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We report an interesting buffer electric relaxation time tuning technique, coupled with a glutaraldehyde cross-linking cell fixation reaction, which allows for sensitive dielectrophoretic analysis and discrimination of bovine red blood cell (bRBC) starvation age. The buffer composition is selected such that two easily accessible dielectrophoretic crossover frequencies (cof) exist. Low concentration glutaraldehyde fixation was observed to produce a threefold decrease in the higher cof with a comparable increase in the lower cof also witnessed. More importantly, increased glutaraldehyde fixation concentration significantly increased the higher cof by a factor found to be sensitive to the bRBC starvation age. 相似文献
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We introduce a method for improved dielectrophoretic (DEP) discrimination and separation of viable and nonviable yeast cells. Due to the higher cell wall permeability of nonviable yeast cells compared with their viable counterpart, the cross-linking agent glutaraldehyde (GLT) is shown to selectively cross-link nonviable cells to a much greater extent than viable yeast. The DEP crossover frequency (cof) of both viable and nonviable yeast cells was measured over a large range of buffer conductivities (22 μS∕cm–400 μS∕cm) in order to study this effect. The results indicate that due to selective nonviable cell cross-linking, GLT modifies the DEP cof of nonviable cells, while viable cell cof remains relatively unaffected. To investigate this in more detail, a dual-shelled oblate spheroid model was evoked and fitted to the cof data to study cell electrical properties. GLT treatment is shown to minimize ion leakage out of the nonviable yeast cells by minimizing changes in cytoplasm conductivity over a large range of ionic concentrations. This effect is only observable in nonviable cells where GLT treatment serves to stabilize the cell cytoplasm conductivity over a large range of buffer conductivity and allow for much greater differences between viable and nonviable cell cofs. As such, by taking advantage of differences in cell wall permeability GLT magnifies the effect DEP has on the field induced separation of viable and nonviable yeasts. 相似文献
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目的:使用免疫BMPs,对含有大肠杆菌O157的粪便标本,进行目标病原菌的捕获和洗净,建立一种快速纯化细菌PCR反应模板的方法。方法:将大肠杆菌O157抗体接种于BMPs表面,制成免疫BMPs。使用免疫BMPs,对含有不同浓度大肠杆菌O157的粪便标本,进行目标病原菌的捕获、分离和洗净,再使用洗净的病原菌制备PCR反应模板。作为对照,对含有不同浓度大肠杆菌O157的粪便标本,常规进行增菌培养及鉴别培养后,制备细菌的PCR反应模板,或粪便标本直接制备PCR反应模板。结果:使用免疫BMPs处理后制备PCR反应模板,与通过常规增菌培养和鉴别培养后制备PCR反应模板,PCR检测结果一致。而粪便标本直接取样制备PCR反应模板,大肠杆菌O157低浓度时,PCR检测的阳性率较低。结论:利用免疫BMPs捕获自然标本(粪便)中的目标病原菌,通过分离和洗净,去除标本中存在的PCR反应抑制物等影响因素,可省略增菌培养及鉴别培养,快速纯化目标病原菌的PCR反应模板。 相似文献
