Investigation of the cyt gene in Bacillus thuringiensis and the biological activities of Bt isolates from the soil of China

The presence of cyt genes was investigated in 80 type strains of Bacillus thuringiensis and 143 isolates obtained from soil samples of China by PCR amplification using two pairs of primers for the cyt1 and cyt2 genes. Three type strains of serotypes H11ac, H14 and H36, eight isolates belonging to H3, H14, H18 and H21, and one isolate of unknown serotype harbored cyt genes. We also tested the cytolytic activity for mammal cells, the hemolytic activity for sheep erythrocytes and insecticidal activity against mosquitoes of five isolates that contained cyt genes but did not belong to B. thuringiensis serovar israelensis. The protein profiles of the five isolates were different from those of the type strains of B. thuringiensis serovar israelensis, and among the five isolates, only Y-5 showed mosquitocidal activity against larvae of Culex quinquefasciatus. All five of the isolates exhibited hemolytic activity, but only three could cause the cell death of A549 cells. The cytopathological changes induced by NX-4 in some A549 cells were characterized with cell-ballooning.     

Antibacterial, antifungal, and antiviral activities of the lipophylic extracts of Pistacia vera

In the present study, antibacterial, antifungal, and antiviral properties of 15 lipohylic extracts obtained from different parts (leaf, branch, stem, kernel, shell skins, seeds) of Pistacia vera were screened against both standard and the isolated strains of Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Candida albicans and C. parapsilosis by microdilution method. Both Herpes simplex (DNA) and Parainfluenza viruses (RNA) were used for the determination of antiviral activity of the P. vera extracts by using Vero cell line. Ampicilline, ofloxocine, ketoconazole, fluconazole, acyclovir and oseltamivir were used as the control agents. The extracts showed little antibacterial activity between the range of 128-256 microg/ml concentrations whereas they had noticeable antifungal activity at the same concentrations. Kernel and seed extracts showed significant antiviral activity compared to the rest of the extracts as well as the controls.     

Influence of culture conditions on the production of milk-clotting enzyme from Rhizomucor

The ratio of milk-clotting activity to proteolytic activity (MC/PA) was used as an index to determine the quality of milk-clotting enzyme. Solid-state fermentation on wheat bran for 5 days at room temperature gave optimal production for enzyme by Rhizomucor miehei and R. pusillus. A ratio of wheat bran to moisture of 1:0.6 (w/v) gave best results. Adding skim milk powder to the media of R. miehei did not improve the MC/PA ratio but 4% (w/w) of the powder did improve the ratio with R. pusillus. Co-cultivation of R. miehei with R. pusillus did not change the MC/PA ratio.The authors are with the Department of Biotechnogy, Bharathiar University. Coimbatore-641 046 India     

Interplant movement by pear psylla (Homoptera: Psyllidae): Effects of sex ratio and reproductive status

Sticky trap catch of pear psylla,Cacopsylla pyricola Foerster, is male biased during the reproductive generations, but not the diapausing generation. In cage studies, we monitored movement by male and female pear psylla between host plants, and tested whether reproductive and diapausing psylla exhibit similar rates of movement. We also experimentally varied sex ratio to determine whether sex ratio affected movement. Male-biased sex ratios prompted increased movement off of the original host by male psylla of the reproductive generations; no such effect was noted for diapausing insects. We interpret these results to indicate that male movements increased under male-biased conditions due to mate-searching activities. There was also evidence in two experiments that severely male-biased sex ratios prompted movement off of the original host plant by reproductive females; this effect may have been due to harassment of ovipositing females by males.     

