• 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.

• Journal of the Korean Vacuum Society
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• v.15 no.6
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• pp.563-575
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• 2006

Ti(C,N) films are synthesized by pulsed DC plasma enhanced chemical vapor deposition (PEMOCVD) using metal-organic compounds of tetrakis diethylamide titanium at $200-300^{\circ}C$. To compare plasma parameter, in this study, $H_2$ and $He/H_2$ gases are used as carrier gas. The effect of $N_2\;and\;NH_3$ gases as reactive gas is also evaluated in reduction of C content of the films. Radical formation and ionization behaviors in plasma are analyzed in-situ by optical emission spectroscopy (OES) at various pulsed bias voltages and gas species. He and $H_2$ mixture is very effective in enhancing ionization of radicals, especially for the $N_2$. Ammonia $(NH_3)$ gas also highly reduces the formation of CN radical, thereby decreasing C content of Ti(C, N) films in a great deal. The microhardness of film is obtained to be $1,250\;Hk_{0.01}\;to\;1,760\;Hk_{0.01}$ depending on gas species and bias voltage. Higher hardness can be obtained under the conditions of $H_2\;and\;N_2$ gases as well as bias voltage of 600 V. Hf(C, N) films were also obtained by pulsed DC PEMOCYB from tetrakis diethyl-amide hafnium and $N_2/He-H_2$ mixture. The depositions were carried out at temperature of below $300^{\circ}C$, total chamber pressure of 1 Torr and varying the deposition parameters. Influences of the nitrogen contents in the plasma decreased the growth rate and attributed to amorphous components, to the high carbon content of the film. In XRD analysis the domain lattice plain was (111) direction and the maximum microhardness was observed to be $2,460\;Hk_{0.025}$ for a Hf(C,N) film grown under -600 V and 0.1 flow rate of nitrogen. The optical emission spectra measured during PEMOCVD processes of Hf(C, N) film growth were also discussed. $N_2,\;N_2^+$, H, He, CH, CN radicals and metal species(Hf) were detected and CH, CN radicals that make an important role of total PEMOCVD process increased carbon content.

• Korean Journal of Environmental Agriculture
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• v.34 no.2
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• pp.91-97
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• 2015

BACKGROUND: Earthworms are essential detritus feeders that play a vital role in the process of decomposition of organic matter and soil metabolism. The complex process of partial breakdown of organic matter and mixing with mucous and gut microbial flora in the form of earthworm cast results in the reduction of the toxicity. This study focused on the change of cast amount and pollutant contents before and after the eating of the organic waste and upland soil with the two species of earthworm. METHODS AND RESULTS: The two species of earthworms were compared to the cast production. In the upland soil material, the daily amount of worm's cast was 1.42 g in E. andrei and 0.40 g in A. agrestis. In the organic waste material, the cast of E. andrei was 0.78~0.83 g and the cast of A. agrestis. have not been collected because all earthworms died after the treatment. The heavy metals treated in the upland soil were evaluated the impact of the worm excretion. With the E. andrei, the cast production was decreased 0.1~0.8 times in zinc, 0.2~0.5 times in copper, and 0.1~0.7 times in cadmium compared to the control treatment according to the levels of concentration. With A. agrestis, the cast amount was decreased 0.3~1.1 times in zinc, 0.2~0.3 times in copper, and 0.1~2.1 times in cadmium, respectively. The changes of pollutant contents before and after the eating of the organic wastes with E. andrei were studied. In the treatment of the Alcohol Fermentation Processing Sludge and the Fruit Juice Processing Sludge, heavy metal content of the cast was increased 0.7~53.3% compared to the sludge materials. PAHs contents were decreased 50.1% in the cast of the Alcohol Fermentation Processing Sludge and 36.6% in the cast of the Fruit Juice Processing Sludge, respectively. CONCLUSION: In conclusion, although the A. agrestis was bigger than E. andrei in size and weight, the cast amount of A. agrestis was small. The two species of earthworm was less excretion with high concentration of heavy metals. While the heavy metals such as zinc, copper, and cadmium were considerably accumulated in the cast, the total compounds, PAHs were fairly decomposed. There results would provide us for restoring contaminated soil and cleaning organic wastes.