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BackgroundEndometritis is the most common disease of dairy cows and traditionally treated with antibiotics. Lactic acid bacteria can inhibit the growth of pathogens and also have potential for treatment of endometritis. Using PacBio single-molecule real-time sequencing technology, we sequenced the full-length l6S rRNA of the microbiota in uterine mucus samples from 31 cows with endometritis, treated with lactic acid bacteria (experimental [E] group) and antibiotics (control [C] group) separately. Microbiota profiles taken before and after treatment were compared.ResultsAfter both treatments, bacterial species richness was significantly higher than before, but there was no significant difference in bacterial diversity. Abundance of some bacteria increased after both lactic acid bacteria and antibiotic treatment: Lactobacillus helveticus, Lactococcus lactis, Lactococcus raffinolactis, Pseudomonas alcaligenes and Pseudomonas veronii. The bacterial species that significantly decreased in abundance varied depending on whether the cows had been treated with lactic acid bacteria or antibiotics. Abundance of Staphylococcus equorum and Treponema brennaborense increased after lactic acid bacteria treatment but decreased after antibiotic treatment. According to COG-based functional metagenomic predictions, 384 functional proteins were significantly differently expressed after treatment. E and C group protein expression pathways were significantly higher than before treatment (p < 0.05).ConclusionsIn this study, we found that lactic acid bacteria could cure endometritis and restore a normal physiological state, while avoiding the disadvantages of antibiotic treatment, such as the reductions in abundance of beneficial microbiota. This suggests that lactic acid bacteria treatment has potential as an alternative to antibiotics in the treatment of endometritis in cattle.How to cite: Yang L, Huang W, Yang C, et al. Using PacBio sequencing to investigate the effects of treatment with lactic acid bacteria or antibiotics on cow endometritis. Electron J Biotechnol 2021:51. https://doi.org/10.1016/j.ejbt.2021.02.004 相似文献
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溶藻弧菌铁调蛋白基因的克隆、表达及其特征分析 总被引：1，自引：0，他引：1
铁为一切有机体所必需。对铁摄取与利用的调控在细菌生长和铁毒性中起重要作用。在革兰氏阴性细菌中,铁摄取调节蛋白(Fur)主管铁的代谢。溶藻弧菌是人和海洋动物共感染的一种主要病原弧菌。本研究克隆表达了溶藻弧菌的fur基因,并分析了其相关特征。该基因序列全长994 bp,其中包含450 bp的开放读码框,编码150个氨基酸残基的蛋白,推导的相对分子量为16.78 kD.在编码序列的上下游,RBS序列-、10序列-、35序列和二个潜在的转录终止子分别得到确定,但没有发现存在于大肠杆菌fur基因中的"铁盒子"。氨基酸序列的中段从G46位至D104位,为高度保守区域,其中包括含组氨酸丰富铁结合模型(H3-D-H-L-V-C-L-D-C-G)。SDS-PAGE分析表明,fur基因可在大肠杆菌中大量表达,带His-Tag的重组融和蛋白经镍亲和层析得到纯化,Western-blot分析结果显示,Fur蛋白具有免疫反应性。为进一步研究Fur蛋白的结构与功能打下基础。 相似文献
