Phage lytic enzymes: a history

There are many recent studies regarding the efficacy of bacteriophage-related lytic enzymes: the enzymes of ‘bacteria-eaters' or viruses that infect bacteria. By degrading the cell wall of the targeted bacteria, these lytic enzymes have been shown to efficiently lyse Gram-positive bacteria without affecting normal flora and non-related bacteria. Recent studies have suggested approaches for lysing Gram-negative bacteria as well(Briersa Y, et al., 2014). These enzymes include: phage-lysozyme, endolysin, lysozyme, lysin, phage lysin, phage lytic enzymes, phageassociated enzymes, enzybiotics, muralysin, muramidase, virolysin and designations such as Ply, PAE and others. Bacteriophages are viruses that kill bacteria, do not contribute to antimicrobial resistance, are easy to develop, inexpensive to manufacture and safe for humans, animals and the environment. The current focus on lytic enzymes has been on their use as anti-infectives in humans and more recently in agricultural research models. The initial translational application of lytic enzymes, however, was not associated with treating or preventing a specifi c disease but rather as an extraction method to be incorporated in a rapid bacterial detection assay(Bernstein D, 1997).The current review traces the translational history of phage lytic enzymes–from their initial discovery in 1986 for the rapid detection of group A streptococcus in clinical specimens to evolving applications in the detection and prevention of disease in humans and in agriculture.

Induction of Extracellular Lytic Enzymes by <i>Fusarium solani</i>

Fusarium solani is a necrotrophic parasitic fungus that causes wilt in some plants, causing severe economic losses in some areas of the country. The objective of this work was to analyze the induction of extracellular lytic enzymes produced by a strain of F. solani, isolated from a culture of tomato, in Villa de Arista, S.L.P. México. Polygalacturonase activity has a greater induction time at 10 days, and the xylanase has two times higher activity at 8 and 13 days of incubation at 28?C. Also, the xylanase activities A and B were very stable at 4?C. After 7 days of incubation, it has an activity of 100% and 96%, respectively, while polygalacturonase retains 61% of its initial activity. Both activities are better induced with glutamate and urea as nitrogen sources respectively, and both exhibit an initial pH optimum of 5.5. Finally, we didn’t find cellulase activity in the analyzing conditions.

Gamma-glutamyl transferase 5 overexpression in cerebrovascular endothelial cells improves brain pathology,cognition,and behavior in APP/PS1 mice

In patients with Alzheimer’s disease,gamma-glutamyl transferase 5(GGT5)expression has been observed to be downregulated in cerebrovascular endothelial cells.However,the functional role of GGT5 in the development of Alzheimer’s disease remains unclear.This study aimed to explore the effect of GGT5 on cognitive function and brain pathology in an APP/PS1 mouse model of Alzheimer’s disease,as well as the underlying mechanism.We observed a significant reduction in GGT5 expression in two in vitro models of Alzheimer’s disease(Aβ_(1-42)-treated hCMEC/D3 and bEnd.3 cells),as well as in the APP/PS1 mouse model.Additionally,injection of APP/PS1 mice with an adeno-associated virus encoding GGT5 enhanced hippocampal synaptic plasticity and mitigated cognitive deficits.Interestingly,increasing GGT5 expression in cerebrovascular endothelial cells reduced levels of both soluble and insoluble amyloid-βin the brains of APP/PS1 mice.This effect may be attributable to inhibition of the expression ofβ-site APP cleaving enzyme 1,which is mediated by nuclear factor-kappa B.Our findings demonstrate that GGT5 expression in cerebrovascular endothelial cells is inversely associated with Alzheimer’s disease pathogenesis,and that GGT5 upregulation mitigates cognitive deficits in APP/PS1 mice.These findings suggest that GGT5 expression in cerebrovascular endothelial cells is a potential therapeutic target and biomarker for Alzheimer’s disease.

