• Title/Summary/Keyword: Protein-based

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Clinical Analysis According to $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ Expression in Gastric Cancer (위암에서의 $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ 단백 발현)

  • Kim, Sin-Sun;Park, Yong-Geun;Jun, Kyong-Hwa;Jung, Hun;Song, Gyo-Young;Kim, Jin-Joo;Chin, Hyung-Min;Kim, Wook;Park, Cho-Hyun;Park, Seung-Man;Lim, Keun-Woo;Kim, Seung-Nam;Jeon, Hae-Myung
    • Journal of Gastric Cancer
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    • v.6 no.1
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    • pp.36-42
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    • 2006
  • Purpose: The $p21^{Waf1/Cip1}$ protein Inhibits the cell cycle by Inhibiting the phosphorylation at the $G1{\rightarrow}S$ check point, and the $p27^{kip1}$ protein similarly performs the suppressor function by controlling the p27-mediated G1 arrest. In this study, we analysed the clinical status and survival rates in correlations with p21 and p27 expression patterns in gastric cancer. Materials and Methods: Between 1993 and 1997, 192 patients who underwent surgeries in Catholic Medical Center were analysed retrospectively in this study. Immunohistochemical staining was performed and if the nuclei of the tumor cells were stained, we assumed those as positive results. Statistical analysis was based on clinicopathological findings and differences in survival rates. Results: The expression rate of p27 was 28.1% and 15.6% in p21 each. The ratio of T1-2(80.0%) was significantly high in p21 (+), but the ratio of T3-4 (50.6%) was slightly high in p21 (-). There was no statistical significance regarding other factors. The results in p27 was not much different from expression rate of p21 in T-stage. In addition, p27 expression in diffuse type (91.3%) was higher than in intestinal type (62.7%) by Lauren's classification (P<0.05). Also, there was no statistical significance in other factors. In the correlation of p21 and p27, p27 was positive when p21 was positive (53.5%). Conversely, p27 was negative when p21 was negative (76.5%, p<0.05). In the p21 and p27 combination test, there was higher rate of T1-2 (87.5%) in p21 (+)/p27 (+), and higher rate of T3-4 (58.1%) in p21 (-)/p27 (-) (P<0.05). Results showed higher rate of intestinal type (100%) in p21 (+)/p27 (+), and diffuse type (87.0%) was dominant in p21 (-)/p27 (-) (P<0.05) by Lauren's classification. Moreover, there was no statistical significance in the 5-year survival rate in the expression of p21 and p27, and the 5-year survival rate was highest in the case of p21 (+)/p27 (+) without statistical significance. Conclusion: In our study, $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ expressed similar patterns. The expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ affected the degree of invasiveness of the tumor, and. Combined examination result revealed the correlation of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ with Lauren's classification and depth of invasion of the tumor. However, we assumed that little difference between the survival rates depending on expression of $p21^{Waf1/Cip1}\;and\;p27^{kip1}$ has limited their value as predictable prognostic indicators.

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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A Study on the Characteristics of Dietary Behaviors and Food Intake Patterns of University Student according to the Obesity Index (대학생의 비만도에 따른 식생활 특성 및 식이섭취 양상에 관한 연구)

  • Oh Se-In;Lee Mee-Sook
    • The Korean Journal of Food And Nutrition
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    • v.19 no.1
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    • pp.79-90
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    • 2006
  • This study was performed to investigate the dietary behaviors and food intake patterns of university students according to the obesity index(OI). The subjects were 349 students(107 males and 242 females) who were assigned to one of the following groups based on the percentage of ideal body weight: underweight(OI< 90%), normal ($90%{\leq}Ol<10%$) and overweight($OI{\geq}110%$). The dietary behaviors and food intake patterns were evaluated by questionnaires and 24 hour recall method. The results were summarized as follows. The rates of underweight, normal, and overweight students were 33.8%, 61.0%, and 5.2% respectively. The ratios of underweight was higher than overweight, especially in case of female. The 60.46% of subjects responded that they had an irregular eating habits, especially in overweight group(72.22%). The 52.44% of subjects showed skipping mealtime more than one time per day. The overweight group had a tendency to overeat themselves than other groups. The normal group had less unbalanced dietary pattern than the other groups(p<0.0019). Most subjects had a snack(96.27%). The percentage of those who drank and smoked were 86.74% and 19.54%, respectively. The smoking value was significantly higher in the overweight group(p<0.0009). The food consumption frequency by food groups was not different among the groups except instant and fast food. More than 50% subjects consumed fish, legumes & its products, and vegetables everyday. There was significantly less rate of the instant and fast food consumption frequency in the normal group(p<0.0177). The 3/4 subjects that showed the lower consumed level in RDA(< 75%) were under-nutritional state in the Fe and Ca. In case of Ca, there was a higher NAR value in the overweight group(p<0.0257) significantly, and Fe, protein, vitamin $B_1$, vitamin $B_2$, and niacin showed similar tendencies. The INQ of Fe was significantly higher in the overweight group than other groups(p<0.0335).

