• Title/Summary/Keyword: Cell Growth

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Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

The Effect of Weight Ratio of Carbon Source to Nitrogen Source on toe Growth and the Composition of Fatty Acid of Thraustochytrium aureum ATCC 34304 (탄소원과 질소원의 무게비가 Thraustochytrium aureum ATCC 34304의 성장 및 지방산 조성 특성에 미치는 영향)

  • Kim Won-Ho;Park Seung-Hye;Song Sang-Kyu;Bae Kyung-Dong;Hur Byung-Ki
    • KSBB Journal
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    • v.20 no.4
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    • pp.266-270
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    • 2005
  • The effect of the weight ratio of carbon source to nitrogen source on the growth and the composition change of fatty acids of Thraustochytrium aureum ATCC 34304 was investigated. The cell concentration of 5 days' culture increased and then decreased according to the increase of C/N, The ratioes for the maximum biomass were unique respectively and distributed between 1 and 4 with the initial sugar concentration in the range of 5 g/L to 25 g/L. The biomass yield, $Y_{x/s}$ decreased along with the increase of C/N, but maintained a constant value 0.35 between 10 and 20. The composition of myristic acid $(C_{14:0})$, one component of the lipid synthesized by T. aureum, was not affected with the change of C/N, but the composition of palmitic acid $(C_{16:0})$ was around $20\%$ below 4 of C/N and decreased to $15\%$ according to C/N about 4. The compositions of oleic acid $(C_{18:1})$ and linoleic acid $(C_{18:2})$ increased from 0 to $20\%\;and\;7\%$ respectively. The composition of $\gamma-linoleic$ acid $(C_{18:3})$, however, reduced from $5\%\;to\;2\%$. EPA $(C_{20:5})$ and DPA $(C_{22:5})$ showed a tendency of reduction in the weight composition according to the increase of C/N, but DHA $(C_{22:6})$ had a trend maintaining an approximately constant value, around $40\%$, irrespective of the change of C/N.

Apolipoprotein A1 Inhibits TGF-β1-Induced Epithelial-to-Mesenchymal Transition of Alveolar Epithelial Cells

  • Baek, Ae Rin;Lee, Ji Min;Seo, Hyun Jung;Park, Jong Sook;Lee, June Hyuk;Park, Sung Woo;Jang, An Soo;Kim, Do Jin;Koh, Eun Suk;Uh, Soo Taek;Kim, Yong Hoon;Park, Choon Sik
    • Tuberculosis and Respiratory Diseases
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    • v.79 no.3
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    • pp.143-152
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    • 2016
  • Background: Idiopathic pulmonary fibrosis (IPF) is a progressive and lethal lung disease characterized by the accumulation of excessive fibroblasts and myofibroblasts in the extracellular matrix. The transforming growth factor ${\beta}1$ (TGF-${\beta}1$)-induced epithelial-to-mesenchymal transition (EMT) is thought to be a possible source of fibroblasts/myofibroblasts in IPF lungs. We have previously reported that apolipoprotein A1 (ApoA1) has anti-fibrotic activity in experimental lung fibrosis. In this study, we determine whether ApoA1 modulates TGF-${\beta}1$-induced EMT in experimental lung fibrosis and clarify its mechanism of action. Methods: The A549 alveolar epithelial cell line was treated with TGF-${\beta}1$ with or without ApoA1. Morphological changes and expression of EMT-related markers, including E-cadherin, N-cadherin, and ${\alpha}$-smooth muscle actin were evaluated. Expressions of Smad and non-Smad mediators and TGF-${\beta}1$ receptor type 1 ($T{\beta}RI$) and type 2 ($T{\beta}RII$) were measured. The silica-induced lung fibrosis model was established using ApoA1 overexpressing transgenic mice. Results: TGF-${\beta}1$-treated A549 cells were changed to the mesenchymal morphology with less E-cadherin and more N-cadherin expression. The addition of ApoA1 inhibited the TGF-${\beta}1$-induced change of the EMT phenotype. ApoA1 inhibited the TGF-${\beta}1$-induced increase in the phosphorylation of Smad2 and 3 as well as that of ERK and p38 mitogen-activated protein kinase mediators. In addition, ApoA1 reduced the TGF-${\beta}1$-induced increase in $T{\beta}RI$ and $T{\beta}RII$ expression. In a mouse model of silica-induced lung fibrosis, ApoA1 overexpression reduced the silica-mediated effects, which were increased N-cadherin and decreased E-cadherin expression in the alveolar epithelium. Conclusion: Our data demonstrate that ApoA1 inhibits TGF-${\beta}1$-induced EMT in experimental lung fibrosis.