S-烯丙基巯基半胱氨酸促进CD8+T细胞杀伤功能的机制研究

目的探究S-烯丙基巯基半胱氨酸(S-allylmercaptocysteine,SAMC)对CD8^(+)T细胞杀伤鼻咽癌细胞功能的影响及机制。方法将人鼻咽癌细胞HK-1和C666-1与SAMC共培养,分为0、25、50、100μmol/L SAMC组,Western blot检测肿瘤细胞程序性死亡配体-1(programmed cell death-ligand 1,PD-L1)表达。CD8^(+)T细胞分别与HK-1和C666-1细胞以10∶1的比例共培养并加入0、25、50、100μmol/L SAMC,检测CD8^(+)T对HK-1和C666-1的杀伤能力,酶联免疫法(ELISA)检测干扰素(INF-γ)和肿瘤坏死因子-α(TNF-α)浓度,构建鼻咽细胞HK-1的小鼠皮下移植瘤模型,分为对照组、CD8^(+)T细胞组、SAMC组、SAMC+CD8^(+)T细胞组,各组小鼠治疗期间测量瘤体积,治疗结束后取小鼠肿瘤组织,Western blot检测肿瘤组织中PD-L1表达,ELISA检测小鼠血清INF-γ和TNF-α浓度。结果相比于0μmol/L SAMC组,25、50、100μmol/L SAMC组HK-1和C666-1细胞PD-L1表达均显著下调(P<0.05),相比于0μmol/L SAMC+CD8^(+)T细胞组,25、50、100μmol/L SAMC+CD8^(+)T细胞组HK-1和C666-1细胞培养上清中INF-γ和TNF-α浓度均能显著增加,HK-1和C666-1细胞裂解率显著增加(P<0.01)。相比于对照组,CD8^(+)T细胞组和SAMC+CD8^(+)T细胞组小鼠瘤体积和瘤重显著下降(P<0.05),小鼠血清INF-γ和TNF-α浓度显著增加,相比于CD8^(+)T细胞组,SAMC+CD8^(+)T细胞组小鼠瘤体积和瘤重显著下降(P<0.05),小鼠血清INF-γ和TNF-α浓度显著增加(P<0.05),肿瘤组织PD-L1表达显著下调(P<0.01)。结论SAMC可通过抑制人鼻咽癌细胞PD-L1表达而促进CD8^(+)T细胞的杀伤功能。

Lytic Characteristics and Identification of Two Alga-lysing Bacterial Strains

All previously reported bacterial species which are capable of lysing harmful algae have been isolated from coastal environments in which harmful algae blooms have occurred. Due to the low concentration of alga-lysing bacteria in an algal bloom, it is difficult to isolate the alga-lysing bacteria by existing methods. In this paper, two algae-lysing bacterial strains, P01 and P03, have been isolated from a biosystem immobilized on a sponge that was highly effective in removing algae and microcystins. Their lysing modes and effects on Microcystis aeruginosa have been studied. The results show that the degradation processes of these two strains for M. aeruginosa accorded with a first-order reaction model when the chlorophylla concentration was in the range from 0 to 1000 μgL-1. The degradation rate constants were 0.1067, 0.1274 and 0.2792 for P01 and0.0683, 0.0744 and 0.028 97 for P03, when the bacterial densities were 8.6 × 105, 8.6 × 106 and 8.6 × 107cells mL-1 respectively. Moreover, the two bacterial strains had favourable lytic effects not only on M. aeruginosa , but also on Chlorella and Scene-desmus. Their lytic effect on M. aeruginosa did not require physical cell to cell contact, but proceeded by the production of an extracellular product. The bacterial strains were identified as Bacillus species by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.

<i>Staphylococcus aureus</i>Can Produce Catalase Enzyme When Adding to Human WBCs as a Source of H₂O₂Productions in Human Plasma or Serum in the Laboratory

