Buccal Epithelium Research Articles

Роль полиморфизма rs2075575 гена аквапорина 4 в развитии первичной открытоугольной глаукомы

Актуальность. Несмотря на всестороннее изучение патогенеза первичной открытоугольной глаукомы (ПОУГ), механизмы возникновения заболевания до конца не изучены. В настоящее время определена роль аквапоринов в регуляции внутриглазного давления. Были обнаружены мутации, усиливающие и снижающие функции аквапорина 4, описано влияние различных генетических вариантов аквапоринов на величину внутриглазного давления. Цель исследования. Исследовать частоту встречаемости вариантов полиморфизма rs2075575 (C/Т) гена аквапорина 4 у больных ПОУГ. Материалы и методы. В исследовании приняли участие 101 пациент с ПОУГ и 80 здоровых людей (контрольная группа). Возраст исследуемых колебался от 45 до 87 лет. Средний возраст составил 66 лет. Критерием включения в основную группу служил диагноз ПОУГ развитой, далеко зашедшей и терминальной стадий. Критериями включения в контрольную группу служили возраст старше 60 лет, отсутствие глаукомы и выраженной соматической патологии. ДНК выделяли из буккального эпителия. Полиморфизм гена аквапорина 4 rs2075575 определяли методом полимеразной цепной реакции. Результаты. Установлено, что распределение генотипов у пациентов с ПОУГ отличается от группы здоровых лиц. Обнаружено, что генотип СС среди больных глаукомой встречается в 1,8 раза чаще, чем в контрольной группе. Наоборот, генотип СТ выявляется в 1,5 раза чаще в контрольной группе. Отношение шансов (OR) для генотипа СС равно 2,48 (95% CI 1,30-4,74). У генотипа СТ выявляется протекторная роль, OR =0,52 (95% CI 0,28-0,97). Частота встречаемости генотипа ТТ в исследуемых группах не различается. Выводы. Полиморфный вариант rs2075575 (C/Т) гена аквапорина 4 вносит вклад в риск развития ПОУГ в исследованной выборке. Background. Despite a comprehensive study of the pathogenesis of the disease, the mechanisms of primary open-angle glaucoma are not completely clear. Currently, the role of aquaporins in the regulation of intraocular pressure has been determined. Mutations were discovered that enhance and decrease the functions of aquaporin 4. The effect of various genetic variants of aquaporins on the value of intraocular pressure is described. The aim of research. To investigate the variability of the polymorphism of aquaporin 4 rs2075575 (C / T) in patients with glaucoma. Materials and methods. 101 persons with primary open-angle glaucoma and 80 persons without glaucoma (the control group) were examined. The age of the subjects ranged from 45 to 87 years. The average age was 66 years. The criterion for inclusion in the main group was the diagnosis of primary open-angle glaucoma of a developed, distant and terminal stage. The criteria for inclusion in the control group were age over 60 years, the absence of glaucoma, the absence of pronounced somatic pathology. DNAs were extracted from buccal epithelium. The polymerase chain reaction (PCR) method was used to determine the polymorphism of the aquaporin gene 4 rs2075575. Results. There was a significant difference in the distribution of genotypes in the study and control groups. СС genotype among patients with glaucoma occurs 1.8 times more often than in the control group. CT genotype, on the contrary, is 1.5 times more often in the control group. The odds ratio (OR) for this genotype is 2.48 (95% CI 1.30 - 4.74). The CT genotype reveals a protective role, OR = 0.52 (95% CI 0.28 - 0.97). The genotype of TT in the studied groups is slightly different in frequency of occurrence. Conclusion. The frequencies of gene polymorphisms of aquaporin 4 rs2075575 (C/Т) in patients with primary open-angle glaucoma and healthy were diverse. The likelihood of developing primary open-angle glaucoma is increased in carriers of the СС genotype. Genotype CT play a protective role for primary open-angle glaucoma.

Use of integral indices in the assessment of bucсal cytology in health and in the oral cavity pathology.

