Trypanosoma cruzi: Synergistic cytotoxicity of multiple amphipathic anti-microbial peptides to T. cruzi and potential bacterial hosts

The parasite Trypanasoma cruzi is responsible for Chagas disease and its triatomine vector, Rhodnius prolixus, has a symbiotic relationship with the soil bacterium, Rhodococcus rhodnii.R. rhodnii that was previously genetically engineered to produce the anti-microbial peptide, cecropin A was co-infected with T. cruzi into R. prolixus resulting in clearance of the infectious T. cruzi in 65% of the vectors. Similar anti-microbial peptides have been isolated elsewhere and were studied for differential toxicity against T. cruzi and R. rhodnii. Of the six anti-microbial peptides tested, apidaecin, magainin II, melittin, and cecropin A were deemed potential candidates for the Chagas paratransgenic system as they were capable of killing T.cruzi at concentrations that exhibit little or no toxic effects on R. rhodnii. Subsequent treatments of T. cruzi with these peptides in pair-wise combinations resulted in synergistic killing, indicating that improvement of the 65% parasite clearance seen in previous experiments may be possible utilizing combinations of different anti-microbial peptides.     

Redox state and gender differences in vascular smooth muscle cells

Vascular smooth muscle cells (VSMC) have been isolated from male and female rat aorta and studied to assess their susceptibility to ultraviolet radiation-induced oxidative stress. Interestingly, a gender difference, in terms of reactive oxygen species production, was detected in both basal and irradiated VSMC. Namely, VSMC from male rats were more susceptible to radiation-induced stress and easier underwent apoptosis in comparison to cells from female rats. Conversely, the latter, in the same experimental conditions, clearly displayed signs of premature senescence. These results indicate that a sort of "gender memory" can be conserved in VMSC in primary culture.     

凋落物输入对三江平原弃耕农田土壤基础呼吸和活性碳组分的影响

土壤有机碳的积累主要由土壤有机质的输入与输出间的净平衡决定的,植被的恢复和凋落物质的大量输入是土壤恢复的先决条件,凋落物的输入在土壤恢复过程中起着至关重要的作用.通过对不同类型凋落物输入到三江平原弃耕农田后土壤的基础呼吸、溶解有机碳(DOC)和土壤微生物量碳(MBC)的研究表明:相同种类凋落物输入后,输入到土壤总有机碳(TOC)背景值低的凋落物被微生物降解的速率大于TOC背景值高的土壤,TOC较低的土壤能够加快微生物对输入凋落物的分解,不利于有机质的积累;不同类型凋落物的输入使土壤基础呼吸、DOC和MBC等活性组分的生成和降解产生差异,改变了凋落物的降解速率,在三江平原研究的4种主要植被类型中,人工林凋落物最容易降解,小叶章、大豆的降解能力次之,玉米是最难降解的凋落物.     

凋落物化学组成对土壤微生物学性状及土壤酶活性的影响

通过模拟试验的方法研究了单一施加杉木(Cunn inghan ia lancceola ta(L am b)Hook.)叶凋落物,杉木(C.lancceola ta)和桤木(A lnus crem astogyne Burk ill)混合凋落物,杉木(C.lancceola ta)和枫香(L iqu id am ba f orm osana H ance)混合凋落物,杉木(C.lancceola ta)、桤木(A.crem astogyne)、枫香(L.f orm osana)混合凋落物对土壤化学性状和土壤微生物量碳、代谢熵(qCO2)、土壤酶活性的影响。研究结果表明,土壤微生物学性状比土壤化学性状对不同凋落物处理的效应反应更敏感;与单一杉木叶凋落物比较,混合凋落物处理的土壤微生物量碳明显增加,土壤脲酶、蔗糖酶、脱氢酶活性升高;土壤代谢熵(qCO2)和土壤多酚氧化酶活性有下降趋势;另外,研究结果也表明,不同树种的叶凋落物混合对土壤质量的影响存在差异,有桤木叶的混合凋落物对土壤质量的改善效果似乎更明显。     

The transglutaminase activating metalloprotease inhibitor from Streptomyces mobaraensis is a glutamine and lysine donor substrate of the intrinsic transglutaminase

Transglutaminase (TGase) from Streptomyces mobaraensis is an extra-cellular enzyme that cross-links proteins to high molecular weight aggregates. Screening for intrinsic substrates now revealed the dual Streptomyces subtilisin inhibitor-like inhibitor Streptomyces subtilisin and transglutaminase activating metalloprotease (TAMEP) inhibitor (SSTI), equally directed against subtilisin and the TGase activating metalloprotease TAMEP, is both a glutamine and a lysine donor protein. Reactivity of glutamines is lost during culture, most likely by TGase mediated deamidation, and, accordingly, cross-linking only occurred if SSTI from early cultures was used. Interestingly, release of buried endo-glutamines by the lipoamino acid N-lauroylsarcosine could restore SSTI reactivity. Formation of lipoamino acids by Streptomycetes suggests such compounds could also modulate in vivo TGase mediated SSTI cross-linking.     

高铁酸钾对几种常见鱼类病原菌的杀菌效果测定

试验测定了高铁酸钾(K2FeO4)溶液对8种常见鱼类病原菌的MIC(最小抑菌浓度,minimal inhibitory concentration,MIC)和MBC(最低杀菌浓度,minimal bactericidal concentration,MBC),并研究了不同浓度的K2FeO4溶液对几种病原菌的杀灭效果.结果显示,K2FeO4对温和气单胞菌(Aeromonas sobria)、河弧菌(Vibrioflurialis)、弧菌Ⅰ组淡水亚组弧菌(Vibrio group Ⅰ freshwater subgroup)的抑制效果良好;当各试验菌液浓度约为105cfu/mL时,施以K2FeO4溶液作用1h后,对以上两种弧菌亦表现出最优的杀灭效果;基于以上试验结果,选取这两种最为敏感的病原菌测定其长势随消毒剂浓度和时间的变化情况.研究对于使用K2FeO4浸泡消毒鱼体或全池泼洒消毒池塘时具有指导意义,并为使用K2FeO4防治鱼类弧菌病研究工作的展开打下基础.     