• Journal of Applied Biological Chemistry
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• v.58 no.1
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• pp.39-50
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• 2015

Stabilization of heavy metals such as Pb, Cd, Zn, and Cu was evaluated in contaminated soil treated with poultry manure (PM) as well as its biochars pyrolyzed at $300^{\circ}C$ (PBC300) and $700^{\circ}C$ (PBC700) at the application rates of 2.5, 5.0, and 10.0 wt% along with the control, prior to 21-days incubation. After incubation, soil pH was increased from 6.94 (control) to 7.51, 7.24, and 7.88 in soils treated with PM 10 wt%, PBC300 10 wt%, and PBC700 10 wt% treatments, respectively, mainly due to alkalinity of treatments. In the soil treated with PM, the concentrations of the toxicity characteristic leaching procedure (TCLP)-extractable Pb, Cd, Zn, and Cu were increased by up to 408, 77, 24, and 955%, respectively, compared to the control. These increases may possibly be associated with an increased dissolved organic carbon concentration by the PM addition. However, in the soil treated with PBC700, TCLP-extractable Pb, Cd, Zn, and Cu concentrations were reduced by up to 23, 38, 52, and 36%, respectively, compared to the control. Thermodynamic modelling using the visual MINTEQ was done to predict the precipitations of $Pb(OH)_2$, $Cu(OH)_2$ and P-containing minerals, such as chloropyromorphite [$Pb_5(PO_4)_3Cl$] and hydroxypyromorphite [$Pb_5(PO_4)_3OH$], in the PBC700 10 wt% treated soil. The SEM-elemental dot mapping analysis further confirmed the presence of Pb-phosphate species via dot mapping of PBC700 treated soil. These results indicate that the reduction of Pb concentration in the PBC700 treated soil is related to the formations of chloropyromorphite and hydroxypyromorphite which have very low solubility.

• Journal of Dental Rehabilitation and Applied Science
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• v.25 no.3
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• pp.225-235
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• 2009

A common procedure of restoration of endodonticlly treated tooth with fiber-reinforced post is followed by core build-up after post cementation. However, this technique is complex and time-consuming. The aim of this study was to compare fracture strength of premolar, restored with various methods of core fabrications on fiber-reinforced posts and casting metal restoration. Forty five freshly extracted human mandibular premolars were obtained and devided into 5 groups acconding to the type of post and methods of core build-up. In Group A, D.T. $Light-post^{(R)}$ were cemented with $DUO-LINK^{TM}$ and then $LIGHT-CORE^{TM}$ was used for core restoration. In Group B, D.T. $Light-post^{(R)}$ and $DUO-LINK^{TM}$ were used for cementing in the postspace, and $DUO-LINK^{TM}$ was used again for core restoration. In Group C, $Light-post^{(R)}$ bonding and the core build-up were performed simultaneously by using $DUO-LINK^{TM}$. In Group D, $LuxaPost^{(R)}$ was bonded by using $LuxaCore^{(R)}-Dual$. Again, $LuxaCore^{(R)}-Dual$ was used for core restoration. In Group E, $LuxaPost^{(R)}$ bonding and the core build-up were performed simultaneously by using $LuxaCore^{(R)}-Dual$. Axial reduction was formed parallelly as possible and 45 degree bevel was made at buccal occlusal surface. Crowns were fabricated and cemented. Each tooth was embedded in self-curing acrylic resin to the level of 2mm below the CEJ. Specimens were fixed on universal testing machin such that the axis of the tooth was at 45 degree inclination to the horizontal plane, and compressive force was applied at a crosshead speed of 1mm/min until failure occurred. The mean fracture strength was the highest in group A followed by descending order in group B, D, E and C. However, there were no statistically significant differences between groups with regard to the fracture strength. The type of the post or build-up methods of the core does not seem to influence the fracture strength.

• Journal of Dental Rehabilitation and Applied Science
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• v.26 no.3
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• pp.241-251
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• 2010

