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The study of bone density assessment on dental implant sites (임플란트 식립 부위의 골밀도 평가에 관한 연구)

  • Park, Su-Won;Jang, Soo-Mi;Choi, Byoung-Hwan;Son, Han-Na;Park, Bong-Chan;Kim, Chang-Hwan;Son, Jang-Ho;Sung, Iel-Yong;Lee, Ji-Ho; Cho, Yeong-Cheol
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.417-422
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    • 2010
  • Introduction: Bone density is one of the important factors for the long term success of endosseous implants. The bone density varies from site to site and from patient to patient. A preoperative evaluation of the bone density is quite useful to oral surgeons for planning dental implantation. More accurate information on the bone density will help surgeons identify suitable implant sites, thereby increase the success rate of dental implantation. This study examined the correlation between the bone density measured preoperatively by computed tomography (CT) and the implant primary stability measured by resonance frequency analysis. Furthermore, the effects of the implant sites, gender, age and generalized systemic disorder patients on the bone density and primary implant stability were examined. Materials and Methods: One hundred and fourteen patients were selected. None of the patients had undergone a tooth extraction or bone graft history in the previous year. Preoperatively, the patients underwent CT scanning to evaluate the Hounsfield unit (HU), and resonance frequency analysis (RFA) was used to evaluate the implant primary stability at the time of implant installation. All implants were 4.0 mm diameter and 11.5 mm length US II. All patients were recorded and the HU and implant stability quotient (ISQ) value were evaluated according to the sites, gender and age. Results: The highest HU values were found in the mandibular anterior site ($827.6{\pm}151.4$), followed by the mandibular molar site ($797{\pm}135.1$), mandibular premolar site ($753.8{\pm}171.2$), maxillary anterior site ($726.3{\pm}154.4$), maxillary premolar site ($656.7{\pm}173.8$) and maxillary molar site ($621.5{\pm}164.9$). The ISQ value was the highest in the mandibular premolar site ($81.5{\pm}2.4$) followed by the mandibular molar site ($80.0{\pm}5.7$), maxillary anterior site ($77.4{\pm}4.1$), mandibular anterior site ($76.4{\pm}11.9$), maxillary premolar site ($74.2{\pm}14.3$) and maxillary molar site ($73.7{\pm}7.4$). The mean HU and ISQ value were similar in females and males. (HU: P=0.331, ISQ: P=0.595) No significant difference was also found in the age group respectively. However, the correlation coefficients between the variables showed a closed correlation between the HU and ISQ value. Conclusion: These results showed close correlation between the bone density (HU) and primary stability value (ISQ) at the time of implant installation (Correlation coefficients=0.497, P<0.01). These results strengthen the hypothesis that it might be possible to predict and quantify the initial implant stability and bone density from a presurgical CT diagnosis.

Mobility of Nitrate and Phosphate through Small Lysimeter with Three Physico-chemically Different Soils (소형 라이시메터시험을 통한 토양특성에 따른 질산과 인산의 이동성 비교)

