[Risk factors for pancreatitis and pancreatic cancer].

AIM: To identify and compare the frequency of alcohol consumption, tobacco smoking, levels of main macronutrients, vitamins and mineral elements consumption in patients with acute (AP) and chronic pancreatitis (CP) and pancreatic cancer (PC). MATERIALS AND METHODS: At the observational clinical cross-sectional uncontrolled case-study 65 patients with AP or CP (group 1) and 45 patients with PC (group 2) were examined. A survey of patients was carried out: questionnaire on tobacco smoking, a frequency questionnaire on alcohol consumption, a questionnaire for assessing the frequency of food consumption. RESULTS: The frequency of smoking (33.8, 20.0%; p0.05), alcohol consumption 1 times/week during the last year (21.5, 15.6%; p0.05) did not differ significantly between the two groups. The highest consumption rates of total, vegetable, animal protein, total carbohydrates, refined sugar, animal fat, cholesterol, MUFA, dietary fiber, vitamins (-carotene, vitamin B1, B2, C, PP), mineral elements (iron, potassium, calcium, magnesium, sodium, phosphorus) and the daily energy content of the diet were determined in PC than in the AP and CP group. Among patients of group 1, deficient intake of fat-soluble vitamin A (93.3, 54.8%; p=0.009) and vitamin E (80.0, 48.4%; p=0.041) was more common in the subgroup of patients with excretory pancreatic insufficiency than without it, and the chance of having hypercholesterolemia was associated with a deficient intake of vitamin E [Ex(B)=3.3, 95% CI 1.59.3; p=0.027]. CONCLUSION: There were no differences in the frequency of smoking and alcohol consumption between patients with AP and CP and PC. The highest indices of the main macronutrients, daily energy content of the diet, micronutrients (except for vitamins A, E) were found in PC than in the group of patients with AP and CP. Among patients with AP and CP with excretory pancreatic insufficiency, a lower intake of fat-soluble vitamins was noted and associations of hypercholesterolemia with deficient intake of vitamin E were obtained.

Effect of Peptide AEDG on Telomere Length and Mitotic Index of PHA-Stimulated Human Blood Lymphocytes.

We studied the effect of peptide AEDG on telomere length and mitotic index of PHA-stimulated blood lymphocytes from young (18-22 years, N=5) and middle-aged (49-54 years, N=6) men. In the younger age group, no significant changes in the mitotic index were detected, while in the middle-aged group, a decrease in this parameter was found in one case. The relative length of telomeric regions of metaphase chromosomes was evaluated by in situ fluorescence hybridization with DNA probes specific to telomeres. After incubation with peptide AEDG, significant changes in the relative telomere length were found in 7 of 11 individuals (3 cases in the younger age group and 4 cases in the middle age group). Significant increase in telomere length after exposure to peptide AEDG was revealed in 5 cases, including two individuals of the younger age group (by 41 and 55%) and three individuals of the middle age group (by 156, 18, and 76%). In one individual of the younger age group and in one of the middle-age group, a significant decrease in telomere length (by 37 and 15%, respectively) was found. A tendency to normalization of telomere lengths was noted: this parameter increased in individuals with initially lower telomere length relative to the group mean value and decreased in individuals with initially longer telomeres compared to the mean length in the group.

Development of a Bacteriophage Complex with Superhydrophilic and Superhydrophobic Nanotextured Surfaces of Metals Preventing Healthcare-Associated Infections (HAI).

We studied antibacterial properties of organo-inorganic hybrid coatings on the AMg2 aluminum alloy including superhydrophilic and superhydrophobic nanotextured metal substrates with applied bacteriophage particles. Bactericidal activity of surfaces after artificial contamination with a bacterial suspension was evaluated. To increase bactericidal effect of the plates, bacteriophage was sorbed on their surface. In the experiments simulating possible spreading of HAI pathogens, higher bactericidal activity of superhydrophilic surfaces in comparison with superhydrophobic ones. Application of bacteriophage particles did not prevent primary colonization of textured metal surfaces by strains used in the experiment, but in some cases increases their bactericidal activity.

[The development of Endo growth medium for detection and identification of enterobacteria.]

The purpose of study is to develop a traditional Endo's growth medium with the view of suppression of swarming of Proteus vulgaris and Proteus mirabilis. The object of study was a commercial dry Endo's growth medium. The inoculation of reference strains of enterobacteria and clinical sample (feces, urine, phlegm) was implemented according the normative documents. It is demonstrated that brining into Endo's growth medium tryptophan and sodium salts of bile acids in a particular ratio of their concentrations suppressed swarming of Proteus vulgaris and Proteus mirabilis. The obtained results demonstrated that application of enhanced Endo's growth medium significantly increases its differential and diagnostic characteristics.