This study aims at contributing to the restorative dentistry by examining results in the vertical load test of four different low invasive fixed partial dentures. Based on a hypothesis on the right upper first molar is missing, three units of FPDs were made for the second premolar and the second molar abutment. that is, twelve metal dies and FPDs were made for resin bonded FPD and Two Key Bridges and Human Bridge without occlusal rest and Human Bridge with occlusal rest. By using universal test machine, the numerical maximum value were recorded during the vertical load test of each FPDs after the bonding process treated by Maxcem which is resin cement. The failure process and its result of prosthesis were also observed. The maximum load was 7,295 N, 4,729 N, 2,190 N, 3,073 N from groups of resin bonded FPD, Two Key Bridge, Human Bridge without occlusal rest and Human Bridge with occlusal rest respectively. There was a statistical significance among the groups of resin bonded FPD, Two Key Bridge and Human Brides. However, there was no significant difference between Human Bridge without occlusal rest and Human Bridge with occlusal rest. Regarding the failure of prosthesis, the groups of Resin Bonded FPD and Two Key Bridge showed that one of the abutment teeth in the both side of retention part was highly failed earlier than the other one (83.2% and 66.6% respectively). While, Human Bridge without occlusal rest and Human Bridge with occlusal rest showed high percentage of failure in the abutment teeth in the both side of retention part at the same time (91.6% and 58.3% respectively). This study demonstrates that the group of Human Bridges has low resistance to the vertical loads of low invasive FPDs in comparison with the groups of resin bonded FPD and Two Key Bridge. Nevertheless, the maximum occlusal load of the restorative position, resistance to diverse restoration failure, amount of tooth reduction and patients' cooperation should be considered when they are applied in the clinic in order to choose an appropriate restoration for each patient.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2003.10a
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• pp.111-124
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• 2003

Processes that cause immobilization of contaminants in soil are of great environmental importance because they may lead to a considerable reduction in the bioavailability of contaminants and they may restrict their leaching into groundwater. Previous investigations demonstrated that pollutants can be bound to soil constituents by either chemical or physical interactions. From an environmental point of view, chemical interactions are preferred, because they frequently lead to the formation of strong covalent bonds that are difficult to disrupt by microbial activity or chemical treatments. Humic substances resulting from lignin decomposition appear to be the major binding ligands involved in the incorporation of contaminants into the soil matrix through stable chemical linkages. Chemical bonds may be formed through oxidative coupling reactions catalyzed either biologically by polyphenol oxidases and peroxidases, or abiotically by certain clays and metal oxides. These naturally occurring processes are believed to result in the detoxification of contaminants. While indigenous enzymes are usually not likely to provide satisfactory decontamination of polluted sites, amending soil with enzymes derived from specific microbial cultures or plant materials may enhance incorporation processes. The catalytic effect of enzymes was evaluated by determining the extent of contaminants binding to humic material, and - whenever possible - by structural analyses of the resulting complexes. Previous research on xenobiotic immobilization was mostly based on the application of $^{14}$ C-labeled contaminants and radiocounting. Several recent studies demonstrated, however, that the evaluation of binding can be better achieved by applying $^{13}$ C-, $^{15}$ N- or $^{19}$ F-labeled xenobiotics in combination with $^{13}$ C-, $^{15}$ N- or $^{19}$ F-NMR spectroscopy. The rationale behind the NMR approach was that any binding-related modification in the initial arrangement of the labeled atoms automatically induced changes in the position of the corresponding signals in the NMR spectra. The delocalization of the signals exhibited a high degree of specificity, indicating whether or not covalent binding had occurred and, if so, what type of covalent bond had been formed. The results obtained confirmed the view that binding of contaminants to soil organic matter has important environmental consequences. In particular, now it is more evident than ever that as a result of binding, (a) the amount of contaminants available to interact with the biota is reduced; (b) the complexed products are less toxic than their parent compounds; and (c) groundwater pollution is reduced because of restricted contaminant mobility.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.8
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• pp.1248-1256
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• 2014

The purpose of this study was to investigate the mechanisms of behind $SO_2$ formation and elevated cause of reducing power in purple bamboo salt (PBS) along with an analysis of physicochemical properties, content of sulfur compounds, oxidation reduction potential (ORP), mineral contents of salt type (MSS, mudflat solar salt; BS, bamboo salt), and addition of raw bamboo (RB). $SO_2$ content of 630 ppm was detected in PBS. $SO_2$ was not detected in MSS, BS, or RB, whereas $SO_2$ (782 ppm) from $K_2SO_4$ was detected after heating a NaCl, KCl, $MgCl_2$, $MgSO_4$, MgO, $CaCl_2$, $K_2SO_4$, and $FeSO_4$ with RB. $SO_2$ content of BS increased with baking time, and it originated from BSRB1 (13.88 ppm) to BSRB4 (109.13 ppm). $SO_3{^{2-}}$ originated only from MSSRB4 and BSRB2~BSRB4. Sulfate ion content decreased along with increasing $SO_2$ and sulfite ion contents. ORP increased with baking time of MSS and BS, and it was present at higher levels in BSRB4 (-211.40 mV) of BS than MSS. Insoluble content was higher in BS than MSS. Further, Ca, K, and Mg ion contents decreased in MSS and increased in BS with baking time. BSRB4 had 1.4 fold higher levels of Ca, 1.5 fold higher levels of Mg, and 1.8 fold higher levels of K than BS. Li, Al, Mn, Fe, and Sr in MSS as well as Al, Fe, and Ni in BS increased with baking time. Anions (Cl, $NO_3$, and Br) and heavy metals (Pb, Cd, Hg, and As) between MSS and BS were not significantly different. These results suggest that the reducing power of BS was due to $SO_2$ and sulfite ion. To increase the amounts of these compounds and reducing power, higher melting temperature and longer baking time are necessary along with BS, which is created by the addition of RB to roasted salt.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.4 s.54
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• pp.289-293
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• 2005

