In vitro induction of endothelial cell fibrinolytic alterations by Nigella sativa

The effect of Nigella sativa (NS) L. oil (blackseed oil) on the fibrinolytic system of the human umbilical vein (HUV) and human uterine arterial (HUA) endothelial cells (ECs) in culture was studied. Both of them showed a concentration-dependent increase in tissue-type plasminogen activator (t-PA). A maximum effect was achieved with 50 microg oil/ml conditioned medium (CM) (1.3+/-0.15ng/10(4) cells/24h vs. control 0.7+/-0.06ng/10(4) cells/24h, and 0.38+/-0.04ng/10(4) cells/24h vs. control 0.24+/-0.02ng/10(4) cells/24h, for HUVEC and HUA-EC, respectively). At 100 microg/ml, there was a significant change in the amount of t-PA antigen produced by either HUVEC or HUA-EC (1.0+/-0.1 ng/10(4) cells/24 h or 0.28+/-0.02 ng/10(4) cells/24 h) as compared to control CM from cells grown under control conditions, but still less than that recorded at 50 microg oil/ml. Plasminogen activator inhibitor-type 1 increased the CM significantly and concentration-dependently in both cells. For HUVEC, the maximum effect was achieved at a concentration of 100 microg/ml (257.7+/-8.0 ng/10(4) cells/24 h vs. control 72.7+/-3.8 ng/10(4) cells/24 h). HUA-EC showed the maximum effect at a concentration of 100 microg/ml (171.6+/-4.4 ng/10(4) cells/24 h vs, control 53.8+/-3.7 ng/10(4) cells/24 h). This study suggests a role for NS oil in modulating the balance of fibrinolysis/thrombus formation by modulating the fibrinolytic potential of endothelial cells.     

Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia-reperfusion injury in rat hippocampus

It has been previously reported that Nigella sativa oil (NSO) and thymoquinone (TQ), active constituent of N. sativa seeds oil, may prevent oxidative injury in various models. Therefore, we considered the possible effect of TQ and NSO on lipid peroxidation level following cerebral ischemia-reperfusion injury (IRI) in rat hippocampus. Male NMRI rats were divided into nine groups, namely, sham, control, ischemia and ischemia treated with NSO or TQ. TQ (2.5, 5 and 10 mg/kg), NSO (0.048, 0.192 and 0.384 mg/kg), phenytoin (50 mg/kg, as positive control) and saline (10 ml/kg, as negative control) were injected intraperitoneally immediately after reperfusion and the administration was continued every 24h for 72 h after induction of ischemia. The transient global cerebral ischemia was induced using four-vessel-occlusion method for 20 min. Lipid peroxidation level in hippocampus portion was measured as malondialdehyde (MDA) based on its reaction with thiobarbituric acid (TBA) following ischemic insult. The transient global cerebral ischemia induced a significant increase in TBA reactive substances (TBARS) level (p<0.001), in comparison with sham-operated animal. Pretreatment with TQ and NSO were resulted a significant decrease in MDA level as compared with ischemic group (66.9+/-1.5 vs. 297+/-2.5 nmol/g tissue for TQ, 10 mg/kg; p<0.001 and 153.5+/-1.3 nmol/g tissue for NSO, 0.384 mg/kg; p<0.001). Using a reversed-phase HPLC system, the amount of TQ in NSO was also quantified and was 0.58% w/w. These results suggest that TQ and NSO may have protective effects on lipid peroxidation process during IRI in rat hippocampus.     

Elicitation studies in cell suspension cultures of Cannabis sativa L.

Cannabis sativa L. plants produce a diverse array of secondary metabolites. Cannabis cell cultures were treated with biotic and abiotic elicitors to evaluate their effect on secondary metabolism. Metabolic profiles analysed by ¹H NMR spectroscopy and principal component analysis (PCA) showed variations in some of the metabolite pools. However, no cannabinoids were found in either control or elicited cannabis cell cultures. Tetrahydrocannabinolic acid (THCA) synthase gene expression was monitored during a time course. Results suggest that other components in the signaling pathway can be controlling the cannabinoid pathway.     

