Fragments of the Fab-3 and Fab-4 Boundaries of the Drosophila melanogaster Bithorax Complex That Include CTCF Sites Are not Effective Insulators.

The segment-specific regulatory domains of the Bithorax complex (BX-C), which consists of three homeotic genes Ubx, abd-A and Abd-B, are separated by boundaries that function as insulators. Most of the boundaries contain binding sites for the architectural protein CTCF, which is conserved for higher eukaryotes. As was shown previously, the CTCF sites determine the insulator activity of the boundaries of the Abd-B regulatory region. In this study, it was shown that fragments of the Fab-3 and Fab-4 boundaries of the abd-A regulatory region, containing CTCF binding sites, are not effective insulators.

The Piragua Gene Is not Essential for Drosophila Development.

Proteins with clusters of C2H2 zinc finger domains (C2H2-proteins) constitute the most abundant class of transcription factors in higher eukaryotes. N-terminal ZAD (zinc finger-associated domain) dimerization domain has been identified in a large group of C2H2-proteins mostly in insects. The piragua gene encodes one of these proteins, Fu2. We have generated CRISPR/Cas9-mediated deletion of the piragua gene that has no phenotype. We have used φC31-mediated attP/attB recombination to generate a transgenic line expressing Fu2 protein fused with HA epitope. This line will be useful for analysis of DNA binding profile and functions of Fu2 protein.

[Choice of optimal treatment policy in patients with descending aortic dissection]. / Vybor optimal'noi taktiki lecheniia patsientov s rassloeniem niskhodiashchei aorty.

Contemporary approaches to treatment of patients with dissection of the descending aorta (DA), as well as the data of international registries and guidelines are contradictory. The endovascular method of treatment of patients with descending aortic dissection has been recognized to be revolutionary. However, there seems to be no commonly accepted concept concerning appropriate use of currently available techniques of treating patients with DA dissection. This study was aimed at analysing the experience gained in using various methods of treatment of patients with descending aortic dissection (DAD). We retrospectively examined the results of surgical treatment of 78 patients with DAD. Of these, 45 patients were found to have Stanford type B dissection and 33 patients had type A dissection, who underwent treatment during ten years from January 2008. The decision concerning the type of care for patients with DAD (conservative, surgical, endovascular or combined methods of treatment) directly depends on the patient's state stability, technical feasibility of the intended procedure, and the presence of concomitant pathology. The findings obtained in our study demonstrated better immediate outcomes of the endovascular approach compared with the hybrid and surgical ones. In order to achieve complete haemodynamic correction of the pathology, to minimize the risk of aortic remodelling and the development of aorta-related complications, the treatment should not be limited to endoprosthetic repair of the thoracic portion of the DA with a stent graft, but should rather be supplemented with stenting of more distal segments, including with uncoated stents at the level of the visceral branches. Only such staged approach may provide better conditions for obliteration of the aortic false lumen and formation of a 'neo-frame' of the aortic wall.

[Modern approaches to the diagnosis of precancerous pathology and early lung cancer]. / Sovremennye podkhody k diagnostike predopukholevoi patologii i rannego raka legkogo.

AIM: To assess the possibilities of modern diagnostic techniques to diagnose precancerous pathology and early central lung cancer. MATERIAL AND METHODS: Analysis of Russian and foreign publications for precancerous pathology and early lung cancer, results and comparison of various diagnostic techniques. RESULTS: Central lung cancer has a stepwise development with transformation of normal bronchial epithelium to hyperplastic followed by focal metaplasia, dysplasia, cancer in situ (CIS) and microinvasive cancer. Fluorography, chest X-ray, computed tomography, magnetic resonance imaging and sputum cytology examination are used to diagnose lung cancer. However, endoscopy plays a leading role in diagnosis of early central lung cancer which rapidly and effectively detects changes of tracheobronchial tree mucosa at the initial stages of carcinogenesis thanks to the use of modern techniques (spectral, autofluorescent and ZOOM - endoscopy).

NEW OPPORTUNITIES OF THE QUALIFYING ENDOSCOPIC DIAGNOSTICS OF PANCREAS NEOPLASMS.