A Preliminary Study on Depressive Symptoms and Glycemic Controls in Diabetic Patients (당뇨병 환자에서의 우울 및 관련증상에 관한 예비적 연구)

  • Ko, Seung-Hyun;Jeong, Jong-Hyun;Hong, Seung-Chul;Han, Jin-Hee;Lee, Seung-Pil;Ahn, Yoo-Bae;Song, Ki-Ho
    • Korean Journal of Psychosomatic Medicine
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    • v.12 no.2
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    • pp.165-173
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    • 2004
  • Objectives: Diabetes mellitus is a heterogeneous, chronic, progressive disease characterized by hyperglycemia and abnormality in protein, carbohydrate, fat metabolism. Recent studies have reorted two times prevalence of depression in individuals with diabetes compared to individuals without diabetics. This study was designed to investigate glycemic controls, anxiety, alexithymia, stress responses between depressed diabetic patients and non-depressed diabetic patients. Methods The subjects were 60 diabetic patients(mean age : $50.3{\pm}9.7$ years, 31 men and 29 women) who were confirmed to have diabetes depending on the laboratory findings as welt as clinical symptoms at the St. Vincent Hospital Diabetes Clinic, from Mar. 2004 to Sep. 2004. Laboratory test including, blood chemistry. glycated hemoglobin, urinalysis for proteinuria and Korean version of Beck Depression Inventory(BDI), State and Trait Anxiety Inventory(STAI), Toronto Alexithymia Scale(TAS) and Stress Response Inventory(SRI) were used for assessment. Based on BDI scores, all diabetics were divided into 13 depressed-diabetics group(above 20 point) and 47 non-depressed group(below 20 point). We compared demographic data. glycemic controls, STAI, TAS and SRI scores between two groups by independent t-test. Results : 1) Depressed diabetic groups were 13(mean age : $55.4{\pm}7.2$ years, 7 men and 6 women) and non depressed groups were 47(mean age $48.9{\pm}9.8$ years, 24 men and 23 women). In depressed diabetics, compared with non-depressed group, manifested aged(p=0.031), but other demographic data showed no difference between two groups. 2) No significant differences were noted in FBS, PP2h, Hb A1C, total cholesterol, HDL-cholesterol, SGOT/SGPT, BUN levels between depressed and non-depressed groups. But, blood creatine levels of depressed group were significantly increased than non-depressed group(p=0.026). 3) No significant differences were found in the score of STAI, STAI-S, STAI-T, TAS between depressed and non-depressed groups. 4) The SRI scores of depressed groups were significantly higher than non-depressed groups$(59.7{\pm}24.9\;vs.\;31.5{\pm}22.0)(p=0.000)$. Conclusion : The above results suggest that depressed diabetic patients are have more stress responses and higher blood creatine levels. However, there were no differences in laboratory data related to glycemic controls, and anxiety. alexithymia levels between two groups. We suggest that physicians should consider integrated approaches for psychiatric problems in the management of diabetes.

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A Hematological Study on Korean of Rural Community - Correllation on Blood Pressure, Hematocrit- (한국(韓國) 일부(一部) 농촌주민(農村住民)에 대(對)한 혈액학적(血液學的) 고찰(考察) -혈압치(血壓値)와 적혈구용적치관계(赤血球容積値關係)를 중심(中心)으로-)