纳米二氧化钛对生菜的生长效应分析

为了阐明纳米二氧化钛颗粒(TiO₂NPs)对生菜(Lactuca sativa)生长的影响，采用自行设计的水培装置探究不同浓度TiO₂NPs (300~1 200 mg/L)下，生菜生长和生理生化指标的变化。结果表明，300 mg/L TiO₂NPs能促进生菜幼苗的根长、茎长、叶表面积、鲜重和干重；随着TiO₂ NPs浓度增大，生菜的生长指标呈现下降趋势，但仍优于对照组。生菜体内的抗氧化酶(SOD、POD)在低TiO₂ NPs浓度(300 mg/L)时，活性明显下降；随着TiO₂ NPs浓度增大，这两种抗氧化酶活性逐渐增强。因此，生菜对TiO₂NPs胁迫具有浓度依赖性，表现为“低促高抑”，且能够通过抗氧化酶系统来减轻TiO₂NPs伤害。     

Neorhizobium petrolearium OS53联合紫花苜蓿协同修复石油污染土壤研究

【目的】探究Neorhizobium petrolearium OS53与紫花苜蓿协同修复石油污染土壤的机制。【方法】使用Illumina和Nanopore平台对菌株OS53进行全基因组测序,构建菌株基因组完成图,并进行基因预测及功能注释,分析其中与结瘤促生及石油降解相关基因,并通过实验测定菌株OS53产吲哚乙酸(indole acetic acid, IAA)、铁载体、溶磷和解钾等与促生相关的能力。使用试剂盒对联合修复前后土壤中土壤脲酶、脱氢酶、多酚氧化酶和脂肪酶活性及紫花苜蓿的叶绿素、丙二醛、脯氨酸、可溶性蛋白、可溶性糖和超氧化物歧化酶等生理指标进行测定。【结果】菌株OS53的基因组由一个5.56 Mb的环形染色体和2个大小分别为0.92 Mb和0.38 Mb的质粒组成,基因组G+C含量为60.2%,共编码6 968个基因。菌株OS53与N. petrolearium DSM 26482^T的16S rRNA基因序列相似性最高,为99.86%,且在系统发育树上形成稳定分支,表明菌株OS53与N. petrolearium为同一种,因此将OS53命名为N. petrolearium OS53。试验结果表明,菌株OS53具有产IAA能力,并在其基因组中也发现相关基因。在初始石油含量为(4 403.30±222.10) mg/kg时,经过120 d修复,OS53与紫花苜蓿协同修复效率能够达到57.53%,比不接种OS53、仅接种OS53和仅种植苜蓿分别提高了44.26%、41.69%和8.84%。在联合修复体系中,紫花苜蓿叶绿素、可溶性蛋白和可溶性糖的含量有所提高,丙二醛和脯氨酸的含量以及超氧化物歧化酶的活性有所降低,同时土壤中多酚氧化酶、脱氢酶、脂肪酶和脲酶的活性都有所提高。【结论】菌株OS53具有产IAA的能力,并且能够促进紫花苜蓿在石油污染土壤中的生长,进而提高土壤中与石油降解相关部分酶活,最终提高联合修复体系对石油污染土壤的修复效果。     

Cimicifuga racemosa extract BNO 1055 inhibits proliferation of the human prostate cancer cell line LNCaP

Extracts from black cohosh (Cimicifuga racemosa, CR) exert an anti-proliferative action in human breast cancer cell cultures, which has been attributed to an anti-estrogenic effect. However, CR constituents do not bind to either of the known estrogen receptors. Thus, the anti-tumor effect of CR me be mediated by mechanisms not involving these receptors. Polycyclic aromatic hydrocarbons are toxic environmental pollutants, which indirectly act as anti-estrogens by activating the aryl hydrocarbon receptor (AhR). The AhR is widely expressed in mammalian tissues and tumors. A recent screening study demonstrated activation of the AhR by a variety of herbal extracts, among others, CR. Since activation of the AhR causes inhibition of growth of prostate cancer cells, we addressed the question, whether CR may not only inhibit growth of breast cancer--but also of prostate cancer cells. In the AhR ligand assay, the CR extract BNO 1055 reduced tracer binding to 71% of the control demonstrating interaction of constituents of this extract with the receptor. Under basal as well as under estradiol- and dihydrotestosterone stimulated conditions, the CR extract dose dependently inhibited proliferation of LNCaP cells. A significant reduction of cell growth was observed at a concentration as low as 50 ng/ml. Thus, it is demonstrated for the first time that CR compounds potently inhibit the growth of human prostate cancer cells in vitro. This anti-proliferative effect may be mediated via the AhR.     

