Effects of Different Preservation Methods on Activity of Recombinant E. coli

[ Objective] To explore different preservation methods of recombinant E. coli and find out the optimal conditions for preservation. [ Method] The recombinant E. coli DH5cx transformed pcDNA.3 were respectively preserved at 4℃ and -70 ℃, and the activity was determined after dif- ferent time. [ Result] The number of living E. coll with high dilutions preserved at 4 ℃ was gradually increased within the first 7 d, peaked on Day 7, and then gradually decreased. The number of living E. coli, which were preserved in 8% glycerol at -70℃ when OD800 at 0.8, were significantly higher than that of other groups after different preservation time. [ Conclusion] The optimal storage time was 7 d for recombinant E. coli at 4 ℃. For preservation at -70 ℃, the bacteria should be in logarithmic growth phase and preserved in 8% glycerol.

A multi-criteria decision-making approach for comparing sample preservation and DNA extraction methods from swine feces

Molecular microbiological methods, such as competetive PCR, real-time PCR, denaturing gradient gel electrophoresis (DGGE) and large-scale parallel-pyrosequencing, require the extraction of sufficient quantity of high quality DNA from microbiologically and chemically complex matrices. Due to difficulties in the field to standardize/select the optimum DNA preservation-extraction methods in view of laboratories differences, this article attempts to present a straight-forward mathematical framework for comparing some of the most commonly used methods. To this end, as a case study, the problem of selecting an optimum sample preservation-DNA extraction strategy for obtaining total bacterial DNA from swine feces was considered. Two sample preservation methods (liquid nitrogen and RNAlater?) and seven extraction techniques were paired and compared under six quantitative DNA analysis criteria: yield of extraction, purity of extracted DNA (A260/280 and A 260/230 ratios), duration of extraction, degradation degree of DNA, and cost. From a practical point of view, it is unlikely that a single sample preservation-DNA extraction strategy can be optimum for all selected criteria. Hence, a systematic multi-criteria decision-making (MCDM) approach was used to compare the methods. As a result, the ZR Fecal DNA MiniPrepTM DNA extraction kit for samples preserved either with liquid nitrogen or RNAlater? were identified as potential optimum solutions for obtaining total bacterial DNA from swine feces. Considering the need for practicality for in situ applications, we would recommend liquid nitrogen as sample preservation method, along with the ZR Fecal DNA MiniPrepTM kit. Total bacterial DNA obtained by this strategy can be suitable for downstream PCR-based DNA analyses of swine feces.

Effects of Different Preservation Methods on Vitamin C and Nitrate Content in Vegetables

This paper studied and compared the effects of three preservation methods( normal temperature 20℃,fresh-keeping box cold storage 4℃,and fresh-keeping box + fresh-keeping film cold storage 4℃) on six kinds of vegetables. The results showed that the content of vitamin C in the six vegetables was higher in cold storage than in normal temperature storage; the nitrate content was lower in cold storage than in normal temperature storage. In both the normal temperature storage and cold storage,the vitamin C in vegetables declined with the increase of the storage time,while the nitrate content increased with the increase of the storage time. Thus,cold storage has better fresh-keeping effect for vegetables. In cold storage conditions,the vitamin C content was higher in vegetables packaged by fresh-keeping box,and the nitrate content was lower,followed by the fresh-keeping film + fresh-keeping box cold storage. In conclusion,the best preservation method for vegetable is fresh-keeping box cold storage.

Different Preservation Methods for Long Term Maintenance of <i>Haemophilus influenzae</i>

Haemophilus species are Gram-negative coccobacilli that require factor X and factor V for growth. Beyond this, it is a finicky bacterium to culture, and any modification of culture procedures greatly reduces isolation rates. Poor quality of laboratories in developing countries results in its poor isolation rates. This study was done with the objective of finding out the optimal cultural environment and media so that it could be maintained for a longer period in economical settings like ours which was done using H. influenzae ATCC 49,766. In this study, several culture media were tested as a means to preserve H. influenzae ATCC like TSB + glycerol + sheep blood, BHI broth, BHI broth + glycerol, BHI broth+ glycerol + sheep blood, Chocolate agar slant and satellitism plate. Three sets of respective media were inoculated with 18 - 24 hours growth of H. influenzae. They were incubated at 37?oC 48 hours in a candle extinction jar. The media were checked for growth by subculturing them on chocolate agar plates and identified by biochemical reactions. Each set was maintained at 2 oC - 8?oC, -20?oC and at room temperature and checked for the viability 24 hourly by subculturing them on chocolate agar. Results showed best growth of H. influenza on chocolate agar slants for 15 - 20 days, followed by BHI + glycerol + sheep blood broth and satellitism plate for 4 - 6 days followed by BHI broth for 2 - 4 days. There was no growth in TSB + glycerol + sheep blood broth and BHI + glycerol broth media. Present study showed similar results as done by NS Srikanth et al. 2003 with growth on chocolate agar & satellitism plate for 3 - 5 days but no growth in TSB + Glycerol + Sheep blood broth media. Chocolate agar slant is by far the most long term preserving media for H. influenzae. However, growth on BHI broth with various modifications is also showed a good preservation for 3 - 5 days, so with further experiments we can hope to maintain the organism in these media also.