Differential DNA Hydroxymethylation in Human Uterine Leiomyoma Cells Depending on the Phase of Menstrual Cycle and Presence of MED12 Gene Mutations.

Using immunofluorescence with specific antibodies, we analyzed DNA hydroxymethylation in uncultured cells from 25 human uterine leiomyomas considering the menstrual cycle phase during surgery and the presence of MED12 gene mutations. It was found that each tumor node had specific DNA hydroxymethylation level that did not depend on the presence of mutations in MED12 gene, but depended on the phase of menstrual cycle. The degree of DNA hydroxymethylation was significantly lower in cells of leiomyomas excised during the luteal phase compared to the follicular phase (p=0.0431). Hormonal status changing at various phases of menstrual cycle is a factor affecting DNA hydroxymethylation in leiomyoma cells.

[NON-ALCOHOLIC FATTY PANCREATIC DISEASE (NAFPD) AND THE METABOLIC SYNDROME: INITIATING FACTOR OR ADDITIONAL COMPONENT?].

Nonalcoholic fatty pancreatic disease (NAFPD) integrates the spectrum of chronic clinical and morphological pancreatic changes: steatosis and nonalcoholic steatopancreatitis. NAFPD prevalence in USA was 27.8%, in China--12.9-16%. According to our data, 51.8% of patients with chronic pancreatitis was diagnosed MS. Association NAFPD with MS has been confirmed in most studies, the presence of any components of MS increases the prevalence NAFPD by 37 %. In the NAFPD pathogenesis is important not only excessive intake of free fatty acids (FFA), which leads to the pancreatic parenchyma inflammation and fibrosis, but also "glucolipotoxicity" (i.e., the combined toxicity of hyperglycemia and increased FFA level) for ß-cells. It is shown that NAFPD is an initial index ofectopic fat deposition, and the earlier manifestation of MS than NAFLD. Most likely, a stage (or degree) of the MS is usefully to determine as the pancreatic status--its exo- and endocrine functions, and fat deposition. This approach will allow us to develop new therapeutic approaches not only to treatment but also to the primary prevention of metabolic syndrome.

[Genetic aspects of pancreatic cancer].

The purpose of the review--to analyze the basic data on modifiable and genetic risk factors of pancreatic cancer (PC). PC is the most fatal disease that kills about 95% of patients. Among the known risk factors for PC only for smoking, obesity, and family history a positive association with the PC risk in meta-analyzes confirmed. The PC etiology remains unclear, more than 90% of patients acquire it sporadically. Currently, the most significant genes for PC include KRAS2, p16/CDKN2, TP53, SMAD4/DPC4. Mutations in the KRAS noted in 90% of cases of pancreatic ducts adenocarcinoma. p16/CDKN2A mutation is accompanied by a 38-fold increased risk of PC compared with the general population. TP53 mutations are associated not only with carcinogenesis but also PC metastasis, as well as SMAD4/DPC4 mutations. Study of the role of genetic aspects in the PC development is necessary both to identify individuals with high PC risk, as well as for the development of gene-specific treatments, such as inhibitors of proteins, histone deacetylase, and histone acetyltransferase (vorinostat, belinostat, entinostat, panobinostat, curcumin) are in clinical trials.

[Pancreatitis, pancreatic cancer and obesity: hypothesis and facts].

THE PURPOSE OF THE REVIEW: Analyze the basic data on the role of obesity in the pathogenesis of pancreatic cancer (PC) and the modern mechanisms of this association. RECENT LITERATURE DATA: In the European Union and in Russia incidence of pancreatic diseases increases, such pancreatic cancer (PC) ranks 10th among cancer diseases. Obesity is a risk factor for not only of severe acute pancreatitis, but also PC at that independently of diabetes. In a meta-analysis the PC risk in obese increased by 47%, while the person with a central obesity have a higher PC risk compared to those with a peripheral type of obesity (odds ratio = 1,45, 95% CI: 1,02-2,07), but association between BMI and PC risk in this Japanese population may be different from that in Western populations, sometimes inversely. The link between obesity and PC is explained by insulin resistance and hyperinsulinemia: was proved a direct correlation between the level of circulating C-peptide and PC, low levels of serum adiponektin and leptin increase the PC risk. There are also genetic risk factors for PC: a statistically significant interaction between IVS1-27777C> and IVS1-23525A>T genotypes of the FTO gene with obesity and the PC risk: AA genotype in patients with BMI < 25 kg/m2 reduced PC risk by 22%-28% (p < 0,0001), and with BMI ≥ 25 kg/m2 was associated with 54%-60% increased PC risk (p < 0,0015). Lifestyle factors (smoking, consumption of saturated fats, etc.) increase the PC risk.