毛咀地星的抗氧化、抗菌和抗肿瘤活性和化学成分研究

毛咀地星是一种药用蘑菇,为了进一步开发利用毛咀地星,该研究探讨了毛咀地星乙醇提取物的抗氧化、抗菌和抗肿瘤活性,并采用硅胶柱色谱、制备薄层色谱和重结晶等方法,对毛咀地星乙醇提取物进行分离纯化,研究了其化学成分。结果表明:毛咀地星乙醇提取物多酚含量为10.53μg·mg~(-1),DPPH和·OH自由基清除能力的IC50值分别为91.35和148.76μg·m L~(-1);乙醇提取物对金黄葡萄球菌、大肠杆菌和枯草芽抱杆菌均无抑制作用;乙醇提取物在浓度为1 mg·m L~(-1)时,对燕麦镰刀菌的抑制率为36.11%;乙醇提取物在浓度为200μg·m L~(-1)时,对BG-803、NCI-H502和MDA-MB-231肿瘤细胞的抑制率分别为18.87%、17.71%和41.23%。并从乙醇提取物中分离纯化到6个化合物,根据理化性质和波谱数据分析,化合物结构分别鉴定为5α,8α-过氧化麦角甾-6,9(11),22-三烯-3β-醇(1)、5α,8α-过氧化麦角甾-6,22-二烯-3β-醇(2)、D-阿拉伯糖醇(3)、L-谷氨酸(4)、麦芽糖(5)和蔗糖(6),以上化合物均为首次从该菌中分离得到。     

大血藤醇提物抗炎镇痛和止血活性研究初探

为观察大血藤醇提物抗炎、镇痛、止血活性,该文采用75％乙醇提取制备大血藤醇提物(AESC),利用HPLC法测定其绿原酸含量;KM鼠(或新西兰兔)在测定抗炎、镇痛、止血活性时随机分为空白对照组、阳性对照组(云南白药酊组)、AESC组,依次测定其抑制二甲苯致小鼠耳肿胀度作用、痛阈值和兔肝脏局部创面损伤出血的记分分值,分别考...     

Growth of wheat seedlings raised from the grains treated with 28-homobrassinolide

An experiment was conducted under controlled conditions, in which grains of wheat (Triticum aestivum L. cv. Raj-3077) were soaked in 0, 1, 3 and 5 μM aqueous solutions of 28-homobrassinolide (HBR) for 4, 8 and 12 hours. The seedlings raised from the grains pre-treated with HBR possessed significantly higher leaf number, fresh and dry weight·plant⁻¹, nitrate reductase (NR,E.C. 1.6.6.1) and carbonic anhydrase (CA, E.C. 4.2.1.1) activities on 25 and 35 days, after sowing. The soaking of the grains in 3 μM concentration for 8 or 12 hours produced the most vigorous seedlings.     

Variation in the biochemical properties of the Drosophila alcohol dehydrogenase allozymes

Thirteen Drosophila Adh variants have been characterized with respect to gene expression, substrate preference, thermostability, and specific activity. The results suggest that the variants may be grouped into two biochemical classes, typified by the properties of the two most common enzyme forms, ADH-F and ADH-S. Membership of these classes cannot be predicted from electrophoretic mobility, nor is any simple classification possible with regard to the characteristics of level of gene expression (in terms of ADH activity or ADH protein) or thermostability of the gene product.     

Ribonuclease,deoxyribonuclease, and antiviral activity of <Emphasis Type="Italic">Escherichia coli</Emphasis>-expressed <Emphasis Type="Italic">Bougainvillea xbuttiana</Emphasis> antiviral protein 1

A full-length cDNA encoding ribosome-inactivating/antiviral protein from the leaves of Bougainvillea xbuttiana was recently isolated. The coding region of cDNA was cloned and expressed in Escherichia coli, and the protein product was designated as BBAP1 (Bougainvillea xbuttiana antiviral protein 1). BBAP1 showed ribonuclease activity against Torula yeast RNA. It also exhibited depurination activity against supercoiled pBlueScript SK+ plasmid DNA in a concentration dependent manner, and was found to convert nicked circular DNA into linear form only at higher concentration. On bioassay, BBAP1 exhibited antiviral activity against sunnhemp rosette virus infecting Cyamopsis tetragonoloba leaves in which 95% inhibition of local lesion formation was observed.     

