短期连续剪切对光生物反应器内海带配子体细胞生长及其恢复能力的影响

以不同搅拌速率（0～1000r/min）对海带（Laminaria japonica）配子体细胞施加短期（0～60h）连续剪切,卸载剪切力之后细胞静止恢复23.5d,在此期间研究不同搅拌速率分别在连续剪切和恢复期间对细胞生长及其恢复能力的影响。研究结果表明,连续剪切期间,90r/min下细胞叶绿素浓度积累达到最大值2.36mg/L;中高速搅拌速率（270～1000r/min）下叶绿素浓度迅速下降,胞内氮磷池释放,1000r/min下细胞损伤率为静止对照样的18倍。恢复期间,所有组别细胞均呈现较强的恢复能力。此外,在连续剪切实验中,海带配子体细胞内叶绿素浓度比干重更能有效的表达生物量浓度,除细胞损伤率外,磷源释放可以作为判别细胞受损的参考指标之一。     

斜生栅藻对振荡和磷胁迫的生理生化响应

为了探讨水文条件的变化对常见绿藻水华发生的潜在影响,实验以斜生栅藻为材料,研究了不同水流状态及磷浓度对其生长及磷利用策略的影响。实验分别设置静止条件(0 r/min)、低振荡条件(90 r/min)和高振荡条件(120 r/min),同时设置磷限制(0.2 mg/L)和磷充足(2 mg/L)两组磷浓度,整个实验过程为3周。实验过程测定的指标为:比生长速率、胞外碱性磷酸酶活性、叶绿素a含量以及磷吸收动力学参数。实验结果表明:(1)同静止条件相比,低振荡和高振荡条件均能显著降低斜生栅藻的比生长速率,但是两种不同磷浓度下其比生长速率却无显著性差异。(2)在实验第21天时,振荡条件培养下,斜生栅藻的胞外碱性磷酸酶活性显著高于静止条件培养,与此相反,胞内磷浓度却显著高于静止条件。(3)在磷限制条件下斜生栅藻的叶绿素a含量显著降低,同时两种振荡条件培养均使其在磷充足条件下叶绿素a含量降低。(4)通过磷吸收动力学参数的比较,静止条件培养的斜生栅藻对磷的亲和力高于其在振荡条件下。由此可见,斜生栅藻适应于静止无扰动且磷营养丰富的水体,随着水体中磷浓度进一步升高,静止和水流缓慢的水体存在导致像斜生栅藻这一类绿藻发生水华的风险。       

搅拌式光生物反应器培养海带配子体细胞：不同脉冲补料方式的研究

本文以批次培养为对照,研究了七种脉冲补料方式对搅拌式光生物反应器中培养大型褐藻海带配子体细胞生长和培养液内氮磷营养盐消耗的影响,并首次探讨了脉冲补料方式下不同补料时间点和补料量的影响作用。培养条件设定为50 mg DCW (细胞干重) L-1接种密度、培养液为改良的APSW人工海水、光强60 µE m-2 s-1、光周期16/8 h L/D、通气速率和搅拌速率分别为50 mL min-1和100 rpm。结果表明少量补料利于细胞对氮磷的协同吸收,进而利于生物量扩增。当培养液内氮磷富足或耗尽时补料对于生物量大量生产效果甚微,可能由于氮磷吸收变缓、其储存现象显著,或是其吸收协同性降低。文中当细胞生长至对数中期开始频繁补加少量氮磷营养盐,即维持培养液内氮磷浓度在各自起始浓度的1/3至1/2之间,对生物量生产最有效,生物量增长倍数高达12.270倍。     

两阶段搅拌转速控制策略发酵生产聚唾液酸

目的:优化聚唾液酸发酵过程的搅拌转速.方法:比较不同搅拌转速对大肠杆菌Escherichia coli K235分批发酵生产聚唾液酸过程的影响.结果:根据发酵前、后期菌体细胞比生长速率和聚唾液酸比合成速率达到最大值所需搅拌转速的不同,提出了两阶段搅拌转速控制策略:发酵前期(0～15h)控制搅拌转速500r/min,发酵中后期控制搅拌转速700r/min.结论:两阶段搅拌转速控制策略使聚唾液酸产量达到3 966mg/L,比恒定搅拌转速500r/min和700r/min分别提高了31.8%和49.3%.将两阶段搅拌转速控制策略与分批补料发酵技术结合,聚唾液酸产量提高到5 108mg/L,山梨醇的转化率达到0.12g/g.     