Health Preservation Methods Of Tibetan Medicine

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Simultaneous Preservation of Soil Structural Properties and Phospholipid Profiles: A Comparison of Three Drying Techniques

There is a need to simultaneously preserve evidence of interactions between the biological community and soil structural properties of a soil in as near an intact （natural） state as possible. Three dehydration techniques were implemented and assessed for their ability to minimise disruption of both biological and physical properties of the same arable soil sample. Dehydration techniques applied until samples were at constant weight were i） air-drying at 20℃ （AD）; ii） -80 ℃ freeze for 24 h, followed by freeze-drying （-80FD）; and iii） liquid nitrogen snap freeze, followed by freeze-drying （LNPD） and were compared to a moist control. Physical structure was determined and quantified in three dimensions using X-ray computed tomography and microbial phenotypic community composition was assessed using phospholipid fatty acid （PLPA） profiling. This study confirms that any form of dehydration, when preparing soil for simultaneous biological and physical analysis, will alter the soil physical properties, and cause some change in apparent community structure. Freeze-drying （both the LNFD and -80FD treatments） was found to minimise disruption （when compared to the moist control soil） to both the soil physical properties and the community structure and is a preferable technique to air-drying which markedly alters the size and character of the pore network, as well as the phenotypic profile. The LNFD was the preferred treatment over the -80FD treatment as samples show low variability between replicates and a fast turn-around time between samples. Therefore snap freezing in liquid nitrogen, followed by freeze drying is the most appropriate form of dehydration when two sets of data, both physical and biological, need to be preserved simultaneously from a soil core.

Optimal Sample Preservation and Analysis of Cr(VI) in Drinking Water Samples by High Resolution Ion Chromatography Followed by Post Column Reaction and UV/Vis Detection

A recent study by the Environmental Working Group reported the detection of hexavalent chromium (Cr(VI)) in tap water at 31 out of 35 states investigated in the United States. Even though Cr(III) is an essential element for human diet, Cr(VI) is a potential carcinogen. Previous work has clearly identified a linear trend of increasing risk of lung cancer mortality with increasing cumulative exposure to water soluble Cr(VI). Regardless, Cr(VI) is still not regulated or monitored in drinking water in the U.S. There is an existing method (EPA 218.6) for the analysis of Cr(VI), however, this analytical method does not addresses detailed sample preservation techniques and optimization process to achieve lowest detection limit possible. In this study, five buffer solutions with pH of 9 and above were compared to determine the most suitable buffer to preserve Cr(VI) in drinking water samples for an extended period of time. Results showed that the five buffers responded very differently to Cr(VI)-fortified drinking water. The best preserving reagent was found to be Ammonium Hydroxide + Ammonium Sulfate (pH 9.2) and Sodium Carbonate + Sodium Bicarbonate+ Ammonium Sulfate (pH 9.7), whereas a buffer solution with Sodium Hydroxide + Sodium Carbonate (pH 11.5+) resulted in a poor chromatographic resolution. A controlled study with a fortified Cr(III) at 1 ppb was also conducted to ensure no false positive detection of Cr(VI) due to the potential oxidation of Cr(III) during sample storage. The optimal preserving reagent identified from this study was compatible with the existing EPA method 218.6 using ion chroma-tography followed by post column reaction, with a method quantitation limit of 0.020 ppb and matrix spike recovery of ± 10%.

Preservation Property and Decay Kinetic of Polyurethane Immobilized Nitrifying Bacteria Pellets

The preservation methods of polyurethane immobilized nitrifying bacteria pellets which had been enriched in laboratory were provided. Factors such as temperature, pH and light, which affect the nitrification activity of polyurethane immobilized pellets, were investigated. The result showed that dark, deionized water and low temperature is suitable for polyurethane immobilized nitrifying bacteria pellets’ long term preservation.