Rapid lateral diffusion of lectin-labelled glycoconjugates in the human colonic adenocarcinoma cell line HT29

The lateral diffusion of lectin-labelled glycoconjugates was studied in the human colon carcinoma cell line HT29 using fluorescence photobleaching techniques. HT29 cells were grown in either Dulbecco's modified Eagle's medium with glucose (25 mM; DMEM-Glu) or with galactose (25 mM; DMEM-Gal). Cell cultivation in the DMEM-Gal medium was assumed to promote a transformation of the cells to become small-intestinal-like with characteristic microvilli and associated enzymes. The diffusion of glycoconjugates labelled with fluoresceinated Triticum vulgaris agglutinin (Wheat germ agglutinin; WGA), Ricinus communis agglutinin-I (RCA-I), Concanavalia ensiformis agglutinin (ConA), Ulex europaeus agglutinin-I (UEA-I) and Arachis hypogaea agglutinin (PNA) was in all cases rapid, with a diffusion constant (D) ranging between 0.4 and 0.8×10^-8 cm² s^-1. As a comparison the diffusion of the fluorescent synthetic lipid analog diI-C₁₄ was characterized by D=0.8 – 1.0 × 10^–8 cm² s^-1. The diffusion of lectin-labelled surface components could not be related to the presence of microvilli on HT29 cells grown in DMEM-Gal, which ought to yield an apparently lower diffusion rate. The results indicate either that surface glycoconjugates in HT29 cells are dominated by glycolipid, or that the labelled glycoproteins are more or less free to diffuse in the plane of the membrane.     

亚麻荠FUS3转录因子的鉴定及功能分析北大核心CSCD

FUS3转录因子是调控植物种子油脂合成的关键因子。为探讨亚麻荠CsFUS 3基因在脂质合成和积累过程中的作用,该研究对CsFUS 3基因家族进行全基因组鉴定,分析CsFUS 3基因的时空表达模式,并解析CsFUS 3-1和CsFUS 3-2基因在植物油脂合成中的功能,为深入解析CsFUS 3基因在亚麻荠油脂合成中的功能及亚麻荠高油品种选育提供理论基础。结果表明:(1)利用AtFUS3蛋白序列,在亚麻荠基因组数据库中鉴定出2条完整的CsFUS3蛋白序列,分别命名为CsFUS3-1和CsFUS3-2,亚细胞定位发现2个CsFUS3蛋白均位于细胞核。(2)亚麻荠CsFUS3-1和CsFUS3-2蛋白与拟南芥AtFUS3蛋白的亲缘关系最近,具有与拟南芥AtFUS3蛋白相似的理化性质、高级结构以及完整的B3功能域。(3)qRT-PCR结果显示,CsFUS 3-1和CsFUS 3-2基因仅在种子中表达,且随着种子的发育成熟,2个CsFUS 3基因的表达量均呈先增高后降低的变化趋势,并在花后30 d时表达量达到最高。(4)CsFUS3和CsWRI1蛋白互作以及CsFUS 3对OLE和ABI 3基因的转录调控可能是亚麻荠高油性状的关键调控途径。(5)烟草瞬时表达分析表明,与野生型相比,转CsFUS 3-1和CsFUS 3-2基因的烟草叶片总油脂含量分别提高了0.95%和1.12%,表明亚麻荠CsFUS 3基因能够提高烟叶总油脂的合成积累。     

In vitro and in vivo leishmanicidal activity of Dysoxylum binectariferum and its fractions against Leishmania donovani