Contrasting of biological samples for X-ray synchrotron microtomography.

The method of contrasting with iodine ions was developed to obtain high-resolution 3D images of large biological specimens using a synchrotron X-ray microtomography unit. It was shown that the samples (late mouse embryos) treated with 50% Lugol solution with addition of 25% ethanol for 48 h followed by a 48-h washout in phosphate buffered saline had maximum contrast and lowest compression artifacts. Processing of samples by this protocol allowed detecting zones of active proliferation. Incubation of brain samples for 120 h in 7.6% meglumine/sodium diatrizoate without washout ensured the best contrast during myelin identification.

[Abnormal development of gonads of dwarf females and low survival of their offspring as the cause of rarity of resident populations of Atlantic salmon (Salmo salar L.)].

Fecundity and mass of eggs, as well as survival of offspring, of hatchery-reared small (less than 406 g) females of Atlantic salmon are determined. The absolute fecundity and mass of egg of artificially reared fish are sufficiently less than that of anadromous spawners. Small females are characterized by delayed spawning; histological analysis has demonstrated significant asynchrony in oocyte development of some specimens. Survival of offspring of dwarf females at the early stages of development is lower than of anadromous females. Our results agree completely with the published data. It is suggested that dwarf females are not able to withstand intraspecific and interspecific competition because of low reproductive ability. This explains rarity of natural populations of dwarf (landlocked) forms of Atlantic salmon.

Specific changes in c-fos expression and colocalization with DNA in identified neuronal nuclei of edible snail following neurotransmitter application.

The effects of serotonin and glutamate on c-fos expression and c-Fos colocalization with DNA were immunohistochemically studied in defense behavior command neurons R-LPa2-3 in snail Helix lucorum. Simultaneous neurotransmitter application resulted in increased c-Fos-immunoreactivity and colocalization with DNA-specific stain Hoechst 33342 in LPa2 and RPa2 neurons with specific dynamics for each identified cell. In the nuclei of LPa3 and RPa3 neurons, neurotransmitter application did not significant change the c-Fos level. These findings are indicative of specific spatiotemporal changes in c-fos expression and c-Fos colocalization with DNA in investigated neurons of edible snail under the influence of neurotransmitters.

5-Bromo-2'-deoxyuridine impairs long-term food aversion memory in edible snail.

We studied the involvement of DNA synthesis into molecular mechanisms of long-term memory. Nucleoside analogue 5-bromo-2'-deoxyuridine (BrdU) is known to incorporate into synthesizing DNA and prevent subsequent DNA replication from this region. To investigate the effect of BrdU administration on long-term memory, terrestrial gastropods edible snails Helix lucorum were trained in the food aversion paradigm. Single-session training (carrot presentation combined with application of 10% quinine solution, three carrot presentations with 10-min intervals) resulted in the formation of long-term memory that persisted for at least 45° days. BrdU administration (250 mg/kg) 30 min before training impaired long-term memory tested 24 h later. Immunohistochemical study revealed BrdU incorporation in the nuclei of identified neurons of defensive behavior.

Sexual dimorphism of mast cells in red bone marrow in normal rats and rats treated with preparation endorfain.

We studied immunological effects of Endorfain preparation. Daily oral administration of the preparation stimulated the immune system increasing migration and proliferation and accelerating differentiation of mast cells in the red bone marrow of rats with the pronounced sexual dimorphism.

DNA methylation patterns of metaphase chromosomes in human preimplantation embryos.

We performed a stage-by-stage study of DNA methylation patterns in metaphase chromosomes from blastomeres of triploid and abnormal diploid human embryos. QFH-banded homologous parental chromosomes differ in their DNA methylation patterns at the metaphase of the 1st cleavage division. Chromosomes of both parental genomes are gradually demethylated at subsequent cleavages, undergoing hemimethylation in 2-cell embryos. At the 4-cell stage hypomethylated chromosomes initially appear and are further registered until the blastocyst stage. The proportion of hemimethylated and hypomethylated chromosomes varies between the blastomeres since the 4-cell stage with no preference for certain chromosomes to be hemi- or hypomethylated demonstrates random segregation of hypomethylated, undermethylated and methylated chromatids during cell cleavage. By the blastocyst stage the chromosomes acquire band- and, thus, chromosome-specific methylation patterns, with 5-methylcytosine-rich DNA preferentially accumulated in R- and T-bands and in the short arms of acrocentric chromosomes. Thus, demethylaton and remethylation of parental genomes of human embryos proceeds in the same manner from the 1st metaphase stage up to the blastocyst. These processes involve all chromosomes and all bands from each chromosome and lead to establishment of chromosome-specific DNA methylation patterns by the blastocyst stage with no differences between homologous chromosomes.