Nowadays there are many sun protection cosmetics including organic or inorganic UV filter as an active ingredient. Chemically stable inorganic sunsEreen agents, usually metal oxides, we widely employed in high SPF products. Titanium dioxide is one of the most frequently used inorganic UV filters. It has been used as pigments for a long period of cosmetic history. With the development of micronization techniques, it becomes possible to incorporate titanium dioxide in sunscreen formulations without whitening effect and it becomes an important research topic. However, there are very few works related to quantitations of titanium dioxide in sunscreen products. In this research, we analyzed amounts of titanium dioxide in sunscreen cosmetics by adapting redof titration, reduction of Ti(IV) to Ti(III) and reoxidation to Ti(IV). After calcification of other organic ingredients of cosmetics, titanium dioxide is dissolved by hot sulfuric acid. The dissolved Ti(IV) is reduced to the Ti(III) by adding aluminum metals. The reduced Ti(III) is titrated against a standard oxidizing agent, Fe(III) (ammonium iron(III) sulfate), with potassium thiocyanate as an indicator In order to test accuracy and applicability of the proposed method, we analyzed the amounts of titanium dioxide in four types of sunscreen cosmetics, such as cream, make-up base, foundation and powder, after adding known amounts of titanium dioxide $(1{\sim}25w/w%)$. The percent recoveries of the titanium dioxide in four types of formulations were in the range between 96 and 105%. We also analyzed 7 commercial cosmetic products labeled titanium dioxide as an ingredient and compared the results with those of obtained from ICP-AES (Inductively Coupled Plasma-Atomic Emission Spectrometry), one of the most powerful atomic analysis techniques. The results showed that the titrated amounts were well coincided with the analyzed amounts of titanium dioxide by ICP-AES. Although instrumental analytical methods, ICP-MS (Inductively Coupled Plasma-Mass Spectrometry) and ICP-AES, are the best for the analysis of titanium, it is hard to adopt because of their high prices for small cosmetic companies. It was found that the volumetric method presented here gat e quantitative and reliable results with routine lab-wares and chemicals.

• Journal of Mushroom
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• v.19 no.3
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• pp.210-215
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• 2021

For sterilization of microorganisms of the Listeria genus contaminating enoki mushroom, pilot mushroom grower equipped with air sterilization devices were developed. Sterilization experiments were performed using physical and chemical treatments. Internal temperature and humidity were controlled, maintaining 6.62℃±0.30 in the upper shelves, 6.46℃±0.24 in the middle shelves, and 6.48℃±0.25 in the lower shelves. Humidities were 79.97%±4.42, 79.43%±4.06, and 79.94±4.30%, respectively, with a temperature setting of 6.5℃, and a relative humidity of 75%. A suitable enoki mushroom cultivation stage for air sterilizer application was during the growth stage, with temperature in the 6.5~8.5℃ range, and humidity of 70~80%. At these same internal conditions, the ozone concentration in the mushroom cultivator was found to be 160 ppb during ion-cluster generator operation. After physical sterilization, the Listeria innocua survival rate was 0.1 to 0.9% using ion cluster sterilization, and 9.3 to 10.6% using UV air sterilization. The Listeria innocua survival rates on different materials were 9.3~10.6% on the metal specimen, and 9.9~16.2% on the plastic wrapper. The survival rate was particularly high on the rough side of the plastic wrapper. Ion cluster air sterilization is a labor-saving and effective method for suppressing the occurrence of Listeria bacteria on mushroom growers walls and shelves. For the plastic wrapper, chemical sterilization is more effective than physical sterilization.