[RNP-complex stoichiometry of Bombyx mori nuclear polyhedrosis virus polyhedra].

By gel-filtration through Sephacryl S-300 it was shown that RNP A complex present in polyhedra of Bombyx mori nuclear polyhedrosis virus has molecular weight (M(w)) about 700 kDa. It was shown that RNP A with M(w) 788 kDa is composed of two polyhedrin 13S-associates with M(w) 342 kDa, two p14 polypeptide with M(w) 14 kDa, two 21 kDa small non-coded RNAs and two 17 kDa small non-coded RNAs. The model of RNP A formation from components making it is proposed. The complex role in the course of polyhedron formation and its role in the course of infection are discussed.

[The search of microRNA genes encoded in antiparallel chains of Autographa californica nuclear polyhedrosis virus genome regions, including most late and complementary genes].

It was shown using bioinformatic approach by analysis of alternative transcriptes secondary structure, that A. californica nuclear polyhedrosis virus gene ph encoded two mature miRNAs and three potential miRNAs. Gene orf1629 complementary to gene ph did not encode miRNAs and pre-miRNA-Cs. Gene p10 encodes mature and potential miRNA. Gene p74 located on complementary chain encodes three mature miRNAs.

[The search of miRNA genes in Bombyx mori nuclear polyhedrosis virus genomes regions complementary to the latest genes].

The search of miRNA genes in Bombyx mori nuclear polyhedrosis virus genome region complementary to very late genes has been carried out. The search miRNA algorithm in silico was developed by us. It was shown that NPV B. mori genome region containing orf4 gene complementary to ph gene encodes the potential miRNA. NPV B. mori genome region containing p74 gene complementary to p10 gene encodes mature miRNA and potential miRNA. The genome region containing orf1629 encodes two small non-coding RNAs complementary to orf 5'-end of polyhedrin miRNA. From obtained results it is proposed that two small noncoding RNAs complementary to regions of polyhedrin miRNA are included in polyhedra.

[Small RNAs in Bombyx mori nuclear polyhedrosis virus polyhedra form complexes with polypeptide p14 and polyhedrin].

It was shown that two small RNAs about 65 and 55 nucleotides long included in NPV B. mori polyhedra form with polypeptides p29 and p14 specific RNP-complexes with molecular weights of 50 and 31 kDa, respectively. Both complexes form high-molecular weight complex with polyhedrin. Origin and nature of p29 and p14 polypeptides are discussed.

[Complete amino acid sequence of catalase from the fungus Penicillium vitale]. / Polnaia aminokislotnaia posledovatel'nost' katalazy griba Penicillium vitale.

The polypeptide sequence of the unmodified catalase from Penicillium vitale containing 696 amino acid residues was deduced. The sequences of 76 tryptic peptides of the unmodified catalase, 63 tryptic peptides of the catalase with modified Lys residues, 48 peptides resulting from catalase cleavage by the Staphylococcus aureus V8 protease, and 9 fragments obtained by BrCN-treatment were considered, and a comparison with the sequences of other catalases was made.

[Comparative study of collagen in hypertrophic and keloid cicatrix]. / Sravnitel'noe issledovanie kollagena gipertroficheskikh i keloidnykh rubtsov.

The relative percentage of type III to type I collagen, the ratio of alpha 1(I) to alpha 2(I) collagen chains and the ratio of dimers (beta-chains) to monomers (alpha-chains) in type I collagen have been measured in solubilized collagen fractions from keloids and hypertrophic scars aged about 1-7 years after burn. In tissue samples the content of crosslink with structure of pyridinoline were analyzed and expressed as mole of crosslink per mole of collagen. A comparison between hypertrophic and keloid scars has shown that the young (about 1 year) hypertrophic scars have higher ratios alpha 1(I)/alpha 2(I) and beta 11 + beta 12/alpha 1(I) + alpha 2(I). The increased proportion of beta-chains in the younger hypertrophic scars approximated to the same level as in normal skin within 7 years after lesion. There was no decrease in the ratio dimers/monomers of collagen type I with age of keloid scar tissue. The relative amount of collagen type III in young keloid scars was found decreased as compared to age-matched hypertrophic scars (13 +/- 3 and 17 +/- 7 respectively), but was similar in the older scars. The average pyridinoline crosslinks content per mole of collagen in keloids was 2 times as high as in hypertrophic scars. The data support the suggestion that general, as well as peculiar disturbances of collagen metabolism are involved in the development of keloids and hypertrophic scars.