Physicochemical properties and biological activities of polysaccharides from the peel of Dioscorea opposita Thunb.extracted by four different methods

Polysaccharides are the important biologically active components found in the peel of Dioscorea opposita Thunb.(DTTP).The influences of 4 extraction methods,namely hot water extraction(W),acidic extraction(HA),hot-compressed water extraction(HCW)and enzyme-assisted extraction(EAE),on the yields,physicochemical properties,hypoglycemic and antioxidant activities of polysaccharides from DTTP were studied and compared.Among these DTTP polysaccharides,DTTP-HA was outstanding in several respects.DTTP-HA was the most water soluble;it had the highest total carbohydrate content(85.08%),the highest uronic acid(13.20%)and the highest thermal stability.DTTP-HA and DTTP-W possessed a triple-helix structure.These 4 kinds of polysaccharides have the same types of monosaccharides,but in different molar percentages.Extraction method had a significant impact on the microstructures of the extracted polysaccharides.DTT-HA exhibited irregular structure with many holes.Among the 4 extracted methods,the DTTP-HA and DTTP-W initially exhibited higher hypoglycemic and antioxidant activities.The better bioactivities of DTTP-HA may be related to the above factors.The findings indicated that acid extraction is an effective method to extract polysaccharides with high biological activities from DTTP.

Preparation methods,biological activities,and potential applications of marine algae oligosaccharides:a review

Marine algae are valuable sources of health-promoting molecules that have been consumed by Asians for decades.Among aquatic flora,marine algae stand out in terms of high content of marine algae polysaccharides(MAP)such as carrageenan,alginate,fucoidan,laminaran,agarose,rhamnan,and ulvan.When hydrolyzed,MAP generate marine algae oligosaccharides(MAO),which have attracted interest in recent years due to their superior solubility compared with MAP.Besides,MAO have been demonstrated numerous biological activities including antioxidant,antidiabetic,anti-inflammatory,antimicrobial,and prebiotic activities.Thus,this review summarizes the main chemical classes of MAO,their sources,and the main processes used for their production(i.e.,physical,chemical,and biological methods),coupled with a discussion of the advantages and disadvantages of these methods.Highlights of the biological activities of MAO and their potential applications in food,nutraceutical,and pharmaceuticals would also be discussed and summarized.

Solution Remapping Method with Lower Bound Preservation for Navier-Stokes Equations in Aerodynamic Shape Optimization

It is found that the solution remapping technique proposed in[Numer.Math.Theor.Meth.Appl.,2020,13(4)]and[J.Sci.Comput.,2021,87(3):1-26]does not work out for the Navier-Stokes equations with a high Reynolds number.The shape deformations usually reach several boundary layer mesh sizes for viscous flow,which far exceed one-layer mesh that the original method can tolerate.The direct application to Navier-Stokes equations can result in the unphysical pressures in remapped solutions,even though the conservative variables are within the reasonable range.In this work,a new solution remapping technique with lower bound preservation is proposed to construct initial values for the new shapes,and the global minimum density and pressure of the current shape which serve as lower bounds of the corresponding variables are used to constrain the remapped solutions.The solution distribution provided by the present method is proven to be acceptable as an initial value for the new shape.Several numerical experiments show that the present technique can substantially accelerate the flow convergence for large deformation problemswith 70%-80%CPU time reduction in the viscous airfoil drag minimization.

Preservation and Regeneration of Beijing's Courtyard Houses Based on Typology Methods

Courtyard house is one of the symbols of Beijing's traditional culture.However,illegal self-constructions,over population and the phenomenon of multiple families crowding in one courtyard house have seriously damaged the traditional courtyard houses and urban texture of Beijing.This article insists that due attention should be paid to the traditional cultures of the original residents and their actual requirements in the preservation and regeneration of the courtyard houses.It proposes a new prototype of multi-family courtyard house which might be a feasible mode for the preservation and regeneration of Beijing's traditional courtyard houses.In addition,based on the typology methods,the article analyzes the evolution of multi-family courtyard houses through investigating the layout of courtyards,and brings forward an idea for its preservation and regeneration.Finally,taking some specific areas of Hutongs and courtyard houses as examples,it also presents a simulation model to validate the feasibility of this mode.

