粮油食品防霉微生物筛选及其活性物质初步研究

为开发新型防霉剂,降低粮油食品储藏、运输、流通等过程中霉变对其品质和食用安全的影响,研究筛选获得具有强防霉活性的微生物。采用抑菌圈法,以禾谷镰刀菌（Fusarium graminearum）为指示菌筛选具有防霉活性的菌株。利用形态学观察、生化特征鉴定和16S rDNA基因序列比对进行菌株鉴定;通过不同温度、pH和蛋白酶K处理对防霉活性物质进行初步分析。结果显示：通过初筛获得33株对禾谷镰刀菌具有强抑制活性的菌株,复筛后选择的7株菌归属为芽孢杆菌属（Bacillus）;活性物质分析发现7株菌的活性物质均不耐高温和强酸碱,同时蛋白酶K处理对其防霉效果没有明显影响,初步推断为肽类;通过抑菌谱测定发现菌株ASAG 62对常见霉菌（产黄青霉Penicillium flavum、黑曲霉Aspergillus Niger、赭曲霉Aspergillus ochre、黄曲霉Aspergillus flavus、禾谷镰刀菌F. graminearum）均具有良好的抑制效果。     

Staphylococcus aureus—A Known Opponent against Host Defense Mechanisms and Vaccine Development—Do We Still Have a Chance to Win?

The main purpose of this review is to present justification for the urgent need to implement specific prophylaxis of invasive Staphylococcus aureus infections. We emphasize the difficulties in achieving this goal due to numerous S. aureus virulence factors important for the process of infection and the remarkable ability of these bacteria to avoid host defense mechanisms. We precede these considerations with a brief overview of the global necessitiy to intensify the use of vaccines against other pathogens as well, particularly in light of an impasse in antibiotic therapy. Finally, we point out global trends in research into modern technologies used in the field of molecular microbiology to develop new vaccines. We focus on the vaccines designed to fight the infections caused by S. aureus, which are often resistant to the majority of available therapeutic options.     

L.paraplantiumⅡ32发酵条件的初步研究

从4种培养基中筛选出MRS培养基为L.paraplantiumⅡ32的最适生长培养基。并运用点滴法,通过单因子筛选法,最终确定该菌生长、最大抑菌活性产生的最佳碳源为麦芽糖;氮源为酵母粉,浓度为2.0%;最适的初始pH值范围为5.5~6.5;温度范围为29~31℃。     

Medicated Scaffolds Prepared with Hydroxyapatite/Streptomycin Nanoparticles Encapsulated into Polylactide Microfibers

The preparation, characterization, and controlled release of hydroxyapatite (HAp) nanoparticles loaded with streptomycin (STR) was studied. These nanoparticles are highly appropriate for the treatment of bacterial infections and are also promising for the treatment of cancer cells. The analyses involved scanning electron microscopy, dynamic light scattering (DLS) and Z-potential measurements, as well as infrared spectroscopy and X-ray diffraction. Both amorphous (ACP) and crystalline (cHAp) hydroxyapatite nanoparticles were considered since they differ in their release behavior (faster and slower for amorphous and crystalline particles, respectively). The encapsulated nanoparticles were finally incorporated into biodegradable and biocompatible polylactide (PLA) scaffolds. The STR load was carried out following different pathways during the synthesis/precipitation of the nanoparticles (i.e., nucleation steps) and also by simple adsorption once the nanoparticles were formed. The loaded nanoparticles were biocompatible according to the study of the cytotoxicity of extracts using different cell lines. FTIR microspectroscopy was also employed to evaluate the cytotoxic effect on cancer cell lines of nanoparticles internalized by endocytosis. The results were promising when amorphous nanoparticles were employed. The nanoparticles loaded with STR increased their size and changed their superficial negative charge to positive. The nanoparticles’ crystallinity decreased, with the consequence that their crystal sizes reduced, when STR was incorporated into their structure. STR maintained its antibacterial activity, although it was reduced during the adsorption into the nanoparticles formed. The STR release was faster from the amorphous ACP nanoparticles and slower from the crystalline cHAp nanoparticles. However, in both cases, the STR release was slower when incorporated in calcium and phosphate during the synthesis. The biocompatibility of these nanoparticles was assayed by two approximations. When extracts from the nanoparticles were evaluated in cultures of cell lines, no cytotoxic damage was observed at concentrations of less than 10 mg/mL. This demonstrated their biocompatibility. Another experiment using FTIR microspectroscopy evaluated the cytotoxic effect of nanoparticles internalized by endocytosis in cancer cells. The results demonstrated slight damage to the biomacromolecules when the cells were treated with ACP nanoparticles. Both ACP and cHAp nanoparticles were efficiently encapsulated in PLA electrospun matrices, providing functionality and bioactive properties.     

Rapid,Label-Free Prediction of Antibiotic Resistance in Salmonella typhimurium by Surface-Enhanced Raman Spectroscopy

The rapid identification of bacterial antibiotic susceptibility is pivotal to the rational administration of antibacterial drugs. In this study, cefotaxime (CTX)-derived resistance in Salmonella typhimurium (abbr. CTX^r-S. typhimurium) during 3 months of exposure was rapidly recorded using a portable Raman spectrometer. The molecular changes that occurred in the drug-resistant strains were sensitively monitored in whole cells by label-free surface-enhanced Raman scattering (SERS). Various degrees of resistant strains could be accurately discriminated by applying multivariate statistical analyses to bacterial SERS profiles. Minimum inhibitory concentration (MIC) values showed a positive linear correlation with the relative Raman intensities of I₉₉₀/I₁₃₄₈, and the R² reached 0.9962. The SERS results were consistent with the data obtained by MIC assays, mutant prevention concentration (MPC) determinations, and Kirby-Bauer antibiotic susceptibility tests (K-B tests). This preliminary proof-of-concept study indicates the high potential of the SERS method to supplement the time-consuming conventional method and help alleviate the challenges of antibiotic resistance in clinical therapy.     

