镧对软腐欧文氏菌生长及其胞外酶活性的影响

研究了镧对软腐细菌（Ｅｒｗｉｎｉａｃｈｒｙｓａｎｔｈｅｍｉ，Ｅｃｈ）生长及胞外酶活性的影响。结果表明，Ｌａ３＋浓度为５０、１００、１５０、２００、２５０、３００和３５０ｍｇ／Ｌ时，固体培养中对Ｅｃｈ生长均呈抑制作用，且随浓度提高，抑制作用加强，表现为菌落出现时间比对照延长，７ｄ后的菌落直径比对照减小。液体培养中，浓度＜２００ｍｇ／Ｌ，Ｌａ在２４ｈ内对Ｅｃｈ生长有轻微刺激作用；但随着浓度提高和培养时间的延长，Ｅｃｈ生长明显受到抑制。当Ｌａ３＋浓度＞３５０ｍｇ／Ｌ，Ｅｃｈ生长基本处于停止状态；浓度为２００ｍｇ／Ｌ，对Ｅｃｈ产生胞外酶能力有一定影响，其中对纤维素酶活性增强最明显，其次是蛋白酶，而对果胶酶活性有减弱作用。经稀土处理的无细胞滤液对马铃薯块茎组织的浸离力降低。     

硒的热化学研究：IV.微量热法研究Na2SeO3对微生物生长代谢的抑制

用微量热法研究了对大肠杆菌、产气杆菌、枯草杆菌、黑根霉菌４种菌作用的产热曲线，计算了微生物的生长速率常数ｋ，比较了Ｎａ２ＳｅＯ３对４种微生物作用的半抑制浓度ＩＣ５０，发现Ｎａ２ＳｅＯ３浓度低于１０ｍｇ／Ｌ时，对微生物生长有促进作用，高于此浓度时有抑制作用，对不同微生物生长代谢的抑制情况不同，反映了微生物之间的差异和硒对细胞作用热动力学特征的不同。     

镧对发菜细胞培养及氨基酸成分的影响

用Ｌａ（ＮＯ３）３处理液体培养的发菜细胞，研究镧对发菜细胞的生长及氨基酸成分的影响。结果表明，浓度为１－２００ｍｇ．Ｌ＾－１的Ｌａ（ＮＯ３）３均能促进发菜细胞的生长，以１００ｍｇ．Ｌ＾－１Ｌａ（Ｎｏ３）３作用最明显，     

镧对中国对虾卵子孵化和无节幼体变态的影响

研究了Ｌａ对中国对虾胚能和无节幼体生长的影响。实验结果表明，对胚肥发育期为单，二细胞的卵子，Ｌａ的最佳浓度范围为０．３０－１．８０ｍｇ／Ｌ，提高孵化率３３．％－４９．１％；对发育阶段为多细胞期的卵子，最佳浓度范围为０．９０－１．８０ｍ，ｇ＝Ｌ，提高孵化度１７．１％－２３．５％。     

La对Pb， Cd复合污染大豆幼苗的缓解作用

以盆栽法研究了Ｐｂ，Ｃｄ复合污染对大豆幼苗的影响及Ｌａ对Ｐｂ，Ｃｄ复合胁迫的缓解效应。结果显示，Ｐｂ５００ｍｇ／Ｌ和Ｃｄ１００ｍｇ／Ｌ的混合液严重抑制大豆幼苗代谢与生长。叶面喷施３０ｍｇ／ＬＬａＣｌ３一次，能减轻Ｐｂ，Ｃｄ复合污染造成的伤害。这与Ｌａ能增加大豆幼苗光合速率，提高叶绿素含量与硝酸还原酶活性，降低质膜透性与植株中Ｐｂ，Ｃｄ含量，维持ＴＴＣ还原力等多重作用相关。     

离子色谱法测定NO_2~－时Cl~－的干扰及消除

研究了氯离子对化学抑制型高效离子色谱法测定亚硝酸根时的干扰。当亚硝酸根的浓度大于２．０ｍｇ／Ｌ时，５．０ｍｇ／Ｌ的氯离子不会对亚硝酸根的峰高信号产生明显的干扰；但当亚硝酸根浓度小于０．５０ｍｇ／Ｌ时，即可引起正干扰。该正干扰随着Ｃｌ~－／ＮＯ~－２浓度比的增加而迅速增大，但与流动相的流速（在２．５～３．０ｍＬ／ｍｉｎ之间）无关。还对４种可能消除氯离子干扰的方法进行了讨论。     

