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Hexokinase 1 and 2 mediates glucose utilization to regulate the synthesis of kappa casein via ribosome protein subunit 6 kinase 1 in bovine mammary epithelial cells

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Animal Nutrition 2024年第1期16卷 338-349页

作者： Tianyu Yang Jia Guo Han Song Osmond Datsomor Yuhang Chen Maocheng Jiang Kang Zhan Guoqi Zhao Institute of Animal culture collection and Application College of Animal Science and TechnologyYangzhou UniversityYangzhou 225009China

Glucose plays a vital part in milk protein synthesis through the mTOR signaling pathway in bovine mammary epithelial cells(BMEC).The objectives of this study were to determine how glucose affects hexokinase(HK)activity in BMEC and investigate the regulatory effect of HK in kappa casein(CSN3)synthesis via the mechanistic target of rapamycin complex 1(mTORC1)signaling pathway in *** this,HK1 and HK2 were knocked out in BMEC using the CRISPR/Cas9 *** gene and protein expression,glucose uptake,and cell proliferation were *** found that glucose uptake,cell proliferation,CSN3 gene expression levels,and expression of HK1 and HK2 increased with increasing glucose ***,glucose uptake was significantly reduced in HK2 knockout(HK2KO)BMEC treated with 17.5 mM ***,under the same glucose treatment conditions,the proliferative ability and abundance of CSN3 were significantly diminished in both HK1 knockout(HK1KO)and HK2KO BMEC compared with that in wild-type *** further observed that the phosphorylation levels of ribosome protein subunit 6 kinase 1(S6K1)were reduced in HK1KO and HK2KO BMEC following treatment with 17.5 mM *** expected,the levels of glucose-6-phosphate and the m RNA expression levels of glycolysis-related genes were decreased in both HK1KO and HK2KO BMEC following glucose *** results indicated that the knockout of HK1 and HK2 inhibited cell proliferation and CSN3 expression in BMEC under glucose treatment,which may be associated with the inactivation of the S6K1 and inhibition of glycolysis.

关键词： Glucose Hexokinase Milk protein Mechanistic target of rapamycin complex 1 signaling pathway Bovine mammary epithelial cell Kappa casein

Periodic exposure to ambient solar irradiance benefits the growth of juvenile seedlings of Hizikia fusiformis

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Chinese Journal of Oceanology and Limnology 2011年第5期29卷 1009-1014页

作者：单体锋逄少军高素芹 Marine Biological culture collection Centre Institute of Oceanology Chinese Academy of Sciences

In our trials, from 2007 to 2008, of mass production of seedlings of Hizikiafusiformis using synchronization techniques, problems of a ＂dark thalli＂ phenomenon and epiphytes contamination severely threatened the health of juvenile seedlings. In this investigation, we optimized conditions for improving the growth of juvenile seedlings. Seven string collectors were seeded with zygotes and a series of experiments were conducted including direct exposure to solar irradiance, co-culture with Ulva spp. and treatment with sodium hypochlorite. It was found that direct exposure to solar irradiance （maximum： 1 740 μmol photons/（m2.s）） for 2 h per day could efficiently enhance the growth of young seedlings and simultaneously inhibit the growth of epiphytic algae. In this treatment, 50-day old seedlings could reach an average of 0.44 cm in length and possess up to nine leaflets. However, a single treatment with 18-mmol/L sodium hypochlorite for 10 rain severely harmed 15-day old seedlings. In comparison, weekly treatment with 2.2-mmol/L sodium hypochlorite for 10 rain brought no apparent harm to seedlings and eliminated epiphytic algae efficiently. However, this treatment significantly increased the detachment rate of seedlings, Inoculating Ulva spp. onto the collector caused a dramatic decrease in the number of seedlings. However, the growth of the remaining seedlings appeared unhampered. All collectors except the control were daily sprayed with a high pressure water jet from the 84 day post fertilization. From the first day to 50th day, no ＂dark thallus＂ was observed on any of the seven collectors. We believe that well-timed daily exposure to solar irradiance would favor H. fusiformis in its early growing stages.