OBJECTIVE: the aim of study was testing and introduction in clinical practice combination of EUS with sonoelastography, fine-needle biopsy and needle-based confocal laser endomicroscopy in advanced diagnostic of pancreatic disease. MATERIALS AND METHODS: in the period from February 2014 to December 2015, we accumulated experience of EUS-FNA in 72 patients mostly with tumor pathology of the pancreas. In 16 cases we used 19G, in 49-22G, and in 7 patients - 25G needles. In 5 patients we used COOK Echotip Procore needle. In 8 cases we performed confocal laser endomicroscopy through the 19G needle (nCLE) in 3 patients with cystic and in 5 - with solid tumors. In 20 patients, there we used.HITA-CHI-PENTAX sonoelastography was performed. RESULTS: There were no complications in EUS-FNA. In all cases we received pathology verification of disease. nCLE results always confirmed by morphological examination of material. In elastography examination normal pancreatic tissue had a equable green-yellow color, in chronic pancreatitis on the same background there were areas of blue seal parenchyma, in the cases pancreatic adenocarcinoma it was marked predominance of blue color scale, in neuroendocrine tumors - diffuse distribution of green and blue areas. Using nCLE in patients with adenocarcinoma revealed the destruction of certain glands, polymorphic nuclei of epithelial cells, with their enlargement, deformation, loss of polarity. Appliance of Procore needle has its technical features, but allowed us to obtain more material with less bloody - due to fewer number of passes. CONCLUSION: Thus, our study confirms the effectiveness of EUS-FNA with sonoelastography and nCLE. It defines objectives for improvement and expansion of the range for their clinical use.

[ENDOCYTOSCOPY--NOVEL ENDOSCOPIC DIAGNOSTICS APPROACH: PRINCIPLES AND PROCEDURE].

Endocytoscopy is one of the most novel endoscopic diagnostic procedures, providing optical magnification up to 1150 times of gastrointestinal and respiratory tract mucosa. Such approach allows real-time tissue and cellular structure visualization. Endocytoscopy, along with confocal laser endomicroscopy, can be considered as "optical biopsy" in vivo. Of course, endocytoscopy currently is experimental diagnostic method, all available endocytoscopes are prototypes. According to published data, endocytoscopy can be used in precancerous conditions and early intramucosal cancer diagnostics in esophagus, stomach, colon and bronchial tree. Different types of endocytoscopes are used for examinations: some of them are baby-scopes, with fixed magnification 570-1150 times, introduced into accessory channel of the therapeutic parent-endoscope, others--are integrated type, providing scalable magnification from 80 to 380 times. As for traditional pathology ex vivo, for endocytoscopy mucosal cell nuclei stain is needed. For vital staining during endocytoscopy methylene blue, toluidine blue and crystal violet in different concentrations are more often used. In cases of squamous-cell dysplasia or cancer, it is recommended to use 1% methylene blue solution, whereas in intestinal type metaplasia, dysplastic changes and cancer (Barrett's esophagus, P. Correa precancerous cascade, colon adenomas), 1% toluidine blue is preferred. With endocytoscopy, after vital staining, we can visualize and estimate mucosa tissue and cell characteristics: papillae, crypt and gland shapes and sizes, their integrity (tissue markers); cell nuclei size and shape, polarity and nuclear dye intensity (cell markers).

[ENDOCYTOSCOPY--NEW TYPE OF ENDOSCOPIC EXAMINATION OF LOWER GASTROINTESTINAL AND RESPIRATORY TRACT].

Majority of published data describing endocytoscopic examination of upper gastrointestinal tract mucosa, but in recent publications, it is reported, that endocytoscopy is suitable for small bowel, colon, respiratory tract and even peritoneum "optical biopsy". In number of articles possibilities of celiac sprue diagnostics with endocytoscopy is discussed, but small-bowel endocytoscopy is limited, due to absence of endocytoscopes, compatible with enteroscopes. More widely endocytoscopy is used in colon, mostly in lateral-spreading adenomas diagnostics. Prof. S-E. Kudo developed endocytoscopic classification of colonic mucosa changes, used for differential diagnostics and lesion mapping, describing hyperplasia, adenomas with different grades of intraepithelial neoplasia, non-invasive and invasive cancer. Some authors reported about good possibilities of endocytoscopy in inflammatory bowel disease diagnostics. Most of data, related to respiratory tract endocytoscopic examination, focused on precancerous conditions and early pharyngeal and lung cancer, and the preliminary results are promising, but, unfortunately, for now, endocytoscopy in bronchial tree is limited, due to lack of thin endocytoscopes. According to some article data, it is possible to use endocytoscopy not only in gastrointestinal and respiratory tract, but also in optical confirmation of peritoneal tumor dissemination in gastric and ovarian cancer patients, and--in bladder mucosa examination.

[Lung-preserving surgical treatment of patients with bronchial carcinoid].