Background: Staphylococcus aureus is one of the most virulent gram positive bacteria. It produces a lot of toxins and enzymes, most of which are virulent factors. Among the enzyme that produces is the catalase which is very useful in differentiating staphylococci from streptococci [1]. Catalase is nearly ubiquitous among some of organisms that can grow in the presence of oxygen (air). It promotes the conversion of hydrogen peroxide, a powerful and potentially harmful oxidizing agent, to water and molecular oxygen;so the major function of catalase within cells is to prevent the accumulation of toxic levels of hydrogen peroxide formed as a by-product of metabolic processes—primarily that of the electron transport pathway. Objectives: The main aim of this study is to prove that human WBCs can produce H2O2. This H2O2 when reacting with catalase producing S. aureus can easily be degraded to H2O + O2. Methodology: In this study a total of 40 subjects were included. Aliquots of 2.5 ml of venous blood were collected by venous puncture after disinfecting the site of collection with 70% alcohol and the collected blood was drawn into EDITA containers (20 subject) and anticoagulant free containers (other 20 subject), centrifugation for 5 minute at 1500 RPM. The separated sera and plasma were converted to new sterile eppendrof tubes and freezing until used (we leaved the eppendrof tubes that contained sera and plasma at room temperature before using it for DE freezing). Standard catalase producing S. aureus were used by taking 1 colony from Macconkey media by using applicator wooden stick, and inserted in eppendrof tube, then air bubbles would appear to indicate occurrence of the reactions. Results: According to this study, it was proved that WBCs in human plasma or serum can produce H2O2;this H2O2 was reacted with catalase enzyme produce from colony of S. aureus to produce air bubbles and water. There were no differences between using H2O2 or human plasma/serum that contains WBCs to detect and identify S. aureus by both techniques. Conclusion: Based on the results of this study, we can use WBCs that are found in human plasma or serum to identify catalase producing S. aureus.

Functions of three ubiquitin-conjugating enzyme 2 genes in hepatocellular carcinoma diagnosis and prognosis

BACKGROUND Liver cancer ranks the third cause of cancer-related death worldwide.The most common type of liver cancer is hepatocellular carcinoma(HCC).The survival time for HCC patients is very limited by years due to the lack of efficient treatment,failure of early diagnosis,and poor prognosis.Ubiquitination plays an essential role in the biochemical processes of a variety of cellular functions.AIM To investigate three ubiquitination-associated genes in HCC.METHODS Herein,the expression levels of ubiquitin-conjugating enzymes 2(UBE2)including UBE2C,UBE2T,and UBE2S in tumor samples of HCC patients and nontumor controls at the Cancer Genome Atlas(TCGA)database,was comprehensively analyzed.The relationship of UBE2 gene expression level with cancer stage,prognostic outcome,and TP53 mutant status was studied.RESULTS Our results showed that UBE2C,UBE2T,and UBE2S genes were overexpressed in HCC samples compared to non-tumor tissues.Dependent on the cancer progression stage,three UBE2 genes showed higher expression in tumor tissues at all four stages compared to non-tumor control samples.Furthermore,a significantly higher expression of these genes was found in stage 2 and stage 3 cancers compared to stage 1 cancer.Additionally,overexpression of those genes was negatively associated with prognostic outcome and overall survival time.Patients with TP53 mutation showed a higher expression level of three UBE2 genes,indicating an association between UBE2 expression with p53 function.CONCLUSION In summary,this study shed light on the potential roles of UBE2C,UBE2T,UBE2S on diagnostic and prognostic biomarkers for HCC.Moreover,based on our findings,it is appealing to further explore the correlation of those genes with TP53 mutation in HCC and the related mechanisms.

Isolation and molecular identification of cellulolytic bacteria from Dig Rostam hot spring and study of their cellulase activity

