[Reconstruction of total portosystemic shunt into selective portosystemic shunt in a child]. / Rekonstruktsiya total'nogo portosistemnogo shunta v selektivnyi portosistemnyi shunt u rebenka.

To date, side-to-side splenorenal shunt (SRS) and its analogues (splenosuprarenal shunts (SSRS)) are mainly used for portal hypertension. These are total portosystemic shunts characterized by total blood shunt from portal vein into inferior vena cava. The latter is fraught with a significant risk of complications such as pulmonary hypertension, decreased portal liver perfusion, liver failure and hepatic encephalopathy. Prevention of these complications is still an urgent problem in modern surgery. However, we proposed a new method of treatment, i.e. reconstruction of SRS and SSRS into selective shunt. This procedure was performed in 37 patients after 2020. We present laparoscopic reconstruction in an 11-year-old girl with portal hypertension and signs of hepatic encephalopathy identified after previous SSRS.

[Laparoscopic decompression of celiac trunk in children]. / Laparoskopicheskaia dekompressiia chrevnogo stvola pri kompressionnom stenoze u detei.

MATERIAL AND METHODS: For the period 2013-2016 four patients were treated at the Filatov Children's City Clinical Hospital #13. There were 2 children aged 14 years and 2 children aged 17 years. All patients have been diagnosed via anamnesis, complaints, pulse-wave doppler sonography, contrast-enhanced MDCT and angiography. After comprehensive examination 3 patients underwent laparoscopic decompression of celiac trunk. In all cases celiac trunk compression was predominantly caused by median arcuate ligament of the diaphragm combined with neurofibrotic tissue of celiac plexus. RESULTS: All patients were discharged after laparoscopic decompression of celiac trunk. Intra- and postoperative complications, as well as cases of conversion were absent. Mean time of surgery was 65 minutes. In all cases postoperative period was smooth (4 days on the average). Two patients underwent follow-up examination in long-term postoperative period: pulse-wave doppler sonography, contrast-enhanced MDCT and angiography. In both cases reduced severity, incidence and duration of pain syndrome were observed. CONCLUSION: Clinical examples show some problems in diagnosis and treatment of compressive stenosis of celiac trunk due to rarity of pathology especially in childhood. Nevertheless, combination of abdominal ischemia and celiac trunk stenosis confirmed by instrumental diagnosis is indication for surgical treatment.

[Dynamic transperineal ultrasonography as indication criteria for biofeedback therapy in children with dysfunctional voiding].

Disorders of pelvic organs' evacuation function, manifested by difficulty in urination and constipation, in conjunction with incontinence, and childhood stool smearing is an urgent medical and social problem. The study presents results of treatment of 36 children (mean age 7,2 +/- 2,3 years) with non-neurogenic variants of pelvic organ dysfunction. The choice of treatment was based on an attempt to form managed urination. Indications to treatment were defined both by traditional methods, and by transperineal ultrasonography - the method developed by the authors. Increased rear urethrovesical angle at rest and during functional tests with retention and straining, the deviation of the bladder neck and urethra to sacrum, shortening of urethra and bladder neck, lack of bladder neck and urethra movement, or the inability to perform volitional exercises were seen as signs of pelvic floor paradoxical movements. Analysis of the results of clinical trials showed, that the method is a reliable, non-invasive and does not require expert class ultrasound equipment. The treatment consisted of biofeedback therapy sessions performed in outpatient settings. 25 children were found to have positive changes, 7 of them fully recovered from voiding dysfunction.

[Coupling between the hemodynamic parameters and the morphological changes in the kidney in children with congenital hydronephrosis].

The data of renal ultrasonographic and Doppler studies were comparatively assessed in children with congenital hydronephrosis. The degree of renal B-mode ultrasound hemodynamic parameters was used as the basis for grouping of the children. The resistance index (RI) of all branches of renal arteries receives attention. Morphological studies were carried out on renal biopsy specimens from 29 children from different groups and on 12 removed kidneys; the expression of TGF beta1 and alphaSMA was revealed using the streptavidin-biotin-peroxidase method. Morphological changes as hypoplasia or dysplasia became more pronounced from Group 1 to Group 3, RI increasing to peak in Group 3. Vascular changes were confined to compensatory processes following the pattern of remodeling that was manifested by vascular wall thickening and a gradual increase in RI. Failing compensatory processes resulted in the development of renal functional and hormonal decompensation and in the elevation of RI. There was a coupling between the magnitude of morphological changes and RI increases. A set of the findings emphasizes the undoubtedly important role of renal ultrasound study that makes it possible to judge the state of the vascular bed and to suggest renal structural problems in congenital hydronephrosis.