  • Nam, Taik-Sung;Kang, Duk-Yong
    • Journal of agricultural medicine and community health
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    • v.2 no.1
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    • pp.36-42
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    • 1977
  • A study on blood pressure and hematocrit values of 1,559 people in 19 Korean rural areas was carried out in 1974 and the results were analyzed statistically. Obtained as follows: 1. The blood pressure according to sex and age groups (from the twenties to the seventies) was as follows: 1) The blood pressure of male by age group il) In the twenties, M (mean) was 125. 85/74, 15mm/Hg, a (standard deviation) was 15.9/10.2, and ill (standard error) was 1.55/0.99. (2) In the thirties, ${\delta}$ was 123.93/77.19 mm/Hg, a was 14.4/10.8, and m was 1.24/0.93. (3) In the forties, M was 128.44/81.15 mm/Hg, a was 23.9/14.7, and m was 2.16/1.33. (4) In the fifties, M was 128.48/181.24 mm/Hg, a was 24.7/13.9, and m was 2.05/1.16. (5) In the sixties, M was 135.80/81.70 mm/Hg, a was 27.4/18.8, and ${\delta}$ was 2.74/1.88. (6) In the seventies, M was 146.84/83. 16mm/Hg, ${\delta}$ was 24.5/10.0, and m was 5.62/2.30. 2) The blood pressure of female by age group (1) In the twenties, M was 117.89/73.33 mm/Hg, ${\delta}$ was 15.7/12.1, and m was 1.42/1.09. (2) In the thirties, M was 118.04/75.71 mm/Hg, ${\delta}$ was 16.9/13.0, and m was 1.13/0.87. (3) In the forties, M was 120.92/78.17 mm/Hg, ${\delta}$ was 20.9/12.9, and m was 1.42/0.87. (4) In the fifties, M was 122.14/79.55 mm/Hg, ${\delta}$ was 24.2/15.9, and m was 1.63/1.07. (5) In the sixties, M was 131.57/84.29 mm/Hg, ${\delta}$ was 28.4/16.9, and m was 2.58/1.53. (6) In the seven ties, M was 139.62/86, 54 mm/Hg, ${\delta}$ was 22.4/15.7, and m was 4.38/3.09. And the range of systolic blood pressure in male was 70~230 mm/Hg and in female was 80-230 mm/Hg. The range of distolic blood pressure in male was 50~160 mm/Hg and in female was 40~140 mm/Hg. 2. The hematocrit value according to sex and age groups was as follows: 1) The hematocrit values of male by age group (1) In the twenties, M was 42.72%, ${\delta}$ was 3.05, and m was 0.30. (2) In the thirties, M was 41.77%, ${\delta}$ was 3.29, and m was 0.28. (3) In the forties, M was 41.39, ${\delta}$ was 3.86, and m was 0.35. (4) In the fifties, M was 40.12%, ${\delta}$ was 3.65, and m was 0.30. (5) In the sixties, M was 39.88%, ${\delta}$ was 3.81. and m was 0.38. (6) In the seventies, M was 38.47%, ${\delta}$ was 2.27, and m was 0.52. 2) The hematocrit values of female by age group (1) In the twenties, M was 35.40%, ${\delta}$ was 3.37, and m was 0.30. (2) In the thirties, M was 35.50%, ${\delta}$ was 3. 35, and m was 0.22. (3) In the forties, M was 35.75%, ${\delta}$ was 3. 18, and m was 0.22. (4) In the fifties, M was 35.84%, ${\delta}$ was 3.30, and m was 0.22. (5) In the sixties, M was 35.70%, ${\delta}$ was 3.35, and m was 0.30. (6) In the seventies, M was 35.08%, ${\delta}$ was 3.08, and m was 0.60. The range of hematocrit values in male was 23~50% and in female was 18~50% (un associated with age groups). 3. In comparison with the blood pressure and the value of hematocrit of study groups showed that the blood pressure raised higher but the value of hematocrit got lowered on the contrary as the groups are getting older. 4. Total number of patients with hypertension was 165(10.6%) which were consisted with 71 male (11.3%) and 94 female (10.1%). But only two cases of the male patient and one case of the female patient were associated with protein uria. 5. The incidence of anemia by hematocrit values was as follows: 1) The incidence of male anemia patients based on$\leqq$41% ($\leqq$39%). (1) In the twenties, incidence was 43.90% (16.98%). (2) In the thirties, 41.48% (25.93%). (3) In the forties, 42.62% (25.41%). (4) In the fifties, 62.76% (40.69%). (5) In the sixties, 70% (38%). (6) In the seventies, 84.21% (73.68%). 2) The incidence of female anemia patients bailed on $\leqq$35% ($\leqq$34%). (1) In the twenties, incidence was 43.90% (37.39%). (2) In the thirties, 48.21 % (33.93%). (3) In the forties, 43.58% (32.57%). (4) In the fifties, 45.91 % (34.09%). (5) In the sixties, 47.11% (37.19%). (6) In the seventies, 50% (46.15%) 6. The incidence rate of anemia patients with hypertension was highest in the age group 51~60 in male and 31~40 in female.

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A survey on daily physical activity level, energy expenditure and dietary energy intake by university students in Chungnam Province in Korea (충남지역 대학생의 신체활동수준, 에너지소비량 및 에너지섭취량 조사)