The effect of Sargassum fusiforme polysaccharide extracts on vibriosis resistance and immune activity of the shrimp, Fenneropenaeus chinensis

Immunostimulants are valuable for control of shrimp diseases and the immunostimulatory effects of some polysaccharide additives for shrimp have been reported. In this study, the Sargassum fusiforme polysaccharide extract (SFPSE) was assessed as a feed additive when supplemented in the diet (0%, 0.5%, 1.0%, and 2.0%) for juvenile shrimp, Fenneropenaeus chinensis, in order to study the effects of SFPSE on vibriosis resistance and immune activity. Shrimp were cultured in the same pond with cages. The body weight, survival, the cumulative mortality after injection with Vibrio harveyi (30 microl V. harveyi suspension at 9.3 x 10(7) CFU ml(-1) per shrimp), the total haemocyte counts (THCs), the protein concentration and the phenoloxidase (PO) activity in supernatant of haemolymph, the lysozyme (LSZ) and superoxide dismutase (SOD) activity in muscle of the shrimp were assayed after 14 days feeding period. The results indicated that shrimp survival under the stress of V. harveyi was affected by the dietary SFPSE. The shrimp treated with 1.0% and 0.5% SFPSE displayed significantly lower cumulative mortalities after being injected with V. harveyi suspension 24 and 30 h later, respectively, compared with that of the control. However, cumulative mortality of 2.0% SFPSE treatment was not significantly different from that of the control. There was no significant difference of cumulative mortality between 0.5% and 1.0% SFPSE treatment groups. The immune activities of the shrimp also were affected by dosage of dietary SFPSE. The THCs of the shrimp rose with increasing SFPSE dosage. The protein concentration and PO activity in supernatant of haemolymph as well as muscular LSZ activity first rose then dropped with increasing SFPSE dosage. The protein concentration in supernatant of haemolymph appeared a maximum of 167.46 mg ml(-1) in 1.0% SFPSE treatment. The PO activity and LSZ activity reached the peaks as 13.20 U and 3.21 U mgprot(-1) in 0.5% SFPSE treatment, respectively. SOD activity of the shrimp was not significantly affected by dietary SFPSE. It is therefore suggested that oral administration of SFPSE at an optimal level of 0.5% and 1.0% for 14 days effectively improved vibriosis resistance and enhanced immune activity of the shrimp in general.     

Antimicrobial activities of 2-arylthio-N-alkylmaleimides

Summary A variety of 2-arylthio-N-alkylmaleimides were prepared, and their antimicrobial activities were examined. Almost all of these compounds exhibited antibacterial activity against Gram-positive bacteria such asBacillus subtilis andStaphylococcus aureus. Some compounds such as 2-(halogeno-phenyl)-thio-N-methylmaleimides (4, 5, 6, 8 and 10) and 2-(2-carbamoylphenyl)thio-N-methylmaleimide(35) exhibited antibacterial activity againstEscherichia coli. All compounds tested were inactive againstPseudomonas aeruginosa except 2-(2-carbamoylphenyl)thio-N-methylmaleimide(35) which was marginally active. Activities against Gram-positive bacteria were not due to the effect of the substituent on the benzene ring, except in the instances 2-carboxy, 2-carbomethoxy, 2-amino groups and alkyl chains, however, activities against Gram-negative bacteria were due to phenylthio and the alkyl substituents. Some of 2-arylthio-N-alkylmaleimides were examined for their antifungal activities using eight strains of fungi, and they showed activity against these.     

