• Journal of Life Science
• /
• v.29 no.11
• /
• pp.1218-1226
• /
• 2019

The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.

• 한국균학회소식:학술대회논문집
• /
• 2015.05a
• /
• pp.45-46
• /
• 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%.

• Tuberculosis and Respiratory Diseases
• /
• v.58 no.5
• /
• pp.498-506
• /
• 2005

Background : This study compared the bronchodilator efficacy and safety of tiotropium inhalation capsules ($18{\mu}g$ once daily) with a ipratropium metered dose inhaler (2 puffs of $20{\mu}g$ q.i.d.) in patients with chronic obstructive pulmonary disease (COPD). Method : After the initial screening assessment and a two-week run-in period, patients received either tiotropium $18{\mu}g$ once daily or ipratropium $40{\mu}g$ four times daily over a period of 4 weeks in a double blind, double dummy, parallel group study. The outcome measures were the lung function, the daily records of the peak expiratory flow rate (PEFR), the patients' questionnaire, and the use of concomitant salbutamol. The forced expiratory volume in one second ($FEV_1$) and the forced vital capacity (FVC) were measured 5 minutes before inhalation, and 0.5, 1, 2 and 3 hours after inhaling the study drug on days 0, 14 and 28. Result : In 16 centers, 134 patients with a mean (SD) age of 66 (7) years and a predicted $FEV_1$ of 42 (12)% were analyzed. The trough $FEV_1$ response was significantly higher in the tiotropium group than in the ipratropium group after a four-week treatment period. The weekly mean morning PEFR of the tiotropium group was consistently higher than that of the ipratropium group during the 4-week treatment period with differences ranging from 12.52 to 13.88 l/min, which were statistically significant. Tiotropium was well tolerated by the COPD patients during the 4-week treatment period and had a similar safety profile to ipratropium. Conclusion : This study shows that tiotropium administrated once daily has a superior bronchodilator effect with a similar safety profile in treating COPD patients compared with ipratropium, inhaled four times daily.

• Journal of the Korean Wood Science and Technology
• /
• v.13 no.1
• /
• pp.19-62
• /
• 1985