  • Kim, Sun Hyo
    • Journal of Nutrition and Health
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    • v.46 no.4
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    • pp.346-356
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    • 2013
  • This study investigated the daily physical activity level, energy expenditure, energy balance, and body composition and their relationship with university students. The participants were 130 male students ($19.5{\pm}0.5$ yrs) and 139 female students ($19.5{\pm}0.3$ yrs) at a university in Chungnam province. Physical activity level was evaluated by an equation based on 24 hr-activity record and dietary nutrient intake was evaluated using the food record method during a three-day period consisting of two week days and one weekend. Body composition was measured using Inbody 430 (Biospace Co., Cheonan, Korea). As a result, mean body mass index (BMI) of subjects indicated that they had normal weight, however mean body fat ratio was $19.1{\pm}5.4%$ for males and $28.4{\pm}5.0%$ for females, indicating that they had higher than normal weight. Daily mean physical activity level was 1.55 for males and 1.47 for females, which was regarded as 'low active', respectively. Females had more light activity than males (p<0.01). Daily mean energy expenditure was $2,803.5{\pm}788.9$ kcal/d for males and $1,915.4{\pm}510.2$ kcal/d for females (p<0.001). Daily mean dietary energy intake was $2,327.0{\pm}562.5$ kcal/d for males and $1,802.1{\pm}523.6/d$ for females (p<0.001), and daily mean energy balance was $-476.5{\pm}955.9$ kcal/d for males and $-113.3{\pm}728.1$ kcal/d for females (p<0.01). Daily mean dietary intake of protein, vitamins, and minerals, except Ca, satisfied recommended nutrient intake. Daily energy expenditure was positively related to body weight (p<0.01), BMI (p<0.01), and fat free mass ratio (p<0.05), but was negatively related to body fat ratio (p<0.01). In conclusion, subjects had a negative energy balance and low physical activity. They had a normal weight by BMI but had a more fat than normal weight by body fat ratio. This appears to be related to their low physical activity. Thus, nutrition education should be provided for university students in order to increase their physical activity for maintenance of normal weight by body composition and health promotion.

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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Effects of Total Mixed Fermentation Feeds Based on Rice-straw and Six Forage Crops on the Productivity of Holstein Cows (청예사료작물과 볏짚 위주의 완전배합발효사료 급여가 Holstein 착유우의 생산성에 미치는 영향)

  • Lee, H. J.;Kim, H. S.;Ki, K. S.;Jeong, H. Y.;Baek, K. S.;Kim, J. S.;Cho, K. K.;Cho, J. S.;Lee, H. G.;Woo, J. H.;Choi, Y. J.
    • Journal of Animal Science and Technology
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    • v.45 no.1
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    • pp.69-78
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    • 2003
  • This experiment was carried out to evaluate the value of total mixed fermentation feeds(TMFF) as completely mixed ration and to observe the effect of various kinds of TMFF on the palatability, feed intake, and milk performance in Holstein cows. The dry matter (DM) content of TMFF used in the experiment was 23.98-28.42% range, and CP, TDN, ADF and NDF were 16.2${\sim}$19.2%, 58.3-65.1%, 34.4-39.6% and 46.9${\sim}$49.9% levels, respectively. The relative feed value (RFV) in rape-, alfalfa-, grass-, oat-, corn-TMFF groups were 138.6, 133.9, 116.5, 111.8, 111.4 and 108.1, respectively. Among these groups, RFV of rye-TMFF group was lowest. Dry matter disappearance(DMD) showed 0.8${\sim}$.9% to the all kinds of TMFF groups. The pH was 3.89${\sim}$.87 and $NH_3$-N concentration was 6.93-8.66 mg/$d\ell$. The acetic acid concentration in the raw material of TMFF showed low level of 0.19${\sim}$0.57%, lactic acid showed high level of 1.17${\sim}$3.21% and butyric acid was very high as 0.03${\sim}$0.32%. Therefore, these results provide evidence that the quality of TMFF was not so bad. In the daily fresh matter intake on the alfalfa-, grass-, rape-, corn-, oats- and rye-TMFF were showed 62.85, 60.48, 58.04, 57.11, 54.61 and 45.74 kg respectively. All TMFF showed high palatability as daily dry matter intake of 1.95 to 2.90% by body weight of experimental cows. Body condition score(BCS) was gradually increased in during 60 days of the experiment term. Average daily gain(ADG) showed about 140.0${\sim}$326.7g. In alfalfa-TMFF group, the ADG was higher than in the other groups (p<0.05). Also, the increase in BCS was observed in grass-TMFF group (3.07 to 3.34) and rye-TMFF group was decreased in 3.07 to 3.34 (p<0.05). The milk yield appropriately showed a range of 16.16${\sim}$18.95 kg in all groups. Among these groups, alfalfa-TMFF group was highest(P<0.05). Average milk fat contents showed high levels of 4.06${\sim}$4.79% and the level was high in order of rape-, grass-, corn-, alfalfa-, rye- and oats-TMFF. Milk protein was highest in forage-TMFF and level of lactose in milk was approximately 4.56% in overall groups. Solid non fat(SNF) and total solid(TS) contents were 8.75% and 12.8%, respectively. However, milk composition was not significantly affected by TMFF.