To develop a yeast strain of Pichia pastoris producing an antibacterial peptide, we have attempted the expression and secretion of an insect defensin. The nucleotide sequences corresponding to mature defensin were chemically synthesized by 6 oligomers, assembled in vitro and the synthesized gene was identified by nucleotide sequencing. The prepro sequence of yeast mating factor $\alpha$1 and the defensin gene were recombined into a Pichia expression vector, pPIC9K. The resulting plasmid, pPIDE, was transformed into P. pastoris GSl15 and transformants selected on histidine-deficient minimal plates were tested for antibacterial activity against Micrococcus luteus. Four strains with different antibacterial activity were selected for further analysis. Southern hybridization and RT-PCR verified the defensin gene was maintained and transcribed in a host. Four strains were cultivated in YPD broth for 96 hours to compare cell growth and antibacterial activity, They showed no difference in cell growth, however, each strain showed different antibacterial activity pattern with culture time. The maximal activity was about 550 AU/ $m\ell$.
Rock mass classification methods such as RMR, Q system and GSl have been widely adopted with certain modifications for the design of mine openings. The GSI system is the only rock mass classification system that is related to Mohr-Coulomb and Hoek-Brown strength parameters and gives a simple method to calculate the engineering properties of rock masses which can be useful input parameters for a numerical analysis. A detailed surveying for GSI mapping as well as far calculating RMR values was undertaken at Daesung and Pyunghae underground limestone mining sites. RQD values were determined for row locations in these two mining sites. Based on GSI values and intact rock strength properties, the rock mass strength modulus of elasticity as well as the Mohr-Coulomb strength parameter c$_{m}$ and $\phi$$_{m}$ were determined. GSI and RMR are correlated.
Journal of the Korean Society of Food Science and Nutrition
/
v.26
no.4
/
pp.669-674
/
1997
This study was attempted to express human lactoferrin gene that has importance as a functional additive in food industry. Lactoferrin has distinctive antibacterial properties. Also, a number of phy-siological roles have been postulated for the lactoferrin in the modulation of immune and inflamatory responses and as a growth factor. Since it did not show feasible growth inhibition by antimicrobial test against HLF, Pichia pastoris was selected the best lactoferrin expression host. HLF expression plasmid pHIL-SI was integrated into the genomic DNA of P. pastoris GSl15. The integration was confirmed not only with 2.4Kb fragment of HLF gene by PCR(polymerase chain reaction) product, but also with same size of specific signal by southern blotting. Among the various pichica transformants, the JY-1 cell showed a positive response for the expression of HLF by the immunoblotting anaysis. The recombinant HLF protein was started to be secreated at 48hr of culture and reached at the highest secreation level at 96hr.
Menni, Alekhya Chowdary;Radhakrishna, Ambati Naga;Prasad, M. Ghanashyam
Journal of Dental Anesthesia and Pain Medicine
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v.20
no.6
/
pp.397-402
/
2020
Background: Inferior alveolar nerve block (IANB) is the most common, painful, and anxiety-provoking procedure involving needle insertion for anesthetic solution deposition. DentalVibeⓇ (DV) delivers vibration at a sustained frequency as a counter-stimulation to the site of injection, thereby alleviating pain. The aim of this study was to evaluate and compare the effectiveness of DV and lignocaine hydrochloride 2% gel (Lox 2% jelly) in pain reduction during IANB in children. Methods: A split-mouth randomized clinical trial was designed with a sample of 60 children (age, 6 to 12 years) requiring bilateral IANB for various dental procedures; DV was used while administering IANB and Lox 2% jelly was used as the topical anesthetic before administering IANB at subsequent appointments. During both appointments, pain perception was measured using the sound, eye, motor (SEM) scale and Wong-Baker faces pain rating scale (WBFPRS); oxygen saturation (SpO2) and pulse rate were measured using a pulse oximeter before, during, and after the IANB procedure. The obtained values were tabulated and subjected to statistical analysis. Wilcoxon test was used for intergroup comparison, and Friedman test, for intragroup comparison of measured variables at different treatment phases. Results: The medians and interquartile ranges of the WBFPRS scores recorded during the IANB procedure for DV and Lox 2% jelly were 2 (2-4) and 2 (0-2), respectively (P < 0.05). The SEM scale scores, mean SpO2, and pulse rate did not show any significant differences during the IANB procedure between both treatments. Conclusion: Both DV and Lox 2% jelly were found to be effective in pain reduction during IANB in children.