Cytological analysis of buccal epithelium (BE) - the buccal cytogram allows to select about 20 parameters reflecting cytogenetic, inflammatory, proliferative and other disorders in epithelial cells used for biomonitoring and laboratory diagnostics. In order to simplify and increase the information content of this study, we conducted a comparative analysis of various integral indices when interpreting the buccal cytogram. 40 practically healthy volunteers, 35 patients with chronic periodontitis, 22 patients with chronic gingivitis and 20 patients with partial loss of teeth were examined. The obtained data revealed an increase in the degree of disorders of proliferation and apoptosis, their ratio as the inflammation increased from gingival tissue lesions to inflammation periodontal tissue. In assessing the condition of the mucous membranes in chronic gingivitis, the apoptosis index turned out to be more informative.In chronic periodontitis the index of cytogenetic disorders is indicative. In this category of patients, the accumulation index of cytogenetic disorders did not have any advantages over the others, and the reparative index was less informative.

Chromosome fragility in the buccal epithelium in patients with Fanconi anemia.

Fanconi anemia (FA) is a rare genome instability syndrome characterized by progressive bone marrow failure and predisposition to cancer, especially head and neck squamous cell carcinoma. Surgical resection is the standard of care for solid tumors, as patients with FA do not tolerate genotoxic chemotherapies or radiation, leading to poor prognosis. It is therefore imperative to develop chemoprevention strategies such as the identification of novel biomarkers to detect the formation of the tumor before its emergence and to use them in clinical trials aimed to counteract genome instability of patients with FA in tissues at risk. Micronuclei (MN) are chromosome fragments that are left behind in anaphase and appear in daughter cells as small additional nuclei. In this work, we analyzed MN frequencies in exfoliated buccal cells from 40 patients with FA and 24 controls. We found that MN frequency was significantly increased in the FA cohort indicating that we can detect chromosome fragility in patients with FA in basal conditions and in a tissue that is divided in vivo. Consequently, the MN assay in exfoliated buccal cells of patients with FA could be used in cancer risk studies and clinical trials aimed to identify cancer chemopreventive drugs.

Transbuccal delivery of benznidazole associated with monoterpenes: permeation studies and mechanistic insights

Benznidazole (BZN) represents the only drug currently available for the treatment of Chagas disease in most endemic countries. When administered orally, high doses are required due to its extensive hepatic metabolism and its toxicity represents the main reason for treatment withdrawals. Because of these complications, transbuccal administration of BZN was investigated. This route avoids the first-pass hepatic metabolism and presents high permeability, with direct access to the systemic circulation. BZN was applied on porcine buccal mucosa after pretreatment with pure eugenol, carvacrol or limonene. Thermal (DSC) and spectroscopic (FT-IR) analyzes were performed to investigate the mechanisms of drug absorption enhancement. The permeability coefficient values of BZN increased 2.6, 2.9 and 4.9-fold after pretreatment with eugenol, carvacrol and limonene, respectively. The lag time, in turn, was shortened in the pretreated samples. The DSC and FT-IR analyzes suggested that transport of BZN through the buccal mucosa is associated with log P and size of monoterpenes. Limonene, the most effective absorption enhancer, contributed to greater interaction with non-polar domains of the buccal epithelium. Overall, BZN showed to be efficiently transported through the buccal route, but in vivo pharmacokinetic studies should be performed to confirm these findings.

Foregut Development and Metamorphosis in a Pyramidellid Gastropod: Modularity and Constraint within a Complex Life Cycle.

Pyramidellids are tiny ectoparasitic gastropods with highly derived feeding structures for piercing and sucking. We attempted to resolve homology controversies about unique pyramidellid feeding structures by examining foregut development through larval and metamorphic stages, using sections for light and electron microscopy. We anticipated that, like many marine invertebrate larvae, post-metamorphic structures would differentiate extensively in late larvae to speed metamorphic transition. Previous studies of gastropods suggested that development of juvenile feeding structures in larvae was facilitated by foregut subdivision into dorsal and ventral developmental modules, and spatial uncoupling of these modules may have facilitated adaptive radiation in neogastropods. Observations of Odostomia tenuisculpta suggested that the stylet may be derived from cuticle-secreting buccal epithelium surrounding the proximal end of the salivary duct, whereas the stylet sheath could be either a derived jaw or a radular tooth. The anterior half of the remarkable buccal pump of these euthyneuran gastropods develops from the larval esophagus, which is unorthodox compared to caenogastropods, where extensive post-metamorphic specialization of a dorsal module component has not been previously described. The introvert tube develops from pouches of the distal larval esophagus and may actually be an eversible oral tube rather than an acrembolic proboscis. Minimal differentiation of presumptive juvenile foregut structures occurred during the larval stage of O. tenuisculpta, when compared to other gastropods. The stylet, stylet sheath, and buccal pump may be incompatible with functioning of the larval esophagus; thus, an explosive period of morphogenesis is necessary at metamorphosis. Although dorsal and ventral modules were recognizable during the development of O. tenuisculpta, we failed to find evidence that this modularity facilitated the extreme evolutionary remodeling of post-metamorphic feeding structures.