Antinociceptive, anti-inflammatory and acute toxicity effects of Zhumeria majdae extracts in mice and rats

Antinociceptive and anti-inflammatory effects and acute toxicity of aqueous infusion and ethanolic maceration extracts of the aerial parts of Zhumeria majdae were studied in mice and rats. Antinociceptive activity was determined using hot-plate and writhing tests. The effect of the extracts against acute inflammation was studied by acetic acid increased vascular permeability and xylene-induced ear edema in mice. The activity of the extracts against chronic inflammation was assessed using the cotton pellet test in rats. LD50 values of the infusion and maceration extracts were 3.09 g/kg body wt., and 3.94 g/kg body wt., respectively. Phytochemical screening of the extracts indicated the presence of flavonoids and tannins. In the hot-plate test, the intraperitoneal injection of both extracts showed significant and dose-dependent antinociceptive activity in mice. Naloxone, an opioid antagonist, on pretreatment inhibited the antinociceptive activity of the extracts. The extracts exhibited antinociceptive activity against acetic acid-induced writhing, which was partially blocked by naloxone. Both extracts showed significant effect against acute inflammation induced by acetic acid in mice. In the chronic inflammation test, efficacy of the extracts was similar to that of baclofen and dexamethasone in rats. It is concluded that the aqueous infusion and ethanolic maceration extract of the aerial parts of Zhumeria majdae have antinociceptive effects and this may be mediated by opioid receptors. The extracts also showed anti-inflammatory effects against acute and chronic inflammation.     

喙尾琵琶甲肠道来源链霉菌(Streptomyces sp.)BPA71次级代谢产物的分离鉴定及其生物活性评价

【背景】昆虫是世界上种类最多、肠道菌群资源最丰富且多样的动物类群之一。昆虫肠道微生物具有产生活性次级代谢产物的能力,是活性天然产物的重要来源。【目的】研究药用昆虫喙尾琵琶甲(Blaps rynchopetera)成虫肠道来源链霉菌(Streptomyces sp.) BPA71的次级代谢产物及其生物活性。【方法】利用正相硅胶柱色谱、葡聚糖凝胶Sephadex LH-20柱色谱等方法分离纯化该菌株的发酵粗提物,采用牛津杯法进行抗菌活性追踪,确定抗菌活性部位,通过ESI-MS、NMR等波谱数据分析对化合物结构进行鉴定,采用微量肉汤稀释法测定最低抑菌浓度(minimal inhibitory concentration, MIC),采用MTS法测定抗肿瘤活性。【结果】从Streptomyces sp. BPA71的固体发酵提取物中共分离得到4个已知化合物,通过对比核磁数据确定为糠酸甲酯(1)、吡咯甲酰胺A (2)、吡咯甲酰胺B (3)和吲哚-3-乙酸甲酯(4)。抗菌活性结果显示化合物2具有广谱抗菌活性。此外,化合物2对宫颈癌细胞HeLa、肺癌细胞A549、肝癌细胞SMMC-7721、乳腺癌细胞MDA-MB-231和结肠癌细胞SW480这5株肿瘤细胞均有明显的抑制活性。【结论】喙尾琵琶甲肠道来源Streptomyces sp. BPA71可产生丰富的生物活性物质,该研究结果为进一步挖掘喙尾琵琶甲肠道链霉菌的活性天然产物奠定了基础,同时丰富了人们对喙尾琵琶甲肠道微生物的认识。     

Antimicrobial activities of someN-alkylmaleimides

Summary A variety ofN-alkylmaleimides have been prepared and screened for antimicrobial activity using various fungi, yeasts and bacteria.N-Methyl,N-octyl andN-decylmaleimide showed antifungal activities. All test compounds showed antibacterial activity.     

