Lipid peroxidation cannot be used as a universal criterion of oxidative stress

Oxidative stress is a term used to denote the imbalance between the concentrations of reactive oxygen and nitrogen species and the defense mechanisms of the body. Although it is generally accepted that such an imbalance plays a pivotal role in many pathologies, the term "oxidative stress" remains ill defined. In an attempt to evaluate the relationship between various assays of oxidative stress, we have analyzed the correlations between the results reported in those publications in which "oxidative stress" has been assayed by at least two methods. We found good correlations between the concentrations of several peroxidation products, including malondialdehyde, F2-Isoprostanes, lipid hydroperoxides, conjugated dienes, glutathione and protein carbonyls, but not with other criteria of "individual oxidative status" such as the concentration of antioxidants and products of DNA fragmentation (the "comet" assay). In light of these findings, we divide the assays used for evaluation of "oxidative stress" into the following three categories: (i) assays based on measuring the concentrations of oxidation products of lipids, proteins and DNA, as well as the concentrations of antioxidants, (ii) assays used to evaluate the oxidative and reductive capacity of biological fluids and (iii) assays used to evaluate the ex vivo susceptibility of lipids to oxidation upon their exposure to a source of free radicals. Our analyses demonstrate that oxidative stress cannot be defined in universal terms. Two results are of special interest:1.the commonly used criteria based on lipid peroxidation can not be regarded as a general estimate of the individual "oxidative status".2.the levels of antioxidants exhibit a non-monotonic relation with other criteria for oxidative stress. Further research is required to evaluate the significance of the latter finding.     

The influence of halogenorganic compounds on radiation induced damage of rat eye lens components

In experiments with white rats the damages of eye lens components of the animals are investigated under combined action of halogenorganic compounds (chloroform and sodium trichloroacetate) and radiation (137Cs) in vivo. It was shown that the injection of these compounds into animals amplifies the radiation-induced oxidation of eye lens components. The effect is manifested in the increase in the content of chromophors with fluorescence in the visible range and products of lipid peroxidation, the decrease in the amount of reduced glutathione. The data support the increased risk of development of radiation-induced cataract in the conditions of tehnogenic pollution of the environment.     

Peroxidation of liposomal lipids

Free radicals, formed via different mechanisms, induce peroxidation of membrane lipids. This process is of great importance because it modifies the physical properties of the membranes, including its permeability to different solutes and the packing of lipids and proteins in the membranes, which in turn, influences the membranes’ function. Accordingly, much research effort has been devoted to the understanding of the factors that govern peroxidation, including the composition and properties of the membranes and the inducer of peroxidation. In view of the complexity of biological membranes, much work was devoted to the latter issues in simplified model systems, mostly lipid vesicles (liposomes). Although peroxidation in model membranes may be very different from peroxidation in biological membranes, the results obtained in model membranes may be used to advance our understanding of issues that cannot be studied in biological membranes. Nonetheless, in spite of the relative simplicity of peroxidation of liposomal lipids, these reactions are still quite complex because they depend in a complex fashion on both the inducer of peroxidation and the composition and physical properties of the liposomes. This complexity is the most likely cause of the apparent contradictions of literature results. The main conclusion of this review is that most, if not all, of the published results (sometimes apparently contradictory) on the peroxidation of liposomal lipids can be understood on the basis of the physico-chemical properties of the liposomes. Specifically: (1) The kinetics of peroxidation induced by an “external” generator of free radicals (e.g. AAPH) is governed by the balance between the effects of membrane properties on the rate constants of propagation (k _p) and termination (k _t) of the free radical peroxidation in the relevant membrane domains, i.e. in those domains in which the oxidizable lipids reside. Both these rate constants depend similarly on the packing of lipids in the bilayer, but influence the overall rate in opposite directions. (2) Peroxidation induced by transition metal ions depends on additional factors, including the binding of metal ions to the lipid–water interface and the formation of a metal ions-hydroperoxide complex at the surface. (3) Reducing agents, commonly regarded as “antioxidants”, may either promote or inhibit peroxidation, depending on the membrane composition, the inducer of oxidation and the membrane/water partitioning. All the published data can be explained in terms of these (quite complex) generalizations. More detailed analysis requires additional experimental investigations. Dedicated to Prof. K. Arnold on the occasion of his 65th birthday.     