  • Han, Kyung-Hwa;Ro, Hee-Myong;Cho, Hyun-Jun;Kim, Lee-Yul;Hwang, Seon-Woong;Cho, Hee-Rae;Song, Kwan-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.4
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    • pp.260-266
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    • 2008
  • Small lysimeter experiment under rain shelter plastic film house was conducted to investigate the effect of soil characteristics on the leaching and soil solution concentration of nitrate and phosphate. Three soils were obtained from different agricultural sites of Korea: Soil A (mesic family of Typic Dystrudepts), Soil B (mixed, mesic family of Typic Udifluvents), and Soil C (artificially disturbed soils under greenhouse). Organic-C contents were in the order of Soil C ($32.4g\;kg^{-1}$) > Soil B ($15.0g\;kg^{-1}$) > Soil A ($8.1g\;kg^{-1}$). Inorganic-N concentration also differed significantly among soils, decreasing in the order of Soil B > Soil C > Soil A. Degree of P saturation (DPS) of Soil C was 178%, about three and fifteen times of Soil B (38%) and Soil A (6%). Prior to treatment, soils in lysimeters (dia. 300 mm, soil length 450 mm) were tabilized by repeated drying and wetting procedures for two weeks. After urea at $150kg\;N\;ha^{-1}$ and $KH_2PO_4$ at $100kg\;P_2O_5\;ha^{-1}$ were applied on the surface of each soil, total volume of irrigation was 213 mm at seven occasions for 65 days. At 13, 25, 35, 37, and 65 days after treatment, soil solution was sampled using rhizosampler at 10, 20, and 30 cm depth and leachate was sampled by free drain out of lysimeter. The volume of leachate was the highest in Soil C, and followed by the order of Soils A and B, whereas the amount of leached nitrate had a reverse trend, i.e. Soil B > Soil A > Soil C. Soil A and B had a significant increase of the nitrate concentration of soil solution at depth of 10 cm after urea-N treatment, but Soil C did not. High nitrate mobility of Soil B, compared to other soils, is presumably due to relatively high clay content, which could induce high extraction of nitrate of soil matrix by anion exclusion effect and slow rate of water flow. Contrary to Soil B, high organic matter content of Soil C could be responsible for its low mobility of nitrate, inducing preferential flow by water-repellency and rapid immobilization of nitrate by a microbial community. Leached phosphate was detected in Soil C only, and continuously increased with increasing amount of leachate. The phosphate concentration of soil solution in Soil B was much lower than in Soil C, and Soil A was below detection limit ($0.01mg\;L^{-1}$), overall similar to the order of degree of P saturation of soils. Phosphate mobility, therefore, could be largely influenced by degree of P saturation of soils but connect with apparent leaching loss only more than any threshold of P accumulation.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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The Neutralization Treatment of Waste Mortar and Recycled Aggregate by Using the scCO2-Water-Aggregate Reaction (초임계이산화탄소-물-골재 반응을 이용한 폐모르타르와 순환골재의 중성화 처리)

  • Kim, Taehyoung;Lee, Jinkyun;Chung, Chul-woo;Kim, Jihyun;Lee, Minhee;Kim, Seon-ok
    • Economic and Environmental Geology
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    • v.51 no.4
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    • pp.359-370
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    • 2018
  • The batch and column experiments were performed to overcome the limitation of the neutralization process using the $scCO_2$-water-recycled aggregate, reducing its treatment time to 3 hour. The waste cement mortar and two kinds of recycled aggregate were used for the experiment. In the extraction batch experiment, three different types of waste mortar were reacted with water and $scCO_2$ for 1 ~ 24 hour and the pH of extracted solution from the treated waste mortar was measured to determine the minimum reaction time maintaining below 9.8 of pH. The continuous column experiment was also performed to identify the pH reduction effect of the neutralization process for the massive recycled aggregate, considering the non-equilibrium reaction in the field. Thirty five gram of waste mortar was mixed with 70 mL of distilled water in a high pressurized stainless steel cell at 100 bar and $50^{\circ}C$ for 1 ~ 24 hour as the neutralization process. The dried waste mortar was mixed with water at 150 rpm for 10 min. and the pH of water was measured for 15 days. The XRD and TG/DTA analyses for the waste mortar before and after the reaction were performed to identify the mineralogical change during the neutralization process. The acryl column (16 cm in diameter, 1 m in length) was packed with 3 hour treated (or untreated) recycled aggregate and 220 liter of distilled water was flushed down into the column. The pH and $Ca^{2+}$ concentration of the effluent from the column were measured at the certain time interval. The pH of extracted water from 3 hour treated waste mortar (10 ~ 13 mm in diameter) maintained below 9.8 (the legal limit). From XRD and TG/DTA analyses, the amount of portlandite in the waste mortar decreased after the neutralization process but the calcite was created as the secondary mineral. From the column experiment, the pH of the effluent from the column packed with 3 hour treated recycled aggregate kept below 9.8 regardless of their sizes, identifying that the recycled aggregate with 3 hour $scCO_2$ treatment can be reused in real construction sites.