Cellulose is the main structural component of lignocellulosic wastes that can be converted to sugars and biofuels by cellulase.Due to wide applications of this enzyme in various industries around the world,cellulase is considered as the third industrial enzyme.The ability of thermophilic bacteria in the production of heat-stable cellulases has made them valuable tools in biotechnology.The aim of this study was isolation and molecular identification of cellulolytic thermophile bacteria from Dig Rostam hot spring and investigating their cellulase activity.Samples were taken from water and sediments of this hot spring,and cellulolytic bacteria were enriched in media containing cellulose as the only carbon source.The bacteria were incubated at 60℃,and single colonies were then isolated on solid media.Congo red assay was used as a quick test for the qualitative screening of cellulase activity.According to these qualitative results,four colonies named CDB1,CDB2,CDB3,and CDB4 were isolated,and their growth curve and some other characteristics were determined by biochemical assays.Moreover,endoglucanase,exoglucanase,and FPase activities of the isolates were investigated quantitatively.Results indicated that CDB1 exhibited the highest endoglucanase(0.096 U/mL)and exoglucanase(0.156 U/mL)activities among other isolates.16S rDNA partial sequencing indicated that CDB1 had 99%similarity to the genus Anoxybacillus,and the other isolates showed the highest similarity to the genus Geobacillus.The cellulase gene of CDB1 isolate with the highest cellulase activity was also cloned,and its sequence is reported for the first time.Further studies on this thermophilic enzyme might be useful for industrial applications.

Salvianolic acid B modulates the expression of drug-metabolizing enzymes in HepG2 cells

BACKGROUND: Enzymes involved in drug and xenobiotic metabolism have been considered to exist in two groups: phase I and phase II enzymes. Cytochrome P450 isoenzymes (CYPs) are the most important phase I enzymes in the metabolism of xenobiotics. The products of phase I metabolism are then acted upon by phase II enzymes, including glutathione S-transferases (GSTs). Herbs that inhibit CYPs such as CYP3A4 or that induce GSTs may have the potential to protect against chemical carcinogenesis since the mutagenic effects of carcinogens are often mediated through an excess of CYP-generated reactive intermediates. This study was designed to investigate the effects of salvianolic acid B (Sal B), a pure compound extracted from Radix Salviae Miltiorrhizae, a Chinese herb, on cell proliferation and CYP1A2 and CYP3A4 mRNA expression in the presence or absence of rifampicin, a potent inducer of CYPs and GST protein expression in HepG2 cells. METHODS: HepG2 cells were incubated with different concentrations of Sal B. Cell proliferation was determined by SYTOX-Green nucleic acid staining. CYP3A4 and CYP1A2 mRNA expression was assayed by real-time PCR. GST protein expression was analyzed by Western blotting. RESULTS: Low concentrations of Sal B (0-20 μmol/L) had no significant effects on cell proliferation, while higher concentrations (100-250 μmol/L) significantly inhibited proliferation in a concentration-dependent manner. Ten μmol/L Sal B, but not 1 μmol/L, down-regulated CYP3A4 and CYP1A2 mRNA expression after 24 hours of incubation, whereas both 1 and 10 μmol/L Sal B down-regulated CYP3A4mRNA expression after 96 hours of incubation; moreover, 1 and 10 μmol/L Sal B inhibited CYP3A4 mRNA expression induced by rifampicin. Both 1 μmol/L and 10 μmol/L Sal B increased GST expression. CONCLUSION: Sal B inhibits CYP3A4 and CYP1A2 mRNA expression and induces GST expression in HepG2 cells.

The Hydroxylation of Vitamin D on C25 in Thyrotoxicosis The Role of the Activity of Microsomal Liver Enzymes

