大气压氮氧等离子体射流灭活表面大肠杆菌及其发射光谱分析研究

研究了氮氧源大气压等离子体射流对表面大肠杆菌灭活作用,分析了氮氧源大气压等离子体射流的光谱性质。结果表明,在放电电压为6.8kV,气体流速为4L?min?1,处理3min时,氮氧比为1:4的大气压等离子体射流对大肠杆菌的灭活率达到98.4%,接近氧气源大气压等离子体射流灭菌效果。通过大气压等离子体射流发射光谱(OES)分析了氮氧源大气压等离子体射流中活性物质,进而解释大肠杆菌微生物灭活机理,认为NO?γ、OI、?OH等活性物质在表面大肠杆菌灭活过程中起到了重要作用。这将为大气压氮氧等离子体射流在环境卫生、微生物灭活等方面的应用研究提供实验基础和技术支持。     

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研究了氮氧源大气压等离子体射流对表面大肠杆菌灭活作用,分析了氮氧源大气压等离子体射流的光谱性质.结果表明,在放电电压为6.8kV,气体流速为4L?min-1,处理3min时,氮氧比为1:4的大气压等离子体射流对大肠杆菌的灭活率达到98.4%,接近氧气源大气压等离子体射流灭菌效果.通过大气压等离子体射流发射光谱(OES)分析了氮氧源大气压等离子体射流中活性物质,进而解释大肠杆菌微生物灭活机理,认为NO-γ、OI、·OH等活性物质在表面大肠杆菌灭活过程中起到了重要作用.这将为大气压氮氧等离子体射流在环境卫生、微生物灭活等方面的应用研究提供实验基础和技术支持.     

Silcone Catheter Sterilization by Microwave Plasma; Argon and Nitrogen Discharge

Sterilization capability of 2.45GHz Electron Cyclotron Resonance plasma system on silicone catheter is investigated for Ar and N2 gas discharges. E.coli, MRSA, Basillus cereus and Bacillus subtilis bacteria infected silicone catheter pieces are tested for sterilization. Basillus subtilis, which is a spore type bacteria, is determined as the most resistant one, and non spore type bacteria, Escherichia coli, is determined as the easiest one. The results show that nitrogen -N2 discharge has better sterilizing effects than Ar discharge by 15% less in time. Doubling the applied microwave source power decreases sterilization time scanty. Sterilization of silicone catheter is less than an hour for above bacteria types. Plasma discharge sterilization preserves the chemical integrity of polymer — based instruments.     

The effect of sonication on microbial disinfection using hypochlorite

Ultrasound alone is capable of killing bacteria when sufficient power is applied but ultrasound at low powers can also be used to improve the effectiveness biocides. In this paper, we explore the effect of the timing of the ultrasonic treatment at 20 and 850 kHz on the biocidal efficiency of sodium hypochlorite solution towards Escherchia coli suspensions. A remarkable frequency effect has been noted. At the lower frequency of 20 kHz the improvement in biocidal activity is greatest when the ultrasound is applied at the same time as the hypochlorite. At the higher frequency of 850 kHz the improvement is best when ultrasound is used as a pre-treatment immediately followed by hypochlorite addition under normal (silent) conditions. The kill rate achieved for pre-treatment using 850 kHz and simultaneous treatment using 20 kHz are very similar. However the former involves less acoustic energy and so is considered to be the more efficient.     