Monitoring of cytogenetic instability by micronuclei assay of both immunocompetent and non-immunocompetent cells in tick-borne encephalitis patients depending on variants of glutathione-S-transferase genes in the genotype

Aim of this study was to study the dynamics of the frequency of cytokinesis-blocked T-lymphocytes with micronuclei in peripheral blood and the frequency of buccal micronucleated epithelium cells for a period of half a year in patients with acute tick-borne encephalitis, depending on burden of active and inactive variants of glutathione-S-transferase genes (GSTM1 and GSTT1) in the patient's genotype. We carried out micronucleus assay in immunocompetent and non-immunocompetent cells in 54 patients with acute tick-borne encephalitis and 35 healthy persons (control) residing in the Tomsk and Tyumen regions. To analyze the frequency of cytokinesis-blocked micronucleated T-lymphocytes was used venous peripheral blood as material for phytohemagglutinin-stimulated cultures, and to study the frequency of buccal micronucleated cells, samples of the buccal mucous membrane epithelial cells were obtained. To carried out both techniques of micronucleus assay, cytological preparations were prepared, which were stained using the Giemsa or Felgen methods. The material for the study was obtained repeatedly during admission of patients to treatment, and also after 1 week, 1, 3 and 6 months. Polymerase chain reaction was used to analyze the alleles of the GSTM1 and GSTT1 genes. As a result of this analysis was found a significant increase in the frequency of micronucleated cells in tick-borne encephalitis patients compared with the control group. In addition, the frequency of cytokinesis-blocked micronucleated T-lymphocytes was increased significantly higher than the one of micronucleated buccal cells. The most significant and prolonged increase in the frequency of micronucleated cells was associated with the mutant inactive variants of the genes GSTM1 (0/0) and GSTT1 (0/0). In the patients with burden the inactive forms of these genes, the cytogenetic instability of the cytokinesis-blocked blood T-lymphocytes could persist for up to six months. In case of buccal cells, the frequency of micronucleated cells was close to the one in the control group as early as 1-3 months after a course of treatment. Conclusion. It was found that the most increased and prolonged frequency of cytogenetically instable cells persisted in cytokinesis-blocked T-lymphocytes of peripheral blood of patients with tick-borne encephalitis who were carriers of the genotype with inactive variants of both GSTM1 (0/0) and GSTT1 (0/0 ) glutathione-S-transferase genes.

Loss of Heterozygosity in the Short Tandem Repeat (STR) Loci Found in Tumor DNA of De Novo Diagnosed ALL Patients As a Factor Predicting Poor Outcome