The leishmanicidal effect of crude ethanolic extract of stem bark of Dysoxylum binectariferum and its fractions has been investigated against Leishmania donovani, the causative agent of visceral leishmaniasis. Ethanolic extract was lethal to promastigotes as well as amastigote forms in macrophage system at the concentration of 100 microg/ml. Chloroform fraction significantly inhibited promastigote multiplication and was also active against amastigotes in infected J774A.1 macrophages at 100 microg/ml. Hexane fraction was moderately active and the other fractions were inactive against both the forms. When tested in vivo in hamsters, ethanolic extract was toxic at 500 mg/kg whereas exhibited marginal activity (67.7+/-5.3%) at 250 mg/kg x 5, p.o. on day 7 post treatment (p.t.) which increases slightly (69+/-4.7) by day 30 p.t. Chloroform and n-hexane fractions exhibited 64.3+/-4% and 47.8+/-4.6% parasite inhibition at the dose of 100 mg/kg x 5 p.o., respectively. The pure compound, rohitukine, obtained from chloroform fraction showed weaker in vitro activity and was ineffective in infected hamsters. The lead potential of this plant need further investigations.     

苗期紫花苜蓿株体对不同地区垂穗披碱草种子萌发生长的化感作用

以发芽率、苗长、根长、苗干重、根干重变化为种子萌发和幼苗生长参数,研究苗期紫花苜蓿(Medicago sativa)植株浸提液对不同地区垂穗披碱草(Elymus nutans)种子萌发生长的化感作用。结果表明:地上部浸提液对LS、DX垂穗披碱草种子发芽率具有明显的促进作用,而对GS、KMX、LKZ、LZ地区垂穗披碱草种子发芽率均表现为抑制作用,其中对GS垂穗披碱草种子发芽率的抑制作用最强,在14.5%和5.5%浓度处理时抑制率分别为57.89%、55.26%;苗长方面,浸提液5.5%浓度对垂穗披碱草苗长的抑制率顺序为:LZNQDXLSKMXQHGSLKZ,14.5%处理时抑制率顺序为:LZKMXLKZNQGSQHDXLS,其中抑制率最高的为LZ垂穗披碱草,在14.5%和5.5%浓度处理时抑制率分别为33.03%、28.97%;根长方面,5.5%处理对垂穗披碱草根长的抑制率顺序为:QHNQLZKMX、GSLKZDX、LS,14.5%处理时抑制率顺序为:GSQH、NQLZLSKMXDXLKZ,其中在高浓度下抑制率最高的为GS垂穗披碱草,抑制率为57.69%;地上部浸提液对LKZ、LZ垂穗披碱草苗干重均具有促进作用,高浓度浸提液对NQ垂穗披碱草苗干重产生促进作用(RI0),而对KMX、DX垂穗披碱草苗干重均表现为抑制作用;根干重方面,浸提液对LS、QH、GS、NQ、LKZ垂穗披碱草根干重均有明显的抑制作用,而对KMX、DX垂穗披碱草根干重产生促进作用,高浓度浸提液对LZ垂穗披碱草的根干重产生促进作用。从根浸提液的作用来看,根浸提液除对LS、DX垂穗披碱草种子发芽率和根干重、GS垂穗披碱草种子发芽率和苗干重及NQ垂穗披碱草根干重具有促进作用外(P 0.05),对其余地区垂穗披碱草的各项指标均有明显的抑制作用(RI0)。所有以上结果表明,紫花苜蓿植株浸提液对垂穗披碱草种子萌发生长的作用具有一定的浓度效应。不同地区垂穗披碱草对紫花苜蓿地上部浸提液的敏感性趋势总体为:GSQHLSKMXNQLKZLZ,最不敏感或有促进作用的是DX垂穗披碱草;对根浸提液的敏感性趋势总体为:QHNQLZKMX,根浸提液对LS、GS、LKZ、DX垂穗披碱草种子萌发生长具有促进作用。紫花苜蓿植株不同部位浸提液对垂穗披碱草种子萌发生长的化感效应顺序为:地上部根。     

Nitric oxide alleviates arsenic toxicity by reducing oxidative damage in the roots of Oryza sativa (rice)

Nitric oxide (NO) is a bioactive gaseous, multifunctional molecule playing a central role and mediating a variety of physiological processes and responses to biotic and abiotic stresses including heavy metals. The present study investigated whether NO applied exogenously as sodium nitroprusside (SNP) has any protective role against arsenic (As) toxicity in Oryza sativa (rice). Treatment with 50 μM SNP (a NO donor) significantly ameliorated the As-induced (25 or 50 μM) decrease in root and coleoptile length of rice. Further, As-induced oxidative stress measured in terms of malondialdehyde (MDA), superoxide ion (), root oxidizability and H₂O₂ content was lesser upon supplementation of NO. It indicated a reactive oxygen species (ROS) scavenging activity of NO. NO addition reversed (only partially) the As-induced increase in activities of antioxidant enzymes – superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and catalase. The study concludes that exogenous NO provides resistance to rice against As-toxicity and has an ameliorating effect against As-induced stress.     