Production and Fuel Properties of Wood Chips from Logging Residues by Timber Harvesting Methods (목재수확 방법에 따른 벌채부산물 목재칩의 생산 및 연료 특성)

  • Choi, Yun-Sung;Jeong, In-Seon;Cho, Min-Jae;Mun, Ho-Seong;Oh, Jae-Heun
    • Journal of Korean Society of Forest Science
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    • v.110 no.2
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    • pp.217-232
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    • 2021
  • This study calculated the productivity and cost of extraction and processing of logging residues by cut-to-length (CTL) and whole-tree (WT) harvesting methods. In addition, the comparative analysis of the characteristics of wood chip fuel to examine whether it was suitable for the fuel conditions of the energy facility. In the harvesting and processing system to produce the wood chips of logging residues the system productivity and cost of the CTL harvesting system were 1.6 Gwt/SMH and 89,865 won/Gwt, respectively. The productivity and cost of the WT harvesting system were 2.9 Gwt/SMH and 72,974 won/Gwt, respectively. The WT harvesting productivity increased 1.3times while harvesting cost decreased by 18.7% compared to the CTL harvesting system. The logging residues of wood chips were not suitable for CTL wood chips based on International Organization for Standardization (ISO 17225-4:2021) and South Korea standard (NIFoS, 2020), but the quality (A2, Second class) was improved through screening operation. The WT-unscreened wood chips conformed to NIFoS standard (second class) and did not conform to ISO but were improved through screening operation (Second class). In addition to the energy facility in plant A, all wood chips except CTL-unscreened wood chips were available through drying processing. The WT-unscreened wood chips were the lowest at 99,408 won/Gwt. Plants B, C, and D had higher moisture content than plant A, so WT-unscreened wood chips without drying processing were the lowest at 57,204 won/Gwt. Therefore, the production of logging residues should improve with operation methods that improve the quality of wood chips required for applying the variable biomass and energy facility.

Construction of Genetic Linkage Map and Identification of Quantitative Trait Loci in Populus davidiana using Genotyping-by-sequencing (Genotyping-by-sequencing 기법을 이용한 사시나무(Populus davidiana) 유전연관지도 작성 및 양적형질 유전자좌 탐색)

  • Suvi Kim;Yang-gil Kim;Dayoung Lee;Hye-jin Lee;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.1
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    • pp.40-56
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    • 2023
  • Tree species within the Populus genus grow rapidly and have an excellent capacity to absorb carbon, conferring substantial ability to effective purify the environment. Poplar breeding can be achieved rapidly and efficiently if a genetic linkage map is constructed and quantitative trait loci (QTLs) are identified. Here, a high-density genetic linkage map was constructed for the control pollinated progeny using the genotyping-by-sequencing (GBS) technique, which is a next-generation sequencing method. A search was also performed for the genes associated with quantitative traits located in the genetic linkage map by examining the variables of height and diameter at root collar, and resilience to insect damage. The height and diameter at root collar were measured directly, while the ability to recover from insect damage was scored in a 4-year-old breeding population of aspen hybrids (Odae19 × Bonghyeon4 F1) established in the research forest of Seoul National University. After DNA extraction, paternity was confirmed using five microsatellite markers, and only the individuals for which paternity was confirmed were used for the analysis. The DNA was cut using restriction enzymes and the obtained DNA fragments were prepared using a GBS library and sequenced. The analyzed results were sorted using Populus trichocarpa as a reference genome. Overall, 58,040 aligned single-nucleotide polymorphism (SNP) markers were identified, 17,755 of which were used for mapping genetic linkages. The genetic linkage map was divided into 19 linkage groups, with a total length of 2,129.54 cM. The analysis failed to identify any growth-related QTLs, but a gene assumed to be related to recovery from insect damage was identified on linkage group (chromosome) 4 through genome-wide association study.

Studies on the Pulping Characteristics of Larchwood (Larix leptolepis Gordon) by Alkaline Process with Additives (첨가제(添加劑) 알칼리 법(法)에 의한 일본 잎갈 나무의 펄프화(化) 특성(特性)에 관(關)한 연구(硏究))