Role of the Nuclear Transcription Factor NF-κB Caused by Acute Hypoxia in the Heart (급성 저산소증 상태에서 심장 내 전사인자 NF-κB의 기능)

  • Joo, Chan Uhng;Juhng, Woo Suk;Kim, Jae Cheol;Yi, Ho Keun
    • Clinical and Experimental Pediatrics
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    • v.45 no.9
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    • pp.1106-1113
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    • 2002
  • Purpose : Nuclear ($factor-{\kappa}BNF-{\kappa}B$) is now recognized as playing a potential role in programmed cell death and the adaptive response to various stress. Cellular hypoxia is a primary manifestation of many cardiovascular diseases. It seems that vascular endothelial growth factor (VEGF) and insulin like growth factor-I(IGF-I) have a function as a protective molecule in the heart against several stress including hypoxia. In this study, the role of $NF-{\kappa}B$ to the cellular response and regulation of protective molecules against the acute hypoxia in the heart was studied. Methods : To cause acute hypoxic stress to the heart, Sprague Dawley rats were exposed to hypoxic chamer($N_2$ 92% and $O_2$ 8%). After the hypoxic exposure, nuclear proteins, total proteins and mRNA were isolated from heart. Translocation of the transcription factors $NF-{\kappa}B$, NF-ATc, AP-1 and NKX-2.5 were evaluated by electrophoretic mobility shift assay(EMSA). The expression of IGF-I and VEGF were studied before and after the hypoxic stress by competitive-PCR, Northern hybridization and Western hybridization. To confirm the role of the $NF-{\kappa}B$ in the heart, the rats also were pretreated with diethyl-dithiocarbamic acid(DDTC) into peritoneal cavity to block $NF-{\kappa}B$ translocation into nucleus. Results : The expression of $NF-{\kappa}B$, AP-1 and NF-ATc were increased by the hypoxic stress. Increased expression of the VEGF and IGF-I were also observed by the hypoxic stress. However, the blocking of the $NF-{\kappa}B$ translocation reduced those expressions of VEGF and IGF-I. Conclusion : These results suggest that $NF-{\kappa}B$ has a protective role against the acute hypoxia through several gene expression, especially VEGF and IGF-I in heart muscle.

Antimicrobial and Anticancer Activity of Korean Traditional Soy Sauce and Paste with Chopi (초피첨가 전통장류의 항균 및 항암활성)

  • Kim, Keun-Ki;Park, Hyean-Cheal;Son, Hong-Joo;Kim, Yong-Gyun;Lee, Sang-Mong;Choi, In-Soo;Choi, Young-Whan;Shin, Teak-Soon
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1121-1128
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    • 2007
  • The fruits of Zanthoxylum piperitum are known as having various physiology vitality, and the abstraction ingredient of the pericarp is also known as having strong antibiotic activities against various bacteria. Therefore, this study was carried out to estimate the effect of physiology vitality when the abstraction ingredient of Z. piperitum was added in soy sauce(Chopi-kanjang) and soybean paste(Chopi-doenjang). For the antibiotic activity against the pathogens of sitotoxism such as Staphylococcus aureus, Salmonella typhimurium, Vibrio parahemolyticus, Escherichia coli 0157:H7, the extracts of the Chopi-kanjang was added 1%, 2%, 4% pericarp of Z. piperitum in the manufacturing process of soy source. According to the results, the growth of E. coli 0157:H7 and V. parahemolyticus were respectively inhibited as 70% and 50% by the Chopi-kanjang added 2% of the ingredient. For the antibiotic effects of the aforementioned Chopi-kanjang against Sal. typhimurium and Sta. aureus, the growth of those pathogens was also inhibited between 40% and 60% according to the manufacturing period of Chopi-kanjang. It was confirmed that the antibiotic activity using the mixture of the abstraction ingredient and Chopi-doenjang was lower than those of Chopi-kanjang. In order to estimate the anticancer activity using by caspase-3 activity, the mixture of the abstraction ingredient of the pericarp of Z. piperitum and Chopi-kanjang was treated to leukemia cells. According to the results, the activities of caspase-3 using the mixture added 1%, 2% and 4% of the abstraction ingredient were respectively increased as much as 4, 12, 15 times comparing with the control which was treated with the soy source only. It could be that the mixture of the abstraction ingredient of the pericarp of Z. piperitum and soy source induced apoptosis, and the mixture of the abstraction ingredient and soybean paste had no effect on the activity of caspase-3. In order to find out the death of the aforementioned cells caused by necrosis or apoptosis, DNA fragmentation in the cell was examined. U-937 cells showed apoptotic DNA fragmentation in the incubation with Chopi-kanjang extract.

