Induction of Vasculo- and Osteogenesis in Spheroids Formed by Adipose-Derived Stromal Cells.

Vascularization of bioengineered bone tissue constructs remains a challenging problem of regenerative medicine. Spheroids generated in 3D culture of adipose-derived stromal cells supplemented with inducing factors demonstrate stable characteristics and express of mesenchymal, endothelial, and osteoblasts markers, and represent a prototype of vascularized microtissue. Adipose-derived stromal cells spheroids induced to both angio- and osteogenic differentiation can be used in development of new innovative technologies for in vitro fabrication of vascularized bioequivalents for repair of large bone defects.

Angiogenic potential of spheroids from umbilical cord and adipose-derived multipotent mesenchymal stromal cells within fibrin gel.

One of the essential goals in regenerative medicine is microvascularization which enables an effective blood supply within de novo constructed tissues and organs. In our study, we used two common multipotent mesenchymal stromal cell (MMSC) sources (subcutaneous adipose tissue and Wharton's jelly of the umbilical cord) where is a subpopulation of endothelial precursors. In the medium supplemented with VEGF, the 3D cultures of UC MMSCs and ADSCs promoted the endothelial cell differentiation. To evaluate their ability to form a capillary-like network, we encapsulated spheroids within non-modified and PEGylated fibrin hydrogels. The PEGylated hydrogel supported better the formation of multibranched cords than the pure fibrin gel. Analysis of tubule growth rate, length, and branching showed that the differentiated ADSCs had higher angiogenic potential than the differentiated hUC MMSCs. Our study can be a basis for the development of new strategies in tissue engineering and treatment of vascular diseases.

The technology of obtaining multipotent spheroids from limbal mesenchymal stromal cells for reparation of injured eye tissues.

It is known that stem and progenitor cells open new possibilities for restoring injured eye tissues. Limbal eye zone, formed mainly by derivatives of neural crest, is the main source of stem cells for regeneration. The current study considers development of innovative technology for obtaining 3D spheroids from L-MMSC. It was shown that under 3D conditions L-MMSC due to compactization and mesenchymal-epithelial transition self-organize into cellular reparative modules. Formed L-MMSC spheroids retain and promote undifferentiated population of stem and progenitor limbal cells, as supported by expression of pluripotency markers - Oct4, Sox2, Nanog. Extracellular matrix synthetized by cells in spheroids allows retaining the functional potential of L-MMSC that are involved in regeneration of both anterior and, probably, posterior eye segment.

[Dynamics of osteogenesis after inoculation of autogenic mesenchymal stem cells of adipose tissue].

Experiment was conducted on 40 rats of Wister line. On the artificially reproduced experimental model autogenic mesenchimal stem cells (MSC) of adipose tissue were inoculated in space between bone autograph of tibia and mandible. MSC wasn't inoculated in the comparison group. Formation of a new bone substance in space between an autograph and mandible bone was observed. It was clear that after 120 days (180 days), there was a statistically significant decline of the area occupied by an immature fibroreticular bone. Described phenomenon, presumably, could be explained as a result of decline of the number of active cells in the population of inoculated MSC according to phenomenon of limited number divisions of cells on telomeres, described by Hayflick L. and Moorhead P.S.

[Correlation of psychoemotional status and adaptation to complete dentures].

Patients with full adentia are characterized by the formation of specific psycho-emotional status. Rational psychotherapeutic support of these patients largely determines the efficiency of dental prosthetic treatment. At the same time, the definition of mental and emotional status is not included in the diagnostic examination protocol. Considering the above the purpose of the study was to evaluate mental and emotional status of patients receiving complete dentures. Prosthetic rehabilitation of 30 patients with complete teeth loss was performed and clinical evaluation and evaluation of mental and emotional status were carried out before and after treatment. Patients with negative experiences of prosthetics showed a higher level of personal and situational anxiety. There was correlation of adaptation to removable dentures and the patient's personality traits. It is determined that emotional instability during treatment tends to decrease affecting the timing of adaptation to complete dentures. It is noted that patients with repeated prosthetics have earlier recovery of coordination ability of the masticatory muscles.

[Cardiovascular disorders in case of acute gastrointestinal bleeding in patients with arterial hypertension].