Introduction: Loss of heterozygosity (LOH) was described for many malignancies including leukemia. Using chromosomal microarray analysis (CMA) we have shown preliminary (Risinskaya et al. HemaSphere, 2019, V.3, No S1, P.782) that LOH in STR loci in tumor cells with the normal karyotype represents the loss of extended chromosomal areas and could be associated with uniparental disomy of the entire chromosomal arm or the entire chromosome. Uniparental disomy results not only in the elimination of the wild type allele, but also may lead to the duplication of the mutant allele of certain candidate tumor gene(s). Retrospective analysis of archival samples have shown that in most cases STR loci absent at the relapse of disease were already lost at the onset of disease. We noted LOH at STR loci is especially frequent for relapsed ALL patients and could be used as prognostic factor for possible poor clinical outcome. The aim: To identify LOH in the blast cells of the patients with ALL at the onset of disease and to analyze therapy outcome relative to the patients without LOH. We expect LOH in the de novo ALL to be an unfavorable factor, and those patients possibly should be treated like high-risk group. Patients and Methods: This study includes a comparative analysis of the STR profiles of the DNA of tumor and normal cells from bone marrow samples on a cohort of 32 patients with de novo diagnosed Ph-negative ALL undergoing treatment according to the "RALL-2016" regimen at the National Research Center for Hematology (Moscow, Russia). Inclusion criteria: de novo diagnosed Ph-negative acute lymphoblastic leukemia (ALL) patients, 18-55 years old, intermediate risk group without MLL translocation t(4;11)(q21;q23), treatment by RALL-2016 protocol. Exclusion criteria: other diagnosis, adult patients older than 55 years, MLL translocation t(4;11)(q21;q23), pretreatment or treatment by other protocol. The presence of blast cells in bone marrow samples was confirmed morphologically. DNA was isolated from patient bone marrow samples taken at different disease stages. Control DNA samples were taken from blood of patients in complete remission and/or from buccal epithelium (buccal swab). STR-profiles were assessed by PCR with COrDIS Plus multiplex kit for amplification of 19 polymorphic STR-markers and amelogenin loci (Gordiz Ltd, Russia). The fragment analysis was performed on ABI3130 Genetic Analyzer. The data processing was accomplished using GeneMapper v.4-0 software. A multivariate survival analysis was used to assess the independent impact of the LOH as risk factor against the background of the abnormal tumor karyotype as standard factor for this cohort of patients. We have chosen Failure Free Survival (FFS) as primary endpoint for short-time follow-up and small patient sample size. Death from any reasons, relapse and second leukemia were chosen as failure events, survival time interval starts from begin of treatment. SAS V9/4 was used for statistical analysis of the data. Results: Of the 32 patients six were found with LOH in the certain STR loci (19%). Of these six, two were resistant to therapy and died from disease progression. One patient with LOH is currently in relapse and one patient with multiple LOH has a second leukemia (Fig.1a). Only two from six patients are "Failure Free" now, whereas only three patients from 26 in group without LOH died (two from infectious complications and one from GvHD after BMT). P value < 0,05 (Fig.1b). Conclusions: We have found statistically significant association of clinical failures with the LOH in STR loci measured at the onset of ALL. We assume that LOH is an unfavorable prognostic factor for de novo ALL patients and could be used for risk stratification and choice of adequate therapy implying allogeneic hematopoietic stem cells transplantation or modern innovative therapy approaches. Disclosures No relevant conflicts of interest to declare.

Impact of Polymorphisms in the Serotonin Transporter Gene on Oscillatory Dynamics during Inhibition of Planned Movement in Children.

The serotonin transporter plays an important role in serotonergic neuromodulation. It is known that polymorphisms in the serotonin transporter gene are linked to the dysregulation of emotions. In the current study, we aimed to investigate the impact of variation in the gene encoding serotonin transporter polymorphism on oscillatory dynamics during inhibition of planned movement in the stop signal paradigm. During performance the stop signal paradigm, electroencephalograms (EEGs) were recorded in 90 healthy Caucasian children (39 girls) from 7 to 12 years. Buccal epithelium probes were taken from all participants. The La, Lg, and S alleles of serotonin transporter-linked polymorphic region (5-HTTLPR) polymorphism were detected in the DNA samples using PCR. LaLa genotype carriers did not differ from LaS\\LgS and LgS\\LgLg\\SS carriers in a number of successful inhibitions of a prepotent response. Carriers of LaLa exhibit higher alpha and beta event-related spectral perturbations (ERSP) in parietal and occipital cortices after the presentation of signal of inhibition of movement than LaS\\LgS and LgS\\LgLg\\SS carriers. Results are consistent with current literature and confirm that S allele carriers are more predisposed to disturbances in cognitive control.

Forensic Spermatozoa Detection.

Semen is crucial evidence for some sex crimes, with its sole confirmation being sperm detection. The success of sperm detection is dependent on all levels of preanalytic and analytic procedures. Specimen collection must be performed by well-trained and competent forensic physicians as well as forensic nurses, with preservation done properly before laboratory transfer. Laboratory procedures should consider archival sperm identification, by visualization, with adequate amounts separated from other cells to obtain male DNA profiles. Differential extraction is robust and accepted as the forensic standard but is time consuming and may result in male DNA loss. Thus, alternative methods and microdevices have been developed. Challenges in sperm isolation from vaginal or buccal epithelium mixes and discrimination in multiperpetrator cases have been overcome by single-cell profiling; however, problems inherent in identical twin discrimination and azoospermia have yet to be solved. Epigenetics and future molecular biomarkers may hold the key; therefore, all laboratory processes must consider DNA and RNA protection. Long-term specimen preservation should be done when possible in light of future confirmatory tests.

