Diversity of Harpin-Like Defense Peptides from Barnyard Grass (Echinochloa crusgalli L.) Seeds.

The results of the study of the structure and function of harpin-like peptides (alpha-harpinins) of the EcAMP group from the barnyard grass (E. crusgalli) seeds and the possibility of their involvement in the innate immunity to biotic stresses are presented.

[Differentiated approach to the treatment of large hernial defects of the anterior abdominal wall in children (clinical observations)]. / Lechenie obshirnykh gryzhevykh defektov perednei briushnoi stenki u detei.

AIM: To present an experience of treatment of children with large hernial defects of the anterior abdominal wall. MATERIAL AND METHODS: This article describes management of 43 children with large (n=31) and giant (n=12) ventral hernia who were at N.F. Filatov Children´s City Hospital #13 for the period 2005-2015. All patients underwent surgical correction using local tissues in 'edge to edge' fashion (n=23), aponeurosis dublication of 'coat floors' type (n=14) and biological implant 'Permacol' (n=6). Etiological causes of ventral hernias in children were comprehensively analyzed. Optimal time for surgical treatment and features of viscero-abdominal disproportion were discussed. CONCLUSION: Early restoration of anatomical and functional structures of the anterior abdominal wall is necessary for good cosmetic result and prevention of recurrent ventral hernia. We presented three clinical examples of commonly used surgical procedures in children including anterior abdominal wall repair using biological material 'Permacol'.

[Expression of plant antimicrobial peptide pro-SmAMP2 gene increases resistance of transgenic potato plants to Alternaria and Fusarium pathogens].

The chickweed (Stellaria media L.) pro-SmAMP2 gene encodes the hevein-like peptides that have in vitro antimicrobial activity against certain harmful microorganisms. These peptides play an important role in protecting the chickweed plants from infection, and the pro-SmAMP2 gene was previously used to protect transgenic tobacco and Arabidopsis plants from phytopathogens. In this study, the pro-SmAMP2 gene under control of viral CaMV35S promoter or under control of its own pro-SmAMP2 promoter was transformed into cultivated potato plants of two cultivars, differing in the resistance to Alternaria: Yubiley Zhukova (resistant) and Skoroplodny (susceptible). With the help of quantitative real-time PCR, it was demonstrated that transgenic potato plants expressed the pro-SmAMP2 gene under control of both promoters at the level comparable to or exceeding the level of the potato actin gene. Assessment of the immune status of the transformants demonstrated that expression of antimicrobial peptide pro-SmAMP2 gene was able to increase the resistance to a complex of Alternaria sp. and Fusarium sp. phytopathogens only in potato plants of the Yubiley Zhukova cultivar. The possible role of the pro-SmAMP2 products in protecting potatoes from Alternaria sp. and Fusarium sp. is discussed.

[Laparoscopic operations for chronic duodenal obstruction in children].

Brief literature review devoted to chronic duodenal obstruction is presented in the article. We described an experience of treatment of 5 children with mechanical form of chronic duodenal obstruction. Duodenal stenosis was observed in 2 patients. Arteriomesenteric compression caused chronic duodenal obstruction in other children. Duodenal obstruction was cured using laparoscopy in all cases. Thus enteroalimentation has been started in early terms and intensive care unit stay and hospital stay was reduced.

[Surgical treatment of Hirschprung's disease total form in children].

It was analyzed the treatment results of 24 patients under the age of 2 years with total aganglionosis for the period from 2000 to 2013. Each of these patients underwent several surgical interventions (on the average 7.8±2.1). All children were operated radically. It was performed ileojejunorectal anastomosis at transitional fold of peritoneum in patients with concomitant short bowel syndrome. Soave's operation was done in 14 patients including by using of laparoscopic technique in 3 cases. Inflammatory complications (paraproctitis) developed in 2 children on maceration background. Perirectal fistula was observed in 1 patient subsequently. In long-term period liquid stool incontinence persisted up to 6 months in 28% of children. In terms of more than 1 year stool incontinence was observed in 12.6% of patients. Increased growth of D-lactate-producing gram-positive anaerobes was revealed during microbiological investigation of feces in 35% of patients in long-term period. Increased growth was accompanied by systemic acidosis and infectious enteritis clinically. Course monthly treatment with antibacterial drugs (Alpha-Normix, Trichopolum, Gentamicin) was prescribed in patients with short bowel syndrome and high frequent of enteritis recurrence. Treatment was used In terms from 6 to 12 months of longer if it was necessary. It was concluded that timely diagnosis and right tactic in neonatal period leads to decrease significantly the number of complications, vain interventions and to improve treatment outcomes. Our experience shows that the best results of surgical treatment were observed in children aged from 2.5-3 years when a child can control urination and defecation. The most mistakes are caused by wrong or failed morphological investigation of large bowel.