Alterations of cytological and karyological profile of human mesenchymal stem cells during in vitro culturing.

Transplantation of human bone marrow mesenchymal stem cells is considered as a promising therapeutic approach to the therapy of many diseases. However, the problem of possible alterations of the properties of mesenchymal stem cells during their expansion in in vitro cultures before transplantation is not solved. In our study, one of two hundred examined cultures of mesenchymal stem cell cultures derived from donors without bone marrow pathologies and developed under standard culturing conditions demonstrated spontaneous disturbances in morphology, proliferation, and karyotype at early passages. The cells of this abnormal culture retained immunophenotype characteristic of normal mesenchymal stem cells, but some of them (15-25%) had numerous numerical and structural chromosome aberrations.

Enhancement of optical transmission capacity of isolated structures in the brain of mature mice.

We developed an efficient method of optical clarification of isolated structures in the brain of mature mice. The method decreases optical density and opens the way to use novel methods of optical tomography in the study of the brain tissue in mature organism. Specimens dehydrated with 2-butoxyethanol, destained with Dent solution, and cleared in benzyl benzoate:benzyl alcohol or in water solution of iothalamate had the lowest optical density. The clearing effect did not directly depend on the refraction index of the composition. Possible mechanisms of increasing optical transmission of myelinated structures in the brain are discussed.

Reaction of Bcl-2-positive splenic cells to glucocorticoids.

Study of the glucocorticoid effects on the counts of Bcl-2-positive cells in various zones of the spleen showed that dexamethasone and prednisolone stimulated migration of apoptosis-resistant cells to the spleen, but their effects on cell distribution in various morphofunctional zones of the spleen were different. The population of Bcl-2-positive cells is divided into morphotypes, differing by location and reaction to glucocorticoids.

Immunohistochemical detection of the activation of CREB and c-Fos transcription factors in the nervous system of the terrestrial snail induced by pentylenetetrazole.

Phosphorylation of CREB transcription factor and expression of the immediate early gene c-fos in the brains of vertebrates play a key role in the molecular genetic mechanisms of long-term neuronal plasticity. The present study identifies the conditions for immunohistochemical detection of pCREB and c-Fos in the nervous system of the mollusk Helix lucorum (Pulmonata: Stylommatophora); activation of these transcription factors was demonstrated after administration of the convulsive agent pentylenetetrazole. Basal pCREB and c-Fos levels in the central nervous system of intact animals were low. Injection of pentylenetetrazole at a dose of 600 mg/kg evoked characteristic stereotypical motor responses, along with sharp reductions in the phosphorylation of CREB1 and expression of the immediate early gene c-fos, this also occurring in identified neurons. Double immunofluorescent labeling of pCREB and c-Fos showed that expression of c-Fos transcription factor was seen only in pCREB-immunoreactive neurons. These data provide evidence that activation of pCREB and c-Fos transcription factors can be used as molecular markers for mapping the processes of neuronal plasticity in the nervous systems of mollusks.

[Immunohistochemical detection of transcription factors CREB and c-Fos activation in the land snail nervous system induced by pentylenetetrazole].

Phosphorylation of transcription factor CREB and expression of immediate early gene c-fos play a key role in molecular mechanisms of long-term neuronal plasticity in the vertebrate brain. Here, we have defined the procedure of immunohistochemical detection of pCREB and c-Fos in the nervous system of the land snail Helix lucorum (Pulmonata: Stylommatophora) and have shown its activation after the convulsant pentylenetetrazole injection. Baseline pCREB1 and c-Fos levels in the intact snail nervous system appeared to be low. In contrast, injection of pentylenetetrazole (600 mg/kg) produced a rapid induction of CREB phosphorylation and c-Fos expression in a wide range of neurons including a number of identified cells. Double immunofluorescence for pCREB and c-Fos showed that c-Fos was always colocalized with pCREB-immunoreactivity, although the latter had a broader pattern. The results suggest that transcription factors CREB and c-Fos can be used as molecular markers for mapping long-term neuronal plasticity in molluscan nervous system.