Preparation, Physicochemical Evaluation and Characterization of Mucoadhesive Buccal Gels Impregnated with Benzydamine Hydrochloride for the Effective Treatment of Aphthous Stomatitis: Effect of Different Grades of HPMC Polymer on In vitro and Ex vivo Performance

Background: Drug delivery across the buccal mucosal epithelium membrane is one of the promising routes to treat various recurrent ailments of the oral cavity. Aphthous stomatitis is an inflammatory oro-mucosal disorder associated with mucous membranes of mouth, cheek, lips, tongue or gingival region. Benzydamine Hydrochloride was designated as a drug of choice by virtue of its anesthetic, antimicrobial, analgesic, anti-inflammatory and antibacterial action. The medication administration through the buccal route is very testing because of restricted absorption area, shorter residence time and movements of the target region. <p></p> Objective: To prepare mucoadhesive buccal gel of Benzydamine Hydrochloride and assess the effects of various HPMC polymer on in vitro and ex vivo performance with the assumption to extend the residence period and achieve maximum drug release in a sustained manner at the target region. <p></p> Methods: Mucoadhesive buccal gels were formulated by utilizing various HPMC grades that served as rate controlling and mucoadhesive polymer. A total of 24 preparations were formulated and subjected to physicochemical evaluation and characterization. <p></p> Results: It was found that the physicochemical parameters varied according to the polymer type and concentration used. Eight formulations were exposed to ex vivo study by virtue of maximum in vitro permeation and mucoadhesion properties. Because of higher ex vivo drug permeation and mucoadhesion, F24 was considered as the final optimized formulation. FTIR and DSC established compatibility between the drug and excipient. The amorphous nature of the drug within the optimized formulation was further unveiled by XRD study. <p></p> Conclusion: The developed buccal gel has a great prospective in contrast to marketed conventional preparation for treating aphthous stomatitis.

The study of the frequency of buccal epithelium cells with nuclear in athletes in dependence on the place got in the competition

Cytogenetic homeostasis ofathletes involved in hand-to-hand armyfighting, depending on the outcome of the fight and the level of aggressiveness has been studied. Status of the genetic apparatus in athletes was evaluated in buccal epithelium with the use of micronucleus test, which is widely usedfor the determination of the influence of various factors on the genetic stability of the organism. Psychological testing of athletes was executed with the use of Spielberger’s State-Trait Anger Expression Inventory scale, Bass-Darky questionnaire for the diagnostics of aggression and hostile reactions, Eysenck Personality Questionnaire test for the evaluation ofpsychic conditions and method of U.P. Ilyin and P.A. Kovalev for the identification of the propensity to conflictness and aggressiveness (positive and negative) as the personal characteristics. The place taken by the athlete on the competition was shown to have an impact on the frequency of buccal epithelium cells with nuclear abnormalities (micronuclei, perinuclear vacuoles, notches, protrusions "tongue" and "broken egg”). Losers have nuclear aberrations more than winners. At that in athletes with higher aggressiveness and its attendant psychological characteristics the influence of the outcome of the competition on the number of cells with aberrations core is more pronounced.

Cytogenetic indices of buccal epithelium in schoolchildren residing in territories with different levels of the air pollution

In the article there are contained the results of the study of ambient air of the regional center, urban-type settlement, and buccal epithelium in 87 schoolchildren aged of 11-13 years. Schoolchildren residing in the regional center were shown to be more susceptible to the exposure to such toxicants as carbon monoxide, nitrogen oxide, fluorhydric acid, chlorine and its compounds, sulfur dioxide, formaldehyde contained in the ambient air. In this group of students if compared with students who live in the village, there were revealed more pronounced cytogenetic deteriorations in oral mucosal epitheliocytes: cells with micronuclei, different forms of protrusion of the core, dual-core cell mitosis, kariokynesis, apoptotic bodies significantly more common occur, that can speak about both not only of cytogenetic instability, but and the degree of environmental genotoxicity, in particular, ambient air. There was noted the tendency to increase the level of cytogenetic damage and destruction of the nucleus in the group of girls in comparison with boys.

