1.Quantification of antigen of Mycoplasma capricolum subsp. capripneumoniae by optical assay.
Jiazhen GE ; Pengcheng GAO ; Tongtong TIAN ; Xiaoni WU ; Qianqian LI ; Kexin TIAN ; Guodong SONG ; Fuying ZHENG ; Yuefeng CHU
Chinese Journal of Biotechnology 2023;39(12):4874-4886
Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.
Humans
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Animals
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Goats
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Cetrimonium
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Mycoplasma
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Polysaccharides
2.Interaction of nonionic surfactant AEO9 with ionic surfactants.
Journal of Zhejiang University. Science. B 2005;6(6):597-601
The interaction in two mixtures of a nonionic surfactant AEO9 (C12H25O(CH2CH2O)9H) and different ionic surfactants was investigated. The two mixtures were AEO9/sodium dodecyl sulfate (SDS) and AEO9/cetyltrimethylammonium bromide (CTAB) at molar fraction of AEO9, alpha(AEO9) The surface properties of the surfactants, critical micelle concentration (CMC), effectiveness of surface tension reduction (gamma(CMC)), maximum surface excess concentration (Gamma(max)) and minimum area per molecule at the air/solution interface (A(min)) were determined for both individual surfactants and their mixtures. The significant deviations from ideal behavior (attractive interactions) of the nonionic/ionic surfactant mixtures were determined. Mixtures of both AEO9/SDS and AEO9/CTAB exhibited synergism in surface tension reduction efficiency and mixed micelle formation, but neither exhibited synergism in surface tension reduction effectiveness.
Cetrimonium Compounds
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analysis
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chemistry
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Colloids
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analysis
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chemistry
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Complex Mixtures
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analysis
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chemistry
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Ions
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Phase Transition
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Sodium Dodecyl Sulfate
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analysis
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chemistry
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Solutions
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Surface Tension
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Surface-Active Agents
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analysis
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chemistry
3.Purification of large-scale plasmid DNAs by selective precipitation with cetyltrimethylammonium bromide.
Quan ZHANG ; Kexiang YU ; Weifeng YUAN ; Fangming XUE ; Huaichang SUN ; Hongfei ZHU
Chinese Journal of Biotechnology 2008;24(12):2117-2121
Following Escherichia coli lysis with alkali, cetyltrimethylammonium bromide (CTAB) was directly titrated into the supernatant. An easy and feasible technology for plasmid purification was established with the optimized proportion between the quantity of CTAB and plasmid, combined with the specific solution for DNA release and TritonX-114 for endotoxin removal. Quality detection showed that the purified plasmid was free of contamination of host RNA. The host bacterial genomic DNA, endotoxin and bacterial protein were less than 10 microg, 50 EU and 10 microg per mg plasmids, respectively. The ratio of OD260/OD280 was between 1.75-1.85. Eighty percent of the prepared plasmids were presented in the supercoiled form. The plasmid purified with this technology can satisfy all criteria stipulated by FDA. The main advantages of the technology include the avoidance of animal-derived enzymes such as ribonucleases A, Proteinase K and toxic reagents like chloroform and phenol. In addition, the technology has low cost and no pollution.
Cetrimonium Compounds
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chemistry
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Chemical Precipitation
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DNA
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genetics
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isolation & purification
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DNA, Bacterial
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genetics
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isolation & purification
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Escherichia coli
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chemistry
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genetics
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Plasmids
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genetics
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isolation & purification
4.Extraction and characterization of total DNA from Dendrobium.
Rui PENG ; Hong-yuan SONG ; Quan-sen LI ; Yu WANG
China Journal of Chinese Materia Medica 2003;28(12):1129-1131
OBJECTIVETo explore the quality of DNA with three methods of DNA extraction and the influence on RAPD-PCR.
METHODthe electrophoresis of total DNA, UV spectrophotometry and RAPD analysis were carried out on DNA extracted from three different methods.
RESULTThe DNA concentration and yields were different, which greatly influenced the result of RAPD-PCR.
CONCLUSIONThe higher quality DNA from Dendrobium can be obtained with the method of CTAB-free extraction medium before total DNA was isolated.