Establishment of Molecular Biological Method for Identification of Bacteria by 16S rDNA and gyrB Gene

[Objectives]The paper was to establish a molecular biological method for identification of bacterial strains.[Methods]The thalli of standard bacterial strains existing in the laboratory were collected and genomic DNA was extracted for amplification of 16S rDNA and gyrB gene.The 16S rDNA and gyrB gene sequences were obtained after sequencing.Sequences were aligned and analyzed via EzBioCloud and NCBI database,and phylogenetic trees were constructed to determine the species relationship of strains.Meantime,they were compared with known strains.[Results]This method could identify 5 standard strains accurately to the species level.The 16S rDNA and gyrB gene sequences were aligned and analyzed in EzBioCloud database and NCBI database.The strain with the max score was consistent with the known strain.And the query cover and ident were both above 99%.[Conclusions]The established molecular biological method for identification of bacterial strains by 16S rDNA and gyrB gene has good accuracy,which effectively solves the problem that the laboratory identification of bacteria relies on traditional methods and the accuracy can not be guaranteed,and further improves the identification ability of laboratory bacterial strains.

Integrated Use of Alternative Methods for Evaluating the Skin Sensitization Potencies of Five Frequently Used Preservatives in Cosmetics

In vitro skin sensitization testing methods based on the adverse outcome pathway(AOP)were used to evaluate the skin sensitization potencies of 5 commonly used preservatives.According to the“2 out of 3”principle of the integrated approaches to testing and assessment(IATA)the direct peptide reactivity assay(DPRA)and the human cell line activation test(h-CLAT)were used to detect the preservatives commonly used in cosmetics,including phenoxyethanol.methyl paraben,propyl paraben,imidazolidinyl urea and DMDM hydantoin.The DPRA and the h-CLA were carried out according to the OEC442C and 442E guidelines,respectively.The results show that.phenoxyethanol and methyl paraben are both negative in DPRA and h-CLAT while imidazolidinyl urea and DMDM hydantoin are both positive in these two tests.Propyl paraben has negative result in DPRA but positive result in h-CLAT.Therefore,imidazolidiny urea and DMDM hydantoin are sensitizers,while phenoxyethanol and methylparaben are non-sensitizers.Taken animal and human data into consideration,it is predicted that propyl paraben should be a non-sensitizer.The combination of DPRA and h-CLAT can make up for the limitations of using a single method,and it is suitable for the preliminary screening of cosmetic raw materials according to skin sensitization.

Analytical methods for determination of terbinafine hydrochloride in pharmaceuticals and biological materials

Terbinafine is a new powerful antifungal agent indicated for both oral and topical treatment of myco- sessince. It is highly effective in the treatment of determatomycoses. The chemical and pharmaceutical analysis of the drug requires effective analytical methods for quality control and pharmacodynamic and pharmacokinetic studies. Ever since it was introduced as an effective antifungal agent, many methods have been developed and validated for its assay in pharmaceuticals and biological materials. This article reviews the various methods reported during the last 25 years.

Effects of Different Application Methods and Application Rates of Biological Nano-Selenium on the Quality of Passion Fruit

[Objectives]To study the effects of different application methods and application rates of biological nano-selenium on the quality of passion fruit.[Methods]In this experiment,passion fruit Tainong 1 was sprayed with biological nano-selenium.The concentration of spraying refers to the recommended application rate of biological nano-selenium manufacturers.Three treatments with different concentrations were carried out:3,4.5 and 6 L/ha biological nano-selenium was sprayed on 100%root,(50%root+50%leaf surface)and 100%leaf surface of passion fruit,respectively.[Results]From the aspects of economic cost and rational utilization of resources,spraying 3 L/ha selenium fertilizer on the leaves of passion fruit played the best role in improving the quality of passion fruit.[Conclusions]To study the selenium-rich production technology of agricultural products and improve the quality of agricultural products is of great significance for improving human health and producing high-quality agricultural products.