The development and evaluation of ultrasound for the treatment of bacterial suspensions. A study of frequency,power and sonication time on cultured Bacillus species

Some species of bacteria produce colonies and spores which agglomerate in spherical clusters (Bacillus subtilis) and this serves as a protection for the organisms inside against biocidal attack. Flocs of fine particles e.g. clay can entrap bacteria which can also protect them against the biocides. It is because of problems such as these that alternative methods of disinfecting water are under active investigation. One such method is the use of power ultrasound, either alone or in combination with other methods. Ultrasound is able to inactivate bacteria and deagglomerate bacterial clusters or flocs through a number of physical, mechanical and chemical effects arising from acoustic cavitation. The aim of this study was to investigate the effect of power ultrasound at different powers and frequencies on Bacillus subtilis. Viable plate count techniques were used as a measure of microbial activity. Results showed a significant increase in percent kill for Bacillus species with increasing duration of exposure and intensity of ultrasound in the low-kilohertz range (20 and 38 kHz). Results obtained at two higher frequencies (512 and 850 kHz) indicated a significant increase in bacteria count suggesting declumping. In assessing the bacterial kill with time under different sonication regimes three types of behaviour were characterized:

• High power ultrasound (lower frequencies) in low volumes of bacterial suspension results in a continuous reduction in bacterial cell numbers i.e. the kill rate predominates.

• High power ultrasound (lower frequencies) in larger volumes results in an initial rise in cell numbers suggesting declumping of the bacteria but this initial rise then falls as the declumping finishes and the kill rate becomes more important.

• Low intensity ultrasound (higher frequencies) gives an initial rise in cell numbers as a result of declumping. The kill rate is low and so there is no significant subsequent decrease in bacterial cell numbers.

     

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基于介质阻挡放电(DBD)原理,研究设计了一种悬浮电极式的低温等离子体发生设备,产生的等离子体温度较低且对人体安全,可直接接触。用所研制的低温等离子体发生设备对大肠杆菌做了灭活实验,实验结果表明,该设备产生的低温等离子体对大肠杆菌作用10s 以上,在等离子体有效作用区内大肠杆菌杀灭率为100%,表明所研制的低温等离子体发生设备具有很好的杀菌消毒效果。     

基于DBD 悬浮电极低温等离子体杀菌消毒仪研制

基于介质阻挡放电(DBD)原理,研究设计了一种悬浮电极式的低温等离子体发生设备,产生的等离子体温度较低且对人体安全,可直接接触。用所研制的低温等离子体发生设备对大肠杆菌做了灭活实验,实验结果表明,该设备产生的低温等离子体对大肠杆菌作用10s 以上,在等离子体有效作用区内大肠杆菌杀灭率为100%,表明所研制的低温等离子体发生设备具有很好的杀菌消毒效果。     

太阳光激发UVC紫外上转换发光材料Y_2SiO_5∶Pr~(3+)的灭菌效果研究

制备了Y_2SiO_5∶Pr~(3+)上转换发光材料,首次实现了太阳光激发下的UVC(220~280 nm)紫外上转换发射。为检测紫外(UVC)上转换发光材料的灭菌效果,从土壤中筛选出铜绿假单胞菌并进行培养。为了便于观察,使用Syto9/PI染色剂对细菌着色。实验结果显示:经过太阳光照射后,附着上转换材料的细菌的死亡率比没有上转换材料的细菌有明显的上升。这说明在太阳光的照射下,上转换材料能够将太阳光转化为紫外线并有效灭菌。     

Discovery of NKT cells and development of NKT cell-targeted anti-tumor immunotherapy

Natural Killer T (NKT) cells are unique lymphocytes characterized by their expression of a single invariant antigen receptor encoded by Vα14Jα18 in mice and Vα24Jα18 in humans, which recognizes glycolipid antigens in association with the monomorphic CD1d molecule. NKT cells mediate adjuvant activity to activate both CD8T cells to kill MHC-positive tumor cells and NK cells to eliminate MHC-negative tumor at the same time in patients, resulting in the complete eradication of tumors without relapse. Therefore, the NKT cell-targeted therapy can be applied to any type of tumor and also to anyone individual, regardless of HLA type.Phase IIa clinical trials on advanced lung cancers and head and neck tumors have been completed and showed significantly prolonged median survival times with only the primary treatment. Another potential treatment option for the future is to use induced pluripotent stem cell (iPS)-derived NKT cells, which induced adjuvant effects on anti-tumor responses, inhibiting in vivo tumor growth in a mouse model.     