Cetrimonium Compounds ; DNA, Plant ; isolation & purification ; Dendrobium ; classification ; genetics ; Electrophoresis ; Plants, Medicinal ; classification ; genetics ; Quality Control ; Random Amplified Polymorphic DNA Technique ; Spectrophotometry, Ultraviolet
5.Study of DNA identification in burned bones.
Jian YE ; An-quan JI ; Xing-chun ZHAO
Journal of Forensic Medicine 2004;20(3):155-159
OBJECTIVE:
For the purpose of solving a problems of DNA testing of burned bones.
METHODS:
We present a novel strategy to obtain DNA from burned bones based on the use of cetyltrimethylammonium bromide (CTAB) lysis buffer and isoamyl alcohol-chlorophorm extraction with subsequent DNA purification using the DNA IQ System.
RESULTS:
The methods were found to be effective in removing the PCR inhibitors from the burned bone. Then the extracted DNA was successfully genotyped by using the florescence labeling STR multiplex method.
CONCLUSION
The results of this research will assist forensic scientists in the identification of DNA from victims whose bodies underwent significant trauma or burning, precluding the utilization of traditional forensic DNA identification techniques.
Bone and Bones/chemistry*
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Burns/metabolism*
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Cetrimonium Compounds
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DNA/isolation & purification*
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DNA Fingerprinting/methods*
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Female
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Forensic Medicine/methods*
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Humans
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Male
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Polymerase Chain Reaction/methods*
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Sensitivity and Specificity
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Tandem Repeat Sequences
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Time Factors
6.Preparation of ATP-2Na loaded liposome and its effect on tissues energy state in myocardial ischemic mice.
Feng-mei PI ; Xi-de TU ; Yue WU
Acta Pharmaceutica Sinica 2010;45(10):1322-1326
The aim of this study is to improve liposome encapsulation efficiency of water soluble drug ATP-2Na with hydrophobic ion pairing method, and evaluate its effect on tissues energy state in myocardial ischemia mice. Ion pair complex of ATP-2Na with HTAB was prepared first; then the liposomes were prepared by ethanol injection method. The size and zeta potential of ATP-2Na liposome were investigated. Its effect on tissues energy state in myocardium ischemia mice was evaluated by detecting ATP-2Na concentration in tissues and blood after injection in comparison to ATP-2Na solution. The diameters and zeta potential of ATP-2Na liposomes were (144.0 +/- 2.7) nm and (+16.2 +/- 1.6) mV, respectively. The encapsulation efficiency was (85.02 +/- 2.31) %. The in vitro drug release pattern from liposomes matches with Weibull equation. Compared with ATP-2Na solution, ATP-2Na liposome increased the ATP concentration of blood in myocardial ischemic mice very significantly; compared with blank, ATP-2Na liposome increased ATP content of myocardium and liver in myocaridal ischemic mice significantly, but ATP-2Na solution didn't show this effect. ATP-2Na liposome might have an advantage in improving tissue energy state on myocaridal ischemic animals.
Adenosine Triphosphate
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administration & dosage
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blood
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metabolism
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Animals
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Cetrimonium Compounds
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chemistry
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Drug Carriers
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Liposomes
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chemistry
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Liver
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metabolism
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Male
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Mice
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Myocardial Ischemia
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blood
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metabolism
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Myocardium
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metabolism
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Particle Size
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Random Allocation
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Surface-Active Agents
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chemistry
7.Sorption of wastewater containing reactive red X-3B on inorgano-organo pillared bentonite.
Journal of Zhejiang University. Science. B 2006;7(4):314-319
Bentonite is a kind of natural clay with good exchanging ability. By exchanging its interlamellar cations with various soluble cations, such as quaternary ammonium cations and inorganic metal ions, the properties of natural bentonite can be greatly improved. In this study, hexadecyltrimethylammonium bromide (HDTMA), CaCl(2), MgCl(2), FeCl(3), AlCl(3) were used as organic and inorganic pillared materials respectively to produce several kinds of Ca-, Mg-, Fe-, Al-organo pillared bentonites. Sorption of reactive red X-3B on them was studied to determine their potential application as sorbents in wastewater treatment. The results showed that these pillared bentonites had much improved sorption properties, and that the dye solutions' pH value had some effect on the performance of these inorgano-organo pillared bentonites. Isotherms of reactive X-3B on these pillared bentonites suggested a Langmuir-type sorption mechanism.