This study was performed to investigate: (i) the bending processing properties of silk worm oak (Quercus acutissima Carr.) and Korean red pine (Pinus densiflora S. et Z.) by boiling and steaming treatments; (ii) the effects of interrelated factors - sapwood and heartwood, annual ring placement, softening temperature and time, moisture content. and wood defects on bending processing properties; (iii) the changing rates of bending radii after release from a tension strap, and (iv) the improving methods of bending process by treatment with chemicals. The size of specimens tested was $15{\times}15{\times}350mm$ for boiling and steaming treatments and $5{\times}10{\times}200mm$ for treatments with chemicals. The specimens were green for boiling treatments and dried to 15 percent for steaming treatments. The specimens for treatments with chemicals were soaked in saturated urea solution, 35 percent formaldehyde solution, 25 percent polyethylene glycol -400 solution, and 25 percent ammonium hydroxide solution for 5 days and immediately followed the bending process, respectively. The results obtained were as follows: 1. The internal temperature of silk worm oak and Korean red pine by boiling and steaming time was raised slowly to $30^{\circ}C$ but rapidly from $30^{\circ}C$ to $80-90^{\circ}C$ and then slowly from $80-90^{\circ}C$ to $100^{\circ}C$. 2. The softening time required to the final temperature was directly proportional to the thickness of specimen. The time required from $25^{\circ}C$ to $100^{\circ}C$ for 15mm-squared specimen was 9.6-11.2 minutes in silk worm oak and 7.6-8.1 minutes in Korean red pine. 3. The moisture content (M.C.) of specimen by steaming time was increased rapidly first 4 minutes in the both species, and moderately from 4 to 20 minutes and then slowly and constantly in silk worm oak, and moderately from 4 to 15 minutes and then slowly and constantly in Korean red pine. The M.C. of 15mm-squared specimen in 50 minutes of steaming was increased to 18.0 percent in the oak and 22.4 percent in the pine from the initial conditioned M.C. of 15 percent The rate of moisture adsorption measured was therefore faster in the pine than in the oak. 4. The mechanical properties of the both species were decreased significantly with the increase of boiling rime. The decrement by the boiling treatment for 60 minutes was measured to 36.6-45.0 percent in compressive strength, 12.5-17.5 percent in tensile strength, 31.6-40.9 percent in modulus of rupture, and 23.3-34.6 percent in modulus of elasticity. 5. The minimum bending radius (M.B.R.) of sapwood and heartwood was 60-80 mm and 90 mm in silk worm oak, and 260 - 300 mm and 280 - 300 mm in Korean red pine, respectively. Therefore, the both species showed better bending processing properties in sapwood than in heartwood. 6. The M.B.R. of edge-grained and flat-grained specimen in suk worm oak was 60-80 mm, but the M.B.R. in Korean red pine was 240-280 mm and 260-360 mm, respectively. Comparing the M.B.R. of edge-grained with flat-grained specimen, in the pine the edge-grained showed better bending processing property than the flat-grained. 7. The bending processing properties of the both species were improved by the rising of softening temperature from $40^{\circ}C$ to $100^{\circ}C$. The minimum softening temperature for bending was $90^{\circ}C$ in silk worm oak and $80^{\circ}C$ in Korean red pine, and the dependency of softening temperature for bending was therefore higher in the oak than in the pine. 8. The bending processing properties of the both species were improved by the increase of softening time as well as temperature, but even after the internal temperature of specimen reaching to the final temperature, somewhat prolonged softening was required to obtain the best plastic conditions. The minimum softening time for bending of 15 mm-squared silk worm oak and Korean red pine specimen was 15 and 10 minutes in the boiling treatment, and 30 and 20 minutes in the steaming treatment, respectively. 9. The optimum M.C. for bending of silk worm oak was 20 percent, and the M.C. above fiber saturation point rather degraded the bending processing property, whereas the optimum M.C. of Korean red pine needed to be above 30 percent. 10. The bending works in the optimum conditions obtained as seen in Table 24 showed that the M.B.R. of silk worm oak and Korean red pine was 80 mm and 240 mm in the boiling treatment, and 50 mm and 280 mm in the steaming treatment, respectively. Therefore, the bending processing property of the oak was better in the steaming than in the boiling treatment, but that of the pine better in the boiling than in the steaming treatment. 11. In the bending without a tension strap, the radio r/t of the minimum bending radius t to the thickness t of silk worm oak and Korean red pine specimen amounted to 16.0 and 21.3 in the boiling treatment, and 17.3 and 24.0 in the steaming treatment, respectively. But in the bending with a tension strap, the r/t of the oak and the pine specimen decreased to 5.3 and 16.0 in t he boiling treatment, and 3.3 and 18.7 in the steaming treatment, respectively. Therefore, the bending processing properties of the both species were significantly improved by the strap. 12. The effect of pin knot on the degradation of bending processing property was very severe in silk worm oak by side, e.g. 90 percent of the oak specimens with pin knot on the concave side were ruptured when bent to a 100 mm radius but only 10 percent of the other specimens with pin knot on the convex side were ruptured. 13. The changing rate in the bending radius of specimen bent to a 300 mm radius after 30 days of exposure to room temperature conditions was measured to 4.0-10.3 percent in the boiling treatment and 13,0-15.0 percent in the steaming treatment. Therefore, the degree of spring back after release was higher in the steaming than in the boiling treatment. And the changing rate of moisture-proofing treated specimen by expoxy resin coating was only -1.0.0 percent. 14. Formaldehyde, 35 percent solution, and 25 percent polyethylene glycol-400 solution found no effect on the plasticization of the both species, but saturated urea solution and 25 percent ammonium hydroxide solution found significant effect in comparison to non-treated specimen. But the effect of the treatment with chemicals alone was inferior to that of the steaming treatment, and the steaming treatment after the treatment with chemicals improved 10-24 percent over the bending processing property of steam-bent specimen. 15. Three plasticity coefficients - load-strain coefficient, strain coefficient, and energy coefficient - were evaluated to be appropriate for the index of bending processing property because the coefficients had highly significant correlation with the bending radius. The fitness of the coefficients as the index was good at load-strain coefficient, energy coefficient, and strain coefficient, in order.