[Recombinant oxyntomodulin].

An artificial gene encoding oxyntomodulin was obtained using chemical and enzymatic methods and cloned into Escherichia coli. A recombinant plasmid was constructed containing a hybrid oxyntomodulin gene and Ssp dnaB intein from Synechocystis sp. The expression of the resulting hybrid gene in E. coli, its properties, and the conditions of its autocatalytic cleavage to oxyntomodulin were studied.

[Recombinant thymosin alpha1]. / Rekombinantnyi timozin alpha1.

An artificial gene encoding thymosin alpha1 was obtained by the chemoenzymatic synthesis and cloned into Escherichia coli. An expressing recombinant plasmid containing the hybrid protein gene, which encodes amino acid sequences of thymosin alpha1 and the Saccharomyces cerevisiae intein Sce VMA, was constructed. The expression of the hybrid protein from the resulting hybrid gene in E. coli, the properties of the resulting hybrid protein, and the conditions for its nonenzymatic cleavage to thymosin alpha1 were studied. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 5; see also http: // www.maik.ru.

[Chemical and chemico-enzymatic synthesis of alpha-thiotriphosphate nucleosides]. / Khimicheskii i khimiko-fermentativnyi sintez alpha-tiotrifosfatov nukleozidov.

New methods of chemical and chemoenzymatic synthesis of nucleoside 5'-thiophosphates and 5'-alpha-thiotriphosphates are developed. The 5'-alpha-thiotriphosphates are used as substrates both in template-dependent enzymatic PCR synthesis and in a T7-RNA transcription polymerase system. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru.

[Methods of preparation of recombinant proteins-cytokines. IV. Renaturation of recombinant human interleukin-3]. / Metody polucheniia recombinantnykh belkov-tsitokinov IV. renaturatsiia rekombinantnogo chelovecheskogo interleikina-3.

Renaturation of recombinant human interleukin-3 produced as inclusion bodies in the transformed cells of Escherichia coli was studied and optimized. Importance was shown of removing from the protein solution the hydrophobic cellular components causing irreversible aggregation of the protein under renaturation conditions. An effect of pH on the secondary structure of the denatured protein was revealed by CD spectroscopy. It was thereby found that at pH 8.5, which is the optimal value for denaturation, the protein has the secondary structure most close to the native one. The isolation according to the scheme proposed allows preparation of interleukin-3 in 50% yield with 99% purity and biological activity 2 x 10(7) U/mg.

[The dependence of the E.coli gene expression level on the structure of the translation initiation region (TIR). IV. Distal complementary TIR interactions with the mRNA coding region]. / Zavisimost' urovnia ékspressii gena v E.coli ot struktury uchastka initsiatsii transliatsii (TIR). IV. Dal'nie komplementarnye vzaimodeistviia TIR s kodiruiushchei chast'iu mRNK.

To evaluate the effect on translation of distal regions of the encoding mRNA part capable of the complementary binding to the ribosome binding site (RBS), a series of plasmids were constructed containing fragments inserted into the il3 gene and determining secondary interactions in mRNA. A comparison of the levels of the in vivo gene expression showed that the complementary interactions of the translation initiation region (TIR) with distal regions of the mRNA encoding part affect translation. The effectiveness of these interactions decreased with an increase in the distance between the RBS and the complementary mRNA region, whereas the secondary structure formed by the TIR and the adjacent mRNA region was more stable despite the presence of regions in mRNA capable of forming energetically more favorable structures involving these elements.

[Construction of artificial genes by PCR methods using the synthetic template]. / Poluchenie nekotorykh iskusstvennykh genov metodom PTsR na sintetiche sko'i matritse.