Heat Shock Treatments Induce the Accumulation of Phytochemicals in Kale Sprouts (열처리에 의한 케일 새싹의 기능성물질 축적)

  • Lee, Min-Jeong;Lim, Sooyeon;Kim, Jongkee;Oh, Myung-Min
    • Horticultural Science & Technology
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    • v.30 no.5
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    • pp.509-518
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    • 2012
  • The objective of this study was to determine the effect of heat shock treatments on the phytochemicals including antioxidants and anticancer materials in kale (Brassica oleracea L. var. acephala) sprouts. In study I, kale sprouts grown under the growing system for four days were soaked at 40, 50, or $60^{\circ}C$ distilled water for 10, 30, or 60 seconds, and in study II, kale sprouts were soaked at $50^{\circ}C$ distilled water for 10, 20, 30, 45, or 60 seconds. After the heat shock treatments, the sprouts were transferred into normal growing conditions and recovered there for two days. Fresh and dry weights, electrolyte leakage, total phenolic concentration, antioxidant capacity, total flavonoid concentration, phenylalanine ammonia-lyase (PAL) activity, and glucosinolates content of the sprouts were measured before and after the heat shock treatments. As a result, there was a significant decrease in the fresh and dry weight of kale sprouts treated with heat shock compared with control at harvest in study I. Especially, heat shock at $60^{\circ}C$ lead to more pronounced growth inhibition compared with heat treatments at 40 and $50^{\circ}C$. Electrolyte leakage by cell collapse was the highest in the sprouts exposed to $60^{\circ}C$ distilled water, which agreed with the growth results. Heat shock at $50^{\circ}C$ significantly induced the accumulation of phenolic compounds. In study II, fresh weight of kale sprouts at $50^{\circ}C$ heat shock showed a significant decrease compared with the control at one and two days after the treatment. However, the decrease was minimal and dry weight of kale sprouts was not significantly different from that in control. In contrast, the heat shock-treated kale sprouts had higher level of total phenolic concentration than control at harvest. Heat shock treatments at $50^{\circ}C$ for 20 seconds or more showed at least 1.5 and 1.2 times higher total phenolic concentration and antioxidants capacity than control, respectively. The change of the total flavonoid concentration was similar with that of antioxidants. PAL activity after 24 hours of heat shock was higher in all the heat shock-treated sprouts than that in control suggesting heat shock may stimulate secondary metabolic pathway in kale sprouts. Seven glucosinolates were identified in kale sprouts and soaking the sprouts with $50^{\circ}C$ water for 20 seconds had a pronounced impact on the accumulation of total glucosinolates as well as two major glucosinolates, progoitrin and sinigrin, at harvest. In conclusion, this study suggests that heat shock using hot water would be a potential strategy to improve nutritional quality of kale sprouts by inducing the accumulation of phytochemicals with antioxidant and anticancer properties.

Gonad Ontogeny in Relation to Somatic Growth in the Brown Croaker Miichthys miiuy (Basilewsky) (민어, Miichtys miiuy (Basilewsky)의 성장과 연관된 생식소 발달)