Evaluation of antimutagenic properties of powdered malt and polymalt extracts with the use of micronucleus test

There were performed studies of the occurrence of cells with pathologies in the buccal epithelium of volunteers who consume drinks based on mixtures ofpowdered malt and polymalt extracts of buckwheat, peas, corn and barley. There was shown their impact on the stability of the genetic material of examined cases. There was established activation of apoptosis, which leads to the elimination of cells with cytogenetic deteriorations. Poliymalt extracts possess protective properties, contribute to the suppression ofprocesses offormation of cells with genetic disorders (micronuclei (from 4.38 ± 0.67 %%, up to 2.53 ± 0.39 %% after intake), protrusions (from 1,98 ± 0,42 %%, up 0,85 ± 0,25 %% after intake), incisures (from 3.34 ± 0.44 %%, up 2.17 ± 0.35 %% after intake), two cores (from 1.63 ± 0.26 %%, up 0.65 ± 0.21 %% after intake) and rid the body of aberrant cells, as evidenced by the increase in the number of cells with karyolysis (up to 5.98 ± 0,91 %%, up 9.55 ± 1.74 %% after intake), karyopyknosis (from 10.71 ± 0.90 %%, up to 11.97 ± 0.85 %% after intake) and perinuclear vacuoles (from 9.24 ± 1.63 %%, up to 12.94 ± 2.57 %% after intake). In women, anti-mutagenic effects ofpolymalt extracts are more pronounced than in men. Antimutagenic effects of extracts can be explained by the properties of contained in them B vitamins and sulfur-containing amino acids (cysteine and methionine).

Cytogenetic status of the residents of the Gydansky Peninsula (Gydan)

The relevance of the study on the Gydansky Peninsula lies in poor knowledge and inaccessibility of the territory, planned intensive industrial development of the Gydansky Peninsula, in 2011 there were received licenses for the exploration of license areas at the peninsula up to 2031. Industrial development will inevitably lead to certain environmental shifts, emission of the harmful substances into biosphere, the accumulation of anthropogenic pollutants in soil and water sources. The proposed development of the territory of the Peninsula Gydan sets the task of assessment of the impact of gas and oil production in conditions of the far North on health, as well indigenous persons as employees recruited to this of the region. One of the informative approaches to the assessment of population health is the assessment of the cytogenetic status with the use of noninvasive analysis of buccal epithelium. The aim of the study is to determine the cytogenetic status of the inhabitants of the village of Antipayuta of the Yamal-Nenets Autonomous Okrug for the assessment of the impact of environmental factors on the health of the population at the present stage of the industrial development of the territory. Samples of buccal epithelium of 81 alien and indigenous people of the Yamal-Nenets Autonomous district were the object of the investigation. There was performed the analysis of indices of cells of buccal epithelium of the residents living in the village in comparison with the control group. The analysis of samples was performed on a Nikon Eclipse E100 microscope. For the assessment of the cytogenetic status of the individual there was used the proposed by Sycheva L. P. (2012-Index of accumulation of cytogenetic damages (Iac). It is shown that the frequency of occurrence of micronucleus and nuclear protrusions does not exceed the performance of the control group. The index of accumulation of cytogenetic damage for the population of the village is 0.78±0.07% and corresponds to a low level. The analysis of indices of buccal epithelial cells of residents living in rural areas was performed in the comparison with the control group The prevalence rate of cells with micronuclei and protrusions were shown to account of 14.6% andfail to exceed indices in the control group. Accumulation index of cytogenetic damages for the village population is 0.78 ± 0.07 % and corresponds to the low level. The amount of cells with micronuclei and protrusions in the current study is by 14.6% lower than the average of the control group. The intensity of proliferative processes and apoptosis in the comparison groups occurs evenly.