  • Lim, Kie-Pyo;Shin, Dong-Sho
    • Journal of the Korean Wood Science and Technology
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    • v.7 no.2
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    • pp.3-30
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    • 1979
  • Larch ($\underline{Larix}$ $\underline{leptolepis}$ GORDON), one of the major afforestation species in Korea in view of its growing stock and rate of growth, is not favored as a raw material for pulp due to its low yield of pulp and difficulties with bleaching arising from the high content of extractives in wood, and the high heartwood ratio and the active phenolics, respectively. The purpose of this study is to investigate the characteristics of firstly pulping with various additives of cellulose protector for the yield of pulp, and secondly bleaching with oxygen for chlotination-alkali extraction of five stage-sequence to reduce chlorine compounds in bleaching effluents. The kraft cooking liquor for five age groups of larchwood was 18 percent active alkali with 25 percent sulfidity and 5 : 1 liquor-to-wood ratio, and each soda liquor for sap-and heart-wood of the 15-year-old larchwood was 18 percent alkali having one of the following cellulose protectors as the additive; magnesium sulfate ($MgSO_4$, 2.5%), zinc sulfate ($ZnSO_4$, 2.5%), aluminium sulfate ($Al_2(SO_4)_3$, 2.5%), potasium iodide (KI, 2.5%), hydroquinone (HQ, 2.5%), anthraquinone (AQ, 0.1%) and ethylene diamine (EDA, 2.5%). Then each anthraquinone-soda liquor for the determination of suitable cooking condition was the active alkali level of 15, 17 and 19 percent with 1.0, 0.5 and 0.1 percent anthraquinone, respectively. The cooking procedure for the pulps was scheduled to heat to 170$^{\circ}C$ in 90 minutes and to cook 90 minutes at the maximum temperature. The anthraquinone-soda pulps from both heartwood and sapwood of 15-year-old larchwood prepared with 0.5 percent anthraquinone and 18 percent active alkali were bleached in a four-stage sequency of OCED. (O: oxygen bleaching, D: chlorine dioxide bleaching and E: alkali extraction). In the first stage oxygen in atmospheric pressure was applied to a 30 percent consistency of pulp with 0.1 percent magnesium oxide (MgO) and 3, 6, and 9 percent sodium hydroxide on oven dry base, and the bleached results were compared pulps bleached under the conventional CEDED (C: chlorination). The results in the study were summarized as follows: 1. The screened yield of larch kraft pulp did not differ from particular ages to age group, but heartwood ratio, basic density, fiber length and water-extractives contents of wood and the tear factor of the pulp increased with increasing the tree age. The total yield of the pulp decreased. 2. The yield of soda pulp with various chemicals for cellulose protection of the 15-year-old larchwood increased slightly more than that of pure soda pulp and was slightly lower than that of kraft pulp. The influence of cellulose protectors was similar to the yield of pulps from both sapwood and heartwood. The effective protectors among seven additives were KI, $MgSO_4$ and AQ, for which the yields of screened pulp was as high as that of kraft pulp. Considering the additive level of protector, the AQ was the most effective in improving the yield and the quality of pulp. 3. When the amount of AQ increased in soda cooking, the yield and the quality of the pulp increased but rejects in total yield increased with decreasing the amount of active alkali from 19 to 15 percent. The best proportion of the AQ seemed to be 0.5 percent at 17 percent active alkali in anthraquinone-soda pulping. 4. On the bleaching of the AQ-soda pulp at 30 percent consistency with oxygen of atomospheric pressure in the first stage of the ODED sequence, the more caustic soda added, the brighter bleached pulp was obtained, but more lignin-selective bleaching reagent in proportion to the oxygen was necessary to maintain the increased yield with the addition of anthraquinone. 5. In conclusion, the suitable pulping condition for larchwood to improve the yield and quality of the chemical pulp to the level for kraft pulp from conventional process seemed to be. A) the selection of young larchwood to prevent decreasing in yield and quality due to the accumulation extractives in old wood, B) the application of 0.5 percent anthraquinone to the conventional soda cooking of 18 percent active alkali, and followed, C) the bleaching of oxygen in atmospheric pressure on high consistency (30%) with 0.1 percent magnesium oxide in the first stage of the ODED sequence to reduce the content of chlorine compounds in effluent.