Value of a newly sequenced bacterial genome

Next-generation sequencing(NGS) technologies have made high-throughput sequencing available to medium- and small-size laboratories, culminating in a tidal wave of genomic information. The quantity of sequenced bacterial genomes has not only brought excitement to the field of genomics but also heightened expectations that NGS would boost antibacterial discovery and vaccine development. Although many possible drug and vaccine targets have been discovered, the success rate of genome-based analysis has remained below expectations. Furthermore, NGS has had consequences for genome quality, resulting in an exponential increase in draft(partial data) genome deposits in public databases. If no further interests are expressed for a particular bacterial genome, it is more likely that the sequencing of its genome will be limited to a draft stage, and the painstaking tasks of completing the sequencing of its genome and annotation will not be undertaken. It is important to know what is lost when we settle for a draft genome and to determine the "scientific value" of a newly sequenced genome. This review addresses the expected impact of newly sequenced genomes on antibacterial discovery and vaccinology. Also, it discusses the factors that could be leading to the increase in the number of draft deposits and the consequent loss of relevant biological information.     

Analysis of silver binding nucleolar organizer regions in exfoliative cytology smears of potentially malignant and malignant oral lesions

Nucleolar organizer regions are nucleolar components that contain proteins that are stained selectively by silver methods; they can be identified as black dots throughout the nucleolus and are known as silver binding nucleolar organizer regions (AgNOR). The number of AgNOR is related to the cell cycle and the proliferative activity of the cells. We investigated AgNOR using exfoliative cytology smears of potentially malignant oral lesions. Eighty individuals were divided into four equal groups: healthy controls, oral leukoplakia, oral submucous fibrosis and oral squamous cell carcinoma. The mean number of AgNOR in each study group gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. The proliferative index was increased in the oral premalignant and malignant patients compared to normal subjects. The mean AgNOR size gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. Spherical shaped AgNOR were most common in controls, whereas large, clustered and kidney shapes were most common in oral squamous cell carcinoma. Multiparameter analysis of AgNOR in oral exfoliative smears is a simple, sensitive and cost-effective method for differentiating premalignant from malignant lesions and can be used in conjunction with routine cytomorphological evaluation.     

A CD8+ T cell heptaepitope minigene vaccine induces protective immunity against Chlamydia pneumoniae

An intact T cell compartment and IFN-gamma signaling are required for protective immunity against Chlamydia. In the mouse model of Chlamydia pneumoniae (Cpn) infection, this immunity is critically dependent on CD8(+) T cells. Recently we reported that Cpn-infected mice generate an MHC class I-restricted CD8(+) Tc1 response against various Cpn Ags, and that CD8(+) CTL to multiple epitopes inhibit Cpn growth in vitro. Here, we engineered a DNA minigene encoding seven H-2(b)-restricted Cpn CTL epitopes, the universal pan-DR epitope Th epitope, and an endoplasmic reticulum-translocating signal sequence. Immunization of C57BL/6 mice with this construct primed IFN-gamma-producing CD8(+) CTL against all seven CTL epitopes. CD8(+) T cell lines generated to minigene-encoded CTL epitopes secreted IFN-gamma and TNF-alpha and exhibited CTL activity upon recognition of Cpn-infected macrophages. Following intranasal challenge with Cpn, a 3.6 log reduction in mean lung bacterial numbers compared with control animals was obtained. Using a 20-fold increase in the Cpn challenging dose, minigene-vaccinated mice had a 60-fold reduction in lung bacterial loads, compared with controls. Immunization and challenge studies with beta(2)-microglobulin(-/-) mice indicated that the reduction of lung Cpn burdens was mediated by the MHC class I-dependent CD8(+) T cells to minigene-included Cpn CTL epitopes, rather than by pan-DR epitope-specific CD4(+) T cells. This constitutes the first demonstration of significant protection achieved by immunization with a CD8(+) T cell epitope-based DNA construct in a bacterial system and provides the basis for the optimal design of multicomponent anti-Cpn vaccines for humans.     

Ultrastructure of L-line lipoblast-like cells during cloning in a medium with a high serum concentration

The ultrastructure of lipoblast-like cells produced by clones of transformed culture of strain in the medium with an increased (60%) concentration of bovine serum was studied. The aim of using the stimulator rich in adipogenous factors was the elucidation of the origin of lipid accumulation in some of cell types during heterogeneous differentiation of clones. In these conditions non-differentiated lipoblasts assumed the fine structure of mature lipid cells. The statement of the marker significance of lipid accumulation in non-differentiated cells of the clones and the conversion of these cells under the influence of non-specific stimulator to the mature lipocytes confirms the data on conservation of over-all cytogenetic potentials and the capacity of their realization in the cells of transformed cultures. It is suggested that the increased concentration of bovine serum in the cultural medium not only stimulates differentiation (by any way determined cells), but is also capable of acting on the stem elements produced by the clone. In that case the influence of the stimulator on the trends of heterogeneous differentiation is not unlikely.