Artificial genes were synthesized by the PCR method. Single-stranded DNA contained in an unpurified mixture of oligodeoxynucleotides after automated synthesis was used as a template. The features of this approach were studied.

[Relation between the level of E. coli gene expression and the structure of translation initiation region (TIR). III. Sites of complementary interaction of TIR with 16S rRNA]. / Zavisimost' urovnia ékspressii gena v E.coli ot struktury uchastka initsiatsii transliatsii (TIR) III. Saity komplementarnogo vzaimodeistviia TIR s 16S rRNK.

Potential sites of the complementary interaction of the translation initiation region (TIR) with 16S rRNA are revealed, and the role of these sites in the gene expression level is studied. The high expression level of a gene depends not only on the complementary interaction of TIR with 16S rRNA in sites proximal to the start codon [anti-Shine-Dalgarno (ASD) (delta G > -8 to -10 kcal/mol) and downstream box (DB)] and located at the -15 to +20 mRNA region but also on complementary interactions in distal sites of the untranslated branch of TIR (mTIR). Among them, the UB (upsteam box) 1 site, complementarily interacting with the exposed 452-490 segment of the 440-490 loop of 16S rRNA, may be located in the -15 to -50 mTIR segment. In the -50 to -70 mTIR segment may be located UB2 and UB3 sites, which interact with the exposed segment 478-488 of the 440-490 loop and segment 520-532 of the 520-540 loop of 16S rRNA, the UB3 site being much more efficacious. The high expression level requires that the total free energy of complementary interactions of UB1, UB2, and UB3 sites with 16S rRNA exceeds -20 kcal/mol.

[Recombinant proteins containing amino acid sequences of two ectatomin chains]. / Rekombinantnye belki, soderzhashchie aminokislotnye posledovatel'nosti dvukh tsepei éktatomina.

Artificial genes for chains A and B of ectatomin, an Ectatomma tuberculatum ant toxin, were obtained by chemical and enzymic synthesis and cloned into new plasmid vectors. Expression plasmids with the genes of hybrid proteins were constructed containing human interleukin-3 or its terminal 63-mer fragment as well as chains A and B of ectatomin, which are linked via a region containing the cleavage site of specific protease, enterokinase (hybrid proteins IL3ETOXA, IL3ETOXB, ILETOXA, and ILETOXB). Escherichia coli producer strains providing a high yield of IL3ETOXA and IL3ETOXB proteins as inclusion bodies were obtained.

[Mechanism of action of the plant hormone jasmonate. 1. Jasomonate-interacting proteins that regulate transcription of the p. pinII potato gene]. / Mekhanizm deistviia rastitel'nogo gormona--zhasmonata. 1. Belki-reguliatory transkriptsii gena p. pinII kartofelia, vzaimodeistviushchie s zhasmonatom.

A fragment containing the regulatory region of the p. pinII gene was isolated from potato DNA by polymerase chain reaction. Interactions of this DNA region with jasmonate determines the transcriptional activation. The isolated DNA fragment was cloned into the pTE2pb plasmid, which was used for preparing an affinity sorbent. Using this sorbent, four proteins were isolated from the total protein capable of desorption at physiological concentration of jasmonate. These proteins are likely to be subunits of two transcription repressors, whereas jasmonate serves as an inducer. Three sequences of the regulatory regions (boxes G, I, and III) are binding sites for repressors; similar sequences were found in various plant genes activated by jasmonate.

[Recombinant proteins containing oxytocin oligomeric sequences]. / Rekombinantnye belki, soderzhashchie oligomernye posledovatel'nosti oksitotsina.

Expression plasmids were constructed with genes encoding the ILOX3, ILOX6, and ILOX9 recombinant proteins, which contain the C-terminal fragments of trimer, hexamer, or nonamer of oxytocinoyl-Lys. Upon expression in E. coli, all three genes yielded inclusion bodies containing protein products of similar length and heterogeneous in the C-terminal region. It is likely that in the case of the ilox3 gene, the obtained protein mixture includes the full-length product of translation with the C-terminal lysine. In the case of the ilox6 and ilox9 genes, the protein products are formed as the result of a site-specific proteolysis in the regions between the second and the fourth oxytocin units.