氯化钐对体外培养甲状腺细胞的影响

应用细胞培养、电镜及放射免疫等技术，研究了不同剂理氯化钐对体外培养的猪甲状腺细胞形态与功能的影响。结果表明，ＳｍＣｌ３浓度为０．００１和０．０１ｍｍｏｌ／Ｌ，测得四碘甲状原氨酸（Ｔ４）数值较对照组明显升高，其摄碘量较对照组增多；电镜下甲状腺细胞呈功能旺盛相；当ＳｍＣｌ３浓度为０．１ｍｍｏｌ／Ｌ时，Ｔ４数值较对照组降低，其摄碘量减少，电镜下甲状腺细胞呈功能不活跃状态。由此可见，ＳｍＣｌ３浓度为０．０     

芦荟大黄素的电化学研究

用单扫示波极谱法研究了芦荟大黄素的电化学行为。在５％Ｎａ２ＣＯ３＋５％ＮａＯＨ（９＋１）底液中，芦荟大黄素于－０．８４Ｖ（Ｐ１）和－０．９７Ｖ（Ｐ２）处产生两个示波极谱峰，利用Ｐ１可测定芦荟大黄素，线性范围为 ０．１１－２４ｍｇ／Ｌ和３．０－２８．６ｍｇ／Ｌ，检测限为０．０６ｍｇ／Ｌ。将该法应用于中药大黄中的芦荟大黄素的测定，结果满意。另外，讨论了芦荟大黄素清除由邻苯三酚自氧化产生的超氧自由基（Ｏ２）的作用。     

离子色谱法测定NO_2~－时Cl~－的干扰及消除

 研究了氯离子对化学抑制型高效离子色谱法测定亚硝酸根时的干扰。当亚硝酸根的浓度大于２．０ｍｇ／Ｌ时，５．０ｍｇ／Ｌ的氯离子不会对亚硝酸根的峰高信号产生明显的干扰；但当亚硝酸根浓度小于０．５０ｍｇ／Ｌ时，即可引起正干扰。该正干扰随着Ｃｌ~－／ＮＯ~－２浓度比的增加而迅速增大，但与流动相的流速（在２．５～３．０ｍＬ／ｍｉｎ之间）无关。还对４种可能消除氯离子干扰的方法进行了讨论。     

固定化米曲霉催化拆分丙氨酸消旋体的反应特性

研究了溶液ｐＨ、搅拌强度，催化剂用量、底物浓度及产物浓度对固定化米曲霉催化拆分Ｎ－乙酰－ＤＬ－丙氨酸反应的影响。结果表明，低底物浓度（Ｓ〈０．２ｍｏｌ／Ｌ）时反应符合Ｍ－Ｍ方程，底物浓度Ｓ〉０．２ｍｏｌ／Ｌ时，有底物抑制现象，当产物浓度Ｐ〉１０ｍｍｏｌ／Ｌ时反应存在产物抑制现象。     

普通小球藻对CTAC污染胁迫的响应及耐受能力研究

研究了不同初始浓度的十六烷基三甲基氯化铵(CTAC)对普通海水小球藻叶绿素a、可溶性蛋白含量、抗氧化酶活性及超微结构的影响。通过实验探究了CTAC对小球藻的毒性作用及小球藻对CTAC污染胁迫的响应及耐受能力,结果表明,CTAC浓度达到0.6 mg·L^-1时,细胞开始受到氧化损伤,但可以通过抗氧化酶清除细胞内产生的活性氧;当浓度达到0.9 mg·L^-1时,细胞生长完全受到抑制;当浓度大于1.2 mg·L^-1时,出现负增长。CTAC胁迫浓度高于0.9 mg·L^-1时,小球藻细胞的超微结构受到严重破坏。因此,阳离子表面活性剂CTAC对小球藻表现出较强毒性,但小球藻通过抗氧化酶对氧化应激的调控亦表现出对CTAC具有一定的耐受能力。     

Two-dimensional electrophoresis of Cereus peruvianus (Cactaceae) callus tissue proteins

Two-dimensional electrophoresis of Cereus peruvianus callus tissues grown in culture media containing two different 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin combinations was used to identify minor differences in polypeptide composition of these cell clones. Altered expression during growth in the two 2,4-D and kinetin combinations was apparent for 13 polypeptides when calluses in the two media were compared. The number of proteins with differential expression (presence or absence of specific spots) was higher in callus tissues cultured in the 4.0 mg/L 2,4-D and 8.0 mg/L kinetin combination than in callus tissues cultured in the 4.0 mg/L 2,4-D and 4.0 mg/L kinetin combination. The present results show that the callus tissues maintained at 4.0 mg/L 2,4-D and 8.0 mg/L kinetin can be used as a matrix for in vitro selection programs.     