Vitamin D3 after its entrance in the organism undergoes hydroxylation on C-25 carbon atom by the action of microsomal liver enzymes giving the metabolite 25 hydroxyvitamin D3 (25OHD3). The function of microsomal liver enzymes is influenced in some specified states by hormones or drugs. It has approved that thyroxin is a potent stimulator of these enzymes while allopurinol suppresses their function. The aim of this issue is to examine 25OHD3 plasma levels in thyrotoxic subjects and in those pretreated with allopurinol on the base of the afford mentioned data. In a first phase 25OHD3 plasma levels were estimated in thyrotoxic subjects against euthytoid healthy controls. In a second phase lmg vitamin D3 was injected intravenously (i.v.) in thyrotoxic subjects and in healthy euthyroid controls. 25OHD3 plasma levels were measured before and in post injection period in six hours intervals for 48 hours. In a third phase a couple of subjects one thyrotoxic and one euthyroid healthy control pretreated both with allopurinol injected lmg of vitamin D3 i.v. In all studied subjects 25OHD3 plasma levels were measured before and in post injection period in six hours intervals for 48 hours. The pre and post injection 25OHD3 plasma levels measured the size of activity of liver enzyme responsible for bioactivation of vitamin D3. In the first phase was indicated that 25OHD3 plasma levels were lower in thyrotoxic subjects comparing with that of euthyroid healthy controls (p 3 in thyrotoxic subjects was 2,5 to 8 times faster comparing with euthyroid healthy controls. In the third phase was shown that allopurinol decreases the activity of liver enzymes function as regard the bioactivation of vitamin D3. The bioactivation of vitamin D3 is accelerated in thyrotoxicosis compared with that in euthyroid state. This phenomenon produces low 25OHD3 plasma levels in thyrotoxic subjects which initially may be normal or slightly increased depended from the vitamin D3 status in the thyrotoxic subjects. By continuous stimulatory action of increased thyroid hormones on liver enzymes the 25OHD3 plasma levels earlier or later decline in levels of hypo-or avitaminosis D3. The previously described biological events may explain the decreased intestinal calcium absorption of vitamin D3 and the osteomalacic component found in a percentage of thyrotoxic bone histology. For the blocking effects of allopurinol on liver enzymes function and possibly of other pharmaceutical products in relation to vitamin D3 bioactivation, available data are still lacking.

设计改造羧酸还原酶合成医药中间体(S)-2-氨基丁醇

(S)-2-氨基丁醇同时具有羟基及氨基官能团,是多种重要药物分子的关键手性中间体,生物合成(S)-2-氨基丁醇尚缺少有效的酶元件。以塞格尼氏菌(Segniliparus rugosus)来源的羧酸还原酶SrCAR为研究对象,通过对实验室已有SrCAR突变文库进行筛选测试,结合活性位点共进化分析,同时利用组合活性中心饱和突变策略(Combinatorial active-site saturation test,CAST)构建新的突变体文库,经测试最终获得优势突变体XH7(G430V/E533F/A627N)。该突变体催化底物N-Boc-(S)-2-氨基丁酸到醛产物的活性(kcat/Km)较野生型SrCAR提高2.1倍,热熔值(Tm)提升2.3℃。进一步通过引入荧光假单胞菌(Pseudomonas fluorescens)来源的醇脱氢酶PfADH,可还原N-Boc-(S)-2-氨基丁醛到醇产物。XH7和PfADH的双酶共表达体系反应5 h即可将20 mmol/L底物实现几乎完全转化,转化率达到99%,并经脱Boc保护与分离纯化获得终产物(S)-2-氨基丁醇,得率为60%。通过分子动力学模拟解析最优突变体活性及热稳定性提高的分子机制,为SrCAR酶设计改造提供新的研究思路,拓展酶法合成(S)-2氨基丁醇的生物酶工具箱,可为类似高附加值医药中间体的生物合成提供理论和实践指导。

Inhibition of beta-site amyloid precursor protein-cleaving enzyme and beta-amyloid precursor protein genes in SK-N-SH cells