  • Park, In-Seok;Seol, Dong-Won;Im, Soo-Yeon;Park, Min Ouk;Hur, Woo June;Cho, Sung Woan;Song, Young-Chae;Kim, Jea-Soo;Jo, Hyo-Jae;Noh, Choong Hwan;Choi, Hee Jung
    • Korean Journal of Ichthyology
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    • v.19 no.2
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    • pp.107-111
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    • 2007
  • Sex differentiation of the brown croaker Miichthys miiuy (Basilewsky) is described from hatching to the 120th day post-hatching (dph) (water temperature $24^{\circ}C$). Primordial germ cells (PGCs) were observed on the 20th dph (10.4 mm total length (TL), 0.14 g body weight (BW), and began to protrude into the peritoneal cavity from the 40th dph (19.4 mm TL, 0.39 g BW). On the 65th dph (31.3 mm TL, 0.93 g BW, $1,560D^{\circ}$ (degree-days)), initial ovarian differentiation was identified by the PGCs with condensed chromatin, and their transformation into meiotic oocytes. By the 120th dph (4.60 mm TL, 1.38 g BW, $2,880D^{\circ}$), the oocytes were in the perinucleolus stage and had increased from 20 to $40{\mu}m$ in diameter. While ovaries gradually grew after sex was differentiated, testes continued to multiply from the 65th dph. On the 80th dph (37.9 mm TL, 1.39 g BW, $1,920D^{\circ}$), the beginning of testis lobule formation was indicated by the occurrence of spermatogonial cysts enveloped by somatic cells in some of the testes. On the 120th dph, the testis lobules of some of the fish contained all germ cell stages through to the spermatocytes. Therefore, the sex differentiation type of the brown croaker is identified as gonochoristic.

Effects of Growth and Cellular Tissue under Abnormal Climate Condition in Chinese Cabbage (이상기상 조건이 배추의 생육 및 세포조직에 미치는 영향)

  • Lee, Sang Gyu;Choi, Chang Sun;Choi, Jun Myung;Lee, Hee Ju;Park, Suhyoung;Do, Kyung Ran
    • Journal of Bio-Environment Control
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    • v.22 no.2
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    • pp.87-90
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    • 2013
  • The average annual and winter ambient air temperatures in Korea have risen by $0.7^{\circ}C$ and $1.4^{\circ}C$, respectively, during the last 30 years. Due to climate change, the occurrence of abnormal weather conditions has become more frequent, causing damage to vegetable crops grown in Korea. Hot pepper, chinese cabbage and radish, the three most popular vegetables in Korea, are produced more in the field than in the greenhouse. It has been a trend that the time for field transplanting of seedlings is getting earlier and earlier as the spring temperatures keep rising. Seedlings transplanted too early in the spring take a longer time to resume the normal growth, because they are exposed to suboptimal temperature conditions. This experiment was carried out to figure out the change of cellular tissue of chinese cabbage under the condition of low temperature to provide the information regarding the coming climatic change, on the performance of 'Chunkwang' chinese cabbage during the spring growing season. In our study, plant height, number of leaf, chlorophyll and leaf area was lower at the open field cultivation than heating house treatment after transplanting 50 days. Especially in fresh weight, compared with heating treatment, open field and not heated treatment were notably low with the 1/3 level. Of damage symptoms due to low temperature cabbage leaves about 10 sheets when $-3.0^{\circ}C$ conditions in chinese cabbage was a little bit of water soaking symptoms on the leaves. $-7.4^{\circ}C$ under increasingly severe water soaking symptoms of leaf turns yellow was dry. Microscopy results showed symptoms of $-3.0^{\circ}C$ when the mesophyll cell of palisade tissue and spongy tissue collapse, $-7.4^{\circ}C$ palisade tissue and spongy tissue was completely collapsed. The result of this study suggests that the growers should be cautioned not to transplant their chinese cabbage seedlings too early into the field, and should be re-transplanting or transplanting other plants if chinese cabbage are exposed to suboptimal temperature conditions ($-3.0^{\circ}C$ or $-7.4^{\circ}C$).

Laboratory-scale fruiting body formation of Pleurotus ostreatus using the petri dish culture (느타리의 기내 자실체 형성 및 그 유도조건에 관한 연구)