Sesbania drummondii cell cultures: ICP-MS determination of the accumulation of Pb and Cu

Sesbania cell cultures grown in the presence of different concentrations of Pb (0-1000 mg/L) and Cu (0-500 mg/L) were assayed for growth, metal accumulations and activities of antioxidative enzymes: superoxide dismutase (SOD), catalase (CAT) and guaiacol peroxidase (GPX). These cultures tolerated Pb up to a concentration of 500 mg/L, registering a fresh weight growth of 500% in 3 weeks. At the same time, cultures registered a growth of 200% in 3 weeks at a Cu concentration of 100 mg/L showing less tolerance than Pb. However, Sesbania cells accumulated more Cu than Pb, as determined by ICP-MS, at all the treatments tested. Cu accumulation reached 3000 mg/kg (dry weight) at a Cu treatment of 100 mg/L, while Pb accumulation was only over 150 mg/kg (dry weight) at 500 mg Pb/L. Metal accumulations were positively correlated with induction of SOD and CAT activities in both the metal treatments. SOD activity of callus was 105 U/mg (fresh weight) at a Pb treatment of 500 mg/L and the corresponding Pb accumulation of 160 mg/kg (dry weight), while the activity rose to 300 U/mg (fresh weight) at a Cu treatment of 100 mg/L and the corresponding Cu accumulation of 3000 mg/kg (dry weight). The pattern of GPX activities was, however, different, particularly in Pb treatments where activities declined with increasing concentrations of Pb in the cells as well as growth medium. This study shows how Sesbania cells withstand heavy metal stress by induction of antioxidative enzyme activities.     

Growth kinetics and diosgenin estimation from callus cultures of Costus speciosus (Koen. ex. Retz.)

The present investigation reports the growth kinetics and diosgenin accumulation in callus cultures of Costus speciosus. Effect of explants, media and plant growth regulators was evaluated with respect to callus induction and growth. Out of the two explants viz pseudostem and seed, pseudostem showed maximum callus induction frequency of 90% on MS medium. The fresh weight of callus was maximum (9-folds) on 28th day on 1.0 mg/L picloram containing medium. The callus obtained was white compact hard (WCH). For growth kinetics study pseudostem derived callus was transferred on different media supplemented with 1.0 mg/L picloram. All phases of growth were seen in callus inoculated on all the three media except the absence of stationary phase on MS and SH media. MS medium proved to be the best for maximum biomass accumulation (9-fold) on 28th day of culture and callus in post-exponential phase showed maximum diosgenin accumulation (33 ppm).     

Production and optimisation of rosmarinic acid by Satureja hortensis L. callus cultures

In this study, production and optimisation of rosmarinic acid, a phenolic acid and an economically important metabolite, was investigated in the callus cultures established from the mature seeds of Satureja hortensis L. (summer savory) plant. Gamborg's B5 basal medium, supplemented with indol butyric acid (IBA) (1.00 mg L(-1)), N6-benzyl aminopurine (6-BA) (1.00 mg L(-1)) and sucrose (2.5%, w/v), was employed for the establishment and maintenance of the callus cultures. Applications were individually prepared by preparing the media containing different IBA/6-BA combinations and sucrose concentrations. All of the applications were carried out in the continuous dark. In the applications, where the effects of IBA/6-BA combinations on the growth and rosmarinic acid accumulation were assayed (1-15 applications), the highest biomass yield was obtained from the medium supplemented with 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA. In the case of the rosmarinic acid accumulation, an opposite relationship was determined between the growth and rosmarinic acid production. While the highest biomass yield was obtained from the medium containing 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA, the highest rosmarinic acid accumulation was obtained from the medium supported with 1.00 mg L(-1) IBA and 1.00 mg L(-1) 6-BA. In the applications where the effects of sucrose concentrations on the growth and rosmarinic acid accumulation were examined, the highest biomass yield was obtained from the medium which is supplemented with 5.0% (w/v) sucrose. In this category, the highest rosmarinic acid accumulation was obtained from the medium which is supported with 3.0% (w/v) sucrose. According to the experiments carried out with the wild S. hortensis, it is found to have 25.02+/-1.21 mg g(-1) rosmarinic acid. No differentiation was observed in any callus during the course of this study.     