BACKGROUND： Previous studies have demonstrated that Piper futokadsura stem selectively inhibits expression of amyloid precursor protein （APP） at the mRNA level. In addition, the piperlonguminine （A） and dihydropiperlonguminine （B） components （1 ： 0.8）, which can be separated from Futokadsura stem, selectively inhibit expression of the APP at mRNA and protein levels. OBJECTIVE： Based on previous findings, the present study investigated the effects of β-site amyloid precursor protein cleaving enzyme （BACE1） and APP genes on the production of β-amyloid peptide 42 （Aβ42） in human neuroblastoma cells （SK-N-SH cells） using small interfering RNAs （siRNAs） and A/B components separated from Futokadsura stem, respectively. DESIGN, TIME AND SETTING： A gene interference-based randomized, controlled, in vitro experiment was performed at the Key Laboratory of Cardiovascular Remodeling and Function Research, Ministries of Education and Public Health, and Institute of Pharmacologic Research, School of Pharmaceutical Science ＆ Department of Biochemistry, School of Medicine, Shandong University between July 2006 and December 2007. MATERIALS： SK-N-SH cells were provided by Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai, China; mouse anti-human BACE1 monoclonal antibody was purchased from R＆D Systems, USA; mouse anti-human APP monoclonal antibody was purchased from Cell Signaling Technology, USA; and horseradish peroxidase （HRP）-conjugated goat anti-mouse IgG was provided by Sigma, USA. METHODS： The human BACE1 cDNA sequence was obtained from NCBI website （www.ncbi.nlm.nih.gov/sites/entrez）. Three pairs of siRNAs, specific to human BACE1 gene, were synthesized through the use of Silencer pre-designed siRNA specification, and were transfected into SK-N-SH cells with siPORT NeoFX transfection agent to compare the effects of different concentrations of siRNAs （10-50 nmol/L） on SK-N-SH cells. Futokadsura stem was separated and purified with chemical methods, and the crystal was composed of A/B components, with an A to B ratio of 1：0.8. The A/B （1 ： 0.8） components were added to the SK-N-SH cells at different concentrations （13.13, 6.56, and 3.28 mg/mL）. MAIN OUTCOME MEASURES： Using RT-PCR and Western blot methods, BACE1 and APP expression at mRNA and protein levels was detected in SK-N-SH cells following treatment with different siRNAs and concentrations of Futokadsura stem-separated A/B components, respectively. Altered Aβ42 secretion by SK-N-SH cells was determined by ELISA. RESULTS： BACE1 mRNA and protein levels were significantly suppressed by 40 and 50 nmol/L siRNAs at 48 hours post-transfection. A/B components （1 ： 0.8）, which were separated from Futokadsura stem, selectively inhibited mRNA and protein expression of APP in SK-N-SH cells. Aβ42 secretion by SK-N-SH cells was significantly decreased following treatment with siRNAs or A/B components. CONCLUSION： Inhibition of BACE1 and APP genes by various materials and methods efficiently decreased production of Aβ42.

周氏啮小蜂CcGSTS1基因的克隆·蛋白表达纯化及酶学特征分析

鉴定一个编码周氏啮小蜂谷胱甘肽S-转移酶序列的全长基因CcGSTS1,系统发育分析发现,该基因与丽蝇蛹集金小蜂NvGSTS6基因具有高同源性。体外表达、纯化的重组CcGSTS1可催化CDNB与GSH结合,最适反应pH为7.0。该试验为进一步研究周氏啮小蜂CcGSTS1基因功能奠定基础。

固定化脂肪酶手性拆分(R,S)-1-苯基乙醇研究进展

(R,S)-1-苯基乙醇是药物合成与精细化学品的重要中间体,可经固定化脂肪酶不对称催化获得光学纯1-苯基乙醇。本文分别从脂肪酶类型、溶剂工程、固定化技术三方面综述了国内外固定化脂肪酶手性拆分(R,S)-1-苯基乙醇的研究进展,并指明该领域中亟待解决的关键问题及解决思路,以期为后续固定化脂肪酶应用于手性拆分(R,S)-1-苯基乙醇提供参考。

镍、锌、铝对羊角月芽藻(Selenastrum capricornutum)生长及酶活性影响研究

研究了不同浓度镍、锌、铝对常见绿藻羊角月芽藻（Ｓ．ｃａｐｒｉｃｏｒｎｕｔｕｍ）生长速度、蛋白质含量、ＡＴＰ水平、葡萄糖－６－磷酸脱氢酶（Ｇ６ＰＤＨ）、酸性磷酸酶及硝酸还原酶活性的影响．结果表明３种金属离子在所试浓度范围内对羊角月芽藻的生长速度均有抑制作用，但单位藻培养物中蛋白质含量却随着金属离子浓度的增加而增加；高浓度金属离子对酶活性有明显抑制作用；藻细胞中ＡＴＰ水平随着金属离子浓度的增加而下降．提出金属离子对藻类生物产生影响的机理可能是：高浓度金属离子的存在，打破了生物最佳的各种营养元素生物可利用性的平衡．并讨论了藻类生长及生化活性对金属离子反应的相关性．