Pluronic和纤维素类对杂交瘤细胞保护特性的研究

利用鼠鼠杂交增2F7细胞(分泌IgG2a单抗)研究了Pluronic F-68、甲基纤维素、羧甲基纤维素及聚醚多元醇与杂交瘤细胞生物相容性及添加限制浓度;研究了添加浓度对葡萄糖的利用及氨的生成影响}在高速搅拌、高剪切力下考查添加剂的保护效果。结果表明,O·05—0·10％(w／V)Pluronic F-68、O．10—0．20％(w／V)甲基纤维素能较好地保护杂交瘤细胞;高浓度Pluronic F-68增加了葡萄糖的比消耗速率及氨的比生成速率;高浓度甲基纤维素增加了氨比生成速率;羧甲基纤维素添加浓度低于0．1％不影响细胞生长,也无保护作用,羧甲基纤维素不影响细胞对葡萄糖的比消耗速率,但增加了氨比生成速率,聚醚多元醇分解细胞。在1．5升GemlliGen生物反应器中,培养基添加0．10％Pluronic F-68、搅拌转速70r／min下细胞正常生长。     

光照与氮磷营养盐的协同作用对海带幼苗释放可溶性有机碳的影响

文章以海带(Saccharina japonica)幼苗为实验材料, 对比了其在自然海水+50%海面光强、自然海水+100%海面光强、营养盐加富海水+50%海面光强、营养盐加富海水+100%海面光强4种条件下的溶解有机碳(DOC)释放速率, 以揭示光照与营养盐的协同作用对大型藻类释放DOC的影响机制。研究发现, 在自然海水条件下, 50%与100%海面光强照射下海带幼苗DOC释放速率分别为(11.67±3.07)和(22.65±4.58) μmol/(g·h), 随着光强增加, 释放DOC速率显著提升(P＜0.05), 氧气净释放速率显著增大(P＜0.05), 且二者光谱斜率差异显著(P＜0.05); 在营养盐加富海水条件下, 50%与100%海面光强照射下海带幼苗DOC释放速率分别为(30.88±7.96)和(39.03±14.78) μmol/(g·h), 随着光强增加, 释放DOC速率提升不显著(P＞0.05), 氧气净释放速率显著提高79.24%(P＜0.05), 二者光谱斜率差异不显著(P＞0.05)。结果表明, 在寡营养条件下(自然海水), 海带幼苗释放DOC速率与光照强度呈正相关, 表现为“溢出”机制; 在富营养条件下, 海带幼苗释放DOC速率与光照强度不相关, 表现为“扩散”机制。海带幼苗释放DOC同时受到“溢出”和“扩散”两种机制的调控, 何种机制占主导取决于营养水平。     

培养方式对真皮组织体外构建的影响

采用静态培养和转瓶培养方式分别构建真皮组织,考察培养方式和搅拌转速对细胞在三维支架材料中增殖、代谢、分布的影响。结果表明,由转瓶培养方式构建的细胞-材料复合物,其最终细胞密度和细胞比生长速率均明显高于静态培养（14.2~27.6×106 cells/cm3 vs 10.1×106 cells/cm3和0.145~0.262 d-1 vs 0.111 d-1）,而转速达80 r/min的转瓶尤其突出;静态培养的细胞-材料复合物内部细胞稀少,且分布不均匀,转瓶培养的细胞-材料复合物在材料表面和内部细胞密度都有所提高,分布情况也得到改善,且80 r/min转瓶培养的组织其细胞密度和分布均优于10 r/min和40 r/min转瓶培养。转瓶培养在其转速达到一定强度时能明显提高细胞在支架中的增殖速率,缩短培养时间,并有效改善细胞在支架内的分布,是一种理想的培养方式。     