[Dependence of the level of gene expression in E. coli on the structure of the translation initiation segment (TIR)]. / Zavisimost' urovnia ékspressii gena v E. coli ot struktury uchastka initsiatsii transliatsii (TIR).

The expression levels of genes that are transcribed to give mRNAs with identical leader sequences and even with identical extended coding regions may differ considerably. In order to determine the mechanism of this phenomenon, secondary structures of some mRNAs synthesized from a series of expression plasmids were studied. It was shown that the effect of the mRNA secondary structure in the translation initiation region on the initiation efficiency is due not only to the hairpin formation in this region but also to long-range interactions. When complementary structures tighter than those resulted from the interaction of regions SD, UB1, UB2, and DB with 16S rRNA are formed, the efficiency of the translation initiation and, consequently, the expression level decrease.

[Dependence of the level of gene expression in E. coli on the structure of the translation initiation segment (TIR). I. Primary structure of TIR]. / Zavisimost' urovnia ékspressii gena v E. coli ot struktury uchastka initsiiatsii transliatsii (TIR). I. Pervichnaia struktura TIR.

The comparison of expression levels of two genes-interleukin-3 (il3) and epidermal growth factor connected to leader peptide of OmpF (lompegf)-was carried out using specially constructed plasmids contained various structures of translational enhancers. It was shown that besides already known binding sites from mRNA translation initiation region (TIR) to 16S rRNA (SD, UB1 and DB), there is additional binding site of TIR disposed from -30 to -60 nt upstream of start codon AUG (UB2) having significant increasing effect on translation initiation and correspondingly on expression level.

[Cloning the repeat sequence of the oxytocin gene]. / Klonirovanie povtoriaiushcheisia posledovatel'nosti gena oksitotsina.

Cloning of a synthetic gene of the oxytocin trimer is accompanied by deletions, caused by nicks in the plasmid DNA. Use of covalently closed circular double-stranded DNA greatly reduces the number of the deletions.

[Recombinant interleukin-3 expression system in E. coli]. / Sistemy ékspressii v E. coli rekombinantnogo interleikina-3.

Plasmid pTOTE2IL3 (III) has been constructed, expressing an artificial human interleukin-3 (hIL3) gene under conditions of the induced protein biosynthesis. Levels of the recombinant protein synthesis have been compared in several E. coli strains containing expression plasmids pTE2IL3 (I) (constitutive biosynthesis) and (III) (induced biosynthesis). Optimal combinations of the expression plasmids and the bacterial strains are of importance. A simple and effective method has been elaborated for isolation, purification and renaturation of the recombinant protein accumulated in inclusion bodies.

[Isolation of recombinant interleukin-3 produced by E. coli]. / Vydelenie rekombinantnogo interleikina-3, produtsiruemogo E. coli.

A synthetic gene coding for human interleukin-3 (hIL3) was cloned in the plasmid pTE2IL3, the gene expression being controlled by the phage fd PVIII promotor and the phage T7 gene 10 translational enhancer. Under constitutive biosynthesis conditions in E. coli, the accumulation of recombinant hIL3 (in the inclusion bodies) was up to 30-40% of the total cell protein. An effective procedure of the hIL3 isolation is suggested. The hIL3 was solubilized in 5 M guanidinium chloride, renaturated and purified to homogeneity by a single chromatographic step. The protein's yield was 34 mg/g wet cells. The isolated hIL3 showed a specific biological activity.

[Chemical-enzymatic synthesis of the amplifier of the T7 phage gene 10 and function of its structural elements]. / Khimiko-fermentativnyi sintez usilitelia transliatsii gena 10 faga T7 i funktsiia élementov ego struktury.

The translational enhancer (TREN) sequence of the phage T7 gene 10 (in full and also its proximal or distal parts) have been obtained by chemical-enzymatic synthesis and cloned into the plasmids immediately before the human interleukin 3 (hIL3) artificial gene. Expression levels of the hIL3 gene in E. coli in these constructions show that the region controlling the specific activity is placed in distal part of TREN more than 40 nucleotides upstream from the initiation codon.