Transformation of group A streptococci by electroporation

Abstract The introduction, via electroporation, of free plasmid DNA into three strains of Streptococcus pyogenes is described. The method is very simple and rapid and efficiencies vary from 1 × 10³ to 4 × 10⁴ per μg of DNA. The method was also used to introduce an integrative plasmid and transformants were obtained, albeit at a somewhat lower frequency (2 × 10²). Some of the plasmids used in this study are derivatives of the Lactococcus lactis subsp. cremoris Wg2 plasmid pWV01. These broad host range vectors replicate in Gram-positives as well as Gram-negatives (viz. Escherichia coli ). Here we show that they also replicate in S. pyogenes and S. sanguis .     

Selective N-heterylazimine inhibition of reactions catalyzed by rat liver glutathione transferase]

Three reactions (nucleophile substitution, thiolysis and N-deoxygenation) catalyzed by rat liver glutathione transferase have been studied using several N-heterylazimine inhibitors. The inhibitors are sharply different in their effectiveness in the transferase reactions. Their efficiency depends on their structure. The mechanism which underlies the found regularities is suggested.     

Caudate-cortical connections in the cat brain

By means of optical and electron microscopic methods, the cortical fields from the ipsilateral hemisphere was analysed in the cat after electrocoagulation of the dorsal part in the nucleus caudatus (NC). Degenerating axonal preterminals and terminals were detected in the preparations impregnated after Nauta--Gygax and Wiitanen's methods and in electronograms. To exclude degeneration of cortical and projective thalamic fibrillae from the great number of regenerated conductors, additional operations were performed on the same cats--thalamic and ventral NC nuclei were damaged and coagulative electrode was inserted into the dorsal NC a month before the last operation.     

Thermal stability of rhodopsins and opsins in warm- and cold-blooded vertebrates

Thermal stability of rhodopsins and opsins has been studied in endothermic (sheep, cattle, pig, rat) and ectothermic (frog) animals under two different conditions -- in the intact photoreceptor membranes (PM) and after substitution of the lipid surrounding of rhodopsins by molecules of a detergent Triton X-100. Lipid composition of PM in these animals was also studied, as well as the effect of proteases (pronase and papaine) upon thermal stability of rhodopsins in PM and in 1% Triton X-100 solutions. The thermal resistance of rhodopsins in PM was found to vary in the animals used to a great extent. The maximal differences in thermal stability of rhodopsins in ecto- and endothermic animals were due to the properties of photoreceptor protein itself, whereas in ectothermic animals they resulted mainly from differences in the lipid composition of PM. PM of endothermic animals differ from those of ectothermic ones by a lower content of polyenoic fatty acids and by a higher amount of phosphatidyl ethanolamine. The thermal stability of rhodopsins is not due to rhodopsin molecule as a whole, and depends mainly on its part which is directly bound to 11-cis retinal, located in hydrophobic region of PM and inaccessible to protease attack.     

Motor alimentary conditioned reflexes and delayed reactions in dogs following destruction of the caudate nucleus]

The characteristics of formation and achievement of motor conditioned reactions were studied in seventeen dogs before and after the lesion of head or body of the caudate nucleus. It has been shown that the degree of higher nervous activity disturbances depends on the localization of injuries in the nucleus, and on the complexity of the tests used. The caudate nucleus exerts a modulating effect on the parameters of motor conditioned reflexes and takes a direct part in the structure of delayed reflexes. Lesion of the caudate nucleus body is attended with more serious disturbances of higher nervous activity than lesion of its head.     

Relation of the individual specificity of emotional autoregulation controlled by EEG feedback to the influence of a genetic factor as revealed by a twin study

On the model of biofeedback connection by the given parameters of the EEG alpha-rhythm the dynamics of bioelectrical and vegetative characteristics and correlative connections between them were compared in the pairs of mono- and dizygotic twins. More likeness in the dynamics of the studied parameters was found in the pairs of genetically identical monozygotic twins. There was a maximum increase of the objective similarity characteristics in the moment of activity realization, judging by the change of alpha-rhythm parameters. The obtained results testify to genetic determination of individual specific styles in organization of complex integrative processes of the higher nervous activity.     

What Is Wrong with Enterococcal Probiotics?

Probiotics and Antimicrobial Proteins - Enterococci, being the common part of indigenous human microbiota and present in numerous fermented food products and probiotics, are often positioned as...     

Use of Corynebacterium pseudodiphtheriticum for elimination of Staphylococcus aureus from the nasal cavity in volunteers exposed to abnormal microclimate and altered gaseous environment

Effectiveness of saline suspension of Corynebacterium pseudodiphtheriticum containing spray was assessed in a 30-days chamber experiment modeling the effects of hyperthermia and polluted air on humans. Spray was targeted at eliminating Staphylococcus aureus from the nasal cavity of human subjects. Three of four volunteers became S. aureus carriers in the course of the experiment, and one was a chronic carrier of methicillin-resistant S. aureus (MRSA). Spray application eradicated S. aureus in three subjects and reduced its presence in the MRSA carrier. Results of the study suggest that C. pseudodiphtheriticum can be used for control of S. aureus in the nasal environment. However, further investigations are required.     

Real-time measurements of dark substrate catalysis

We have developed a novel procedure to monitor the real-time cleavage of natural unmodified peptides (dark substrates). In the competition-based assay, the initial cleavage rate of a fluorogenic peptide substrate is measured in the presence of a second substrate that is not required to exhibit any optical property change upon cleavage. Using a unique experimental design and steady-state enzyme kinetics for a two-substrate system, we were able to determine both Km and k(cat) values for cleavage of the dark substrate. The method was applied to HIV-1 protease and to the V82F/I84V drug resistant mutant enzyme. Using two different substrates, we showed that the kinetic parameters derived from the competition assay are in good agreement with those determined independently using standard direct assay. This method can be applied to other enzyme systems as long as they have one substrate for which catalysis can be conveniently monitored in real time.