Dual Elicitation for Improved Production of Withaferin A by Cell Suspension Cultures of Withania somnifera

High yielding transformed callus culture of W. somnifera was established by infecting hypocotyls with Agrobacterium tumefaciens MTCC-2250. Maximum withaferin A content of 0.0875 mg/g dry cell weight and transformation efficiency of 80% were obtained. Confirmation of transformation was done on the basis of the presence of the ags gene by using polymerase chain reaction. Various abiotic elicitors (arachidonic acid, methyl jasmonate, calcium chloride, and copper sulfate) and biotic elicitors (cell extracts and culture filtrates of Alternia alternata, Fusarium solani, and Verticilium dahaliae) were tested at different concentrations to enhance withaferin A production in suspension culture of transformed cells. Maximum enhancements of 5.4 times and 9.7 times, respectively, were obtained when copper sulfate (100 microM) and the cell extract of V. dahaliae (5% v/v) were added separately to suspension cultures. The dual elicitation strategy by the combined addition of these two elicitors resulted in 13.8-fold enhancement of withaferin A content in comparison to control cultures (2.65 mg/L). The present study indicates the potential of this biotechnology-based methodology for the large-scale production of withaferin A.     

Optimization of culture parameters for production of podophyllotoxin in suspension culture of Podophyllum hexandrum

The root explants of the germinated seedlings of Podophyllum hexandrum were grown in MS medium supplemented with indole acetic acid (IAA) (2 mg/L) and activated charcoal (0.5%), and healthy callus culture was obtained after incubation for 3 wk at 20°C. The cultivation of plant cells in shake flask was associated with problems such as clumping of cells and browning of media, which were solved by the addition of pectinase and polyvinylpyrrolidone. The effect of major media components and carbon source was studied on the growth and podophyllotoxin production in suspension culture. It was found that glucose was a better carbon source than sucrose and that NH₄ ⁺:NO₃ ⁻ ratio (total nitrogen concentration of 60 mM) and PO₄ ³⁻ did not have much effect on the growth and product formation. The relative effect of culture parameters (inoculum level, pH, IAA, glucose, NH₄ ⁺:NO₃ ⁻ ratio, and PO₄ ³⁻) on the overall growth and product response of the plant cell suspension culture was further investigated by Plackett-Burman design. This indicated that inoculum level, glucose, IAA, and pH had significant effects on growth and production of podophyllotoxin. To identify the exact optimum concentrations of these parameters on culture growth and podophyllotoxin production, central composite design experiments were formulated. The overall response equations with respect to growth and podophyllotoxin production as a function of these culture parameters were developed and used to determine the optimum concentrations of these parameters, which were pH 6.0, 1.25 mg/L of IAA, 72 g/L of glucose, and inoculum level of 8 g/L.     

Scarlet Flax Linum grandiflorum (L.) In Vitro Cultures as a New Source of Antioxidant and Anti-Inflammatory Lignans

In vitro cultures of scarlet flax (Linum grandiflorum L.), an important ornamental flax, have been established as a new possible valuable resource of lignans and neolignans for antioxidant and anti-inflammatory applications. The callogenic potential at different concentrations of α-naphthalene acetic acid (NAA) and thidiazuron (TDZ), alone or in combinations, was evaluated using both L. grandiflorum hypocotyl and cotyledon explants. A higher callus induction frequency was observed on NAA than TDZ, especially for hypocotyl explants, with a maximum frequency (i.e., 95.2%) on 1.0 mg/L of NAA. The presence of NAA (1.0 mg/L) in conjunction with TDZ tended to increase the frequency of callogenesis relative to TDZ alone, but never reached the values observed with NAA alone, thereby indicating the lack of synergy between these two plant growth regulators (PGRs). Similarly, in terms of biomass, NAA was more effective than TDZ, with a maximum accumulation of biomass registered for medium supplemented with 1.0 mg/L of NAA using hypocotyls as initial explants (DW: 13.1 g). However, for biomass, a synergy between the two PGRs was observed, particularly for cotyledon-derived explants and for the lowest concentrations of TDZ. The influence of these two PGRs on callogenesis and biomass is discussed. The HPLC analysis confirmed the presence of lignans (secoisolariciresinol (SECO) and lariciresinol (LARI) and neolignan (dehydrodiconiferyl alcohol [DCA]) naturally accumulated in their glycoside forms. Furthermore, the antioxidant activities performed for both hypocotyl- and cotyledon-derived cultures were also found maximal (DPPH: 89.5%, FRAP 866: µM TEAC, ABTS: 456 µM TEAC) in hypocotyl-derived callus cultures as compared with callus obtained from cotyledon explants. Moreover, the anti-inflammatory activities revealed high inhibition (COX-1: 47.4% and COX-2: 51.1%) for extract of hypocotyl-derived callus cultures at 2.5 mg/L TDZ. The anti-inflammatory action against COX-1 and COX-2 was supported by the IC₅₀ values. This report provides a viable approach for enhanced biomass accumulation and efficient production of (neo)lignans in L. grandiflorum callus cultures.     