Aluminum Compounds
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chemistry
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Bentonite
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chemistry
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Calcium Chloride
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chemistry
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Cations
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Cetrimonium Compounds
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chemistry
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Chlorides
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chemistry
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Coloring Agents
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chemistry
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Ferric Compounds
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chemistry
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Hydrogen-Ion Concentration
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Magnesium Chloride
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chemistry
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Naphthalenes
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chemistry
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Triazines
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chemistry
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Waste Disposal, Fluid
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methods
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Water Pollutants, Chemical
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isolation & purification
8.Improvement on the activity of microbial transglutaminase with Streptomyces hygroscopicus by the addition of surfactant CTAB.
Li CHENG ; Dong-Xu ZHANG ; Guo-Cheng DU ; Jian CHEN
Chinese Journal of Biotechnology 2007;23(3):497-501
Effect of CTAB addition on the accumulation of microbial transglutaminase (MTG) with Streptomyces hygroscopicus was investigated. The results showed that the addition of CTAB enhanced MTG accumulation, and the optimal addition time and concentration of CTAB were 32 h and 1%. The maximum MTG activity in the culture broth was 5.04 u/mL and increased by 21.8% compared with the control. With the addition of CTAB, pro-MTG was activated to become MTG. CTAB could enhance the production of pro-MTG by relieving the product inhibition, and the accumulation of MTG was improved.
Bacterial Proteins
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metabolism
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Cetrimonium Compounds
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pharmacology
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Dose-Response Relationship, Drug
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Electrophoresis, Polyacrylamide Gel
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Enzyme Activation
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drug effects
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Enzyme Precursors
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metabolism
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Industrial Microbiology
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methods
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Streptomyces
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drug effects
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enzymology
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metabolism
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Surface-Active Agents
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pharmacology
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Time Factors
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Transglutaminases
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metabolism
9.Residual activity of cetrimide and chlorhexidine on Enterococcus faecalis-infected root canals.
Carmen María FERRER-LUQUE ; María Teresa ARIAS-MOLIZ ; Matilde RUÍZ-LINARES ; ; Pilar BACA
International Journal of Oral Science 2014;6(1):46-49
Effective final irrigation regimen is an important step in order to achieve better disinfection and ensure residual antimicrobial effects after root canal preparation. The aim of this study was to compare the residual antimicrobial activity of 0.2% cetrimide, and 0.2% and 2% chlorhexidine in root canals infected with Enterococcus faecalis. Biofilms of E. faecalis were grown on uniradicular roots for 4 weeks. After root canal preparation, root canals were irrigated with 17% ethylenediaminetetraacetic acid (EDTA) to remove the smear layer. The roots were randomly divided into three experimental groups (n=26) according to the final irrigating solution: Group I, 5 mL 0.2% cetrimide; Group II, 5 mL 0.2% chlorhexidine; and Group III, 5 mL 2% chlorhexidine. Samples were collected for 50 days to denote the presence of bacterial growth. The proportion of ungrown specimens over 50 days was evaluated using the nonparametric Kaplan-Meier survival analysis. Differences among groups were tested using the log-rank test and the level of statistical significance was set at P<0.05. The highest survival value was found with 2% chlorhexidine, showing statistically significant differences from the other two groups. At 50 days, E. faecalis growth was detected in 69.23% specimens in Groups I and II, and in 34.61% specimens of Group III. There were no significant differences between 0.2% cetrimide and 0.2% chlorhexidine. Final irrigation with 2% chlorhexidine showed greater residual activity than 0.2% chlorhexidine and 0.2% cetrimide in root canals infected with E. faecalis.
Anti-Infective Agents, Local
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administration & dosage
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therapeutic use
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Bacterial Load
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drug effects
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Biofilms
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drug effects
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Cetrimonium Compounds
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therapeutic use
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Chlorhexidine
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administration & dosage
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therapeutic use
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Dental Pulp Cavity
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microbiology
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Edetic Acid
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therapeutic use
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Enterococcus faecalis
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drug effects
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Gram-Positive Bacterial Infections
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drug therapy
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Humans
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Microscopy, Electron, Scanning
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Root Canal Irrigants
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administration & dosage
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therapeutic use
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Root Canal Preparation
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methods
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Smear Layer
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Time Factors