3-O-methoxyimino group inhibits interactions between progestins and blood transcortin.

The interactions between E- and Z-isomers of 3-O-methoxyimino-pregn-4-ene-20-one and its 17α-hydroxy derivative and transcortin from human blood were investigated. The substitution of the progesterone 3-oxo group for a 3-O-methoxyimino group was shown to diminish the affinity of the steroid for transcortin by approximately one order of magnitude irrespective of the substituent's orientation. The data suggests that progesterone derivatives substituted thereby must have higher bioavailability compared to progesterone and must not significantly affect the biodynamics of glucocorticoid in vivo.

Changes in levels of gene expression in human aortal intima during atherogenesis.

Changes in the contents of 36 mRNAs species related to lipid turnover, inflammation, metabolism and the action of sex hormones in samples of aortal intima along the "intact tissue - lesions of type I - lesions of type II - lesions of type Va" sequence were analyzed using quantitative PCR. The expression of several mRNAs coding for components of the vesicular transfer and lipid turnover machinery was found to be resistant to atherogenesis or even decline in the course of atherogenesis. Decrease in expression was also recorded for steroid sulfatase, androgen receptor, and low density lipoprotein receptor mRNAs. However, the contents of the majority of other mRNA species increased gradually during disease progression. The earliest changes found as early as in lesions of type I were characteristic for estrogen sulfotransferase, apolipoprotein E, scavenger receptor SR-BI, collagen COL1A2, as well as chemokine CCL18 mRNAs. The contents of several mRNAs in intact tissue and atherosclerotic injuries had gender differences. Additionally, responses of two mRNAs, for aromatase and sterol regulatory element binding protein 2, to atherosclerotic lesion were also sex-differentiated. The contents of the majority of analyzed mRNAs in peripheral blood monocyte-derived macrophages were higher than in intact aorta. The correlations found in atherosclerotic lesions between mRNA species that predominant in macrophages and those expressed at comparable levels in macrophages and intact aorta or mainly in aorta suggest that the observed rise in the content of the majority of mRNAs during atherogenesis is determined by increase in expression in resident cells. The data suggest that the revealed absence of homeostatic regulation of expression of a number of genes associated with vesicular transfer and lipid turnover can serve as one of the reasons for lysosomal function insufficiency that leads to foam cell formation in atheroma. The observed sex differences in expression of a number of mRNAs suggest that estrogens in women perform their atheroprotective effects starting with predisposition to the disease and finishing with advanced stages of the pathologic process.

Approaches to the design of selective ligands for membrane progesterone receptor alpha.

A number of progesterone derivatives were assayed in terms of their affinity for recombinant human membrane progesterone receptor alpha (mPRα) in comparison with nuclear progesterone receptor (nPR). The 16α,17α-cycloalkane group diminished an affinity of steroids for mPRα without significant influence on affinity for nPR, thus rendering a prominent selectivity of ligands for nPR. On the contrary, substitution of methyl at C10 for ethyl or methoxy group moderately increased the affinity for mPRα and significantly lowered the affinity for nPR. A similar but even more prominent effect was observed upon substitution of the 3-oxo group for the 3-O-methoxyimino group. A significant preference towards mPRα was also rendered by the 17α-hydroxy group and additional C6-C7-double bond. The data suggest that the modes of ligand interaction with mPRα and nPR in the C3 region of the steroid molecule are different. One can speculate that combination of the above substitutions at C17, C10, C6, and C3 may give ligand(s) with high specificity towards mPRα over nPR.

Coordination in gene expression during atherogenesis.

General tendencies in the regulation of gene expression during atherogenesis were investigated using correlation analysis for 34 mRNA species of several functional groups. The contents of mRNA were measured by quantitative PCR in samples of human aortal intima containing no lesions or atherosclerotic lesions of types I (initial lesions), II (fatty streaks), and Va (fibroatheromas). The coupling between mRNA contents in lesions and the same mRNAs in intact tissue was found to descend in the course of the disease progression. The data are in accordance with the opinion that successive morphologic types of atherosclerotic lesions correspond to steps of atherogenesis. In addition, the contents of individual mRNA species could correlate with each other within the given sample type, the extent of this coupling rising along with the disease progression. The exception from this rule was a collapse in coupling for several functional groups of mRNA in lesions of type I. This collapse could indicate special position of these lesions in pathogenesis. Statistically significant correlations between mRNAs found in samples of all four types comprised in total about 50% of all possible correlations. 66% of these correlations were conservative, i.e. observed in at least two sample types. By coupling-strength, the studied mRNAs could be divided into four clusters whose composition significantly varied along with the disease progression. The disease progression was also associated with decline in number of regulatory factors that determine coordination in expression of the analyzed genes.