AIM: To investigate the blood pressure changes and electrocardiogram parameters for the 1st day after acute gastrointestinal bleeding in patients with arterial hypertension. MATERIAL AND METHODS: Indexes of blood pressure monitoring and electrocardiogram in case of acute gastrointestinal bleeding in patients with arterial hypertension were studied. 45 patients with arterial hypertension and acute gastrointestinal bleeding, 63 patients in acute stage of bleeding without hypertension and 48 patients suffering from hypertension without gastrointestinal bleeding were studied. Parallel simple open controlled investigation included blood pressure monitoring and Holter monitoring. RESULTS: Circadian dynamics of blood pressure in patients with arterial hypertension on the background of gastrointestinal bleeding was noted by reduced variability and insufficient reduction of pressure at night. Diastolic pressure decreased in accordance with severity of blood loss. Holter monitoring revealed high incidence of supraventricular ectopic activity, sinus arrhythmia and significant elongation of the ventricular electrical systole.

3D-technology of the formation and maintenance of single dormant microspheres from 2000 human somatic cells and their reactivation in vitro.

We developed an original reproducible 3D-technology for preparation of single dormant microspheres consisting of 2000 somatic cells. The dynamics of microsphere assembly from mesenchymal and epithelial cells of retinal pigment epithelium was traced using time-lapse microscopy: formation of a loose aggregate over 24 h followed by its gradual consolidation and formation of a compact viable microsphere with a diameter of 100-150 µ by day 7. The cell number in the formed microspheres remains unchanged. Reactivation observed upon fusion of epithelial and/or mesenchymal microspheres results in the formation of a united compact microtissue. The fusion dynamics reproduces spherogenesis irrespective of the initial amount of co-cultured microspheres. Reactivation via two-step induced angiogenesis opens new prospects for production of vascularized microspheres and microtissues.

[Dynamics of formation of bone tissue in rats under the influence of new xenotransplantat "Norian"].

In experiments on rats CD comparative studies have been carried out which is the effect of osteoplastic material NORIAN CRS, the rate of bone regeneration after mandibular dental implants (titanium screw) and the combined effect of these processes NORIAN CRS protein and protein (Emdogein). Functionality and maturity of the newly formed bone tissue was evaluated by histological and biochemical methods. The results suggest that the use of osteoplastic material NORIAN CRS is the 120 days leading to almost complete regeneration of bone tissue. It was also found that the combined use of NORIAN CRS and protein Emdogein impractical because adding Emdogein not accelerate bone regenerative process, but in the early postoperative period may cause an inflammatory reaction.

Radiological investigations at the "Taiga" nuclear explosion site, part II: man-made γ-ray emitting radionuclides in the ground and the resultant kerma rate in air.

Samples of soil and epigeic lichens were collected from the "Taiga" peaceful nuclear explosion site (61.30°N 56.60°E, the Perm region, Russia) in 2009 and analyzed using high resolution γ-ray spectrometry. For soil samples obtained at six different plots, two products of fission ((137)Cs and (155)Eu), five products of neutron activation ((60)Co, (94)Nb, (152)Eu, (154)Eu, (207)Bi) and (241)Am have been identified and quantified. The maximal activity concentrations of (60)Co, (137)Cs, and (241)Am for the soils samples were measured as 1650, 7100, and 6800 Bq kg(-1) (d.w.), respectively. The deposit of (137)Cs for the top 20 cm of soil on the tested plots at the "Taiga" site ranged from 30 to 1020 kBq m(-2); the maximal value greatly (by almost 3 orders of magnitude) exceeded the regional background (from global fallout) level of 1.4 kBq m(-2). (137)Cs contributes approximately 57% of the total ground inventory of the man-made γ-ray emitters for the six plots tested at the "Taiga" site. The other major radionuclides -(241)Am and (60)Co, constitute around 40%. Such radionuclides as (60)Co, (137)Cs, (241)Am, and (207)Bi have also been determined for the epigeic lichens (genera Cladonia) that colonized certain areas at the ground lip produced by the "Taiga" explosion. Maximal activity concentrations (up to 80 Bq kg(-1) for (60)Co, 580 Bq kg(-1) for (137)Cs, 200 Bq kg(-1) for (241)Am, and 5 Bq kg(-1) for (207)Bi; all are given in terms of d.w.) have been detected for the lower dead section of the organisms. The air kerma rates associated with the anthropogenic sources of gamma radiation have been calculated using the data obtained from the laboratory analysis. For the six plots tested, the kerma rates ranged from 50 to 1200 nGy h(-1); on average, 51% of the dose can be attributed to (137)Cs and 45% to (60)Co. These estimates agree reasonably well with the results of the in situ measurements made during our field survey of the "Taiga" site in August 2009.