  • Joh, Joong-Ho;Chu, Kyo-Sun;Kim, Beom-Gi;Kong, Won-Sik;Yoo, Young-Bok;Lee, Seung-Jae;Cho, Bong-Gum;Lee, Chang-Soo
    • Journal of Mushroom
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    • v.2 no.1
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    • pp.15-20
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    • 2004
  • Pleurotus ostreatus, the oyster mushroom, is one of the most widely cultivated and important edible mushrooms in the world. In order to study the developmental process of P. ostreatus and its regulatory mechanism, a new culturing method needs to be established for inducing the fruiting body and sporulation in the laboratory. In this study, we have examined whether the fruiting body of P. ostreatus can be formed on the plastic petri dish which are commonly used for cell culture in the laboratory. The strain was cultured on $60{\times}15mm$ plastic petri dish with potato dextrose agar media at $28^{\circ}C$ for mycelial growth and then at $18^{\circ}C$ for the formation of primordia and fruiting bodies within plant growth chamber. The development of primordia into fruiting bodies was achieved on cultured dishes under air ventilation. At the primordia stage, the normal formation of fruiting body was blocked by sealing the plastic dish with parafilm. The periods requiring for the formation of primordia and fruiting bodies were examined on the dish culture. About 96% and 76% of cultured samples formed primordia and fruiting bodies under the optimal conditions during ten weeks of culture, respectively. These culturing periods, however, were changed by the mechanical injury treatment to mycelia. As other factors affecting the fruiting body formation, the effects of light and cold shock have been tested. No fruiting formation was observed on the cultured dishes under the dark. The cold shock treatment by storing cultured dishes for one day at $4^{\circ}C$ did not have any significant effects in the fruiting body formation. Spores of fruiting bodies acquired from the petri dishes could be germinated on culture media at $28^{\circ}C$. These results suggest that the fruiting bodies of P. ostreatus can be formed on the experimental petri dish and this dish-culturing method is useful for understanding of the developmental process of P. ostreatus in the laboratory. Furthermore, the dish-culturing method is able to shorten the life cycle of P. ostreatus without requiring large area and expensive device.

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An Initiative Study on Relationship between Algal Blooms and Asian Dust for Regulation of Algal Blooms (조류 성장 억제를 위한 녹조 및 적조 발생과 황사의 상관관계 초기적 연구)

  • Kim, Tai-Jin;Jeong, Jaechil;Seo, Rabeol;Kim, Hyung Moh;Kim, Dae Geun;Chun, Youngsin;Park, Soon-Ung;Yi, Sehyoon;Park, Jun Jo;Lee, Jin Ha;Lee, Jay J.;Lee, Eun Ju
    • KSBB Journal
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    • v.29 no.4
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    • pp.285-296
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    • 2014
  • Although the problems of the algal blooms have been world-widely observed in freshwater, estuary, and marine throughout the year, it is not yet certain what are the basic causes of such blooms. Consequently, it is very difficult to predict when and where algal blooms occur. The constituents of the Asian dust are in a good agreement with the elements required for the algal growth, which suggests some possible relationship between the algal blooms and the Asian dust. There have been frequently algal blooms in drinking water from rivers or lakes. However, there is no any algal blooms in upwelling waters where the Asian dust cannot penetrate into the soil due to its relatively weak settling velocity (size of particles, $4.5{\pm}1.5{\mu}m$), which implies the possible close relationship of the Asian dust with algal blooms. The present initiative study is thus intended firstly in Korea to illustrate such a relationship by reviewing typical previous studies along with 12 years of weekly iron profiles (2001~2012) and two slant culture experiments with the dissolved Asian dust. The result showed bacterial suspected colonies in the slant culture experiment that are qualitatively in a good agreement with the recent Japanese studies. Since the diatoms require cheap energy (8%) compared to other phytoplankton (100%) to synthesize their cell walls by silicate, the present results can be used to predict algal blooms by diatoms if the concentrations of iron and silicate are available during spring and fall. It can be postulated that the algal blooms occur only if the environmental factors such as light, nutrients, calm water surface layer, temperature, and pH are simultaneously satisfied with the requirements of the micronutrients of mineral ions supplied by the Asian dust as enzymatic cofactors for the rapid bio-synthesis of the macromolecules during algal blooms. Simple eco-friendly methods to regulate the algal blooms are suggested for the initial stage of blooming with limited area: 1) to cover up the water surface with black curtain and inhibit photosynthesis during the day time, 2) to blow air (20.9%) or pure oxygen into the bottom of the water and inhibit rubisco for carbon uptake and nitrate reductase for nitrogen uptake activities in algal growth during the night, 3) to eliminate the resting spores or cysts by suction of bottom sediments as deep as 5 cm to prevent the next year germinations.