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超嗜热菌Aquifex aeolicus 亮氨酰-tRNA 合成酶(αβ-LeuRS) 是已知的唯一的双肽链LeuRS。通过αβ-LeuRS和大肠杆菌LeuRS相应肽段的重组和重组酶性质的研究发现，αβ-LeuRS的α亚基C末端36个氨基酸对于αβ-LeuRS的氨基酰化活力至关重要。αβ-LeuRS的两个亚基人为融合得到的单亚基酶SLeuRS-αβ，具有完全的野生型双亚基酶活力，其热稳定性显著增强。同时，通过亚基重组揭示了LeuRS 对于不同种属的tRNALeu的识别元件位于α亚基内，而其中的CP1 结构域不仅是LeuRS 行使编校功能的结构基础，对于LeuRS识别不同种属来源的tRNALeu 也是十分重要的。通过不同来源的LeuRS一级结构比较，克隆了单独的CP1结构域组成的肽(CP1),发现唯有A. aeolicus αβ-LeuRS 的CP1对错误的氨基酰化产物具有体外编校功能。它与其它3种来源于细菌LeuRS的CP1都能够编校误氨基酰化的古老形式的亮氨酸小螺旋 (minihelixLeu)。通过同源序列比较发现，在αβ-LeuRS的CP1内存在一个由20个氨基酸组成的特异的结构模体，它对CP1发挥体外编校功能必不可少，引入原本不具有体外编校功能的E. coli的CP1 后则可使其获得编校功能，但是该模体不影响全酶的编校功能。αβ-LeuRS中与tRNA结合有关的亚基－β亚基也可以赋予E. coli CP1编校功能。这些研究结果充分表明，来源于古老的超嗜热菌A. aeolicus 的αβ-LeuRS 保留了进化的遗迹－具有编校活力的CP1和与其编校功能密切相关的由20个氨基酸组成的特异的结构模体，它们在进化过程中随着其它结构域招募进合成酶中而逐渐失去了对全酶编校功能的作用。这一发现有力地支持了aaRS通过纳入新的结构域以加速进化过程的理论，并且也从不同角度为aaRS/tRNA共进化理论提供了可靠的依据。 相似文献
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Rapid concentration and detection of bacteria in integrated chips and microfluidic devices is needed for the advancement of lab-on-a-chip devices because current detection methods require high concentrations of bacteria which render them impractical. We present a new chip-scale rapid bacteria concentration technique combined with surface-enhanced Raman scattering (SERS) to enhance the detection of low bacteria count samples. This concentration technique relies on convection by a long-range converging vortex to concentrate the bacteria into a packed mound of 200 μm in diameter within 15 min. Concentration of bioparticle samples as low as 104 colony forming units (CFU)∕ml are presented using batch volumes as large as 150 μl. Mixtures of silver nanoparticles with Saccharomyces cerevisiae, Escherichia coli F-amp, and Bacillus subtilis produce distinct and noticeably different Raman spectra, illustrating that this technique can be used as a detection and identification tool. 相似文献
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为了研究4条人工合成抗菌肽的抑菌和溶血作用。在大肠杆菌悬浮液和绵羊血红细胞悬浮液中,分别加入不同溶度的人工合成抗菌肽,观察其抑菌效果和溶血作用。同时,用圆二色谱仪测定抗菌肽的二级结构。在4条人工合成抗菌肽中,抗菌肽CM-S的抑菌效果为最佳,最小抑菌溶度为25μg/mL,而LB平板菌落的杀菌溶度为50μg/mL;在对绵羊血红细胞的溶血试验中,CM-S在50μg/mL时无溶血作用,100μg/mL时只表现出轻微溶血特性;用圆二色谱仪测定CM-S的二级结构,在2.5%SDS溶液中,208 nm和222 nm出现典型α-螺旋特征峰。人工合成抗菌肽CM-S,含有17个氨基酸,呈典型α-螺旋结构,对大肠杆菌的抑杀效果显著,且溶血性很低,有良好的临床药物开发前景。 相似文献
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Sajitha GR Jose R Andrews A Ajantha KG Augustine P Augusti KT 《Indian journal of clinical biochemistry : IJCB》2010,25(3):280-288
Daily feeding of drinking water containing lead acetate (160 mg/l) or 10% alcohol by volume or a combination of both to rats
for a month produced certain deleterious effects through oxidative stress. Both heavy metal lead and alcohol are capable of
doing such damages. The deleterious alterations observed were in the parameters of blood, serum and tissues, viz; Hb, Pb,
proteins, lipids, lipid per oxidation, Vitamins C and E levels and enzyme activities of AST, ALT, and catalase. Simultaneous
feeding of either of the two antioxidants garlic oil (GO) and vitamin E at equal doses of 100 mg/kg/day, to the rats counteracted
the deleterious effects of the above two chemicals significantly. The maximum damage was brought about by feeding of drinking
water containing both lead acetate and alcohol. The protective effects of GO and Vitamin E were not significantly different.