关键词： Hizikiafusiformis solar irradiance dark thalli epiphytes resistance

Butyrate inhibits the bovine rumen epithelial cell proliferation via downregulation of positive regulators at G0/G1 phase checkpoint

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BIOCELL 2022年第7期46卷 1697-1704页

作者： KANG ZHAN MAOCHENG JIANG TIANYU YANG ZIXUAN HU GUOQI ZHAO Institute of Animal culture collection and Application College of Animal Science and TechnologyYangzhou UniversityYangzhou225009China

Short-chain fatty acids(SCFAs)butyrate promote the postnatal rumen epithelial development and maturation in ***,molecular mechanisms of effects of butyrate on the bovine rumen epithelial cells(BRECs)proliferation remain ***,purpose of this study was to investigate the effects of butyrate on the expression of genes and proteins at G0/G1 and S phase of BRECs *** results showed that BRECs treated with butyrate inhibited(P<0.05)the proliferation of BRECs,relatively to *** cytometric assays revealed that butyrate triggers the BRECs cycle arrest at the G0/G1 ***-PCR analyses of mRNA level of genes involved in the G0/G1 phase of cell cycle showed that butyrate significantly upregulated(P<0.001)the expression of mRNA encoding p21^(Cip1)compared with control group,but it decreased(P<0.05)the mRNA levels of cyclin D1 and CDK4 genes at G0/G1 phase checkpoint compared with ***,Western blot also revealed that butyrate downregulated the expression of cyclin D3,CDK6,p-Rb,and E2F1 proteins involved in the modulation of G0/G1 phase of cell *** conclusion,our results demonstrated that butyrate inhibits the proliferation of BRECs via downregulation of positive regulators at G0/G1 phase checkpoint.

关键词： Butyrate Bovine rumen epithelial cells Proliferation G0/G1 phase

Nonculturability of the pathogenic Vibrio parahaemolyticus in live culture of Grateloupia turuturu is associated with bacterial attachment to the algal thalli

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Acta Oceanologica Sinica 2010年第6期29卷 92-103页

作者： LIU Feng PANG Shaojun Marine Biological culture collection Centre Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China Graduate University of Chinese Academy of Sciences Beijing 100049 China

The invasive red alga Grateloupia turuturu Yamada could turn Vibrio parahaemolyticus into nonculturable state in live algal *** order to elucidate the mechanism of such an effect,a series of culture experiments were performed in this investigation based on three hypothesized causes,namely bacterial attachment,production of reactive oxygen species （ROS） and the discharge of water soluble secondary metabolic *** results reveal that attachment to the thallus surface of *** was the major reason for the decrease of *** in *** investigations show that *** attachment to the surface of algal thallus in live cultures of seaweeds was a common ***,the disappearance of the culturability of *** occurred only on the thallus of *** over 72 h among all six algal species *** microscopic scanning shows that most of *** attached to *** changed from the initial normal bacilli to coccoid-shape after 72 *** enclosure experiments by enclosing the algal thallus in tubes demonstrate that the nonculturability of *** in the water of live culture of *** occurred after the physical contact of the *** to the *** capacity of *** in affecting the culturability of *** was not influenced after inhibition of photosynthesis by treatment of 3（3,4dichlorophenyl）-1 ,1dimethyl urea （DCMU） at non-lethal *** of reactive oxygen species after addition of live culture of bacteria was excluded by on-line analyzing the oxidation of dichlorohydrofluorescein （DCFH） to dichlorofluorescein （DCF） in the presence of peroxidase on a VersaFluor fluorometer.