[Metal-dependent collagenolytic activity in primary foci and metastatically involved lymph node in melanoma]. / Metallozavisimaia kollagenoliticheskaia aktivnost' v pervichnykh ochagakh i metastaticheski porazhennykh limfaticheskikh uzlakh melanom.

Collagenolytic activity was estimated in primary focuses of melanoma, in preparations of normal skin surrounding the tumor, in benign pigmentary neoplasms (nevus) as well as in normal and metastatic lymph nodes. Both metal-dependent and -independent collagenolytic activity was significantly increased in tumoral tissues as compared with that of healthy tissues. At the same time, the collagenolytic activity was considerably lower in benign pigmentary neoplasms than that of melanoma primary focuses. Importance of the collagenolytic enzymes in the processes of melanoma invasion and metastatic spreading is discussed.

The amino acid sequence determination of a granulin and polyhedrin from two baculoviruses infecting Agrotis segetum.

The amino acid sequence of Agrotis segetum Granulosis virus (AsGV) granulin and A. Segetum nuclear polyhedrosis virus (AsNPV) polyhedrin was determined by sequencing tryptic and chymotryptic peptides from reduced and carboxymethylated proteins and tryptic fragments of oxidized and maleylated granulin. The comparison of the established peptide structures with the primary structures of other occlusion body proteins from related baculoviruses was also used for the polypeptide chains' reconstruction. The polypeptide chains of AsGV granulin and AsNPV polyhedrin comprise 247 and 246 amino acid residues, respectively. The proteins possess a high percentage of homology in their primary structures (63%).

[Age changes in the level of pyridinoline cross-linking and ratio of soluble and insoluble collagen in human rib collagen]. / Vozrastnye izmeneniia soderzhaniia piridinolinovykh sshivok i otnosheniia rastvorimogo i nerastvorimogo kollagena v rebernom khriashche cheloveka.

The changes in the content of mature crosslinks with pyridinoline structure and soluble/insoluble collagen ratio in the costal cartilage tissue of human beings aged from 1 month to 57 years were found to be age-dependent. The effect of the pyridinoline crosslink content on the soluble/insoluble collagen ratio in human costal cartilage tissue may constitute no less than 67% of the total influence of the sum of all factors. The pronounced nonlinearity of the studied dependencies points to a possible involvement of a factor(s) other than the pyridinoline crosslink content.

[Comparative characteristics of oxyproline excretion in urine in children with post-burn hypertrophic and keloid scars]. / Sravnitel'naia kharakteristika ékskretsii oksiprolina s mochoi u detei s posleozhogovymi gipertroficheskimi i keloidnymi rubtsami.

Catabolism of collagen was studied in 25 children with keloid and hypertrophic scars by means of evaluation of total and polypeptide-bound hydroxyproline excreted with urine. Formation of postburn pathological scars led to an increase in excretion of collagen degradation products, the rate of the increase depended on area and age of scar. In hypertrophic cicatrization rate of collagen metabolites excretion correlated with the scar age reversely, demonstrating the tendency to decrease of the scar collagen metabolites with its ageing. Excretion of collagen metabolites with urine in patients with keloid scars depended on area of the scar and only slightly on its age. Besides, in patients with keloids rather high, statistically distinct correlation (r = 0.855) was found between the ratio macrophages/neutrophils under studies using "skin window" as well as between the ratio of hydroxyproline content in the polypeptide fractions with molecular mass above 1,500 and in the fraction containing oligopeptides and free hydroxyproline. This correlation was not found in patients with hypertrophic scars. Interaction of macrophages, neutrophils and, probably, eosinophiles appears to have a definite importance in regulation of collagen degradation, impairment of which may be substantial for scars formation.

