Construction of a hybrid col E1 plasmid carrying the gene for bacteriophage lambda repressor.

A biologically active hybrid DNA molecule was constructed from plasmid Col E1 and the Eco R1 fragment of lambda DNA containing the gene for lambda repressor. The presence of this gene in the hybrid molecule was demonstrated genetically. The hybrid plasmid contains two closely located targets for restriction endonuclease Hind 111 in the integrated fragment. Thus, the plasmid may be used as a vector not only for Eco R1 fragments but also for Hind 111 fragments.     

Prevalence of widespread <Emphasis Type="Italic">BRCA1</Emphasis> gene mutations in patients with familial breast cancer from St. Petersburg

Ten variants different from the canonical nucleotide sequence (GenBank, U14680) has been identified when studying the mutation spectrum in gene BRCA1. Six of them (5382insC, 2963del10, 3819del5, 3875del4, 2274insA, and R1203X) cause premature termination of protein synthesis, thus predisposing to breast cancer. A missense mutation E1250K is presumed to be a factor of predisposition to cancer. We classified three variants of nucleotide sequence found in a number patients as DNA polymorphisms S694S, L771L, and E1038G. The 5382insC and 3819del5 mutations have been recorded in four and two families, respectively. Five of the mutations detected have not been found in Russia before. However, all mutations except for 2963del10 have been found in other populations of the world, which indicates their long evolutionary history. Two mutations found in patients from St. Petersburg (5382insC and 3875del4) have also been found in oncological patients from other regions of the Russian Federation.Translated from Genetika, Vol. 41, No. 3, 2005, pp. 405–410.Original Russian Text Copyright © 2005 by Grudinina, Golubkov, Tikhomirova, Brezhneva, Hanson, Vasilyev, Mandelshtam.     

Mitochondria-targeted plastoquinone derivatives as tools to interrupt execution of the aging program. 4. Age-related eye disease. SkQ1 returns vision to blind animals

Mitochondria-targeted cationic plastoquinone derivative SkQ1 (10-(6′-plastoquinonyl) decyltriphenylphosphonium) has been investigated as a potential tool for treating a number of ROS-related ocular diseases. In OXYS rats suffering from a ROS-induced progeria, very small amounts of SkQ1 (50 nmol/kg per day) added to food were found to prevent development of age_induced cataract and retinopathies of the eye, lipid peroxidation and protein carbonylation in skeletal muscles, as well as a decrease in bone mineralization. Instillation of drops of 250 nM SkQ1 reversed cataract and retinopathies in 3-12-month-old (but not in 24-month-old) OXYS rats. In rabbits, experimental uveitis and glaucoma were induced by immunization with arrestin and injections of hydroxypropyl methyl cellulose to the eye anterior sector, respectively. Uveitis was found to be prevented or reversed by instillation of 250 nM SkQ1 drops (four drops per day). Development of glaucoma was retarded by drops of 5 μM SkQ1 (one drop daily). SkQ1 was tested in veterinarian practice. A totally of 271 animals (dogs, cats, and horses) suffering from retinopathies, uveitis, conjunctivitis, and cornea diseases were treated with drops of 250 nM SkQ1. In 242 cases, positive therapeutic effect was obvious. Among animals suffering from retinopathies, 89 were blind. In 67 cases, vision returned after SkQ1 treatment. In ex vivo studies of cultivated posterior retina sector, it was found that 20 nM SkQ1 strongly decreased macrophagal transformation of the retinal pigmented epithelial cells, an effect which might explain some of the above SkQ1 activities. It is concluded that low concentrations of SkQ1 are promising in treating retinopathies, cataract, uveitis, glaucoma, and some other ocular diseases. Electronic Supplementary Material  Supplementary material is available for this article at and is accessible for authorized users. Published in Russian in Biokhimiya, 2008, Vol. 73, No. 12, pp. 1641–1654.     

Partial molar volumes and adiabatic compressibilities of unfolded protein states

We determined the partial molar volumes, V degrees , and adiabatic compressibilities, K degrees (S), of N-acetyl amino acids with neutralized carboxyl termini, N-acetyl amino acid amides, and N-acetyl amino acid methylamides between 18 and 55 degrees C. The individual compounds in the three classes have been selected so as to collectively cover the 20 naturally occurring amino acid side chains. We interpret our experimental results in terms of the volumetric contributions and hydration properties of individual amino acid side chains and their constituent atomic groups. We also conducted pH-dependent densimetric and acoustic measurements to determine changes in volume and compressibility accompanying protonation of the aspartic acid, glutamic acid, histidine, lysine, and arginine side chains. We use our resulting data to develop an additive scheme for calculating the partial molar (specific) volume and adiabatic compressibility of fully extended polypeptide chains as a function of pH and temperature. We discuss the differences and similarities between our proposed scheme and the reported additive approaches. We compare our calculated volumetric characteristics of the fully extended conformations of apocytochrome c and apomyoglobin with the experimental values measured in water (for apocytochrome c) or acidic pH (for apomyoglobin). At these respective experimental conditions, the two proteins are unfolded. However, the comparison between the calculated and experimental volumetric characteristics suggests that neither apocytochrome c nor apomyoglobin are fully unfolded and retain a sizeable core of solvent-inaccessible groups.     