Isolated bronchus resection for central cancer was performed in 25 patients including preoperative bronchoscopic removal of exophytic tumor in 20 (80%) observations in thoracic department of P. Gertsen Moscow Research Cancer Institute. According to morphological study typical carcinoid was diagnosed in 23 (92%) patients, atypical - in 2 (8%) cases. All patients underwent conventional mediastinal lymphadenectomy. Postoperative complications after bronchus resection developed in 6 (33.3%) patients. There were no deaths. Overall 5- and 10-year survival was 100% and 96% respectively. The authors consider that by strict indications combination of endoscopic removal with isolated bronchus resection preserves all pulmonary parenchyma without prejudice for surgical radicalism.

[THE FIRST CLINICAL EXPERIENCE IN THE CELIAC PLEXUS BLOCKADE UNDER ENDOSCOPIC ULTRASOUND NAVIGATION IN PATIENTS WITH PANCREATIC CANCER].

OBJECTIVE: Implementation into clinical practice and getting our own data of the effectiveness of EUS-CPN in chronic pain syndrome associated with pancreatic cancer in the palliative treatment. MATERIALS AND METHODS: In the period from October 2014 to May 2015 in P.A. Herzen Moscow Research Oncological Institute--filial FGBI "NMIRC" gained the first clinical experience in the celiac plexus blockade under endoscopic ultrasound navigation in 10 patients with pain associated with pancreatic cancer. The study group included 6 women and 4 men aged 54-83 years. In 2 of 10 cases out celiac ganglia were located and injections made in side them. In eight cases, the blockade was performed after injection into the tissue surrounding the celiac trunk. In most cases, we used standard FNA-needle with diameter 22G. Blockade performed by a combination of local anesthetic (3-4 ml of 0.5% bupivacaine) and 14-20 ml of 96% ethanol. RESULTS: There were no complications after EUS-BCS. 9 of 10 patients (90%) had a significant (2-4 points on a visual analogue scale) reduction of pain after 1-3 days after the procedure. These patients had a significant reduction in the frequency and dosage of analgesics. Four patients required repeat of anesthesia after 2 months--the same with a marked positive effect. One patient, in spite of the double execution of EUS-CPN did not note reduction in pain intensity. CONCLUSION: EUS-CPN procedure is highly effective, very simple and safe. It provides a high quality of life and of social adaptation oncology patients. It should be more widely used in clinical practice.

[Confocal laser endomicroscopy principles and performing algorithm in gastric mucosa examination].

Accuracy of endoscopic examination in early gastric cancer and precancerous conditions diagnostics for many years depended only on quality of biopsy. That's why, risk of overlooking gastric focal carcinoma, particularly-- multiple, was relatively high. Last couple of years new endoscopic method--confocal laser endomicroscopy (CLE) was released for commercial use. This approach provides real-time information about morphology of gastric mucosa during endoscopic examination. CLE is a variation of confocal microscopy--morphologic technique, providing examination of thick specimens or live tissue. CLE system is a single-channel fluorescence microscope, used in endoscopy, where confocal probe incorporated into endoscope or mounted into accessory channel. For proper results of CLE intravenous administration of fluorescence agent is needed. In our study in P.A. Herzen Moscow Cancer Research Institute we have used 10% fluorescein sodium, due to acriflavine use is prohibited in Russian Federation. In 157 patients with suspected early gastric cancer mean time of CLE was 24 ± 3.5 min. In all cases descriptive images were acquired. Mean amount of endomicrosocpic images in one patient was as high as 162 ± 8.3.

[Early gastric cancer and precancerous conditions diagnostics with confocal laser endomicroscopy].

AIM: To develop endomicroscopic criteria of early gastric cancer and precancerous conditions METHODS: 157 suspected for early gastric cancer patients were included in our study. In all cases complex endoscopic examination (narrow-band imaging NBI-HD endoscopy, endosonography (EUS) and endoscope-based confocal laser endomicroscopy (CLE) with precise biopsy) was performed. CLE images compared to pathology data. RESULTS: Precise criteria of intestinal metaplasia in CLE was presence of oval gray goblet cells in epithelium. Gastric adenoma with moderate dysplasia appeared on CLE images as glandular structures in mucosa surface layers. The signs of high-grade dysplastic changes in adenoma were appearing of different shape and size cells with lost polarity. Well-differentiated carcinoma featured destruction of some glands and forming dark cell clusters. Revealed criteria of poor-differentiated carcinoma was total disintegration of glandular structures. Overall accuracy of CLE in early gastric cancer and precancerous conditions diagnostics reached 95,6% (P < 0.001). Selected accuracy in different condition was: 100% in intestinal metaplasia identifying, 86.1%--in adenoma with moderate dysplasia, 96.2% in high-grade dysplasia and cancer in situ, and 100%--in poor-differentiated gastric carcinoma types). CONCLUSION: As the result of our study we have developed precise endomicroscopic criteria of intestinal metaplasia in stomach, gastric adenoma with moderate and high-grade dysplasia and early gastric cancer--as well-differentiated, as poor-differentiated.