The mechanism of actions of the Vitamin E and GO is probably due to their efficiency as detoxifying agents and antioxidants,
to scavenging free radicals as well as an independent action of GO on the removal of lead salt as lead sulfide. 相似文献
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M. Skolimowski M. Weiss Nielsen F. Abeille P. Skafte-Pedersen D. Sabourin A. Fercher D. Papkovsky S. Molin R. Taboryski C. Sternberg M. Dufva O. Geschke J. Emnéus 《Biomicrofluidics》2012,6(3)
A modular microfluidic airways model system that can simulate the changes in oxygen tension in different compartments of the cystic fibrosis (CF) airways was designed, developed, and tested. The fully reconfigurable system composed of modules with different functionalities: multichannel peristaltic pumps, bubble traps, gas exchange chip, and cell culture chambers. We have successfully applied this system for studying the antibiotic therapy of Pseudomonas aeruginosa, the bacteria mainly responsible for morbidity and mortality in cystic fibrosis, in different oxygen environments. Furthermore, we have mimicked the bacterial reinoculation of the aerobic compartments (lower respiratory tract) from the anaerobic compartments (cystic fibrosis sinuses) following an antibiotic treatment. This effect is hypothesised as the one on the main reasons for recurrent lung infections in cystic fibrosis patients. 相似文献
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抗生素除了大量用于人类疾病的治疗外,还作为饲料添加剂被广泛应用于动物养殖业。微生物的抗生素耐药性就是指微生物能够在抗生素存在的情况下生长和繁殖。抗生素耐药性是环境微生物固有的,即所谓的内在抗性,但是人类大量使用抗生素带来的抗生素抗性基因的扩散和传播普遍存在,且已开始威胁到全球人群的健康。微生物对抗生素的抗性主要有3个机制:(1)抗生素的外排;(2)抗生素的降解或修饰;(3)抗生素作用位点的保护。大量研究表明,抗生素的使用和抗生素抗性的蔓延呈现良好的相关性,而且环境微生物的抗性可以通过基因横向转移向人类致病菌扩散,最终可能导致超级细菌的爆发,直接影响人类健康。为了应对全球性的抗生素抗性问题,必须加强:(1)全球抗生素使用和环境排放的监管政策和管理体系;(2)建立快速和透明的抗生素耐药性监测体系,使其涵盖医院、养殖业、污水处理厂等;(3)建立抗生素药物创新基金,通过政府和企业的联合,加快新型药物的研制;同时加强知识产权保护,使新药创制走上可持续之路;(4)加强抗生素耐药性相关的基础与应用研究,包括耐药性发生和传播的生态学机制,消除和缓解耐药性发生和传播的环境技术及其系统解决方案等,包括改进污水处理厂的处理工艺,削减出水中抗性基因和抗性菌的比例;(5)加强抗生素耐药性的科普宣传,提高全社会对耐药性的认知能力,从而在源头上有效控制抗生素在农业和医疗方面的滥用及其环境污染。 相似文献
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M. S. Ghadge A. V. Sirsat M. S. Bhansali L. J. Desouza P Jagannath 《Indian journal of clinical biochemistry : IJCB》2001,16(1):60-64
Serum levels of leucine amino peptidase (LAP) was studied along with bilirubin, aspartate transaminase (AST), alanine transaminase
(ALT), gamma glutamyl transpeptidase (GGT), alkaline phosphatase (ALP) and the ratio of AST/ALT and GGT/AST in 25 healthy
subjects and 52 patients with hepatobiliary malignancies of which 12 were with hepatocellular carcinoma, 12 with liver metastasis,
6 with obstructive jaundice, 9 with carcinoma of gall bladder, 6 with carcinoma of pancreas and 7 with periampullary carcinoma.
24 Of the 52 patients studied had jaundice and 28 were without jaundice.
LAP as compared to the other enzymes AST, ALT, GGT, ALP and AST/ALT ratio and GGT/AST ratio showed 100% elevation in obstructive
jaundice, carcinoma of gall bladder and pancreas and periampullary carcinoma, 91.7% elevation in hepatocellular carcinoma
and 83.3% elevation in liver metastasis. On comparing the levels of these enzymes in non jaundiced and jaundiced groups, LAP
was elevated in both jaundiced and non jaundiced groups in 95.8% and 92.9% cases respectively whereas the other enzymes AST
showed increase from 67.9% to 100%, ALT from 21.4% to 83.3%, GGT from 71.4% to 95.4% and ALP from 82.1% to 100% in non jaundiced
and jaundiced groups respectively indicating that LAP rises in hepatic dysfunction due to hepatobiliary malignancy whereas
the other liver function enzymes showed increased hepatic dysfunction due to hepatobiliary malignancy with the onset of jaundice
thereby indicating that LAP is a better indicator of hepatobiliary malignancy as compared to other enzymes.
The quantitative methods used for determination are reliable, accurate, simple, rapid and cost effective and therefore have
better application in a clinical setting. 相似文献
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O. Yassine C. P. Gooneratne D. Abu Smara F. Li H. Mohammed J. Merzaban J. Kosel 《Biomicrofluidics》2014,8(3)
This study describes the development and testing of a magnetic microfluidic chip (MMC) for
trapping and isolating cells tagged with superparamagnetic beads (SPBs) in a microfluidic
environment for selective treatment and analysis. The trapping and isolation are done in two
separate steps; first, the trapping of the tagged cells in a main channel is achieved by soft
ferromagnetic disks and second, the transportation of the cells into side chambers for isolation is
executed by tapered conductive paths made of Gold (Au). Numerical simulations were performed to
analyze the magnetic flux and force distributions of the disks and conducting paths, for trapping
and transporting SPBs. The MMC was fabricated using standard microfabrication processes. Experiments
were performed with E. coli (K12 strand) tagged with 2.8 μm SPBs.