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Studies on the Physical and Chemical Denatures of Cocoon Bave Sericin throughout Silk Filature Processes (제사과정 전후에서의 견사세리신의 물리화학적 성질변화에 관한 연구)

  • 남중희
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.21-48
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    • 1974
  • The studies were carried out to disclose the physical and chemical properties of sericin fraction obtained from silk cocoon shells and its characteristics of swelling and solubility. The following results were obtained. 1. The physical and chemical properties of sericin fraction. 1) In contrast to the easy water soluble sericin, the hard soluble sericin contains fewer amino acids include of polar side radical while the hard soluble amino acid sach as alanine and leucine were detected. 2) The easy soluble amino acids were found mainly on the outer part of the fibroin, but the hard soluble amino acids were located in the near parts to the fibroin. 3) The swelling and solubility of the sericin could be hardly assayed by the analysis of the amino acid composition, and could be considered to tee closely related to the compound of the sericin crystal and secondary structure. 4) The X-ray patterns of the cocoon filament were ring shape, but they disappeared by the degumming treatment. 5) The sericin of tussah silkworm (A. pernyi), showed stronger circular patterns in the meridian than the regular silkworm (Bombyx mori). 6) There was no pattern difference between Fraction A and B. 7) X-ray diffraction patterns of the Sericin 1, ll and 111 were similar except interference of 8.85A (side chain spacing). 8) The amino acids above 150 in molecular weight such as Cys. Tyr. Phe. His. and Arg. were not found quantitatively by the 60 minutes-hydrolysis (6N-HCI). 9) The X-ray Pattern of 4.6A had a tendency to disappear with hot-water, ether, and alcohol treatment. 10) The partial hydrolysis of sericin showed a cirucular interference (2A) on the meridian. 11) The sericin pellet after hydrolysis was considered to be peptides composed with specific amino acids. 12) The decomposing temperature of Sericin 111 was higher than that of Sericin I and II. 13) Thermogram of the inner portioned sericin of the cocoon shell had double endothermic peaks at 165$^{\circ}C$, and 245$^{\circ}C$, and its decomposing temperature was higher than that of other portioned sericin. 14) The infrared spectroscopic properties among sericin I, II, III and sericin extracted from each layer portion of the cocoon shell were similar. II. The characteristics of seriein swelling and solubility related with silk processing. 1) Fifteen minutes was required to dehydrate the free moisture of cocoon shells with centrifugal force controlled at 13${\times}$10$^4$ dyne/g at 3,000 R.P.M. B) It took 30 minutes for the sericin to show positive reaction with the Folin-Ciocaltue reagent at room temperature. 3) The measurable wave length of the visible radiation was 500-750m${\mu}$, and the highest absorbance was observed at the wave length of 650m${\mu}$. 4) The colorimetric analysis should be conducted at 650mu for low concentration (10$\mu\textrm{g}$/$m\ell$), and at 500m${\mu}$ for the higher concentration to obtain an exact analysis. 5) The absorbing curves of sericin and egg albumin at different wave lengths were similar, but the absorbance of the former was slightly higher than that of the latter. 6) The quantity of the sericin measured by the colorimetric analysis, turned out to be less than by the Kjeldahl method. 7) Both temperature and duration in the cocoon cooking process has much effect on the swelling and solubility of the cocoon shells, but the temperature was more influential than the duration of the treatment. 8) The factorial relation between the temperature and the duration of treatment of the cocoon cooking to check for siricin swelling and solubility showed that the treatment duration should be gradually increased to reach optimum swelling and solubility of sericin with low temperature(70$^{\circ}C$) . High temperature, however, showed more sharp increase. 9) The more increased temperature in the drying of fresh cocoons, the less the sericin swelling and solubility were obtained. 10) In a specific cooking duration, the heavier the cocoon shell is, the less the swelling and solubility were obtained. 11) It was considered that there are differences in swelling or solubility between the filaments of each cocoon layer. 12) Sericin swelling or solubility in the cocoon filament was decreased by the wax extraction.. 13) The ionic surface active agent accelerated the swelling and solubility of the sericin at the range of pH 6-7. 14) In the same conditions as above, the cation agent was absorbed into the sericin. 15) In case of the increase of Ca ang Mg in the reeling water, its pH value drifted toward the acidity. 16) A buffering action was observed between the sericin and the water hardness constituents in the reeling water. 17) The effect of calcium on the swelling and solubility of the sericin was more moderate than that of magnecium. 18) The solute of the water hardness constituents increased the electric conductivity in the reeling water.

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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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