Effect of sex hormones on levels of mRNAs coding for proteins involved in lipid metabolism in macrophages.

The effects of sex hormones estradiol (E2), testosterone (Te), and 5α-dihydrotestosterone (DT) on cholesterol accumulation induced by modified low density lipoproteins (LDL) in macrophages differentiated from human peripheral blood monocytes and on the levels of mRNAs coding for proteins involved in lipid metabolism have been studied. All three hormones at physiological concentrations (1 nM) are capable of reducing cholesterol accumulation in cells. The treatment of cells with modified and native (not inducing cholesterol accumulation) LDL results in similar alterations in the expression of several mRNAs aimed primarily at homeostatic regulation of lipid metabolism. These alterations depend on the sex of macrophage donors and in some cases are even reversed in cells obtained from male and female donors. The cells not treated with modified LDL have no significant gender differences in the expression of the examined mRNAs. Hormones, either independently or in combination with the modified LDL, influence the levels of some mRNAs, and each hormone shows an individual range of effects. Correlation analysis of changes in mRNA content in the cells showed that the hormones may interfere with coordination of gene expression. Hormone action leads to: (1) reduced coupling of the content of individual mRNAs with their initial levels in the control cells; (2) reduced coupling of different mRNA levels; (3) regrouping of mRNAs between the clusters; and (4) changes in the number of factors that determine the correlation links between mRNAs. The data show that sex hormones may have impact on the level of expression of certain genes and, in particular, on the coordination of gene expression in macrophages.

[The magnetic foreign bodies of the gastrointestinal tract in children].

The authors own the experience of treatment of 23 children with magnetic foreign bodies (MFB) of the gastrointestinal tract. They share the possibilities of diagnostics and surgical treatment of children on the vivid clinical examples. By MFB in the stomach, which do not leave the gastrointestinal tract naturally, the use of fibroesophagoduodenoscopy is recommended. The laparoscopy is helpful in case of the uncomplicated MFB in the intestine. The X-ray control of the complete foreign bodies evacuation is necessary.

Contents of mRNAs encoding endosome/lysosome components in normal human aorta and in stage II of atherogenesis: a hidden regulation.

Contents of mRNAs encoding endosome/lysosome components EEA1, Rab5a, Lamp1, Lamp2, p62 (SQSTM1), and CD63 were measured by quantitative PCR and compared in intact fragments of human aorta and in aorta fragments with atherosclerotic lesions of stage II (fatty streaks) of the same donors. During atherogenesis an increase was detected only in the level of p62 mRNA but not in other mRNAs. Nevertheless, correlation analysis revealed a profound rearrangement of inter-gene correlations: only 30% of correlations found in the fatty streaks coincided with the correlations in normal fragments. Thus, new constellations were formed in fatty streaks concurrently with disappearance of correlations between mRNAs under study and mRNAs encoding factors of lipid accumulation, reverse cholesterol transfer, and some lipid sensors/transcription regulators of lipid metabolism.

Lipid signaling in the atherogenesis context.

Normally macrophages localized in the arterial vessel wall perform the "reverse transfer" of cholesterol, which includes endocytosis of low density lipoproteins (LDL), cholesterol transfer to newly formed high density lipoprotein particles, and their following elimination by the liver. The homeostatic function of macrophages for cholesterol involves a system of lipid sensors. Oxysterol sensors LXRs, oxysterol and cholesterol sensors INSIG and SCAP acting through controlled transcription factors SREBP, as well as sensors for oxidized fatty acids and their derivatives, PPAR, are the best studied. Activation of LXR and PPAR is also accompanied by inhibition of macrophage functions related to inflammation. Accumulation of oxidized and otherwise modified LDL in the subendothelial space induced by endothelium injury, infection, or other pathogenic factors instead of stimulation of the homeostatic functions of macrophages leads to their weakening with a concurrent increase in the inflammatory potential of these cells. These shifts seem to drive the transformation of macrophages into foam cells, which form the core of sclerotic plaques. The intervention of another lipid sensor, TLR4, can trigger such a radical change in the functional activity of macrophages. The interaction of modified LDL with this signaling receptor results in inhibition of the homeostatic oxysterol signaling, induction of additional LDL transporters, and activation of the phagocytic function of macrophages. The re-establishment of cholesterol homeostasis under these circumstances can be achieved by administration of LXR and PPARgamma agonists. Therefore, it is urgent to design ligands with reduced side effects.

[Isolation of the lipid-transporting protein Ns-LTP1 from seeds of the garden fennel flower (Nigella sativa)].

A novel lipid-transporting protein (Ns-LTP1) has been isolated from seeds of the garden fennel flower Nigella sativa. The molecular mass, N-terminal amino acid sequence, and amino acid composition of the protein have been determined. Ns-LTP1 has a molecular mass of 9602 Da and contains eight cysteine residues which form four disulfide bridges. The protein is capable of suppressing the development of some phytopathogenic fungi and oomycetes.