北桑寄生产黄酮内生菌的分离鉴定及其抗氧化和抑菌活性评价

【背景】北桑寄生(Loranthus tanakae)植物中黄酮类化合物成分含量较高,生物活性较好。由于植物资源短缺,导致其生物活性成分的研究受限。【目的】从北桑寄生中分离、筛选和鉴定产黄酮类化合物的内生菌,并评估其体外抗氧化和抑菌活性。【方法】以北桑寄生枝条为研究对象分离内生菌,通过显色反应和薄层色谱法(thin-layer chromatography,TLC)筛选产黄酮类化合物的内生菌,并采用NaNO₂-Al (NO₃)₃比色法测定其总黄酮含量。利用清除自由基和抑菌的体外实验来初步评估产黄酮内生菌的生物活性。【结果】共筛选出4株产黄酮内生菌,其中3株为内生真菌,1株为内生细菌,通过形态学和分子生物学方法将其分别鉴定为Botryosphaeria sp.(ZC020)、Phoma sp.(ZZ105)、Nemania sp.(ZS042)和Pseudomonas sp.(ZC026)。4株菌中ZC020和ZS042的总黄酮含量较高,分别为(44.58±0.72) mg/L和(31.98±0.18) mg/L (P<0.05,n=3),并且ZS042可产生与北桑寄生植物相同类型的黄酮类化合物。ZC026和ZS042表现出优异的抗氧化活性,其1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)和2,2''-联氮-二(3-乙基苯并噻唑啉-6-磺酸)二铵盐[2,2''-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt,ABTS]自由基清除率分别为72.85%±0.66%(ZC026)、57.01%±0.89%(ZS042)和85.36%±0.75%(ZC026)、88.17%±0.15%(ZS042)(P<0.05,n=3)。ZZ105对革兰氏阳性菌和革兰氏阴性菌均有抑制作用。【结论】从北桑寄生枝条中分离出4株产黄酮类化合物的内生菌,其中有一株为内生细菌。这项工作为北桑寄生黄酮类化合物的生产提供了新的资源,也为其他珍稀药用植物和材料的开发和保护提供了参考。     

Antioxidant and antiviral activities ofEuphorbia thymifolia L.

The antioxidant and antiviral activities ofEuphorbia thymifolia L. (Euphorbiaceae) were investigated in this study. The results showed that all of the fractions (MeOH, CHCl₃, EtOAc, n-butanol and water) and pure compounds (3-O-galloyl-4,6-(S)-HHDP-D-glucose, rugosin B and 1,3,4,6-tetra-O-galloyl-K--D-glucose)tested possessed antioxidant activities, with the exception of the organic aqueous fraction in the anti-lipid and anti-super-oxide formation assays. The range of IC₅₀ of anti-lipid formation, anti-superoxide formation and free radical scavenging assays for all fractions and pure compounds were 2.81–7.63, 0.03–2.18 and 0.013–2.878 mg/ml, respectively. Electron spin resonance studies showed that water extract and pure compounds ofE. thymifolia exhibited superoxide radical and hydroxyl radical scavenging activities. Besides antioxidant activities, 3-O-galloyl-4,6-(S)-HHDP-D-glucose and EtOAc fraction also showed anti-HSV-2 activity. Thus,E. thymifolia was concluded to possess antioxidant and anti-HSV-2 activities.     

Antiviral and antimicrobial activities of three sesquiterpene lactones from Centaurea solstitialis L. ssp. solstitialis

Three sesquiterpene lactones (centaurepensin=chlorohyssopifolin A, chlorojanerin and 13-acetyl solstitialin A) isolated from the aerial parts of Centaurea solstitialis L. ssp. solstitialis (Asteraceae) were investigated for antimicrobial and antiviral activities. For the antimicrobial activity assessment, both standard and isolated strains of Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Candida albicans and C. parapsilosis were employed by the microdilution method. Herpes simplex type-1, a DNA virus, and Parainfluenza, an RNA virus, were employed for the determination of the antiviral activity of these three sesquiterpene lactones using Vero cell lines. Ampicilline, ofloxocine, ketoconazole, fluconazole, acyclovir and oseltamivir were used as the reference drugs. 13-Acetyl solstitialin A displayed remarkable antibacterial activity against isolated strains of E. faecalis at 1 μg/ml concentration, which was close to the effective concentrations of ampicillin. The same compound also showed significant activity against the DNA virus, being as potent as the reference compound acyclovir at maximum and minimum concentrations of 16–<0.00006 μg/ml. This is the first report showing that 13-acetyl solstitialin A possesses significant antiviral activity.     