In vitro transformation of paralytic shellfish toxins in the clams Mya arenaria and Protothaca staminea

Dissected tissues of two clam species, the Pacific littleneck, Protothaca staminea, and soft-shell, Mya arenaria, were evaluated for in vitro conversion of paralytic shellfish poisoning (PSP) toxins. Tissue homogenates were incubated with purified PSP toxins to determine the time-course of toxin conversion. The effects of boiling and addition of a natural reductant (glutathione) on toxin conversion were also assessed. For P. staminea, the digestive gland showed the greatest capacity for biotransformation, followed by gill, but mantle, adductor muscle, and siphon tissues exhibited very low conversion. In this species, the production of decarbamoyl derivatives was much greater from low potency N-sulfocarbamoyl toxins than from carbamate analogues. Decarbamolyation exhibited apparent specificity for α-epimers of all toxin substrates and this reaction was inhibited by boiling. Glutathione-mediated desulfation was tissue specific and had apparent specificity for β-epimers. These observations on P. staminea suggest that the above reactions are enzyme-mediated. In contrast, there was little toxin conversion in M. arenaria homogenates, but even this low activity was heat-labile and thus likely enzyme-mediated.     

H-NS蛋白对副溶血弧菌hcp1的转录调控

【目的】研究调控子H-NS对副溶血弧菌T6SS1结构蛋白基因hcp1的转录调控机制。【方法】利用Western blot检测Hcp1蛋白在野生株(WT)和hns基因敲除株(Δhns)中表达水平的差异。提取WT和Δhns的总RNA,采用实时定量RT-PCR的方法验证H-NS对hcp1的转录调控关系。进而采用引物延伸实验研究hcp1的转录起始位点,并根据产物的丰度判断H-NS对hcp1的调控关系。PCR扩增hcp1的整个启动子区DNA序列,并纯化His-H-NS蛋白,通过凝胶阻滞实验(EMSA)验证His-H-NS对hcp1启动子区是否具有直接的结合作用。【结果】Western blot和实时定量RT-PCR结果显示H-NS能抑制hcp1的表达;引物延伸结果显示hcp1只有一个转录起始位点T(–62)(翻译起始位点为+1),且其转录活性是H-NS和σ54依赖性的;EMSA实验表明H-NS对hcp1的启动子区具有直接的结合作用。【结论】H-NS能直接结合到hcp1启动子区而抑制其转录表达。     

In vitro glucose uptake activity of Aegles marmelos and Syzygium cumini by activation of Glut-4, PI3 kinase and PPARγ in L6 myotubes

The purpose of the present study is to investigate the effect of methanolic extracts of Aegles marmelos and Syzygium cumini on a battery of targets glucose transporter (Glut-4), peroxisome proliferator activator receptor gamma (PPARgamma) and phosphatidylinositol 3' kinase (PI3 kinase) involved in glucose transport. A. marmelos and S. cumini are anti-diabetic medicinal plants being used in Indian traditional medicine. Different solvent extracts extracted sequentially were analysed for glucose uptake activity at each step and methanol extracts were found to be significantly active at 100ng/ml dose comparable with insulin and rosiglitazone. Elevation of Glut-4, PPARgamma and PI3 kinase by A. marmelos and S. cumini in association with glucose transport supported the up-regulation of glucose uptake. The inhibitory effect of cycloheximide on A. marmelos- and S. cumini-mediated glucose uptake suggested that new protein synthesis is required for the elevated glucose transport. Current observation concludes that methanolic extracts of A. marmelos and S. cumini activate glucose transport in a PI3 kinase-dependent fashion.     

Glutamate-induced cell death in neuronal HT22 cells is attenuated by extracts from St. John''s wort (Hypericum perforatum L.)