Estimation of the Biological Methods of Assessing Soil N-Supplying Capacity in Calcareous Soil

Although many biological methods are used to determine soil nitrogen supplying capacity, there are certain differences in the results for different types of soils and various ways of measurement due to the complexity of soil N conformation, the high variance of soil and microorganism, and the difference of environment. Therefore, it is not clear about which biologic incubation method is better for calcareous soil. In this study, pot experiments were performed by using 25 different calcareous surface soil samples on the Loess Plateau and taking the N uptake of wheat and corn with leaching soil initial nitrate and without leaching in pot experiments as the control to investigate the difference of eight biological incubation methods for reflecting soil nitrogen supply capacity. The eight biological methods are waterlogged incubation, aerobic incubation for 2 weeks and for 4 weeks, dry-wet alternation aerobic incubation for 2 weeks, long-term alternate leaching aerobic incubation （and N mineralization potential, No）, short-term leaching aerobic incubation, microbial biomass carbon （Bc）, and microbial biomass nitrogen （BN） method, respectively. Among these methods, the dry-wet alternation aerobic incubation and aerobic incubation for 4 weeks were the modification of the method of aerobic incubation for 2 weeks according to the actual farmland moisture. The results showed that the correlation coefficients between these methods and crop uptake N with leaching soil initial nitrate were 0.530, 0.700, 0.777, 0.768, 0.764 （and 0.790, No）, 0.650, 0.555, and 0.465, respectively （r0.05 = 0.369, r0.0l = 0.505）. While without leaching soil initial nitrate, their coefficients were 0.351, 0.963, 0.962, 0.959, 0.825 （and0.812, No）, 0.963, 0.289, and 0.095, respectively （r0.05 = 0.369, r0.01 = 0.505）. In conclusion, excluding the soil initial nitrate, the correlation coefficients between the eight methods and crop uptake N were, from high to low, N0, aerobic incubation for 4 weeks, dry-wet alternation aerobic incubation for 2 weeks, and long-term alternate leaching aerobic incubation, while including the soil initial nitrate the correlation coefficients between them increased significantly and the values were all beyond 0.950 for these four methods, including aerobic incubation for 2 weeks and for 4 weeks, dry-wet alternation aerobic incubation for 2 weeks and short-term leaching aerobic incubation. The waterlogged incubation method, Bc and BN in the calcareous soil, had lower correlation coefficient with crop uptake nitrogen compared with other methods. Thus, dry-wet alternation aerobic incubation for 2 weeks was a better index for evaluating calcareous soil N supply capacity due to some other methods having disadvantages and not suitable for the actual farmland characteristics.

Prevention and Control Effects of New Biological Formulations Sprayed by Different Methods on Proceras venosatus (Walker) and Tryporyza intacta Snellen in Middle and Late Stages

To screen new biological formulations, accurate and efficient application technology, field test was conducted with 72% Bacillus thuringiensis(Bt)·monosultap WP, 8% lambda-cyhalothrin·emamectin benzoate SA and 3.6% lambda-cyhalothrin·Bt SA through manual spraying and unmanned aerial vehicle spraying. Test results and comprehensive evaluation analysis demonstrated that 72% Bt·monosultap WP and 8%lambda-cyhalothrin·emamectin benzoate SA by manual spraying and unmanned aerial vehicle spraying had good prevention and control effect on strains and internodes damaged by Tryporyza intacta Snellen in middle and late stage, which were ideal new biological formulations with high effi-ciency and low risk for prevention and control of T. intacta in middle and late stage, and could be popularized in sugarcane area. 72% Bt·monosultap WP at the dose of 3 000 g/hectare and 8% lambda-cyhalothrin·emamectin benzoate SA at the dose of 750 mL/hectare could be sprayed in mid September at the peak occurrence period of the forth and fifth generations of T. intacta and Proceras venosatus. Agents were diluted with 900 kg water per hectare, and manually sprayed with electric knapsack sprayer; or agents were diluted with 15 kg of Haoyang aerial control special addi-tives and water per hectare, and sprayed with unmanned aerial vehicle. The control effect against borer-damaged strain rate was above 81.3% and that against borer-damaged internode rate was above 88.6%.

Clinical Value of Molecular Biological Methods in Respiratory Tuberculosis in Children

The pattern of clinical forms of respiratory tuberculosis in children shows a preponderance of intrathoracic lymph node tuberculosis （89.4%） that is characterized by a complicated process in every third child under present-day conditions. Positive result of PCR closely correlates with the severity and extent of the specific process in children. Real-time PCR （RT-PCR） was ascertain to exhibit the highest sensitivity in detecting Mycobacterium tuberculosis DNA in children with primary generalized tuberculosis （62.5%） and in those with a disseminated specific process （55.6%）, which was much higher than conventional bacteriological study of diagnostic materials. By taking into account the findings, the RT-PCR detection of M. tuberculosis was considered as a substantial criterion for evaluating the magnitude of specific changes and the degree of tuberculosis infection activity in children.