棉铃虫谷胱甘肽S-转移酶的活性分布和发育期变化及植物次生物质的诱导作用

对棉铃虫 Helicoverpa armigera (Hübner)谷胱甘肽S-转移酶（GSTs）发育期活性进 行了跟踪测定。从卵期开始，GSTs活性呈上升趋势，6龄幼虫达到最高，然后逐渐下降，但 化蛹初期仍维持较高水平；用培养基混药法研究发现，芸香苷和2-十三烷酮诱导种群5龄时 活性即达到峰值。6龄幼虫脂肪体GSTs活性最高，中肠次之，头部和表皮最低。用1-氯-2, 4-二硝基苯（CDNB）作底物时，中肠GSTs的 K m值最小。5龄幼虫中肠匀浆液中，GSTs活 性90%以上集中在细胞液层，在微粒体、线粒体和细胞碎片层的分布均不超过5%；芸香苷诱 导种群3龄幼虫微粒体层GSTs活性分布明显比对照种群减少，而上清液层则比对照种群 有所增高。

茜素红S对中华倒刺鲃幼鱼不同组织抗氧化酶活性和脂质过氧化的影响

以中华倒刺鲃幼鱼[平均体长(12.3±0.6)cm,平均体重(41.8±3.6)g]为研究对象,通过测定不同浓度茜素红S溶液(ARS)浸泡24h后肝脏、脑和鳃组织中抗氧化酶(SOD、CAT、GSH-Px)的活性和丙二醛(MDA)含量,评估ARS对中华倒刺鲃幼鱼的生理生化影响。结果表明:除脑组织GSH-Px随着ARS浓度的升高而不断升高外,肝脏、鳃和脑组织的抗氧化酶活性均表现为低浓度被诱导而高浓度受抑制的规律,与ARS浓度呈抛物线型剂量效应关系。肝脏、鳃、脑组织抗氧化酶活性达到最大值所对应的浓度分别是300、300和400 mg/L。除鳃部MDA含量随着茜素红S浓度升高不断升高外,脑和肝脏组织的丙二醛含量随着ARS浓度的升高呈现出先降低后上升趋势。根据试验的结果,推荐120—200 mg/L为ARS染色标记中华倒刺鲃幼鱼(体长10 cm)的适宜浓度。

芽孢皮层裂解酶的提取与SDS-PAGE分析

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日本血吸虫中国大陆株28kDa GST基因在大肠杆菌中的表达

在大肠杆菌ＴＢ１中表达合日本血吸虫中国大陆株２８ｋＤｅ抗原基因的重组质粒，表达产物是融 合蛋白，分子量为３３ｋＤａ。采用谷胱甘肽琼脂糖亲和层析柱纯化表达产物。２、４－二硝基氯苯／谷胱 甘肽分光光度测定法和琼脂糖－淀粉凝胶电泳显示重组抗原具有较高的谷胱甘肽Ｓ－转移酶（ＧＳＴ） 活力。酶联免疫吸附试验（ＥＬＩＳＡ）结果表明该重组抗原可检测到重组２８ｋＤａ ＧＳＴ或日本血吸虫 虫体ＧＳＴ免疫绵羊血清中的特异性抗体；免疫印迹试验（Ｗｅｓｔｅｒｎ Ｂｌｏｔｔｉｎｇ）分析揭示重组抗原的 ３３ｋＤａ抗原分子能被这两种免疫血清识别，证明它具有ＧＳＴ抗原性。

S_(3307)对树木扦插生根的效应及其作用机理研究

研究了不同浓度Ｓ３３０７对雪松、桧柏、龙柏插枝生根的效应，通过与ＰＰ３３３，ＮＡＡ等药剂处理的比较，证明Ｓ３３０７促进插枝生根的高效性及其与ＮＡＡ互作的加成效应。试验结果还进一步证实，Ｓ３３０７可通过抑制插枝中ＧＡｓ的生物合成，提高插枝基部多酚氧化酶活性，降低ＩＡＡ氧化酶活性，改变内源抑制物质和生长物质的比例，从而促进了树木的插枝生根效应。