Role of aminopeptidases in enkephalin catabolism: comparative study of the regional distribution of aminopeptidases and enkephalinase A in the rat brain

In order to elucidate the role of aminopeptidases in enkephalin catabolism in rat brain, the local distribution of two types of cerebral cellular membrane aminopeptidases (puromycin-sensitive and puromycin-insensitive ones) and of the enkephalin system marker, enkephalinase A, was studied. It was found that the distribution patterns of the former enzymes differ essentially from that of enkephalinase A. Study of coupling between the enzymatic activities in different regions of rat brain revealed a strong correlation between the activities of puromycin-insensitive aminopeptidase and enkephalinase A in midbrain (including hypothalamus). It was supposed that in midbrain the role of aminopeptidase M in intrasynaptic inactivation of enkephalins is much more conspicuous than in other regions of rat brain. The puromycin-sensitive aminopeptidase activity does not seem to play a role in enkephalin catabolism.     

Histamine and bradykinin stimulate the phosphoinositide turnover in human umbilical vein endothelial cells via different G-proteins

The G-proteins which regulate hormonal turnover of phosphoinositide (PI) in human umbilical vein endothelial cells have been investigated. A 40-41 kDa doublet present in the membranes of these cells was selectively ADP ribosylated by pertussis toxin (PTx), and this doublet was Gi alpha 2 and Gi alpha 3 according to immunoblotting with specific antisera. By contrast, a doublet of 24-26 kDa proteins in the same membrane preparations was ADP ribosylated by the C3 component of botulinum toxin (BoTx). PTx-dependent ADP ribosylation blocked stimulation of PI turnover by histamine, but did not affect stimulation by bradykinin, whereas BoTx (C2 + C3 components) had the opposite effect. Thus two different groups of G-proteins may be involved in hormone-dependent stimulation of PI turnover in human umbilical vein endothelial cells.     

The anticoagulant properties of the endothelium studied by the standard venous occlusion test]

Venous occlusion (VO) during which thrombin (Th) is postulated to be generated is routinely used for evaluation of fibrinolytic potential of endothelium (E). This study was performed to find out whether VO can also be used for assessment of anticoagulant function of E. VO was performed in 98 male patients (pts) with ischemic heart disease. Levels of protein C (PrC) which is related to Th binding by thrombomodulin and fibrinopeptide A (FpA)--a marker of presence of free Th--were determined together with some other factors of coagulation and fibrinolysis. Differences between pre- and postVO PrC levels fluctuated from -54.8% to +57.3%. According to reaction of PrC to VO pts were divided into 2 groups: 13 pts with increase or no change and 17 pts with decrease (consumption) of PrC. In pts without PrC consumption there was a significant increase in FpA. In pts with PrC consumption FpA was unchanged. In pts with PrC consumption exceeding its median value for this group (14%) PAI-1 antigen level fell significantly (-8.4 + 4%) during VO. Thus PrC consumption after VO indicates that TH is effectively removed from blood stream by endothelial factors. Absence of consumption of PrC is a sign of ineffective anticoagulant function of E. Increase in PrC level during VO in some pts may be due to its escape from tissue depot.     

Use of integrated phytoremediation for cleaning-up of oil-sludge-contaminated soil

The possibility of using multicomponent systems, including plants, mineral fertilizers, and plant growth promoting microorganisms, has been studied in vegetative experiments in order to stimulate phytoremediation of oil-sludge-contaminated soil. Winter rye (Secale cereale L.) was used as the principal phytoremediating plant species, whereas alfalfa (Medicago sativa L.), nitrogen fertilizer (ammonium nitrate), and a PGPR strain (Azospirillum brasilense SR80) were applied as additional components, individually or in various combinations. The obtained data revealed the critical importance of alfalfa for phytoremediation of hydrocarbon-contaminated soil. Application of different multicomponent treatments resulted in approximately 70% reduction of pollutant concentration in soil. The developed technological approaches were successfully tested in the remediation of an ex-oil-sludge pit on the ground of a petroleum refinery.     

Molecular Mechanisms of Disease-Causing Missense Mutations

Genetic variations resulting in a change of amino acid sequence can have a dramatic effect on stability, hydrogen bond network, conformational dynamics, activity and many other physiologically important properties of proteins. The substitutions of only one residue in a protein sequence, so-called missense mutations, can be related to many pathological conditions and may influence susceptibility to disease and drug treatment. The plausible effects of missense mutations range from affecting the macromolecular stability to perturbing macromolecular interactions and cellular localization. Here we review the individual cases and genome-wide studies that illustrate the association between missense mutations and diseases. In addition, we emphasize that the molecular mechanisms of effects of mutations should be revealed in order to understand the disease origin. Finally, we report the current state-of-the-art methodologies that predict the effects of mutations on protein stability, the hydrogen bond network, pH dependence, conformational dynamics and protein function.     

Pressure algometry and enzyme-linked immunosorbent assay of natural antibodies in the evaluation of pain syndrome

The dynamics of possible markers of pain syndrome: pain pressure thresholds of intolerance (PTI) and natural antibodies to mediators of pain processing (nAbs) in blood serum at dorsalgia was studied. We have shown that most men and women had reduced PTIs. Night PTIs were lower than daytime PTIs regardless of the gender. The study of the content of nAbs to opioids and biogenic amines by ELISA have shown a prolonged maintenance of their elevated and high levels that could evoke long-term effect in pain chronization. Thus, the pressure algometry and ELISA of nAbs to pain processing mediators make it possible to assess the individual pain status objectively and, on this bases, to propose personal schemes of treatment.