杜仲对铜胁迫后杜氏盐藻恢复速率的影响

本研究通过检测不同浓度的杜仲药液对铜胁迫下的盐藻细胞数和叶绿素含量,来探讨杜仲对铜胁迫后杜氏盐藻恢复速率的影响。结果表明,适当浓度的杜仲药液会加快盐藻的恢复速率,其中5mg/ml的药液作用最为明显。     

控氧发酵对干酪乳杆菌合成苯乳酸的影响

在静置、搅拌及通气搅拌3种不同控氧条件下,分别用干酪乳杆菌Lactobacillus casei B3发酵及全细胞转化合成了苯乳酸,考察菌体生长、葡萄糖消耗及其发酵与转化合成苯乳酸的规律。结果表明:在转速100 r/min的搅拌条件下,L.casei B3发酵合成苯乳酸的浓度比静置发酵条件下提高了41.4%;但在空气流量2 L/min及转速100r/min的通气搅拌下,发酵合成苯乳酸的浓度较静置发酵时下降了60.3%;以8 g/L苯丙酮酸为底物,以相应静置、搅拌及通气搅拌条件下所得的菌体为全细胞催化剂转化合成苯乳酸,其摩尔转化率分别为67.2%、62.7%和35.9%。此结果说明:适度的搅拌促进了发酵过程的底物和产物传质,但充足或过量供氧会影响细胞内的转化合成酶系,不利于苯乳酸的全细胞转化合成。     

桑沟湾春季叶绿素a浓度分布及其影响因素

依据2014年5月走航和定点连续调查资料,分析了桑沟湾叶绿素a的空间分布及昼日变化特征,结合理、化环境因素的相关性分析,探讨影响叶绿素a浓度的主要因素。(1)走航调查的结果显示,桑沟湾春季叶绿素a浓度较低,叶绿素a浓度范围为0.11—1.40μg/L,平均为(0.64±0.36)μg/L。叶绿素a浓度从湾内向湾外逐步降低,贝类区混养区海带区外海区;湾内表层叶绿素a浓度均高于底层,而湾外的非养殖区则相反(2)网箱区叶绿素a浓度最高,日平均为1.70μg/L,显著高于其它3个区;海草区最低,为0.57μg/L,与海带养殖区无显著性差异,显著低于贝类养殖区和网箱区。叶绿素a浓度的总体趋势为:网箱区贝类区海带区和海草区。而且,不同养殖区域,叶绿素a浓度昼夜变化规律各不相同,反映了养殖活动的影响。海草区白天表层高于底层,而夜间则相反;网箱区底层均高于表层;贝类区表层均高于底层;海带区表底层叶绿素a浓度呈现出升降交替的规律。(3)叶绿素a浓度与硅酸盐、水温显著正相关,与其他环境因子,如氨氮、亚硝酸盐、磷酸盐等无显著相关性。硅酸盐和温度可能为影响桑沟湾春季浮游植物生长的主要限制性因素。(4)桑沟湾春季浮游植物生长受多重因素的限制,湾内营养盐浓度与叶绿素a浓度并未呈现显著的规律性,营养盐的上行控制和贝类摄食的下行控制均能影响浮游植物的生长。     