[Hormonal mechanisms of sex differentiation of the liver: the modern concepts and problems].

The level of thousands of genes expression in the liver is differentiated on the basis of sexual dimorphism that affects the frequency of appearance of different pathological forms. The main hormonal factors of the liver's sex differentiation are sex steroids and growth hormone. The impulsive and close to continuous secretion character of growth hormone in male and female individuals may have effects on masculinization or feminization processes, accordingly. The mechanism of decoding the growth hormone's secretion pattern by liver cells is not known. Some genes in the liver with the expression of sex differentiated genes, have so called memory of gender, which is created, probably, during early postnatal ontogenesis with involvement of both androgens and growth hormone. The physical transporter of this memory is not known. The possible molecular mechanisms of various effects based on sex differentiation in liver have been described in this survey, including unique cases of determining the role of the growth hormone's pattern and permissive function of the growth hormone concerning the direct effect of sex steroids to hepatocytes.

[The purification and characterization of a novel lipid transfer protein from caryopsis of barnyard grass (Echinochloa crusgalli)].

A novel lipid transfer protein called Ec-LTP was isolated from resting caryopsis of weed barnyard grass Echinochloa crusgalli (L.) Beauv.; its molecular weight, amino acid content and N-terminal amino acid sequence were determined. Ec-LTP was a 9150 Da protein, containing eight cysteine residues, which formed four disulfide bonds. The isolated protein could significantly inhibit the development of pathogenic fungi Phytophthora infestans and Helminthosporium sativum, causing the late blight of potato and tomato and the root rot of herbs, respectively.

[Video-assisted surgery of Hirschsprung's disease in children].

Results of the laparoscopic treatment of 36 patients with Hirschsprung's disease were analyzed. The majority of patients were the newborns or before 12 months old. The use of laparoscopy permitted a full-fledged topical diagnostics, aganglionosis zone identification, gut mobilization and voiding. Laparoscopic method also allowed to decrease the ICU stay in 3,1 times and start breastfeeding earlier, lessen the pelvic floor intraoperative traumatization and decrease the level of commissural and inflammatory complications. Good and excellent functional results were achieved in 93,7% of children, stenosis of the anastomotic area was registered postoperatively in 2 patients.

[A cause of the abnormal properties of melt water].

The physicochemical properties and crystal structure of melt water were studied. Melt water was found to be in the active metastable state with the non-equilibrium concentration of hydrogen and hydroxyl. The biological activity of such water is accounted for by its small sizes of supramolecular complexes. Films of structural formations were first obtained and their acoustic emission in the homogeneous medium recorded.

Formation and germination of Phytophthora infestans oospores in different regions of Russia.

On occurrence of oospores (sexual stage), 88 Phytophthora infestans populations were investigated. The populations were collected in 14 regions of Russia. In total, 3677 samples have been checked: 2888--from potato leaflets (55 populations), 344--from tomato leaflets (16 populations), 445--from tomato fruits (17 populations). The oospores have been detected both in potato and tomato leaflets and in fruits with different occurrence--4.4%, 11.9%, and 28.8%. Occurrence of oospores per sample was also maximal in tomato fruits. After overwintering a majority of oospores (50-90%) kept their viability. Sometimes, loss of viability of some oospores was caused by the external negative influence. About 25% of oospores formed at crossing between current Russian P. infestans strains were capable to germinate. Treatment with negative temperature (-12 degrees C) or Trichoderma suspension increased percentage of germinated oospores. In some field trials, the oospores caused affection of quite many of potato plants. However, in other trials only particular plants or no plants were affected. Tomato germlings were affected only under unfavourable conditions.

[Laparoscopic surgery and postoperative adhesions in children].

Data of 568 patients, aged from 3 weeks to 14 years, with acute adhesive bowel obstruction, treated during 1981--2007 yy. are analyzed. Most often previous laparotomy was performed on account of appendicitis, predominately complicated with peritonitis--in 52,2% of patients. Obstruction, caused by intestinal invagination was observed in 19,4%, malformation--9,3%, tumor--2,3%, other--1,7%. The treatment results of appendicular peritonitis and intestinal invagination are studied. Two comparative groups of patients with appendicular peritonitis, 100 children each, were treated laparoscopically and traditionally. The laparoscopic method allows to stop pain and endotoxicosis earlier, faster gastrointestinal function recovery, decrease of complication in the laparoscopical group rates and hospital-stay period on 30,8%. 100 children with intestinal invagination were operated laparoscopically and 50 patients were experienced a traditional <> operation. There were no complications, but 2 cases of postoperative adhesive obstruction in the <> group. The average hospital stay was 2,5 times shortened in the laparoscopical group.