高能电子束辐照模式蛋白辣根过氧化物酶的损伤作用研究

作为一种载能粒子,高能电子束具有能量利用率高、贯穿深度大、方便实用等优点,可以用来杀灭细菌和抑制病原体生长,因而被广泛应用于食品保质储藏和医疗卫生等方面。但是,对于高能电子如何与生物及其生物分子发生作用,以及作用的过程、方式、效果和机理等方面,人们还缺乏深入、细致的了解。本研究利用高能电子束辐照模式蛋白酶辣根过氧化物酶(HRP:horseradish peroxidase),研究了电子束对它的损伤作用和机理。研究发现,电子束损伤HRP 的方式主要通过羟基自由基破坏蛋白质肽链,同时,辐照产生的活性氧物质也会作用于HRP 分子活性中心引起其辅基血红素的损伤。这与我们以前研究的等离子体放电辐照HRP其损伤主要通过H2O2 作用于变性蛋白血红素引起酶失活有所不同。进一步,通过利用活性氧和自由基清除剂的方法,具体分析和阐明了电子束辐射条件下同活性氧和自由基在损伤HRP 过程中所起的破坏作用。As one kind of charged energetic particles, high-energy electron-beam (e-beam) can kill bacteria effectively; and because of its advantages of low-cost and high-efficiency, e-beam has been widely applied in food sterilization and storage industry as well as biomedical areas. However, currently we still lack the in-depth understanding of the mechanism of the interactions between high-energy particles and biological systems. To this end, we thus initiated the study of e-beam induced damage of horseradish peroxidase (HRP: horseradish peroxidase). Our results revealed that the e-beam induced damage of HRP was mainly through the hydroxyl radical attack on the polypeptide chains, and at the same time, the heme active site of HRP was also injured by the reactive oxygen species (ROS) produced by the electron-beam. This hydroxyl radical damage mechanism is different from the hydrogen peroxide damage mechanism that plays the dominant role in non-thermal plasma treatment as we reported previously. Moreover, by using the ROS and free radical scavengers, we analyzed and identified the major factors that contributed to the HRP damages.     

Spectroscopic and antimicrobial studies of polystyrene films under air plasma and He-Ne laser treatment

Polystyrene (PS) films are used in packaging and biomedical applications because of their transparency and good environmental properties. The present investigation is centered on the antifungal and antibacterial activities involved in the film surface. Subsequently, microbial formations were immobilized on the modified PS films. Living microorganisms such as bacteria and yeast were used. Untreated PS films show very fast rate of growth of bacteria within few hours. The study involves developments of polymer surfaces with bacterial growth and further studies after giving antibacterial treatment such as plasma treatment. Major emphasis has been given to study the effect of various parameters which can affect the performance of the improved material. Films were prepared by two methods: plasma treatment under vacuum and under ongoing He-Ne laser source. The parameters such as (1) surface area by contact angle measurements, (2) quality of material before and after treatment by SEM and FTIR spectra and (3) material characterization by UV-vis spectra were studied. It was observed that plasma treatment of the PS material for different processing time improved the surface properties of PS films.      

Phagocytosis of latex beads and bacteria by hemocytes of the triatomine bug Rhodnius prolixus (Hemiptera: Reduvidae)