绿色合成桑叶银纳米粒及其抗菌抗癌活性

为了更好地开发出具有良好物理化学及生物活性的银纳米粒,利用桑叶水提取物,通过绿色方法制得桑叶银纳米粒。以AgNO3浓度、反应温度、桑叶水提物的用量、pH以及反应时间为影响因素,优化桑叶银纳米粒最佳合成条件;通过UV-Vis,SEM及FTIR等对产物进行结构表征;通过测定抑菌圈、最小抑菌浓度和细胞毒实验评价其抗菌及抗癌活性。结果显示,最佳制备条件为：AgNO3浓度5 mmol/L、反应温度35 ℃、桑叶水提液与AgNO3溶液体积比1∶5、反应体系pH 11.0及反应时间6 h。在此条件下制备的桑叶银纳米粒为大小均一的球形,平均粒径（48.78 ± 0.39） nm,电位（-27.8 ± 2.00） mV;相比于桑叶水提物,桑叶银纳米粒对大肠杆菌、铜绿假单胞菌,金黄色葡萄球菌、枯草杆菌及白色念球菌均表现较好的抑菌效果,其最小抑菌浓度分别为12.50、25.00、25.00、100.00和100.00 mg/L;桑叶银纳米粒对人宫颈癌（IC50为60.63 mg/L）,人肝癌（IC50为 26.98 mg/L）和人乳腺癌（IC50为18.65 mg/L）细胞有很好的抑制作用。     

Double-Headed Cationic Lipopeptides: An Emerging Class of Antimicrobials

Antimicrobial peptides (AMPs) constitute a promising tool in the development of novel therapeutic agents useful in a wide range of bacterial and fungal infections. Among the modifications improving pharmacokinetic and pharmacodynamic characteristics of natural AMPs, an important role is played by lipidation. This study focuses on the newly designed and synthesized lipopeptides containing multiple Lys residues or their shorter homologues with palmitic acid (C₁₆) attached to the side chain of a residue located in the center of the peptide sequence. The approach resulted in the development of lipopeptides representing a model of surfactants with two polar headgroups. The aim of this study is to explain how variations in the length of the peptide chain or the hydrocarbon side chain of an amino acid residue modified with C₁₆, affect biological functions of lipopeptides, their self-assembling propensity, and their mode of action.     

The Antimicrobial Peptide Temporin G: Anti-Biofilm,Anti-Persister Activities,and Potentiator Effect of Tobramycin Efficacy Against Staphylococcus aureus

Bacterial biofilms are a serious threat for human health, and the Gram-positive bacterium Staphylococcus aureus is one of the microorganisms that can easily switch from a planktonic to a sessile lifestyle, providing protection from a large variety of adverse environmental conditions. Dormant non-dividing cells with low metabolic activity, named persisters, are tolerant to antibiotic treatment and are the principal cause of recalcitrant and resistant infections, including skin infections. Antimicrobial peptides (AMPs) hold promise as new anti-infective agents to treat such infections. Here for the first time, we investigated the activity of the frog-skin AMP temporin G (TG) against preformed S. aureus biofilm including persisters, as well as its efficacy in combination with tobramycin, in inhibiting S. aureus growth. TG was found to provoke ~50 to 100% reduction of biofilm viability in the concentration range from 12.5 to 100 µM vs ATCC and clinical isolates and to be active against persister cells (about 70–80% killing at 50–100 µM). Notably, sub-inhibitory concentrations of TG in combination with tobramycin were able to significantly reduce S. aureus growth, potentiating the antibiotic power. No critical cytotoxicity was detected when TG was tested in vitro up to 100 µM against human keratinocytes, confirming its safety profile for the development of a new potential anti-infective drug, especially for treatment of bacterial skin infections.     

接枝型水不溶高分子抗菌材料的制备与抗菌特性

以接枝微粒氯甲基聚苯乙烯/硅胶(CMPSt/SiO2)为原料,用三乙胺(TEA)、三丁胺(TBA)及三苯基膦对其进行季铵(QN)化与季鏻(QP)化反应,制备了2种水不溶抗菌复合材料QN-PSt/SiO2与QP-PSt/SiO2;以大肠杆菌为致病菌体,采用平板活菌计数法研究了其抗菌性能及抗菌基团结构与抗菌性能的关系,采用胞外DNA和RNA测定法探索了其抗菌机理. 结果表明,2种材料具有很强的抗菌能力,QP-PSt/SiO2用量15 g/L时与浓度为109 mL-1的菌悬液接触1 min,杀菌率达100%. 影响复合微粒抗菌率的主要因素是抗菌基团的化学结构及其在材料表面的密度,接枝大分子链CMPSt的季铵(鏻)化程度即材料表面的季铵(鏻)基团密度越高,抗菌性能越强;季鏻盐型QP-PSt/SiO2的抗菌性能高于季铵盐型QN-PSt/SiO2;以三丁胺为季铵化试剂制备的QN-PSt/SiO2(TBA)的抗菌性能优于以三乙胺为季铵化试剂制备的QN-PSt/SiO2(TEA). QN-PSt/SiO2与QP-PSt/SiO2的抗菌作用实质是杀菌.