ВОЗРАСТНАЯ ДИНАМИКА СТОМАТОЛОГИЧЕСКИХ И ЛАБОРАТОРНЫХ ПОКАЗАТЕЛЕЙ ЗДОРОВЬЯ У ДЕТЕЙ 5-6, 12, 15 ЛЕТ ГОРОДА ЕКАТЕРИНБУРГА

Subject. The health status of children and adolescents is one of the most acute medical and social problems. It is known, that with dental diseases, changes occur not only in the immunological profile of the oral fluid, but also in morphology of the oral tissues. New approaches to the traditional cytological study of buccal epithelium such as analysis of the cytogram with the isolation of various types of cells, as well as the detection of cytological abnormalities of cells, allows us to evaluate the reactivity of the oral mucosa in pathological processes. According to WHO recommendations (2013), groups of children 5-6, 12, 15 years of age are the global indicator age groups for monitoring disease trends and comparisons on an international scale.
 The objective of the study is to assess the health status of children age 5-6, 12, 15 with non-invasive methods. It is based on the results of a clinical and laboratory examination of 179 children, attending organized children's groups. Children underwent a comprehensive dental examination, which included a questionnaire according to the WHO method, an external examination of the maxillofacial region, an examination of the oral cavity, identification of pathology of hard tooth tissues. 
 Methodology. We studied the change in the dental status of patients, indicators of oral fluid and basal epithelium with age, in order to prognostically use non-invasive assessment methods in a comprehensive health examination, planning and evaluating the effectiveness of prevention programs. 
 Results. It was noted that the dental health status of children 5-6, 12, 15 years old can be assessed as satisfactory, while dental, laboratory and cytological health indicators worsen with age. Non-invasive methods for assessing the dental status of patients can be used in a comprehensive examination of children's health, planning and evaluating the effectiveness of prevention programs.

Nano-enabled liposomal mucoadhesive films for enhanced efavirenz buccal drug delivery

Buccal films (BFs) were prepared using a solvent casting method using the liposomal suspension as the dispersing medium. Optimization of some physical properties of the films containing different amounts of polymers was done using digital Vernier calliper and digital weighing balance. The physiochemical properties of the best optimized properties were characterized using Differential scanning calorimetry (DSC), X-ray diffraction (XRD), Fourier transfer infrared spectroscopy (FTIR), Transmission Electron Microscopy (TEM), Energy dispersive X-ray spectroscopy (EDS), and Scanning Electron Microscopy (SEM). Permeation study of the BFs composed of Carbopol (CP) alone and CP to Pluronic 127 (PF127) demonstrated better bio-adhesive properties than the films made of other polymers such as HPMC (hydroxyl propyl methyl cellulose) and HPMC-PF127. These CP based BFs (without and with PF127) exhibited good film thickness 0.88 ± 0.10 and 0.76 ± 0.14 mm, with weight uniformity 68.22 ± 1.04 and 86.28 ± 2.16 mg, satisfactory flexibility values 258 and 321, and slightly acidic pH 6.43 ± 0.76 and 6.32 ± 0.01. The swelling percentage was found to be 50% for CP and 78% for CP-PF127. The cumulative amount of drug that permeated through the buccal epithelium after 24 h was about 66% from CP and 75% from CP-PF127.

Buccal epitelium cytogenetic status in schoolchildren living in high and middle latitudes.

The Russian northern regions development is associated with the extraction and processing of natural resources, which leads to environmental pollution and makes the task of sanitary-hygienic monitoring relevant. Buccal micronucleus cytome assay is one of the toxicological methods for human population studies. Studies on the inter-latitudinal comparison of Buccal micronucleus cytome assay are lacking, therefore there are difficulties in comparing the results obtained in the Russian Arctic with data from more southern regions. The aim of this study is to compare cytogenetic abnormalities in the buccal epithelium in two groups of older schoolchildren living in high and middle latitudes. The study was conducted in the city of Apatity (Murmansk region, 67 ° 34'03 ″ N, 33 ° 23'36 ″ E) and the city of Serpukhov (Moscow region, 54 ° 54 '56 ″ N, 37 ° 24 '40 "E). A total of 61 children were examined: 41 children from the Apatity and 20 children from the Serpukhov (16-18 years old). The Buccal micronucleus cytome assay was carried out according to an international protocol. It was shown that the average frequency values of the cells with micronucleus in the comparison groups of schoolchildren living in high and middle latitudes did not significantly differ and did not exceed the values for the average population norm. The frequency of cells with nuclear buds and two nucleus was significantly higher in the group of schoolchildren living in middle latitudes, which, in turn, is compensated by a higher rate of elimination of cells with impaired. Therefore, when comparing Buccal micronucleus cytome assay data, it is quite possible not to take into account the breadth of the studied groups.