关键词： Grateloupia turuturu attachment Vibrio parahaemolyticus nonculturability effective quantum yield

Temperature and light tolerance of representative brown,green and red algae in tumble culture revealed by chlorophyll fluorescence measurements

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Acta Oceanologica Sinica 2008年第4期27卷 137-146页

作者： PANG Shaojun SHAN Tifeng Marine Biological culture collection Center Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China Graduate University of Chinese Academy of Sciences Beijing 100049 China

Laminaria japonica, Undaria pinnatifida, Ulva lactuca, Grateloupia turuturu and Palmaria palmata are suitable species that fit the requirements of a seaweed-animal integrated aquaculture system in terms of their viable biomass, rapid growth and promising nutrient uptake rates. In this investigation, the responses of the optimal chlorophyll fluorescence yield of the five algal species in tumble culture were assessed at a temperature range of 10 - 30℃. The results revealed that Ulva lactuca was the most resistant species to high temperature, withstanding 30℃ for 4 h without apparent decline in the optimal chlorophyll fluorescence yield . While the arctic alga Palmaria palmata was the most vulnerable one, showing significant decline in the optimal chlorophyll fluorescence yield at 25℃ for 2 h. The cold-water species Laminaria japonica, however, demonstrated strong ability to cope with higher temperature （24 -26℃ ） for shorter time （within 24 h） without significant decline in the optimal chlorophyll fluorescence yield . Grateloupia turuturu showed a general decrease in the optimal chlorophyll fluorescence yield with the rising temperature from 23 to 30℃ , similar to the temperate kelp Undaria pinnatifida. Changes of chlorophyll fluorescence yields of these algae were characterized differently indicating the existence of species-unique strategy to cope with high light. Measurements of the optimal chlorophyll fluorescence yield after short exposure to direct solar irradiance revealed how long these exposures could be without significant photoinhibition or with promising recovery in photosynthetic activities. Seasonal pattern of alternation of algal species in tank culture in the Northern Hemisphere at the latitude of 36°N was proposed according to these basic measurements.

关键词： seaweeds chlorophyll fluorescence measurements Pulse Amplitude Modulation temperature tolerance integrated aquaculture

Purification and molecular weight distribution of a key exopolysaccharide component of Bacillus megaterium TF10

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Journal of Environmental Sciences 2018年第1期30卷 9-15页

作者： Lingling Wang Longfei Wang Qingshan Shi Hanqing Yu State Key Laboratory of Applied Microbiology Southern china Guangdong Provincial Key Laboratory of Microbial Culture Collection and ApplicationGuangdong Institute of Microbiology Department of Chemistry University of Science & Technology of China

Extracellular polymeric substances（EPS） are organic metabolic compounds excreted by microorganisms. They largely impact microbial aggregate structures and *** polysaccharides（EP） in EPS are responsible for the formation of microbial aggregates. In this work, we successfully separated and characterized EP from EPS of the bacterium Bacillus megaterium TF10. Extraction of EP from EPS was optimized using Sevag＇s reagent. Chemical characteristics, functional groups, and molecular weight（MW） distribution of EP were compared with the harvested EPS and soluble microbial products（SMP）. We found that the polymers of lower MW and free proteins were successfully removed by Sevag＇s reagent. The higher MW components of EPS were predominantly polysaccharides,while the polymers of lower MW tended to secrete to the supernatant and were described as SMP. A part of the proteins in the EP was polysaccharide-bonded. Our results can be further used in elucidating the complex flocculation mechanisms in which EP play a major role.

关键词： Extracellular polysaccharides(EP) Extracellular polymeric substances(EPS) Soluble microbial products(SMP) Molecular weight(MW) distribution

Liquid-nitrogen cryopreservation of three kinds of autotrophicbioleaching bacteria

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中国有色金属学会会刊：英文版 2008年第6期18卷 1386-1391页

作者：吴学玲辛晓红蒋莹梁任星袁鹏方呈祥 Key Laboratory of Biometallurgy of Ministry of Education School of Minerals Processing and BioengineeringCentral South University china Center for Type culture collection College of Life ScienceWuhan University

Three kinds of autotrophic bioleaching bacteria strains,including mesophilic and acidophilic ferrous ion-oxidizing bacteria Acidithiobacillus ferrooxidans(***),mesophilic and acidophilic sulfur-oxidizing bacteria Acidithiobacillus thiooxidans(***),and moderately thermophilic sulfur-oxidizing bacteria.4cidianus brierleyi,were cryopreserved in liquid nitrogen and their ferrous ion-or sulfur-oxidizing activities were investigated and compared with the original *** results revealed that ferrous ion/sulfur oxidation activities of the strains were almost equal before and after *** was used as cryoprotective *** conclusion,liquid-nitrogen cryopreservation is a simple and effective method for autotrophic bioleaching microorganisms.