Purification and structural characterization of a novel antibacterial peptide from Bellamya bengalensis: activity against ampicillin and chloramphenicol resistant Staphylococcus epidermidis

Increasing tendency of clinical bacterial strains resistant to conventional antibiotics has being a great challenge to the public's health. Antimicrobial peptides, a new class of antibiotics is known to have the activity against a wide range of bacteria resistant to conventional antibiotics. An antimicrobial peptide of 1676 Da was purified from Bellamya bengalensis, a fresh water snail, using ultrafiltration and reversed phase liquid chromatography. The effect of this peptide on Staphylococcus epidermidis resistant to ampicillin and chloramphenicol was investigated; the MIC and MBC values were 8 μg/ml and 16 μg/ml, respectively. Complete sequence of the peptide was determined by tandem mass spectrometry (MS/MS). Further, peptide net charge, hydrophobicity and molecular modeling were evaluated in silico for better understanding the probable mechanisms of action. The peptide showed the specificity to bacterial membranes. Hence, this reported peptide revealed a promising candidate to contribute in the development of therapeutic agent for Staphylococcal infections.     

The mode of action of cytotoxic and antitumor 1-nitroacridines. III. In vivo interstrand cross-linking of DNA of mammalian or bacterial cells by 1-nitroacridines

To define a critical lesion in presumable target DNA cause in vivo by the antitumor and cytotoxic 1-nitroacridines, Ehrlich ascites tumor (Eat) cells implanted into mice, HeLa cells grown in monolayer culture or Bacillus subtilis SB 1058 strain cells were exposed to Ledakrin [Nitracrine; 1-nitro-9-(3'-dimethylamino-n-propylamino)acridine], its non-antitumor congeners, or mitomycin C tested for comparison; total intracellular DNA was isolated from control or treated cells and denatured by heat, alkali or formamide, after which the chemically-induced fraction of interstrand cross-linked DNA molecules was assessed by thermal denaturation-renaturation curve analysis, hydroxylapatite column chromatography, or partitioning in a Dextran T500-polyethylene glycol 6000 biphasic system. Ledakrin, as compared to mitomycin C, was a very effective cross-linking agent, inducing one covalent cross-link per approx. 20 X 10(3) (B. subtilis), 56 X 10(3) (HeLa) or 80 X 10(3) (Eat) DNA base pairs. The first cross-links were introduced in B. subtilis cell genomes at minimal bactericidal concentrations of Ledakrin of mitomycin C. Ledakrin failed to induce discernible cross-linking of bihelical DNA when complexed with in cell-free system. Unlike the other two 1-nitroacridines which cross-linked DNA of cultured HeLa or B. subtilis cells, the non-antitumor 2-, 3- or 4-nitroacridines did not cause such effect and diminished cross-linking by Ledakrin or mitomycin C. Hence, upon metabolic activation in mammalian or bacterial cell Ledakrin and, most probably other 1-nitroacridines, become very effective DNA cross-linking agents and such effects appear to be responsible for the antitumor and potent cytotoxic activities of 1-nitroacridines.     

Aggregation of Staphylococcus aureus following treatment with the antibacterial flavonol galangin

AIM: The flavonol galangin, an antimicrobial constituent of the traditional medicines propolis and Helichrysum aureonitens, is being assessed as part of an ongoing investigation into the antibacterial activity of flavonoids. The present study sought to establish whether galangin has any aggregatory effect on bacterial cells. METHODS AND RESULTS: In preparatory time-kill assays, 50 microg ml(-1) of galangin was found to reduce colony counts of c. 5 x 10(7) CFU ml(-1)Staphylococcus aureus NCTC 6571 by approximately 15 000-fold during 60 min of incubation. Subsequent light microscopy studies demonstrated significant increases in the number of large clusters of bacterial cells in populations treated with the flavonol. CONCLUSION: Data presented here show that galangin causes aggregation of bacterial cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that galangin causes bacterial cells to clump together may implicate the cytoplasmic membrane as a target site for this compound's activity. More importantly, this observation indicates that decreases in CFU numbers detected in time-kill and minimum bactericidal concentration (MBC) assays in previous investigations were at least partially attributable to this aggregatory effect. This raises the possibility that galangin is not genuinely bactericidal in action, and calls into question the suitability of time-kill and MBC assays for determining the nature of activity of naturally occurring flavonoids.     

银杏外种皮提取物对病原菌﹙Cylindrocladium colhounii﹚的抑制作用

采用菌丝生长抑制法,研究了银杏外种皮3种不同提取物对病原菌（Cylindrocladium colhounii）的抑制作用并测定了这些提取物的MIC/MBC。结果表明,在相同实验天数内,抑菌效果（最小抑菌率）由高到低依次为银杏外种皮乙醇提取物（37.4%）、石油醚提取物（23.7%）和新鲜外种皮汁液（18.4%）。当提取物的浓度提升到一定程度时,三者的抑菌率均可达到100%;三种提取物的MIC/MBC分别为：86.25/86.25,172.5/276,293.25/345 mg·mL^-1。可见银杏外种皮具有明显抑制该病原菌生长的作用,且抑菌成分在乙醇中的溶解度较大。这为进一步研究该抑菌成分乃至生物农药的开发奠定了基础。