Nitrogen (N) is a vital element in growing crops and is essential for improving the yield and quality of crops. Thus, N fertilizer is the most widely used fertilizer and the primary N input source in soil-crop systems. Inorganic fertilizers such as urea are known to improve crop productivity and increase soil fertility. However, application with excessive amounts can interfere with crop growth and accelerate soil acidification. For these reasons, the use of organic fertilizers, which mainly contain organic nitrogen, has gradually increased worldwide. Therefore, this study evaluated the effects of N fertilizer on the growth of Chinese cabbage including its functional compounds glucosinolates (GSLs). For the cultivation of Chinse cabbage, inorganic fertilizer was used for urea, and organic fertilizers were divided into conventional and biochar-based fertilizers. The growth parameters of Chinese cabbage treated by organic fertilizers was better than those of the inorganic fertilizers. Additionally, it was found that their co-application was more efficient. However, their GSL contents were lower with the application of the organic fertilizers. The characteristics of the experimental soil also changed according to the type, amounts and co-application of fertilizers. Therefore, this study presents the basis for an eco-friendly method that can increase the functionality and productivity of Chinese cabbage compared to conventional cultivations.
In this study, we analyzed the changes in glucosinolate content and gene expression in TO1000DH3 and Early big seedling upon methyl jasmonate (MeJA) treatment. Analysis of glucosinolate contents after MeJA treatment at $200{\mu}M$ concentration showed that the total glucosinolate content increased by 1.3-1.5 fold in TO1000DH3 and 1.3-3.8 fold in Early big compared to those before treatment. Aliphatic glucosinolates, progoitrin and gluconapin, were detected only in TO1000DH3, and the changes in the content of neoglucobrassicin were the greatest at 48 hours after MeJA treatment in TO1000DH3 and Early big. The transcriptomic analysis showed that transcripts involved in stress or defense reactions, or those related to growth were specifically expressed in TO1000DH3, while transcripts related to nucleosides or ATP biosynthesis were specifically expressed in Early big. GO analysis on transcripts with more than two-fold change in expression upon MeJA treatment, corresponding to 12,020 transcripts in TO1000DH3 and 13,510 transcripts in Early big, showed that the expression of transcripts that react to stimulus and chemical increased in TO1000DH3 and Early big, while those related to single-organism and ribosome synthesis decreased. In particular, the expression increased for all transcripts related to indole glucosinolate biosynthesis, which is associated with increase in glucobrassicin and neoglucobrassicin contents. Upon MeJA treatment, the expression of AOP3 (Bo9g006220, Bo9g006240), TGG1 (Bo14804s010) increased only in TO1000DH3, while the expression of Dof1.1 (Bo5g008360), UGT74C1 (Bo4g177540), and GSL-OH (Bo4g173560, Bo4g173550, Bo4g173530) increased specifically in Early big.
The spawning and growth of eightspine stickleback,Pungitius sinensis kaiborae (Tanaka, 19151, were investigated in the Chavang Stream from March 1990 to february 1991 as a part of study on its life history. Nest building, spavming and parental behaviors were similar to those known on the sticklebacks of Europe, North America and Japan. Bht f sinensis kaibarae in the Chayang Stream, peculiarly, showed only superficial gluing behavior in nest building phase and made two nurseries iust behind the nest in Parental Phase. The individuals with fully mature eggs were found from late February to late June. The fecundity of an adult female was about 21 to 110 (mean 47.07) eggs and the diameter of kllv matured ovarian eggs ranged 1.3 to 1.5 mm. The smallest female with fully mature eggs was 25.8 mm in body length and had 21 eggs. The relationship between the number of mature eggs in ovary and body length was No. of eggs = 2.857BL -55.134 (r = 0.890). The number of deposited eggs in a nest was 27 to 637 and thew were composed of several clusters of various developmental stages. The distribution pattern of gonadsomatic index (GSl) against the body length shown that in females 1+ year-old fishes mainly participate in spawning but in males 11 and 2+ year-old fishes equally participate. From the fluctuat지n pattern of GSI and the number of mature eggs in ovary, it was assumed that the spawning season was from February to June and the peak of spawning was early March to late April. The newly hatched vouns was found at late April and their body length was about 10 mm. The vouns (O+) grew rapidly until late August. Then their srowth was slowed down and finallY ceased in midautumn. The young reached about 35 mm BL in their first year of life. The halting of growth was lasted to next Sune when the spawning season would be nearly closed. They (1+) began to grow asain from late june, grew rapidly until tate september and reached about 415 mm BL. Then there was no more grouvth until to die as 2+ yearold fish. Therefore it was assumed that the life span of P. sinensis kaibarae in the Chavang Stream would be about ko years. The relationship between the body length and the body weight was logBW = 2.9541ogBL -4.802 (r = 0.998).