关键词：液态氮超低温保存生长活性细菌

The protective roles of tea tree oil extracts in bovine mammary epithelial cells and polymorphonuclear leukocytes

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Journal of Animal Science and Biotechnology 2020年第4期11卷 1117-1128页

作者： Kang Zhan Tianyu Yang Baobao Feng Xinyu Zhu Yinyin Chen Yongjiu Huo Guoqi Zhao Institute of Animal culture collection and Application College of Animal Science and TechnologyYangzhou UniversityYangzhou 225009China Wuxi Chenfang Biotechnology Co. Ltd.WuxiChina

Background: Tea tree oil(TTO) plays an important role in antibacterial activity and alleviating the inflammatory responses. Bovine mammary epithelium and polymorphonuclear leukocytes(PMNL) can actively respond to bovine mastitis infection. However, regulatory effects of TTO extracts on the innate immune response of bovine mammary epithelial cells(BMECs) and PMNL remain not reported. Therefore, aim of the study was to evaluate the effects of TTO extracts on the m RNA levels of the genes involved in the innate immune response of BMECs and ***: Our results demonstrated that addition of 0.025% and 0.05% TTO increased the proliferation of BMECs, and significantly enhanced(P < 0.05) the viability of BMECs exposed to Staphylococcus aureus(S. aureus). An inhibitory effect was observed against the growth of S. aureus by TTO incubation. The 0.05% TTO reduced S. aureus biofilm formation, association and invasion of S. aureus to BMECs, and changed the morphological and structural features of S. aureus. The proinflammatory cytokines IL-1β, IL-6, and TNF-α were decreased(P < 0.001) by the incubation of TTO. Interestingly, the expression of IL-8 known for PMNL chemotactic function was elevated(P < 0.05) by 0.05%TTO treatment. Consistently, 0.05% TTO increased the migration of PMNL in S. aureus-exposed BMECs when compared with S. aureus treatment alone(P < 0.05). In addition, PMNL incubated with 0.05% TTO decreased the levels of NFKB inhibitor alpha(NFKBIA) and TNF-α.Conclusions: Our results indicate that use of TTO can relieve the BMECs pro-inflammatory response caused by *** and promote the migration of PMNL to mount the innate immune responses, and it may be novel strategy for the treatment of bovine mastitis caused by S. aureus.

关键词： Bovine mammary epithelial cells Mastitis PMNL Staphylococcus aureus Tea tree oil

Circadian rhythms in the growth and reproduction of the brown alga Undaria pinnatifida and gametogenesis under different photoperiods

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Acta Oceanologica Sinica 2007年第4期26卷 104-111页

作者： ZHANG Zhihuai PANG Shaojun Qingdao Jinying Ocean Science Technology Development Co Ltd Qingdao 266520 China Marine Biological culture collection Centre Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China

Circadian growth rhythm of the juvenile sporophyte of the brown alga Undaria pinnatifida was measured with the computer-aided image analysis system in constant florescent white light under constant temperature （ 10 ℃ ）. The growth rhythm persisted for 4 d in constant light with a free-running period of 25.6 h. Egg release from filamentous gametophytes pre-cultured in the light - dark regime was evaluated for six consecutive days at fixed time intervals in constant white light and 12 h light per day. Egg release rhythm persisted for 3 d in both regimes, indicating the endogenous nature. Temporal scale of egg release and gametogenesis in 18, 16, 12 and 8 h light per day were evaluated respectively using vegetatively propagated filamentous gametophytes. Egg release occurred 2 h after the onset of dark phase and peaked at midnight. Evaluation of the rates of oogonium formation, egg release or fertilization revealed no significant differences in four light-dark regimes, indicating the great plasticity of sexual reproduction. No photoperiodic effect in gametogenesis in terms of oogonium formation and egg release was found, but fertilization in short days was significantly higher than in long days. Results of this investigation further confirmed the general occurrence of circadian rhythms in intertidal seaweed species.

关键词： Undaria pinnatifida circadian rhythm egg release gametogenesis