Effect of Aeration on Production of Anticancer Lignans by Cell Suspension Cultures of Linum album

The effects of aeration within the range of 0.2-0.5 vvm on transformed and high yielding cell cultures of Linum album were investigated in a 5-L stirred tank bioreactor equipped with low shear Setric impeller. The kinetics of cell growth, substrate utilization, and production of lignans, namely, podophyllotoxin and 6-methoxypodophyllotoxin, were established. Maximum biomass of 23.2 g/L and lignan accumulation levels of 176.3 mg/L podophyllotoxin and 10.86 mg/L 6-methoxypodophyllotoxin were obtained with initial air flow rate of 0.3 vvm. Specified oxygen demand of cells was estimated to be 1.35 g O(2)/g biomass. The optimum oxygen transfer coefficient was found to be 16.7 h(-1) (,) which corresponded to aeration rate of 0.3 vvm. The effect of minimum dissolved oxygen (DO) concentration was investigated with respect to biomass and lignan production by comparing identically aerated and agitated bioreactor cultivations at dissolved oxygen concentrations of 10%, 30%, and 50%. Cell growth and podophyllotoxin accumulation were not affected significantly at these DO levels, but 6-methoxypodophyllotoxin production was enhanced when cells were cultivated at 30% DO level. The maximum volumetric productivities of 18.2 mg/L day and 3.2 mg/L day for podophyllotoxin and 6-methoxypodophyllotoxin, respectively, were obtained. These results establish the key role of oxygen on mass scale production of anticancer lignans by cell cultures of L. album. It may serve as a suitable parameter for scale-up.     

Cellular compatibility of biomineralized ZnO nanoparticles based on prokaryotic and eukaryotic systems

Zinc oxide nanoparticles (NPs) with the size of ～100 nm were prepared via a facile biomineralization process in the template of silk fibroin (SF) peptide at room temperature. These ZnO NPs have shown the remarkable behavior of low toxicity to gram-positive bacteria (Staphylococcus aureus, Staphylococcus agalactiae), gram-negative bacteria (Escherichia coli), and eukaryotic cells (mouse L929 fibroblasts). Bacteriological testing indicated that ZnO NPs presented a 50% inhibitory effect on Streptococcus agalactiae at the concentrations of >100 mM, whereas at the same concentrations, the growth of Staphylococcus aureus and Escherichia coli were hardly inhibited. On the other hand, a remarkable proliferation of Staphylococcus aureus or Escherichia coli was observed at the concentrations of ZnO NPs <50 mM. Moreover, the cytotoxicity test demonstrated that ZnO NPs mineralized with SF peptide possessed a low toxicity to mouse L929 fibroblasts. The SF peptide coated on the surface of ZnO NPs permitted greater adhesion and consequently greater proliferation of mouse L929 fibroblasts. Besides, from TEM micrographs of the cell ultrastructure, endocytosis of NPs into the cytoplasm can be detected and the ultrastructure of the cell underwent few changes. The cell membrane retained integrity, euchromatin dispersed homogenously inside the cytoplasm, the mitochondrial architecture remained intact, and no intracellular vacuoles were observed. High-resolution transmission electron microscopy images and selected area electron diffraction patterns of ultrathin cell sections indicated that the crystal structure of NPs was not damaged by the organelle or cytoplasm. All these observations indicated that ZnO NPs mineralized with the SF peptide possess good cytocompatibility.