The reproduction of marbled sole Limanda yokohamae, caught near around the southeastern coast of Korea, from December 1983 to November 1984, was investigated based on such annual variations as gonadosomatic index(GSL), gametogenesis, reproductive cycle, spawning number, hepatosomatic index (HSI), and fatness. GSI began to increase in the autumn season with the onset of shorter day length and colder water temperature, and reached the maximum value in December with the shortest day length and the lowest temperature over the year. The gonad activated the proliferation of oogonia and spermatogonia in June, reached the mature stage in October, ripe in December, and spawning from the end of December to January. After spawning, it showed the resting stage which gonad remained regressive and suppressive from February to May. In addition, the adult individuals observed discharged eggs only once during their spawning period. At yolk globular stage, the substance of vitellogenin synthesized from the liver was considered to participate in the active yolk accumulation of oocytes. Marbled sole was concluded to be a typical winter spawning species in that such environmental factors as short day length and low water temperature were closely related with the gametogenesis, the stimulation of oocyte maturation, and were also affecting the ovulation.
A new soybean cultivar 'Daewang' was released by Agricultural Research & Extension Services, Gangwon Province in 2008. The Goals of breeding the variety was for large seed size, high yield, lodging tolerance and resistance to diseases such as soybean mosaic virus (SMV) and bacterial pustule. 'Daewang' was derived from the cross of GWS91, which has yellow seed coat, large seed size and late maturity, and 'Seokryang-putkong', which has large seed size and early maturity. The preliminary, advanced and regional yield trials for evaluation and selection were carried out from 2002 to 2007. It has a determinate growth habit with purple flower, yellow seed coat, yellow hilum and large seed weight (33.6 g per 100 seeds). The maturity date of 'Daewang' is 16 days later than the 'Taekwang'. It has a high content of total isoflavone ($1,851{\mu}g/g$). The average yield of 'Daewang' was 2.68 MT/ha in the regional yield trials (RYT) carried out in four locations in Gangwon province from 2004 to 2007 which was 4 percent higher than the check cultivar 'Taekwang'.
Jo, Ji-Young;Sung, Ho-Young;Chun, Jin-Hyuk;Park, Jong-Seok;Park, Sang-Un;Park, Young-Jun;Kim, Sun-Ju
Korean Journal of Environmental Agriculture
/
v.37
no.3
/
pp.197-206
/
2018
BACKGROUND: The present study aimed to examine the crops capable of growing and adapting to the external environment and various stresses of reclaimed agriculture land for the development of high value-added agricultural utilization technology based on reclaimed land through standardization and empirical study of cultivation environment for cultivating crops. METHODS AND RESULTS: Two crops namely turnips and beets were selected for the salt tolerance test of soil environmental conditions on reclaimed land. Turnip and beet seedlings were planted on the soil collected at the 'Seokmun' reclaimed land. There are five treatments such as non-treatment, 1.0, 2.0 (control), 4.0 and $8.0dS{\cdot}m^{-1}$ of EC. The contents of betacyanin in beet roots was highest in control and decreased with increasing salt concentration. The GSL contents in the turnip roots waswere highest at EC 2.0 and decreased with increasing salt concentration, whereas those in turnip leaves waswere high both in the non-treated control and atthe EC 1.0-treatment. But, tThere was, however, no statistical differences among the treatments. CONCLUSION: The degree of salt tolerance of crops was examined, and the limit EC iswas expected to be $3.0{\sim}4.0dS{\cdot}m^{-1}$ as reported to date. If the soil improvement is performed and irrigation systems are used in the actual reclaimed land, the EC of supplied irrigation will be low, and desalination effecttreatment by the lower EC of the supplied irrigation on the soil will lead to more favorable soil condition of the rhizosphere and cultivation environment offor the crops than those in the port experiment. Therefore, monitoring the salinity, water content and ground water level will enable prediction of the rhizosphere environment, and setting up irrigation management and supplying irrigation will lead to crop cultivation results that are close to normal.
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