Agitation,aeration and perfusion modules for cell culture bioreactors

For an optimized bioreactor design which is adapted to the cultivation of sensitive animal cells different modular bioreactor components for gentle agitation, sufficient aeration and long-term perfusion were developed and investigated with respect to their suitability from laboratory to production scale. Aeration systems have been designed for both shear sensitive cells and cells which tolerate bubbles. The systems are based on either membranes for bubble-free aeration or stainless steel sparger systems. They were characterized by determination of their oxygen transfer capacity and optimized in cultivation processes of different cell lines under process conditions such as batch and perfusion mode.Different impellers for suspension cells and cells grown on carriers were investigated for their suitability to ensure homogeneous gentle mixing. A large pitch blade impeller as well as a novel 3-blade segment impeller are appropriate for homogeneous mixing at low shear rates. Especially with the 3-blade segment impeller fluid mechanical stress can be reduced at a given stirrer speed which is advantageous for the cultivation of cells attached to microcarriers or extremely shear sensitive suspension cells. However, our results indicate that shear sensitivity of animal cells has been generally overestimated.Continuous perfusion of both suspension cell cultures and cells cultivated on microcarriers could be successfully performed over extended periods of time using stainless steel spinfilters with appropriate pore sizes and systems based on microporous hydrophilic membranes. Spinfilters are suitable cell retention systems for technical scale bioreactors allowing continuous perfusion cultures of suspension cells (pore size 10 to 20 m) as well as anchorage dependent cells grown on microcarriers (pore size 75 m) over six weeks to 3 months.Applying the developed modules for agitation, aeration and perfusion process adapted bioreactor set-ups can be realized which ensure optimum growth and product formation conditions in order to maximize cell and product yields.     

高温胁迫下三唑酮对黄瓜幼苗某些生理性质的影响

研究了三唑酮浸种对黄瓜幼苗抗热性的影响。结果表明,高温胁迫下黄瓜幼苗子叶叶绿素a、b及可溶性蛋白质含量下降,Chla／b比值、游离脯氩酸和可溶性糖含量增加。20mg／L三唑酮浸种抑制了黄瓜幼苗子叶的叶绿素a、b和可溶性蛋白质含量的下降及可溶性糖含量的增加,但脯氨酸含量降低,Chla／b比值上升。恢复温度后子叶的叶绿素a、b及可溶性蛋白、脯氨酸和可溶性糖含量与胁迫后相比均有不同程度的下降,但三唑酮处理下降的幅度低于对照。综上结果,三唑酮增强了黄瓜幼苗的耐热性。     

Development of Oxidative Stress in the Cells of Chlorella stigmatophoraand Their Bleaching by the Glycolate Pathway Inhibition under Salinization Conditions

The changes in chlorophyll and proline contents, the rate of photosynthetic oxygen evolution, the activity of superoxide dismutase (SOD), and NADPH concentration in the cells of Chlorella stigmatophoraButcher were followed under the conditions when the glycolate pathway was inhibited by 10 mM -hydroxy-2-pyridinemethane sulfonate + 10 mM isonicotinoyl hydrazine or with 425 mM NaCl. The oxidative stress exerted by the inhibition of the glycolate pathway developed in three phases. At the first phase (15 to 60 min), the photosynthetic rate slightly increased, chlorophyll and proline contents and SOD activity declined, whereas NADPH pool increased considerably. At the second phase (1 to 7 h), in addition to a short-period chlorophyll accumulation, proline synthesis enhanced together with SOD activity, whereas NADPH pool decreased. Degradation processes dominated the third phase (24 to 72 h): cells absorbed rather than evolved oxygen, and the contents of proline, chlorophyll, and NADPH as well as the SOD activity decreased dramatically. The authors conclude that the inhibition of photorespiration in chlorella cells disrupted the functions of the electron transport chain (ETC); as a result, the oxygen stress developed, and cells became bleached. The accumulation of free proline in the cells under salinization provides a mechanism subduing ETC overreduction, and photorespiration is a component of the cell antioxidant system.     

