A simple titration method for determining Chlamydia infectivity using an image analyzing system

In the biological study of Chlamydia, it is very important to determine the infectivity titer of the organism. For many years researchers used the serial dilution method to determine this titer. This method consists of diluting the material to be examined, inoculating suitable dilutions into susceptible cell cultures and cultivating them. The number of inclusions formed in host cells can be calculated with the naked eye under a microscope. The precision and accuracy of this method, however, depend on the number of inclusions per field and the number of fields counted. In this report, we present a simple and rapid method for counting a large number of inclusions using an image analysis system and an appropriate number of samples, and propose a sampling method based on a statistical analysis of the data obtained with 84 microscopic fields.     

A new simple method for determination of erythrocyte filtrability.

A new simple filtration technique designed for measuring red cell filtrability in the routine laboratory use was developed. The suspension of the whole blood in saline (1:20,000 dilution) was processed on the Sartorius filter membranes, pore size 8 micron. The percentage of passed erythrocytes indicating red cell filtrability was determined. The suitability and perspective applicability of this method for studying various hematological disorders is proposed.     

A simple spectroscopic method for studying erythrocyte ghost resealing.

A novel spectroscopic method is described for following the kinetics of resealing of hemolysed erythrocyte ghosts. The procedure is based on the broadening of the EPR spectrum of nitroxyl radicals by paramagnetic ions. The method is used to study the effect of Ca2+, Mg2+ and dimethonium ion on the kinetics of resealing.     

A simple and fast method for determining colony forming units

Aims: To develop a flexible and fast colony forming unit quantification method that can be operated in a standard microbiology laboratory. Methods and Results: A miniaturized plating method is reported where droplets of bacterial cultures are spotted on agar plates. Subsequently, minicolony spots are imaged with a digital camera and quantified using a dedicated plug‐in developed for the freeware program Image J. A comparison between conventional and minicolony plating of industrial micro‐organisms including lactic acid bacteria, Eschericha coli and Saccharomyces cerevisiae showed that there was no significant difference in the results obtained with the methods. Conclusions: The presented method allows downscaling of plating by 100‐fold, is flexible, easy‐to‐use and is more labour‐efficient and cost‐efficient than conventional plating methods. Significance and Impact of the Study: The method can be used for rapid assessment of viable counts of micro‐organisms similar to conventional plating using standard laboratory equipment. It is faster and cheaper than conventional plating methods.     

A simple method for determining specific esterase activity in tissue extracts.

A simple procedure for estimating specificB-naphthyl esterase enzyme content and activity in small amounts of crude tissue extract is described. Esterase activity is determined by quantitative densitometry of histochemically stained acrylamide gels. Activity measured this way is linear with the extract volume applied. Esterase content is determined by isotopic labeling with a stoichiometric covalently binding enzyme inhibitor, diisopropylfluorophosphate; followed by electrophoresis and gel slice counting.     

A simple method for determining the boundary layer resistance in leaf cuvettes

Abstract. A new method is described for determining the boundary layer resistance over wet filter paper exposed within a leaf cuvette, based on the energy balance of the filler paper. The boundary layer resistance is calculated by an iterative procedure from measurements of the relative humidity and temperature of the air in the cuvette. Comparisons between the new and the conventional method, involving measurement of the filter paper temperature, show close agreement.
To simplify the method further, a graph has been constructed for the relationship between boundary layer resistance and cuvette relative humidity at temperatures from 15 to 35°C, determined at one value of the ratio of the flow rate through the cuvette to the filter paper area.
An analysis of errors suggests that the new method is less sensitive than the conventional to errors in temperature and humidity.     

A simple matrix method for determining flux control coefficients in complex pathways

Flux control coefficients express in quantitative terms the extent to which the steady state flux through a metabolic pathway is controlled by a particular parameter. Enzyme flux control coefficients can be calculated using matrix algebra methods which express the control coefficients in terms of parameters which can be determined experimentally (enzyme elasticities, flux ratios, metabolite ratios). This paper describes an algorithm based on a 'constraint' matrix which enables expressions for enzyme control coefficients to be written for pathways of any complexity.     