Effect of photosynthetically active radiation, salinization, and type of nitrogen nutrition on growth of Salicornia europaea plants

Effects of various combinations of nutrient solution salinity (0.3, 171, and 342 mM NaCl), photosynthetically active radiation (PAR) of 600 or 1150 μmol/(m² s), and type of nitrogen nutrition (amide-N or nitrate-N) on the productivity and the content of accumulated mineral nutrients and free amino acids were studied in Salicornia europaea plants. At PAR of 600 μmol/(m² s), plant productivity increased with elevation of salinity level; at 1150 μmol/(m² s), the maximum productivity was observed in the plants grown at 171 mM of NaCl. The content of free amino acids in shoots, regardless of PAR, decreased with growing salinity level, whereas Na content, on the opposite, increased. Glutamic acid, rather than proline, was shown to be the main organic osmolyte in this plant species. Comparison of the productivity of plants grown on solutions with amide (urea) or nitrate nitrogen showed that higher biomass accumulation was achieved in the former case.     

Correction of immunity disorders, treatment of intestinal infections and disbiosis in children (clinical review)

Acute viral and bacterial intestinal infections in children provoke the Tn2 immune response, resulting in development of severe and complicated forms of the disease and sustained by the disbiotic disturbances due to unnecessarily prolonged use of antibacterial drugs. Cycloferon, an early inductor of interferon-1 and -2, was shown to be safe and efficient in the complex therapy of the intestinal infections. It promoted generation of the Th2 immune response and decrease of the repeated isolation of the pathogen with normalization of the disease clinical signs. The host intestinal microflora was normalized and the level of the opportunistic organisms decreased.     

见血封喉乳汁脂溶性成分及其抗氧化活性研究

首次采用气相色谱.质谱联用技术(GC-MS)对见血封喉(Antiaris toxicaria(Pers.)Lesch.)乳汁的脂溶性成分进行了分析,共鉴定了27个化学成分,占其总量的91.7%.用清除DPPH自由基能力的方法测定了见血封喉乳汁脂溶性部位的抗氧化活性,结果显示出一定的抗氧化活性,SC50值为500μg ml-1.     

Methemoglobin Reductase of Bacteria and Bacteroids <Emphasis Type="Italic">Bradyrhizobium lupini</Emphasis>: Purification and Properties

Reductase capable of reducing hemoglobin-like proteins was isolated from nodule bacteria Bradyrhizobium lupini and bacteroids of lupine root nodules. It is similar in some properties to many known methemoglobin reductases reducing animal and plant hemoglobins. It is a NADH-dependent FAD-containing flavoprotein with molecular weight of 87 kDa without metals. The presence of such enzymes in prokaryotes could be an explanation for the physiological activity of both bacterial and eukaryotic hemoglobins expressed in bacterial cells.     

Conformational “Fingerprint” of the Angiotensin-Converting Enzyme

The angiotensin-converting enzyme (ACE) is a zinc-dependent metalloproteinase widely occurring in the organism; it metabolizes many peptides and plays a key role in blood pressure regulation and vascular remodeling. This enzyme is expressed as a type-1 membrane glycoprotein on the surface of endothelial and epithelial cells, but is also found in a soluble form in biological fluids. In this study, we used purified ACE from lungs, which is mainly produced by endothelial cells of lung capillaries; ACE from heart, produced by endothelial heart cells and, probably, by myofibroblasts; and ACE from seminal fluid, produced by the epithelial cells of the prostate and epididymis. The pattern of binding of a set of 17 mAbs to different conformational epitopes on the surface of two domains of the human ACE significantly differed for ACEs from different organs. This pattern (the conformational “fingerprint” of ACE) reflects the local conformation of the surface of a particular ACE. The differences in the conformational fingerprints of ACEs expressed by different cell types, or even by similar cells but in different organs, can be explained by the posttranslational modification of ACE protein in these organs and, primarily, different glycosylation of N-glycosylation sites Asn25, Asn117, Asn289, Asn666, Asn685, and Asn731. The mass spectrometry of tryptic hydrolyzates of ACEs isolated from different human organs made it possible to reveal, in the composition of different ACEs, N-glycosylation sites that are really occupied by glycans, namely, Asn in positions 82, 117, 416, 648, 666, 685, and 731 in ACE from seminal fluid; Asn in positions 117, 648, 666, and 685 in ACE from lungs; and Asn in positions 117, 480, 666, and 685 in ACE from heart. Differences in the plausible structures of glycans in ACE, in particular, at the Asn666 N-glycosylation site were demonstrated, which can explain the differences in the efficiency of binding of mAbs to ACE from different organs.     

A study of the functional activity of macrophages of peritoneal exudate of mice exposed to low-intensity laser radiation in vitro and in vivo

The effect of low-intensity laser radiation generated by semiconductor devices in the red (650 nm) and infrared (850 nm) regions of the spectrum in vitro and in vivo on the phagocytic activity and synthesis of proinflammatory cytokines by peritoneal macrophages during the phagocytosis of bacterial cells has been studied. A culture of the clinical strain of the enteropathogenic bacterium Escherichia coli was used as an object. The radiation dose was varied by changing the power and duration of exposure. The results obtained indicate that infrared low-intensity laser radiation has a stimulating effect on the phagocytic activity of macrophages. It was shown that the effect of low-intensity laser radiation on the activity of the phagocytic process, the enhancement of the adhesion of bacteria by macrophages, killing of bacteria, and the production of proinflammatory cytokines is dose-dependent. The exposure to the rays of the red region of the spectrum on phagocytizing macrophages induced a decrease in their activity; as the dose was increased, the destruction of cells was registered.