Effect of phosphorus and temperature on chlorophyll <Emphasis Type="Italic">a</Emphasis> contents and cell sizes of <Emphasis Type="Italic">Scenedesmus obliquus</Emphasis> and <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis>

An experiment was carried out to evaluate the effects of phosphorus concentration (1, 4 and 10 mg l⁻¹) and temperature (15 and 25°C) on chlorophyll a (chl a) contents and cell size/volume of green alga Scenedesmus obliquus and blue green alga Microcystis aeruginosa. Long-term field data from Lake Taihu, a large, shallow eutrophic lake between Jiangsu and Zhejiang Provinces, China, was also used to evaluate the effect of temperature on the model between chl a and total phosphorus (TP). The chl a content of both algae increased with an increase in phosphorus concentration and temperature. Temperatures showed a significantly different effect on chl a content of S. obliquus at a phosphorus concentration of 10 mg l⁻¹, whereas there was no significant difference at the two lower phosphorus levels. For M. aeruginosa, temperatures presented significantly different effects on the chl a contents at three phosphorus concentrations. Chl a content of neither alga presented an interaction between the nutrient and the temperature. Long-term field data from Lake Taihu also indicated that the addition of temperature to the model increased predictability of chl a by TP. The length/diameter and volume of both algae were greater at the lower temperature and phosphorus concentration. Moderate negative correlations were observed between algal size, volume, and chl a content. Our results suggest that phosphorus concentration and temperature could change chl a contents and size in species-specific algal cells and that temperature should be considered when building the model of TP and chl a concentration.     

CO2和O3浓度倍增及其复合作用对大豆叶绿素含量的影响

利用开顶箱(OTC)法研究了在CO2和O3浓度倍增及其复合作用下,大豆叶片叶绿素含量及叶绿素a／b值的变化规律。结果表明,不同生育时期大豆叶片中叶绿素含量不同,Chla、Chlb和ChlT都表现出低．高一低的趋势,而且不同处理间变化不同步。不同处理间比较,O3处理的植株叶绿素含量下降最为明显,其次是复合处理的影响,而CO2浓度倍增对提高叶片叶绿素含量有一定的作用。Chla/b呈下降趋势,受CO2倍增影响最明显,有利于提高作物的光合性能。     

Adaptation of the thylakoid membranes of pea chloroplasts to light intensities. I. Study on the distribution of chlorophyll-protein complexes

The effect of light intensity (16 h white light and 8 h dark) during growth of pea plants at 20°C on the chlorophyll composition and on the relative distribution of chlorophyll amongst the various chlorophyll-protein of pea thylakoids was studied. The chl a/chl b ratios increased from 2.1 to 3.2 as light intensity during growth varied from 10 to 840 Em^-2 s^-1. This function can be described by two straight lines intersecting at a transition point of approximately 200 Em^-2 s^-1. Similar discontinuities in the responses were observed in the changes in the relative distribution of chlorophyll amongst the various chlorophyll-protein complexes. This demonstrates that the chl a/chl b ratio of the various thylakoids is a good indicator of changes in the relative distribution of chlorophyll. As the chl a/chl b ratio decreased, the amount of chlorophyll associated with photosystem I complexes decreased, that with photosystem II core reaction centre complex was halved, and that with the main chl a/b-proteins of the light-harvesting complex was markedly increased.Abbreviations chl chlorophyll - PS photosystem - SDS sodium dodecyl sulphate - Tricine N-tris (hydroxymethyl) methylglycine     

PHOTOACCLIMATION AND GROWTH RATE RESPONSES OF ULVA ROTUNDATA (CHLOROPHYTA) TO INTRADAY VARIATIONS IN GROWTH IRRADIANCE1

A vegetative clone of the chlorophyte macroalga Ulva rotundata was maintained in an outdoor continuous flow system under nutrient sufficient conditions and various light regimes. Step changes between 9 and 100% incident irradiance (I^o) were employed to simulate cloud passage. Temporary (1–4 h) midday (I^o)perturbations evoked net changes in growth rate (μ) and chlorophyll (chl) content. Under I_o alternating at various periodicities from 15 min to 7 h, net μ was the average of the μ under steady state 9 and 100% I^o, regardless of periodicity. However, the μ in alternating light was considerably less than μ under steady state 55% I^o(? 9%+ 100%/2), as expected based on the nonlinear shape of the μ-I relationship. Unlike μ, chl content depended primarily on the total daily irradiance, probably clue to the slower response of chl compared to photosynthetic rate. On time scales ≥ one day, chl was linearly correlated with light-regulated daily μ under both steady state and intraday fluctuating irradiance, consistent with photosynthetic feedback regulation of chl concentration.     