Insect circulating hemocytes are primarily responsible for the immune defense against parasites and pathogens. Here, we have analyzed phagocytosis of both biotic (bacteria) and abiotic (latex) particles by circulating hemocytes of 5th-instar nymphs of the triatomine bug Rhodnius prolixus. The following hemocyte types were identified: prohemocytes, plasmatocytes, granulocytes, oenocytoids and adipohemocytes. There was a considerable change in the relative percentage of plasmatocytes and prohemocytes in the hemolymph after challenge with both latex beads and bacteria. Granulocytes and oenocytoids also change their relative percentage in response to latex bead and Staphylococcus aureus, respectively. No significant change was observed in adipohemocytes at any time or treatment. Our data demonstrated that plasmatocytes were the only cell type involved in phagocytosis of foreign particles. As in mammal cells, phagocytosis by both zipper and trigger mechanisms were observed for the uptake of latex beads and bacteria. Neither melanization nor micro-aggregation was observed towards latex particles or Escherichia coli. On the other hand, R. prolixus produced a strong melanization reaction against S. aureus, thus showing that differences exist in the responses to E. coli and to S. aureus. Ultrastructural changes observed in plasmatocytes, adipohemocytes and oenocytoids suggest that these hemocyte types are directly involved in the immune defense of R. prolixus against foreign particles.     

Surface modification of multi-walled carbon nanotubes by O2 plasma

The surface modification of multi-walled carbon nanotubes (MWCNTs) by O₂ plasma was carried out in this study. In order to achieve a relatively homogeneous treatment of MWCNTs powder, a rotating barrel fixed between the two discharge electrodes was used. The effect of plasma treatment parameters, such as power, time, and positions of samples (inside and outside the barrel), on the morphology and structure of MWCNTs surface was systematically analyzed by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM). The results showed that the direct discharge (outside the barrel) could result in not only a quick grafting of polar functional groups but also an easy damage of MWCNTs after longer time, particularly under intensive power. It was found that the surface of MWCNTs powder might be changed in three steps—expansion (loosed structure formed), peel off and oxidization with increasing of treatment time during the irradiation. In this way, a complete purification of MWCNTs powder could be finished within 30 min via plasma treatment. Our work suggested that plasma treatment could be a simple and nonpolluting method for a large scale purification of MWCNTs.     

Surface diffuse discharge mechanism of well-aligned atmospheric pressure microplasma arrays

A stable and homogeneous well-aligned air microplasma device for application at atmospheric pressure is designed and its electrical and optical characteristics are investigated. Current-voltage measurements and intensified charge coupled device(ICCD) images show that the well-aligned air microplasma device is able to generate a large-area and homogeneous discharge at the applied voltages ranging from 12 kV to 14 kV, with a repetition frequency of 5kHz, which is attributed to the diffusion effect of plasma on dielectric surface. Moreover, this well-aligned microplasma device may result in the uniform and large-area surface modification of heat-sensitive PET polymers without damage, such as optimization in hydrophobicity and biocompatibility. In the biomedical field, the utility of this well-aligned microplasma device is further testified. It proves to be very efficient for the large-area and uniform inactivation of E. coli cells with a density of 103/cm~2 on LB agar plate culture medium, and inactivation efficiency can reach up to 99% for 2-min treatment.     

Ultrasound pasteurization: the effects of temperature, soluble solids, organic acids and pH on the inactivation of Escherichia coli ATCC 25922

The objectives of this research were to study the effects of temperature, sugar concentration (8, 12, and 16 g/100ml), organic acids (citric and malic acids) and pH (2.5 and 4.0) on ultrasound pasteurization. The model organism used for the research was Escherichia coli ATCC 25922, and ultrasound treatment times were conducted to achieve a 5 log (base 10) reduction. Ultrasound generates heat, therefore the study involved removing the heat using a jacketed beaker with refrigerant (-30 degrees C) to maintain processing temperature at or below 30 degrees C and eliminate the thermal inactivation effects. Overall, ultrasound increased the sensitivity of E. coli to thermal inactivation. The presence of soluble solids had a protective effect where the sonication time requirement increased. Similar to heat sensitivity, the lower pH environment resulted in E. coli having less resistance to sonication. The type of organic acid had the least significant effect on ultrasound inactivation of E. coli.     