[Choice of the operation for perforated ulcers of the stomach and duodenum].

Actual questions of diagnostics and treatment of 776 patients with perforated gastroduodenal ulcers are discussed. The most informative method of the diagnosis, particularly covered perforation, is laparoscopy. Decision on the operation must be individually grounded. Radical operations were performed in 599 patients with 2% postoperative lethality. Lethality after suturing of the perforation was 10.3%, after resection operations--4.1%, after suturing the perforation in combination with selective proximal vagotomy--0.3%.

Radiological investigations at the "Taiga" nuclear explosion site: Site description and in situ measurements.

In the summer of 2009, we performed a field survey of the "Taiga" peaceful underground nuclear explosion site, the Perm region, Russia (61.30° N, 56.60° E). The explosion was carried out by the USSR in 1971. This paper provides an extended summary of the available published data on the "Taiga" experiment. A detailed description of the site is illustrated by original aerial and ground-level photos. A large artificial lake (700 m long and 350 m wide) currently occupies the central area of the experimental site. The ground lip surrounding the lake is covered by a newly grown mixed forest. In situ measurements, performed in August 2009, revealed elevated levels of the γ-ray dose rate in air on the banks of the lake "Taiga". Two hot spots were detected on the eastern bank of the lake. The excess of the γ-ray radiation is attributable to the man-made radionuclides (60)Co and (137)Cs. The current external γ-ray dose rate to a human from the contaminations associated with the "Taiga" experiment was between 9 and 70 µSv per week. Periodic monitoring the site is recommended.

[Surgical tactics and causes of death in ulcer gastroduodenal bleedings].

4212 patients with ulcer gastrointestinal bleedings were treated during 1990-2007 years. 364 patients had gastric ulcers, other patients had duodenal ulcers. 1819 patients (43.2%) were operated. 418 patients (23%) were operated due to ineffective endoscopic hemostasis and recurrent bleedings. 409 patients (22.5%) underwent urgent operations because of risk for recurrent bleeding. Elective operations were carried out to 1358 patients (74.7%). Postoperative lethality amounted 5.8% including 19.1% lethality after emergency operations, 4.2% lethality - after urgent operations and 0.8% lethality - after delayed operations. The general hospital lethality amounted 3.3%. Lethality can be decreased by extension of indications for urgent operations, preventing recurrent bleedings.

[Relict microorganisms of cryolithozone as possible objects of gerontology].

Permafrost is widely distributed in the Northern Hemisphere, and its age reaches hundreds of thousands and millions of years. Permafrost contains alive microorganisms which are not frozen due to relatively high temperature of the environment (-2...-8 degrees C), but the microorganisms are immobilized and therefore aged probably similar to the age of permafrost. Longevity of the relict microbial cells is related obviously to their mechanism of protection against heat, radiation, free radicals and other damaging agents. A strain of Bacillus sp. was isolated from permafrost aged of about 3 million years, 16S rDNA sequence was identified and preliminary testing of bacterial culture on Drosophila melanogaster and mice was made. Immune stimulation and improvement of physical condition were observed, and that, together with the age of the microbial cells, presents the relict microorganisms as objects of gerontology.

[Experience with using a one-layer uninterrupted suture in abdominal surgery].

The authors made an analysis of using a one-layer uninterrupted suture with a synthetic absorbable thread in forming gastrointestinal, intestinal, duodenojejunal and pancreatojejunal anastomoses. The operation technique and early complications are described showing advantages of the one-layer uninterrupted suture with a synthetic thread.

[Impact of xenotransplantation of multipotent mesenchymal stem cells of the bone marrow on the recovery of the structural organization and functional activity of the retina in its damage].

