A cautionary note on the use of stable transformed cells

Gene transfection and ectopic expression is a widely used method in experimental biology. In the present report, we would like to point out that this approach may, in certain circumstances, lead to a modification of the transfected cell phenotype. Indeed, we observed that after transfection of bcl-2 gene in the neuronal PC12 cell line some of the selected clones have lost their neuronal and catecholaminergic characteristics, i.e. TH expression and ability to grow neurites in response to NGF. Thus, the resistance of some PC12-Bcl-2 clones against neurotoxic insults may not necessarily reflect the potential benefit afforded by Bcl-2 expression. We therefore encouraged authors to verify cell phenotype after stable transfection to avoid misinterpretation of their results.     

A note on the use of multiple linear regression in molecular ecology

Multiple linear regression analyses (also often referred to as generalized linear models – GLMs, or generalized linear mixed models – GLMMs) are widely used in the analysis of data in molecular ecology, often to assess the relative effects of genetic characteristics on individual fitness or traits, or how environmental characteristics influence patterns of genetic differentiation. However, the coefficients resulting from multiple regression analyses are sometimes misinterpreted, which can lead to incorrect interpretations and conclusions within individual studies, and can propagate to wider‐spread errors in the general understanding of a topic. The primary issue revolves around the interpretation of coefficients for independent variables when interaction terms are also included in the analyses. In this scenario, the coefficients associated with each independent variable are often interpreted as the independent effect of each predictor variable on the predicted variable. However, this interpretation is incorrect. The correct interpretation is that these coefficients represent the effect of each predictor variable on the predicted variable when all other predictor variables are zero. This difference may sound subtle, but the ramifications cannot be overstated. Here, my goals are to raise awareness of this issue, to demonstrate and emphasize the problems that can result and to provide alternative approaches for obtaining the desired information.     

A note on the use of a pocket-calculator to work out MPN-values

The possibilities of computing MPN-values were evaluated; and a program was developed to run on a small programmable pocket-calculator. The mean introduction time of the data amounts to 9.4 s and the time needed to key in a significant number and to calculate the MPN-value can be estimated by the equation: Time (s) = (32/REF) + 92.64. Introducing a REF-value of one results in a time of two minutes, which is too long to drop the use of tabulated values. The agreement between calculated and tabulated figures is good.     

A note on the use of the Catalasemetre in assessing the quality of milk

The Catalasemetre, for assessing the quality of raw and pasteurized milk, has been studied. No correlation was found between catalase activity and bacterial counts for farm bulk tank milks within the range 5.2 ± 10²-5.4 ± 10⁵ cfu/ml. Similarly, no relation was observed between catalase activity and somatic cell counts of milk (range of counts from 0.08 to 3.5). However, the catalase activity and bacterial count of pasteurized milks which had been pre-incubated at 21.C for 25 h in the presence of crystal violet-penicillin-nisin to inhibit Gram-positive bacterial growth were significantly related. Thus, the use of this pre-incubation procedure coupled with the Catalasemetre to estimate bacterial growth, has potential in assessing the keeping quality of pasteurized milk samples within 25.5 h of production. Results on the thermostability of native milk catalase are also presented.     

A cautionary note on the use of certain restriction endonucleases with methylated substrates

Methylation of adenine and cytosine residues in DNA isolated from common strains of Escherichia coli K-12 can render that DNA resistant to cleavage by certain restriction endonucleases at those sites at which the recognition sequence for such an endonuclease overlaps (but does not include) a sequence recognized by methylases specified by the dam or dcm gene.     

A note on the use of dinitrosalicylic acid for determining the products of enzymatic reactions

Summary Factors contributing to non-linearity of enzyme assays incorporating detection of reaction products using dinitrosalicylic acid (DNS) are discussed. The common practice of diluting reaction products before quantification of reducing compounds is shown to be one cause of non-linearity. Insufficiency of substrate is also an important contributory factor in most modern versions of the method, although the original procedure was correctly designed to ensure a linear reaction. The inherent inaccuracy involved in expression of the results of non-linear reactions in units implying linearity (katals or International Units) is emphasized.     

A note on the use of metal species in microbiological tests involving growth media

The feasibility of using traditional growth media for biological testing of metal species, for example as potential microbiocides, was investigated. Significant interactions between both of the representative metal species studied, Cu²⁺ and FeEDTA, and the test media were found. It is recommended that the use of growth media for tests on metal species should be avoided.