Low temperature argon plasma sterilization effect on Pseudomonas aeruginosa and its mechanisms

In the present work, low temperature argon plasma was used to sterilize Pseudomonas aeruginosa on PET sheets. The discharge conditions were as follows: power of 30 W and gas flux of 20 cm³/min, and the pressure in the reaction chamber was 6.2 Pa. In a self-designed plasma reaction equipment, active species (electron, ion, radical and UV light, etc.) were separated effectively, and the discharge zone, afterglow zone and remote zone were plotted out in the plasma field. For a treatment time of 60 s, the germicidal effects were 3.94, 3.86 and 2.93 respectively in the three zones. The results show that argon plasma can effectively sterilize P. aeruginosa in a short time. Before and after plasma treatment, cell morphology was studied via scanning electron microscopy (SEM). Cell wall or cell membrane cracking was tested to determine the content of nucleic acid in the bacteria suspensions using ultraviolet spectroscopy. To test the content of protein, a coomassie light blue technique was used. The results show that plasma activity cracks the cell wall and cell membrane, resulting in cellular content leakage. In addition, the results from electron spin resonance (ESR) spectroscopy and double Langmuir electron probe show that electrons, ions and argon free radicals play important roles in sterilization in the discharge zone, but only argon free radicals act to sterilize bacteria in the afterglow zone and the remote zone.     

Microorganism response to atmospheric pressure helium plasma DBD treatment

The effect of an atmospheric pressure plasma dielectric barrier discharge on some microorganisms was investigated, due to the possible extent of plasma applications in controlling microbial contamination. The present investigation involved both Gram-positive and Gram-negative germs – bacteria and yeasts – treated in helium plasma (asymmetric dielectric barrier discharge) for various time durations between 25 s and 100 s. Circular growth inhibition zones were measured for two distances between the discharge electrodes, the correlations with the plasma treatment time being analyzed. Differences between the sensitivities of the four tested microorganisms to helium plasma in open atmosphere were discussed.     

Correlations between microstructure of plasma-modified gold nanoclusters and their optical properties

Gold nanoclusters with diameters up to 50 nm were grown in sandwich structures consisting in 15 nm of plasma deposited silicon nitride, 1 nm of gold grown by sputtering and 15 nm of plasma deposited silicon nitride (SiN/Au/SiN). Previous to the last step, ammonia plasma treatments of the gold surface were carried out with time as the main variable. The resulting structures were analyzed by high resolution transmission electron microscopy and spectroscopic ellipsometry. As a result of plasma treatments, island-like structures of as-grown gold clusters evolve to near spherical-shape features with decreasing diameter as the plasma treatment time rises. Ellipsometric spectra were modeled based on the Bruggeman effective medium approximation and the influence of size and shape of nanoparticles on the optical properties were calculated.     

Lasing from Escherichia coli bacteria genetically programmed to express green fluorescent protein

We report on lasing action from colonies of Escherichia coli bacteria that are genetically programmed to synthesize the green fluorescent protein (GFP). When embedded in a Fabry-Perot type cavity and excited by ns-pulses of blue light (465 nm), the bacteria generate green laser emission (～520 nm). Broad illumination of pump light yields simultaneous lasing over a large area in bacterial colonies.     

原核表达过程中非目标蛋白质识别与确认的方法:NRSF/REST蛋白功能结构域ZnF2-8原核表达过程中β-内酰胺酶的确认

大肠杆菌常被用来大量快速制备重组蛋白质。但是,在原核表达目标蛋白质时非目标蛋白质经常会意外表达。有时这些非目标蛋白质也非常有使用价值,但是最终确认这些非目标蛋白质的过程昂贵又及其耗时。基于此,该文发展了一个新的基于二维核磁共振波谱技术、X-单晶衍射技术、结合其他生物化学方法,确认在原核表达神经限制性沉默因子 NRSF/REST 蛋白（该蛋白能够特异性识别神经限制性沉默因子 RE1 dsDNA及神经限制性激活因子dsRNA,以调节神经元干细胞的发育）功能结构域ZnF2-8时非目标蛋白b-内酰胺酶(b-lactamase)。