The Baseline Level of Cytogenetic Damage in Lymphocytes and Buccal Epitheliocytes of Lung Cancer Patients

The results of the study of chromosomal aberrations and micronuclei in blood lymphocytes, as well as micronuclei in buccal epithelium cells of 50 untreated men with lung cancer (LC) and 62 control donors, are presented. Using a variety of cytogenetic tests, a significant unidirectional increase in the base level of genetic instability in the cells of the patients was detected compared to the control. It was shown that, in LC patients, the elimination of damaged cells occurs predominantly by necrosis, whereas the frequencies of cells at the apoptosis stage do not differ in patients and control. The age of the patient, smoking status, type, and stage of LC do not have a significant impact on the performance of the baseline level of genetic damage. This fact suggests that the cytogenetic instability of the somatic cells of untreated LC patients is a constant sign that characterizes the genome response to the effect of the oxidative stress accompanying the neoplastic process.

Unidirectional drug release from 3D printed mucoadhesive buccal films using FDM technology: In vitro and ex vivo evaluation

Oromucosal delivery of active pharmaceutical ingredients provides an attractive alternative route of administration, due to avoidance of the first pass effect and improved patient compliance. In the current work, fused deposition modelling (FDM) 3D printing was investigated as an additive manufacturing approach for poly(vinyl alcohol)-based mucoadhesive films, enabling unidirectional drug release. For this purpose, chitosan was incorporated as a permeation and mucoadhesion enhancer whereas ethylcellulose and commercial wafer sheets were evaluated as backing layers. The formulated films were initially assessed for structural integrity and dose uniformity. Solid-state characterization of the films, including thermal methods (DSC, TGA), diffraction (XRPD) and Raman spectroscopy, was implemented to characterize the physicochemical properties of the produced polymeric filaments and buccal films. The mechanical properties of the products were investigated by instrumented indentation and tensile tests. Evaluation of buccal films was assessed in vitro, to study the effect of backing-layer type on hydration capacity of the films, diffusion of the drug throughout the restricting layer and release profiles in simulated saliva. The ex vivo performance of the manufactured products, associated with the presence of chitosan, was investigated by textural analysis for mucoadhesion properties, whereas permeation studies and histological studies were performed across porcine buccal epithelium. The results demonstrated that FDM printing is a timesaving and versatile approach in the context of manufacturing multi-layered mucoadhesive buccal films, providing unidirectional release properties.

Expression of galectin 9 mRNA in lactose maldigestion and children’s obesity

OBJECTIVES AND STUDY: To investigate the association of mRNA expression of galectin - 9 (Gal - 9) and lactose maldigestion in children’s obesity with different genotypes of the lactase gene (LCT).METHODS: The study involved 85 children with obesity (BMI>97th percentile) aged 6-18 years. The study defined genotypes of LCT (material for investigation - venous blood), included expression of Gal - 9 mRNA (material for investigation - buccal epithelium) by real-time polymerase chain reaction analysis, and utilized the study of lactose maldigestion by Hydrogen breath testing. The first group of observations was presented by 44 children with genotype C/C -13910, the second group consisted of 41 children with phenotypically identical genotypes C/T -13910 and T/T -13910, p>0.05.RESULTS: Lactose maldigestion in children with genotype C/C -13910 averaged 36.05±4.73 ppm, in children with genotypes C/T -13910 - 22.61±4.1 ppm (p<0.05) and with genotype T/T -13910 - was absent (p<0.05). The average expression level of Gal - 9 mRNA in children with genotype C/C -13910 was 564.6±35.8 relation units (RU) ∆mRNA Gal - 9/mRNA actin and was 61.02±15.8 RU ∆mRNA Gal - 9/mRNA actin, p<0.01 in children with genotypes C/T and T/T -13910. The lowest mean level of expression of Gal - 9 mRNA (42.47±13.4 RU ∆mRNA Gal - 9/mRNA actin) was recorded in the subgroup of children with genotype C/C -13910 and lactose maldigestion (n=22). Whereas the largest mean level of expression of Gal - 9 mRNA was recorded in the subgroup of children with the C/C -13910 and without lactose maldigestion (n=22) - 1086.73±51.2 relation units ∆mRNA Gal - 9/mRNA actin, p<0.01.CONCLUSION: The expression level of Gal - 9 mRNA depends on lactose maldigestion in children with genotype C/C -13910