The ratio of total sugar to titrable acid (sugar/acid: the grade of sweet taste) were lower than foreign products . 5. From the titration curves of fruit juices the characteristic of neutralization of juices could be grouped in three types, and other values, that is, pH, the content of total acid, the percentage of combined acid, the ratio of total acids in edible part and rind, the content of sugar, and the grade of sweet taste were also devided into the same three categories. 6. The contents of macroelements were different along to the each part of fruit. The content, in the seed were high and the ones in the rind were low. The contents of each element were in the order of $K_2O>N>P_2O_5{\gtrless}CaO{\gtrless}MgO$ in the edible part, $K_2O>N>CaO>P_2O_5{\gtrless}MgO$ in the rind, $N>K_2O>P_2O_5>CaO>MgO$ in the seed. The content of potassium was especially high in Marumeru and Hakyul and the content of calcium in citrus fruit was higher than others.

  • PDF

Lipopolysaccharide-induced Synthesis of IL-1beta, IL-6, TNF-alpha and TGF-beta by Peripheral Blood Mononuclear Cells (내독소에 의한 말초혈액 단핵구의 IL-1beta, IL-6, TNF-alpha와 TGF-beta 생성에 관한 연구)

  • Jung, Sung-Hwan;Park, Choon-Sik;Kim, Mi-Ho;Kim, Eun-Young;Chang, Hun-Soo;Ki, Shin-Young;Uh, Soo-Taek;Moon, Seung-Hyuk;Kim, Yang-Hoon;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
    • /
    • v.45 no.4
    • /
    • pp.846-860
    • /
    • 1998
  • Background: Endotoxin (LPS : lipopolysaccharide), a potent activator of immune system, can induce acute and chronic inflammation through the production of cytokines by a variety of cells, such as monocytes, endothelial cells, lymphocytes, eosinophils, neutrophils and fibroblasts. LPS stimulate the mononucelar cells by two different pathway, the CD14 dependent and independent way, of which the former has been well documented, but not the latter. LPS binds to the LPS-binding protein (LBP), in serum, to make the LPS-LBP complex which interacts with CD14 molecules on the mononuclear cell surface in peripheral blood or is transported to the tissues. In case of high concentration of LPS, LPS can stimulate directly the macrophages without LBP. We investigated to detect the generation of proinflammatory cytokines such as interleukin 1 (IL-1), IL-6 and TNF-$\alpha$ and fibrogenic cytokine, TGF-$\beta$, by peripheral blood mononuclear cells (PBMC) after LPS stimulation under serum-free conditions, which lacks LBPs. Methods : PBMC were obtained by centrifugation on Ficoll Hypaque solution of peripheral venous bloods from healthy normal subjects, then stimulated in the presence of LPS (0.1 ${\mu}g/mL$ to 100 ${\mu}g/mL$ ). The activities of IL-1, IL-6, TNF, and TGF-$\beta$ were measured by bioassaies using cytokines - dependent proliferating or inhibiting cell lines. The cellular sources producing the cytokines was investigated by immunohistochemical stains and in situ hybridization. Results : PBMC started to produce IL-6, TNF-$\alpha$ and TGF-$\beta$ in 1 hr, 4 hrs and 8hrs, respectively, after LPS stimulation. The production of IL-6, TNF-$\alpha$ and TGF-$\beta$ continuously increased 96 hrs after stimulation of LPS. The amount of production was 19.8 ng/ml of IL-6 by $10^5$ PBMC, 4.1 ng/mL of TNF by $10^6$ PBMC and 34.4 pg/mL of TGF-$\beta$ by $2{\times}10^6$ PBMC. The immunoreactivity to IL-6, TNF-$\alpha$ and TGF-$\beta$ were detected on monocytes in LPS-stimulated PBMC. Some of lymphocytes showed positive immunoreactivity to TGF-$\beta$. Double immunohistochemical stain showed that IL-1$\beta$, IL-6, TNF-$\alpha$ expression was not associated with CD14 postivity on monocytes. IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$mRNA expression were same as observed in immunoreactivity for each cytokines. Conclusion: When monocytes are stimulated with LPS under serum-free conditions, IL-6 and TNF-$\alpha$ are secreted in early stage of inflammation. In contrast, the secretion of TGF-$\beta$ arise in the late stages and that is maintained after 96 hrs. The main cells releasing IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ are monocytes, but also lymphocytes can secret TGF-$\beta$.

  • PDF

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
    • /
    • 2016.05a
    • /
    • pp.19-20
    • /
    • 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.

  • PDF