[The connection between the decomposition of mature collagen and the level of natural cytotoxicity in patients with focal scleroderma]. / Sviaz' mezhdu raspadom zrelogo kollagena i urovnem estestvennoi tsitotoksichnosti u bol'nykh ochagovoi sklerodermiei.

Increased rate of pyridinoline excretion with urine was detected in patients with focal sclerodermia: the ratio pyridinoline/creatinine was equal to 0.094 +/- 0.049 as compared with that of 0.073 +/- 0.049 in controls. Concentration of mature collagen cross-links involving pyridinoline structure in urine of patients with focal sclerodermia did not depend on the patients age and the disease duration. However, this pattern correlated distinctly with clinical form of the disease (0.073 +/- 0.049 in local form and 0.013 +/- 0.017 in generalized form of sclerodermia) as well as correlated reversely with the rate of activity of natural killer cells (r = -0.977).

[Excretion of pyridinoline with urine in children with Ehlers-Danlos syndrome and Marfan's syndrome and the effect of beta blocker and complex vitamin therapy on it]. / Ekskretsiia piridinolina s mochoi u detei s sindromami Elersa-Danlo i Marfana i vliianie na nee terapii beta-adrenoblokatorom i kompleksom vitaminov.

Excretion of pyridinoline and polypeptide-bound hydroxyproline with urine was studied in 27 children with hereditary impairment of connective tissue. At the same time, effects of beta-adrenoblocking agents and vitamin complex, prescribed during preoperation period before thoracoplasty in hereditary chest deformation, were investigated. Clinical efficiency of the treatment depended distinctly on the initial value of ratios pyridinoline/creatinine and polypeptide-bound hydroxyproline/creatinine. Total positive effect of the therapeutic course, considering also echocardiographic examination and postoperational complications, was observed in 44% of patients with Ehlers-Dunlop syndrome and in 75% of patients with Marfan syndrome.

[Physiological changes in pyridinoline excretion in urine depending on age and sex]. / Fiziologicheskie izmeneniia ékskretsii piridinolina s mochoi v zavisimosti ot vozrasta i pola.

Natural physiological alterations in excretion of pyridoline (total hydroxypyridoline and lysyl pyridoline) with urine were studied in children and adults depending on age (1-86 years old) and sex. Age-dependent alterations of pyridoline excretion with urine had apparently four phases: an increase, a decrease, stabilization and secondary increase. Distinct dissimilarity between men and women in the rate of pyridoline excretion was found only in the age group of 16-21 years old.

Characterization of costal cartilage collagen in funnel chest.

Parallel measurements of pyridinoline content, the ratio of soluble:insoluble collagen, and the percentage of endogenous collagenolysis were made in samples of costal cartilage from forty-five children with funnel chest (FC) and from twenty-two control children. From an analysis of the influence of different factors, such as FC type (isolated or syndromic), a diagnosed syndrome, extent of FC depression (second or third), and age, two biochemical variants of FC-variants a and b, which were not related to the presence or absence of a known concurrent syndrome, were distinguished. These variants differed from each other in all the parameters under study. Variant a occurred about five times less frequently than b, and was characterized by a pyridinoline content of about 50% of that of b, an elevated soluble:insoluble collagen ratio, and an increased percentage of endogenous collagenolysis compared to controls. For variant b, the pyridinoline content fell within normal limits, but the soluble:insoluble collagen ratio, and the percentage of endogenous collagenolysis were below normal. The data suggest that the formation of variant a may be related to defect(s) in collagen crosslinking, whereas the formation of variant b may result from other unknown factor(s) involved in the formation and maturation of costal cartilage.

[Structural characteristics of collagens from the skin and rib cartilage of patients with Ehlers-Danlos syndrome type II]. / Strukturnye kharakteristiki kollagenov kozhi i rebernogo khriashcha bol'nykh s sindromom Elersa-Danlo II tipa]