[Endoscopic stenting of malignant stenosis in the cervical esophagus].

OBJECTIVE: to define the optimal approach for self-expanding metal stents (SEMS) installing in the cervical esophagus in cases of malignant stenosis and to select the optimal types of stents for this tumor location. MATERIALS: From 2004 to 2013 in P.A. Herzen Moscow Cancer Research Institute in 24 patients with cervical esophagus tumor stenosis endoscopic stenting was performed. 70% of installed stents were Choo Cervical (M.I. Tech, Korea), 13%--unspecialized Choo and Hanaro stents (M.I. Tech, Korea), and 17%--Gianturco-Z stents (Wilson-Cook, Ireland). Main contraindication for cervical esophagus stenting was location of the tumor margin less than 1 cm from the upper esophageal sphincter. In 21% of cases, endoscopic stenting was performed under combined fluoro- and endoscopic guidance, in 79%-- using only endoscopic approach. RESULTS: Technical success of stent placement was achieved in all patients; SEMS was installed at the previously defined level. The accuracy of stenting was not depended from the type of guidance procedure. The most intensive pain was observed in cases with "not specialized" stents with diameter of 18, 20 or 22 mm. Average pain level in such patients was 4.54 points (VAS). In the subgroup with "cervical" stents average pain intensity was not exceed 3 points. Chronic pain syndrome lasts lifelong in 17% of patients. CONCLUSION: The procedure of SEMS positioning under endoscopic guidance with individual selection of stent type allows restoring oral nutrition in patients with severe tumor stenosis of the cervical esophagus.

Hamiltonian field theory of ferrohydrodynamics.

A Hamiltonian field theory of ferrohydrodynamics is derived, with dissipation included by use of a Rayleigh dissipation function. It is shown that kinematic assumptions on the behavior of magnetization under displacements of a volume element of fluid leave a certain freedom in the construction of dynamical equations describing the time-dependence of mass density, flow velocity, entropy density, magnetization, and spin density. The convective behavior of magnetization may be characterized by two dimensionless coefficients.

[Prothymosin alpha interacts with C-terminal domain of histone H1 and dissociates p53-histone H1 complex].

A novel mode of the tumor suppressor protein p53 regulation, mediated by recruitment of the linker histone H1 to the promoters of p53 target genes leading to specific repression of p53-dependent transcription, has recently been uncovered. Yet, how this repression could be relieved is not clear. Previously, a histone-binding nuclear protein prothymosin alpha (ProTa) was shown to trigger a p53 response. The histone-binding region of ProTa was found to be essential for this effect, raising a possibility that ProTa stimulates p53-dependent transcription by dissociating the p53-histone H1 repressive complex. Here, we have shown that ProTa interacts with the same C-terminal domain of histone H1 as p53 does and, therefore, ProTa and p53 could compete for binding to histone H1. Furthermore, ProTa, when competent for histone H1 binding, is able to liberate p53 from the histone H1-p53 complex in vitro. In vivo, stimulation of p53-dependent transcription by ProTa correlates with ability of ProTa to interact with histone H1. Ectopic expression of histone H1 or its C-terminal ProTa-binding domain specifically suppresses the stimulating effect of ProTa on transcription of the p53-responsive reporter gene in cultured cells. These results are consistent with the model that ProTa may enhance p53 transcription activity by displacement of histone H1 from p53-H1 repressive complex.

[Endobronchial surgery and photodynamic therapy for the treatment of multiple primary lung cancer].

Endoluminal endoscopic surgery and fotodynamic therapy were used in treatment of 104 patients with multiple primary lung cancer (MPLC), or more exactly, of trachea and lobar and segmental bronchi. Diagnostic division included videobronchoscopy of high resolution in with light and NBI-regimen; autoflourescent and 5-ALA-induced fluorescent videobronchoscopy, endosonography, computed tompgraphy or magnetic resonance imaging of the thorax and epithelial mucine (MUC-1) immunohistochemical analysis of scarificates. Result of treatment strongly depended on the size of primary tumor. Complete regression of cancer was observed for all tumors less then 1 sm in diameter. Endoscopic treatment, including fotodynamic therapy and argon coagulation, proved to be a method of choice in treatment early synchronous or metachronous multiple primary lung cancer in incurable patients.