流体动力对HEK293细胞的细胞团形成及细胞生长和代谢的影响

利用HEK293细胞在悬浮培养中具有聚集成团的体外培养特性,在250mL的Bellco的搅拌培养体系中,以HEK293细胞团的粒径、细胞数、细胞活力、葡萄糖比消耗率(qglc)、乳酸比产率(qlac)和乳酸转化率(Ylacglc)为观察指标,考察HEK293细胞在搅拌速度分别设置为25、50、75和100rmin的培养条件下的细胞团形成、粒径分布以及细胞生长和代谢。HEK293细胞在搅拌速度为50rmin和75rmin培养条件下所形成细胞团的粒径大小适中、离散度小。培养7d后,HEK293细胞团的平均粒径分别为201μm和175μm,其中粒径≥225μm的细胞团所占比例均低于10%;在整个培养过程中,细胞团中的HEK293细胞活力维持在90%以上,qglc、qlac和Ylacglc等反映HEK293细胞代谢的参数保持相对恒定。实验结果提示:合适的搅拌速度所产生的流体动力既可使细胞团的粒径控制在合适的范围内,也可为细胞团中的HEK293细胞提供基本满足其正常生长和代谢需要的物质传递效率。     

The chl a fluorescence intensity is remarkably insensitive to changes in the chlorophyll content of the leaf as long as the chl a/b ratio remains unaffected

The effects of changes in the chlorophyll (chl) content on the kinetics of the OJIP fluorescence transient were studied using two different approaches. An extensive chl loss (up to 5-fold decrease) occurs in leaves suffering from either an Mg(2+) or SO(4)(2-) deficiency. The effects of these treatments on the chl a/b ratio, which is related to antenna size, were very limited. This observation was confirmed by the identical light intensity dependencies of the K, J and I-steps of the fluorescence rise for three of the four treatments and by the absence of changes in the F(685 nm)/F(695 nm)-ratio of fluorescence emission spectra measured at 77K. Under these conditions, the F(0) and F(M)-values were essentially insensitive to the chl content. A second experimental approach consisted of the treatment of wheat leaves with specifically designed antisense oligodeoxynucleotides that interfered with the translation of mRNA of the genes coding for chl a/b binding proteins. This way, leaves with a wide range of chl a/b ratios were created. Under these conditions, an inverse proportional relationship between the F(M) values and the chl a/b ratio was observed. A strong effect of the chl a/b ratio on the fluorescence intensity was also observed for barley Chlorina f2 plants that lack chl b. The data suggest that the chl a/b ratio (antenna size) is a more important determinant of the maximum fluorescence intensity than the chl content of the leaf.     

Fluid-mechanical forces in agitated bioreactors reduce the CD13 and CD33 surface protein content of HL60 cells

Flow cytometry was used to examine the effect of hydrodynamic forces in surface aerated stirred tank bioreactors on the quantity of CD13 and CD33 surface proteins of Hl60 (human promyelocytic leukemia) cells. A step increase in agitation of the 2-L bioreactors from 80 to 400 rpm reduced the apparent growth rate and the average CD13 and CD33 content per HL60 cell. The effects on the two surface proteins were observed within 30-60 min following the increase in the agitation and preceded observed effects on cell growth by at least 10 h. Upon reduction of the agitation rate back to 80 rpm, the CD13 and CD33 content recovered (in ca. 10 h) for CD13 and ca. 29h for (CD33) to the levels of the control culture whose agitation rate was maintained at 80rpm. The CD13 and CD33 cell content was reduced even at agitation rates (270 rpm) that did not affect cell proliferation. Pluronic F68 (a commonly used shear protectant) had a protective effect on the CD33 content per cell of cultures subjected to hydrodynamic injury but no effect on the CD13 cell content. Possible bioprocessing and physiological implications of these findings are discussed (c) 1993 Wiley & Sons, Inc.