The results showed that E. coli can be separated from a sample solution by trapping
them at the disk sites, and then isolated into chambers by transporting them along the tapered
conducting paths. Once the E. coli was trapped inside the side chambers, two
selective treatments were performed. In one chamber, a solution with minimal nutrition content was
added and, in another chamber, a solution with essential nutrition was added. The results showed
that the growth of bacteria cultured in the second chamber containing nutrient was significantly
higher, demonstrating that the E. coli was not affected by the magnetically driven
transportation and the feasibility of performing different treatments on selectively isolated cells
on a single microfluidic platform. 相似文献
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Seema Gupta Rajesh Pandey Ranjan Katyal H. K. Aggarwal R. P. Aggarwal S. K. Aggarwal 《Indian journal of clinical biochemistry : IJCB》2005,20(1):67-71
The present study was conducted to evaluate some of the components of antioxidant defense system and oxidative damage in 20
male patients of alcoholic liver disease (ALD). The results were compared with 20 healthy male smokers and 20 healthy male
non-smokers volunteers. Patients were subjected to detailed clinical examination and laboratory investigations. Blood samples
were collected for estimating reduced glutathione (GSH), total thiols (T-SH) malondialdehyde (MDA), transaminases (AST, ALT),
glutathione-S-transferease (GST) and gammaglutamyl transferase (GGT). Serum aspartate amino transferase (AST)/alanine amino
transferase (ALT) ratio was significantly (p<0.01) reduced in ALD patients as compared to the controls. However, the core
of utility of MDA and GST was found to be significantly (p<0.01) increased in ALD patients compared to controls. There was
a significant negative correlation of MDA with both GSH and TSH. Plasma GGT levels were significantly (p<0.01) increased in
alcoholics and the enzyme showed a significant positive correlation with MDA. These results give enough evidence of increased
oxidative stress and compromised antioxidant defense system in patients with ALD. 相似文献
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BackgroundBiosurfactants are surface active molecules produced by microorganisms which have the ability to disrupt the plasma membrane. Biosurfactant properties are important in the food, pharmaceutical and oil industries. Lactic acid bacteria can produce cell-bound and excreted biosurfactants.ResultsThe biosurfactant-producing ability of three Lactobacillus strains was analyzed, and the effects of carbon and nitrogen sources and aeration conditions were studied. The three species of lactobacillus evaluated were able to produce biosurfactants in anaerobic conditions, which was measured as the capacity of one extract to reduce the surface tension compared to a control. The decreasing order of biosurfactant production was L. plantarum>Lactobacillus sp.>L. acidophilus. Lactose was a better carbon source than glucose, achieving a 23.8% reduction in surface tension versus 12.9% for glucose. Two complex nitrogen sources are required for growth and biosurfactant production. The maximum production was reached at 48 h under stationary conditions. However, the highest level of production occurred in the exponential phase. Biosurfactant exhibits a critical micelle concentration of 0.359 ± 0.001 g/L and a low toxicity against E. coli. Fourier transform infrared spectroscopy indicated a glycoprotein structure. Additionally, the kinetics of fermentation were modeled using a logistic model for the biomass and the product, achieving a good fit (R2 > 0.9).ConclusionsL. plantarum derived biosurfactant production was enhanced using adequate carbon and nitrogen sources, the biosurfactant is complex in structure and because of its low toxicity could be applied to enhance cell permeability in E. coli.How to cite: Montoya Vallejo C, Florez Restrepo MA, Guzmán Duque FL, et al. Production, characterization and kinetic model of biosurfactant produced by lactic acid bacteria. Electron J Biotechnol 2021;53. https://doi.org/10.1016/j.ejbt.2021.06.001 相似文献