[Endolaryngeal surgery and photodynamical therapy of using flexible videoendoscopic devices to treat laryngeal cancer and premalignancy].

Patients with laryngeal cancer (LC) and premalignancy admitted to P.A. Gertsen Moscow Research Oncological Institute were treated by the methods of endolaryngeal surgery making use of flexible videoendoscopic devices, Nd:YAG laser-assisted thermal ablation, argon plasma coagulation, electroresection, and photodynamical therapy. A total of 129 patients were given treatment by these techniques following distant radiotherapy including 23 with grade I-III laryngeal dyskeratosis and dysplasia, 36 with virus-associated recurrent laryngeal papillomatosis, 37 with early cancer (Tis-T1NoMo) in the middle part of the larynx, 17 with a residual tumour, and 16 with mid-laryngeal cancer (T1-2NoMo). The above treatments resulted in complete regression (CR) of dyskeratosis and grade I-III dysplasia in 22 (95%) of the 23 patients and CR of recurrent papilllomatosis in 27 (75%) of the 36 patients. Moreover, complete regression was achieved in 34 (92%) of the 37 patients with laryngeal cancer. It recurred in 3 (9%) of the 34 patients. In all cases, the vocal function was completely restored. In groups with residual and recurring laryngeal tumours, complete remission was documented in 27 (82%) of the 33 patients. LC recurred in three patients within 1-5 years after completion of the treatment.

[Barrett's esophagus: endoscopic and immunomorphological parallels].

Barrett esophagus is the most serious sequela of the gastroesophageal reflux disease being an obligate precancer with a high index of the neoplastic transformation as to an adenocarcinoma esophagus. THE PURPOSE OF THE PAPER: To reveal the extent of the susceptibility to oncogenesis of the Barrett esophagus-patients and to determine high-risk groups. MATERIALS AND METHODS: Our paper has shown the examination results of the 55 Barrett esophagus-patients (29 women and 26 men at the age of 39 to 62 years old), including 40 intestinal metaplasia-patients and 15 patients of the intestinal metaplasia + dysplasia of long clinical course given corresponding correcting cure. In our investigation a DNA-flow cytometry was a method of determining the adenocarcinoma esophagus risk secondary to the Barrett esophagus as well as an index of the proliferation and an index of the aneuploidy were the factors analyzed. THE RESULTS: 1) As the pathosis histology-progresses from metaplasia to dysplasia and adenocarcinoma esophagus the increase in the aneuploidy rate, the proliferation index, and S-cycling state cells portion is observed; 2) the proliferation index is a rather stable factor of the human body susceptibility to oncogenesis and varies only slightly in the course of the cure. The index of the aneuploidy is a labile factor that can be subject to decrease down to normal figures given corresponding cure (p > 0.05); 3) the increased aneuploidy index in the presence of the intestinal metaplasia free of esophagus dysplasia can serve as an objective factor for neoplastic progression.

[New possibilities for the treatment of pharyngeal and tracheal stenoses following resection of an organ and laryngectomy in patients with disseminated laryngeal and laryngopharyngeal cancer].

Results of the treatment of 174 patients with stenosis of larynx and trachea by conservative, surgical, and endoscopic methods at different periods after operation indicate that application of novel endoscopic and physical techniques substantially extends possibilities for the low-invasive correction of postoperative laryngeal and tracheal stenoses.

[Influence of prothymosin alpha and its mutants on activity of the p53 tumor suppressor].

Overexpression of the nuclear oncoprotein prothymosin a enhanced and, in a reciprocal experiment, down-regulation of endogenous prothymosin alpha by RNA interference approach inhibited transcriptional activity of the p53 tumor suppressor in the reporter gene assay. Ectopic expression of prothymosin alpha enhanced not only p53-dependent transcription, but also intracellular level of p53 in HeLa (but not HCT116) cells. Ability to stimulate p53-dependent transcription was lost by C-terminal mutants of prothymosin alpha with impaired nuclear accumulation, but not by N-terminal deletion mutants and by the double mutant of prothymosin alpha with impaired ability to bind Keap1, suggesting that prothymosinalpha-Keap1 interaction is dispensable for p53 response. Our data suggest that the central "acidic" region of prothymosin alpha together with intact nuclear localization signal is responsible for stimulation of p53-dependent transcription. This conclusion was confirmed by the fact that another protein containing long "acidic" region and nuclear localization signal, parathymosin, was able to stimulate transcription of p53-responsive reporter gene.