Collagens were analyzed in skin and rib cartilage of 9 patients with Ehlers-Danlos syndrome of the II type. Electrophoresis and CNBr-peptide mapping showed that extended inserts and deletions as well as rough impairments of post-translation processing were not detected in collagens of the I, II and III types from these patients. In the patients with Ehlers-Danlos syndrome of the II type distinct increase was observed both in the total ratio of collagens III/I (P = 0.95) and in the ratio of intact collagens III/I free of cross-links. A decrease in content of dimers beta 11 and beta 12 was found in two patients. The data obtained suggest that the Ehlers-Danlos syndrome of the II type involved deteriorations in the structure of collagens I responsible for decrease in stability and sometimes for impairments in cross-link formation. Increase in content of collagen II fraction, predisposed to proteolytic hydrolysis of terminal sites, as well as elevated sensitivity of collagen II to pepsin hydrolysis were found in collagens of rib cartilage from patients with the syndrome and with funnel chest deformation. This suggests the lowered stability of collagen II from rib cartilage in funnel chest deformation.

[Changes in the structure of type I collagen and cross-links between type I and type III collagen chains in a patient with funnel chest]. / Narushenie struktury kollagena I tipa i nalichie sshivok mezhdu tsepiami kollagenov I i III tipa u bol'nogo s izolirovannoi formoi kilevidnoi deformatsii grudnoi kletki.

Skin and rib cartilage collagens were studied in patient 1.I.K. with isolated form of pigeon chest as well as in a group of children without any impairments of connective tissue. Distinct decrease in stability of collagen I, an increase in the ration of alpha 1 (I)/alpha 2(I) chains and impairment in formation of beta 12 dimers were detected in the patient with pigeon chest. In the patient skin total ratio between collagens I and III, calculated from a content of BrCN-peptides, was similar to normal level, whereas the proportion was markedly increased between intact molecules of collagens III and I free of cross-links, which was calculated from the ratio of alpha 1(III)/alpha 2(I) chains. Presence of cross-links between alpha 1 (III) and alpha 2 (I) chains as well as between alpha 1 (III) and alpha 2 (I) chains was detected after peptide mapping of polypeptides arranged in the region of beta 11 and beta 12 dimers. All the collagen I preparation, extracted from skin of the patient 1.I.K., contained molecules with unstabilized N-terminal sites. These results suggest that mutation occurred in the N-terminal region of alpha 1(I) chain. Analysis of collagen from the patient 1.I.K. rib cartilage demonstrated a slight decrease in total stability of collagen II as well as elevated concentration of collagen II molecules containing unstabilized N-terminals. Mechanisms responsible for formation of cross-links between polypeptide chains of collagens I and III detected in human skin are discussed.

[Determination of the amino acid composition of cartilage and creatine phosphokinase isoenzymes with the use of phenylthiocarbamyl amino acid derivatives]. / Opredelenie aminokislotnogo sostava khriashchevoi tkani i izofermentov kreatinfosfokinazy s ispol'zovaniem feniltiokarbamil'nykh proizvodnykh aminokislot.

A method for estimation of amino acid composition and collagen-specific amino acids was developed. The assay was based on the precolumn reaction of amino acids with phenylisothiocyanate. The phenylthiocarbamyl derivatives were analyzed by means of reverse-phase chromatography on octadecyl sorbents, which involved gradient elution with ethanol and sodium or ammonium acetate. The method was used for effective separation of collagen-specific and main amino acids in cartilage hydrolysate from healthy persons and of patients with funnel chest and Ehlers-Danlos syndrome and also for determination of amino acid composition in creatine phosphokinase B- and M-subunits.

[Heterogeneity of collagen molecules types I and II according to their resistance to proteolysis]. / Geterogennost' molekul kollagenov I i II tipov po ustoichivosti k proteolizu.

Study of the effects of pepsin treatment on soluble collagens type I of the skin and collagens type II of the costal cartilage of healthy subjects revealed the presence of two classes of molecules differing in the stability of their three-helical structure. In collagen molecules possessing a low stability (their number may amount to 20-30%) within the temperature range of 4-30 degrees C pepsin causes a split-off of N-terminal sites with the formation of short chains, i.e., alpha 1(I), alpha 2(II), and alpha 1(II), whereas at higher temperatures (33 degrees C for collagens type I and 37 degrees C for collagens type II) a complete degradation of these molecules takes place. It was found that collagens types I and II molecules contain a high number of three-helical sites with a high susceptibility to pepsin. The putative functional role of structural heterogeneity of collagen molecules is discussed.