Glutamate-induced cell death of hippocampal HT22 cells is a model system for neuronal disorders due to depletion of glutathione levels and increase of intracellular reactive oxygen species. Standardized extracts of Hypericum perforatum (HPE) contain flavonoids known for antioxidative properties. In the above model, cytoprotective effects at a concentration of 0.05% HPE by attenuation of calcium fluxes and cellular energy statuses are reported.     

紫花苜蓿叶绿体基因组密码子偏好性分析

为分析紫花苜蓿叶绿体基因组密码子偏好性的使用模式,该文以紫花苜蓿叶绿体基因组中筛选到的49条蛋白质编码序列为研究对象,利用CodonW、CUSP、CHIPS、SPSS等软件对其密码子的使用模式和偏好性进行研究。结果表明:(1)紫花苜蓿叶绿体基因的第3位密码子的平均GC含量为26.44%,有效密码子数(ENC)在40.6～51.41之间,多数密码子的偏好性较弱。(2)相对同义密码子使用度(RSCU)分析发现,RSCU>1 的密码子数目有30个,以A、U结尾的有29个,说明了紫花苜蓿叶绿体基因组A或U出现的频率较高。(3)中性分析发现,GC3与 GC12的相关性不显著,表明密码子偏性主要受自然选择的影响; ENC-plot 分析发现一部分基因落在曲线的下方及周围,表明突变也影响了部分密码子偏性的形成。此外,有17个密码子被鉴定为紫花苜蓿叶绿体基因组的最优密码子。紫花苜蓿叶绿体基因组的密码子偏好性可能受自然选择和突变的共同作用。该研究将为紫花苜蓿叶绿体基因工程的开展和目标性状的遗传改良奠定基础。     

AsSlt2基因对茄链格孢细胞壁完整性的调控

【背景】由茄链格孢(Alternaria solani)引起的马铃薯早疫病被普遍认为是马铃薯生产上的第二大叶部病害,在马铃薯各产区普遍发生,给马铃薯生产造成了巨大的经济损失。【目的】明确AsSlt2基因对茄链格孢细胞壁完整性的影响。【方法】在含有刚果红、细胞壁降解酶和十二烷基硫酸钠(sodiumdodecylsulfate,SDS)等细胞壁胁迫的培养基上观察ΔAsSlt2缺失突变株的生长情况,计算相对生长抑制率;通过实时荧光定量PCR (RT-qPCR)方法检测ΔAsSlt2菌株中细胞壁合成相关基因的表达情况;进一步检测ΔAsSlt2细胞壁中几丁质的含量及胞外酶活性。【结果】ΔAsSlt2缺失突变株对SDS、刚果红、细胞壁降解酶等细胞壁胁迫的敏感性增强,在加入细胞壁降解酶后突变株原生质体释放量显著增多;ΔAsSlt2对外源氧胁迫更敏感,突变株胞外过氧化物酶和漆酶活性均显著降低;进一步研究发现,ΔAsSlt2细胞壁中几丁质含量减少,几丁质合成相关基因与漆酶合成相关基因的表达量均明显降低。【结论】AsSlt2基因在茄链格孢细胞壁的完整性及抵御外界胁迫方面发挥重要作用。     

Differential induction of three pathogenesis-related genes, PR10, PR1b and PR5 by the ethylene generator ethephon under light and dark in rice (Oryza sativa L.) seedlings

Ethylene has been shown to be involved in triggering pathogenesis-related (PR) gene expression mainly in dicotyledonous species; however, less attention has been devoted identifying and characterizing PR genes in rice (Oryza sativa L.), a monocot and a model of cereal crop genera. Here, we demonstrate that ethylene induces at least three important rice PR genes, the PR10, PR1 basic (PR1b), and PR5 genes in rice (cv. Nipponbare) seedling leaf, upon treatment with the ethylene generator, ethephon (ET), in a light-, time- and dose-dependent manner. Induction of these PR genes was partially blocked by cycloheximide (CHX), a eukaryotic cytoplasmic protein synthesis inhibitor, which indicates an involvement of cytoplasmic de novo protein synthesis in their induction. These results clearly indicate a dynamic role for ethylene in PR genes induction in rice.