Role of EDTA in a simple method for determining toxicity using a bacterial indicator organism

A method to determine toxicity using a bacterium as the indicator organism previously developed (Botsford 1998) perceives most divalent cations as being toxic. Mercury is perceived as the most toxic, followed by cadmium, zinc and copper. It was found that adding 2.5 m EDTA to the reaction would relieve the toxicity of the 15 divalent cations tested. This effect does not appear to be simple chelation. One micromolar EDTA eliminated the toxicity of 1.6 m calcium or 0.006 m mercury. Thirty-six chemicals were tested for their toxicity in the presence and absence of 2.5 m EDTA and 25 ppm calcium. Twenty-one were less toxic and two of these, p-aminobenzoic acid and tetrachloroethylene would no longer appear to be toxic according to the assay when these additions were present. Six chemicals had the same toxicity with and without the additions. Nine chemicals were more toxic when the EDTA and calcium were present. This experiment was repeated with six chemicals and ten times the EDTA concentration and ten times the calcium concentration. The toxicity with 10× was compared with the toxicity with 1× the additions. The toxicity of 4 of the six chemicals changed with the higher concentration of EDTA and calcium when the absorbancy values observed in samples with the lower levels were compared with samples with the higher levels. Obviously before EDTA can be added to mitigate the toxicity of divalent cations, it must be determined how much EDTA is required to eliminate the toxicity by the ions present in the sample. Alternatively, if the nature of the contaminating organic chemical is known, it can be determined what the effect of EDTA and the divalent cation present is on the apparent toxicity of the compound.     

A simple novel method for determining mortality rates in HIV treatment programs worldwide

In this Perspective, Gregory Bisson discusses a new article by Matthias Egger and colleagues that introduces a simple method for HIV treatment programs to more accurately estimate the proportion of patients dying in the first year after ART initiation.     

A simple method for quantifying high density antigens in erythrocyte membrane by flow cytometry

RBC flow cytometric analysis is usually used to quantify antigen content. Calibration systems enable antigen content determination by relating mean fluorescence intensity with the number of bound antibody molecules (equivalent to the number of antigen molecules). For that reason, antibodies must be used at saturating concentration, which may lead to agglutination when working with high density antigens. Then, forward scattering, side scattering and fluorescence will be increased, thus obtaining wrong results. In this work, the simple Langmuir adhesion model was applied. Flow cytometry was used to quantify GPA, a transmembrane protein present at high density on RBC. The fluorescence intensity of samples at different anti-GPA sub-saturating concentrations was measured. Sometimes, agglutinates were present and two peaks of fluorescence were observed, the principal one corresponding to isolated cells and the secondary one corresponding to agglutinated cells. In those cases, the principal peak was taken into account for the analysis. The GPA antigen content obtained for nine analyzed samples ranged from 3 to 13 x 10(5) sites per cell, which is similar to those values found in literature. Therefore, the Langmuir adsorption model enables us to determine the antigen content for the anti-GPA/GPA system on RBC membrane. This model could be used to quantify high density antigens in RBC and in other cells.     

A simple, rapid, and sensitive method of determining CSF IgG, using latex

A simple, rapid and sensitive method of determining cerebrospinal fluid IgG is presented. The procedure depends upon the fact that spinal fluid gamma globulin, and two of its components (IgG and IgM) will precipitate latex particles in proportion to their concentration, and that the optical density of the supernatant solution containing unprecipitated latex particles after centrifugation is inversely proportional to the concentration of these immunoglobulins. The method is sensitive to 50 nanograms, is inexpensive, requires 0.1 ml. or less of spinal fluid, and can be performed in minutes. Preliminary studies show that it compares favorably with results obtained by radial immunodiffusion.     

A simple method of determining zearalenone in cereals by liquid chromatography

Summary The paper described a simplified method for the separation, identification and determination of zearalenone by liquid chromatography in cereals. The cereal medium is dried, homogenised and extracted with ether. From the ether extract zearalenone is transferred into ethyl acetate, whereafter the sample is analysed using an external standard. The detection limit of the method is 500 g/kg. Recoveries of zearalenone varied from 94.5% to 96.2%.The conditions of the liquid chromatography are described.