Among the potential sources of cells for transplantation, the most promising autologous material for cell therapy and tissue engineering are multipotent mesenchymal stem cells (MMSC). The authors carried out an experiment dealing with MMSC xenotransplantation into the rabbit suprachoidal space on a model of laser retinal coagulation. For objective assessment of the magnitude and pattern of changes in the functional activity of different retinal cellular elements and the efficiency of MMSC transplantation, they performed electroretinographic and histological studies. The study indicated that MMSC transplantation into the suprachoroidal space is a safe and effective procedure for cell incorporation in order to treat retinal diseases. Electroretinography provided evidence that single MMSC transplantation better preserved the function of photoreceptors (a-wave). Histomorphometric studies demonstrated that the inserted cells stimulated reparative processes in the laser-induced damaged retina.

[Restriction endonuclease Asi256I recognizing the nucleotide sequence 5'-GATC-3'].

A strain producing a restriction endonuclease was isolated from soil samples and identified as the Arthrobacter sp. strain Ck256. The enzyme produced by this strain was termed Asi2561. The isolation procedure for this enzyme was described, and the optimal conditions for its function were determined. It was shown that the restriction endonuclease Asi256I is a true isoschizomer of MboI, it has a temperature optimum of 6 degrees C, and can be used in molecular-biological and genetic-engineering studies performed at low temperatures.

[The new eubacterium Roseomonas baikalica sp. nov. isolated from core samples collected by deep-hole drilling of the bottom of Lake Baikal].

Microbiological analysis of samples of sedimentary rocks from various eras of the geological history of the Baikal rift has enabled us to isolate a large number of microorganisms that can be classified into new, previously undescribed species. The present work deals with the identification and study of the morphological, biochemical, and physiological properties of one such strain, Che 82, isolated from sample C-29 of 3.4-3.5 Ma-old sedimentary rocks taken at a drilling depth of 146.74 m. As a result of our investigations, strain Che 82 is described as a new bacterial species, Roseomonas baikalica sp. nov., belonging to the genus Roseomonas within the family Methylobacteriaceae, class Alphaproteobacteria.

[Consumption of hydrocarbons by psychrotolerant degrader strains].

Oil-oxidizing microorganisms have been sampled in various regions of Siberia and used in strain associations, which degrade n-alkanes of oil from various fields by 64-92% after 6 days of growth in a wide temperature range. These strains are salt-tolerant and psychrotolerant. They are compatible with aboriginal soil microflora. Promising results have been obtained in experiments on growing plants on oil-polluted soil purified with a biodegrader of this series.

Cell biology of fetal tissues and fundamental medicine.

During the recent decade the interests of specialists in experimental mammalian embryology were focused on studies of signal networks, regulatory programs, phenotype and behavior of embryonic and mesenchymal stem cells in culture, which triggered more active use of pluripotent stem cells in cell technologies. Laboratory preparation of primary induction tissue (extraembryonic endoderm, mesoderm, hypoblast, nodule, notochord, primary streak) remains impossible, but primary and secondary organizers and antagonist tissues are essential for simulation of axial development. Only in these sophisticated heterogeneous cell system will it be possible to simulate the real physiology and bioinformatics of the embryonic leaflets, their staged transformation into definitive organs. At present, mammalian and human fetal tissues remain the carriers of unique information of the middle and late embryogenesis. For this reason, functional and therapeutic activities of fetal cells and tissues are intensely studied in foreign countries; numerous banks of fetal somatic and stem cells are created. Banks of extraembryonic umbilical blood tissue have been created in Russia. However, the biological and information potentials of fetal/embryonic tissue remain not realized at the theoretical and applied levels.

[Surgical treatment of ulcer disease in arterio-mesenterial compression of the duodenum]. / Khirurgicheskoe lechenie iazvennoi bolezni pri arteromezenterial'noi kompressii dvenadtsatiperstnoi kishki.

Experience in surgical treatment of 124 patients with gastric and duodenal ulcers associated with chronic duodenal obstruction due to arterio-mesenterial compression of the duodenum is analyzed. This combination occurred in 12% of all patients with ulcers over 20-year period. Special methods of examination were used for diagnosis: probe duodenography with duodeno-manometry, hepato-bili-scintigraphy. Differential approach to choice of surgical method was developed, and two original methods of surgery are presented: selective proximal vagotomy and resection of the stomach in combination with duodeno-jejuno-anastomosis. Long-term results were studied in 75 patients, and in 93% of them they were assessed as excellent and good. Quality of life of patients after surgery depends on severity of chronic duodenal insufficiency.