徐闻普哥滨珊瑚土曲霉XWC21-10代谢产物活性及其活性物质的研究

为了开发利用海洋活性天然产物,对分离自普哥滨珊瑚的1株土曲霉XWC21-10次级代谢产物的活性及其活性成分进行初步研究。结果表明,该菌株发酵液的4种萃取物对6种指示菌均有明显的抑菌活性,说明具有广谱抑菌作用;菌丝体氯仿萃取物和发酵液乙酸乙酯萃取物对乙酰胆碱酯酶(Ach E)抑制作用最为明显,抑制率分别达到(79.37±1.53)%,(67.04±1.96)%。对具有明显活性的乙酸乙酯萃取物经硅胶柱层析分离得到11个级分。活性跟踪检测组分6、7、8、11活性较好,组分6、7成分相似,其主要含有还原性物质、高级醇、甾类等化合物;而级分11仅含有共轭基团化合物,成分相对简单。因此,该菌代谢产物具有良好的抑菌和抑制乙酰胆碱酯酶活性,活性物质还有待进一步的纯化和确定,该研究可为开发抑菌剂和治疗早老性痴呆症提供基础。     

具核梭杆菌CarRS双组分系统组氨酸激酶CarS的表达、纯化及生化活性测定

具核梭杆菌(Fusobacterium nucleatum)是一种条件致病菌,能够在结直肠癌组织中富集,影响结直肠癌发生发展的多个阶段。双组分系统在病原菌耐药性、致病性相关基因的调控和表达中起重要作用。本文以具核梭杆菌CarRS双组分系统为研究对象,重点对其组氨酸激酶蛋白CarS进行重组表达和性质研究。利用在线软件SMART、CCTOP和AlphaFold2对CarS二级结构和三级结构进行预测,其结果表明CarS蛋白具有2个跨膜螺旋区,包含9个α螺旋和12个β折叠结构;由两个结构域构成,一是位于N末端的跨膜域(氨基酸1–170),另一个是位于C末端的胞内域,胞内域由信号接收域(histidine kinases,adenylyl cyclases,methyl-accepting proteins,prokaryotic signaling proteins,HAMP)、磷酸受体结构域(histidine kinase domain,HisKA)和组氨酸激酶催化结构域(histidine kinase-like ATPase catalytic domain,HATPase_c)组成。由于全长的CarS蛋白未能在宿主细胞中表达,因此根据其二级、三级结构特点,构建了CarS胞内蛋白的融合表达载体pET-28a(+)-MBP-TEV-CarScyto,并在大肠杆菌(Escherichia coli)BL21-CodonPlus(DE3)-RIL中进行过表达。经亲和层析、离子交换层析和凝胶过滤层析,最终获得纯度较高的CarScyto-MBP蛋白,终浓度达20 mg/mL。活性实验结果表明,CarScyto-MBP具有蛋白激酶和磷酸转移酶双活性,麦芽糖结合蛋白(maltose binding protein,MBP)标签对CarScyto蛋白的生物活性无影响。上述结果为深入解析CarRS双组分系统在具核梭杆菌中的生物学功能提供了一定的理论基础。     

Active substances against trypomastigote forms of Trypanosoma cruzi and microorganisms are produced in sequence by Talaromyces flavus

The conditions for the sequential production of antibiotic activity by Talaromyces flavus were determined. The highest level of activity against Trypanosoma cruzi was obtained from the aqueous extract of the Czapeck's fermentative culture after 48 hours, with lysis of 97.58% of the trypomastigote forms of Trypanosoma cruzi (red blood cells remained normal). The antimicrobial activity was detected in the extracts of fermentative cultures from different media just after 144 hours of incubation. Maximum activities against Micrococcus luteus, Staphylococcus aureus and Candida albicans were present in chloroform, butanolic and water extracts, in this order, when Talaromyces flavus was cultivated at pH 5.0. The minimal inhibitory concentration (MIC) of extracts of Takeuchi's cultures were determined.