Buccal Epithelium Research Articles

ХАРАКТЕРИСТИКА РОТОВОЙ ЖИДКОСТИ И БУККАЛЬНОГО ЭПИТЕЛИЯ В ОЦЕНКЕ НЕПЕРЕНОСИМОСТИ КОНСТРУКЦИОННЫХ МАТЕРИАЛОВ

Introduction. The phenomena of intolerance to structural materials often occur in patients using removable dentures made of acrylic plastics, in this regard, the problem of intolerance to structural materials does not lose its relevance. However, today, the lack of knowledge in diagnostics forces us to delve into the search for diagnostic criteria. The current level of development of dentistry determines the need to search for diagnostic criteria that can reliably identify patients with a high risk of developing intolerance to structural materials (SIM) and predict the likelihood of its occurrence.
 This study is devoted to assessing the effectiveness of laboratory diagnostics in case of intolerance to structural materials.
 The aim is to evaluate the effectiveness of some methods of laboratory diagnostics in identifying non-allergic (toxic) etiology of intolerance to structural materials.
 Methodology. 22 patients were examined, of which 14 healthy volunteers and 8 patients with signs of intolerance to structural materials can be distinguished. An analysis of buccal cytograms (buccal epithelium) was carried out, and a study of the oral fluid was also carried out (a study of dry chemistry).
 Results. When studying the buccal cytogram in the main group, an increase in the apoptosis index (1.12) and a decrease in the reparative index (0.67) were found in patients with intolerance to structural materials. When studying the oral fluid (physicochemical study), signs of minimal inflammation activity were revealed in the main group of patients. We observe a decrease in pH (6), an increase in total protein (0.5 g/L) and an increase in leukocytes (65).
 Conclusions. The data obtained indicate that in patients with intolerance to construction materials, there are signs of minimal inflammation when examining the oral fluid, impaired reparative processes and an increasing index of apoptosis when examining the buccal cytogram.

Vegetative, metabolic and immune accompaniments of changes in the electrokinetic index of the buccal epithelium under the influence of therapeutic factors

Background. In previous studies, we have shown that electrokinetic index of buccal epithelium (EKI) correlated with some functional and metabolic parameters. Subsequent studies have shown that сhanges in EKI correlated with changes in some parameters of EEG, HRV, hemodynamics, metabolism, immunity and fecal microbiocenosis. Further research in this direction was continued on a significantly increased contingent of patients and with the involvement of new methods and factors of influence. This message starts the presentation of the obtained results. Material and methods. Under a observations were 44 men (49±15 years) and 30 women (51±13 years) without clinical diagnosis or with chronic pyelonephritis in the phase of remission (23 men). We registered caused by the various therapeutic factors changes in EKI, state of the vegetative and hormonal regulation as well as immunity and metabolism, then calculated relationships between changes. Results. In 49 patients the changes in EKI were in the range of ±2,5%, in 19 people EKI increased by more than 2,5% (M±SD=+4,0±1,6%), while in 9 people decreased by more than 2,5% (-4,2±1,7%). The canonical correlation between changes in EKI, on the one hand, and HRV and immunity parameters, on the other, is moderate: R=0,478; p=0,023. The method of discriminant analysis revealed 10 immune and 6 HRV parameters as well as triglycerides and cholesterol, whose changes are characteristic of multidirectional changes in EKI. Conclusion. Electrokinetic index of buccal epithelium responds to therapeutic factors in different directions, accompanied by characteristic changes in a number of parameters of HRV, immunity and metabolism.

Investigation of the relief geometry of the membrane of human buccal epithelium cells using atomic force microscopy methods

The analysis of the shape of the Brownian relief of the surface of human buccal epithelium cells was carried out using the method of fractal geometry. The scaling coefficients eta, zeta and the fractal dimension Df were used to estimate their actual areas Sc(fact) and linear characteristics lf. Keywords: atomic force microscopy, buccal epithelium, fractal number.

The impact of metabolic disorders on ageing rates in patients with moderate cardiovascular risk

Introduction. Prediction of cardiovascular risk (CVR) and the effectiveness of prevention of metabolic-associated diseases (MAD) is a serious challenge of modern medicine. Recently, for this purpose, it has been proposed to estimate the biological age (BA) of a person for the timely determination of premature aging. Calendar age (CA) is an important risk factor for the development of age-related diseases, including cardiovascular disease (CVD). Biological age (BA) may be more accurate for assessing CVR and predicting early vascular ageing and the development of atherosclerosis. The available data allow us to testify about the relationship between premature aging and the most significant pathogenetic factors of MAD. Purpose: to assess the impact of metabolic disorders on biological age in patients with moderate cardiovascular risk and the significance for the development of premature ageing. Materials and methods. 120 patients with moderate CVR (by SCORE) aged from 32 to 59 years (women n=65, 54.17%) were involved in the examination. They were divided into the main group (n= 67) who additionally had metabolic disorders (overweight/obesity, insulin resistance, dyslipidemia) and a comparison group (n=53) without metabolic disorders. All included patients underwent anthropometric studies with calculation of body mass index (BMI), studies of indicators of carbohydrate metabolism and lipid metabolism. Determination of biological age was carried out according to the methods of A.G. Gorelkin and B.B. Pinkhasov and P.V. Voytenko. Telomere length (TL) of blood leukocytes and buccal epithelium was determined by real-time PCR. Statistical data processing was performed using the Statistica program package, version 10.0. Results and discussion. The BA of normal-weight patients not only did not exceed the CA, but was also lower in both the main group and the comparison group. At the same time, in the overweight patients BA did not differ from CA in both groups, and the presence of obesity led to an acceleration of the rate of ageing in patients of the main group. It was determined that an increase in BMI and waist circumference (WC), hyperinsulinemia, hyperuricemia, and high levels of proatherogenic lipids are associated with an increase in BA regardless of the presence of metabolic disorders and may influence the ageing rate. Also, the presence of overweight and obesity is associated with a decrease in TL and an increase in BA. Conclusions. Assessment of premature ageing allows to identify persons with a high risk of developing age-related diseases at a time when there are no clinical symptoms yet, and to develop measures for timely and effective prevention.

Human buccal epithelial cell envelope reactions to external micromechanical stimuli

The reaction of the shell of an adult living cell of human buccal epithelium to an external micromechanical stimulus was studied. The dependence of the magnitude of the reaction on the level of passive or active stimulation, namely, the level of force exposure, was established. It is shown that the cell can exhibit both active and passive reactions to an external micromechanical stimulus, as well as control the level of exposure to adhesive forces. Keywords: human buccal epithelium cell, shell, micromechanical stimulus, shell elasticity, AFM force spectroscopy.

Исследование геометрии рельефа оболочки клеток буккального эпителия человека методами атомно-силовой микроскопии

The analysis of the shape of the Brownian relief of the surface of human buccal epithelium cells was carried out using the method of fractal geometry. The scaling coefficients ,  and the fractal dimension Df were used to estimate their actual areas Scfact and linear characteristics lf.

BUCCAL CELL MICRONUCLEI AMONG PATIENTS WITH ORAL LEUKOPLAKIA.

The aim: The purpose of research was to evaluate the efficiency of micronucleus test in buccal cells for the diagnosis of oral leukoplakia. Materials and methods: We have conducted a comprehensive clinical and laboratory examination of 155 patients with oral leukoplakia. It was conducted histological examination leukoplakia mucosal sites, to assess the buccal epithelium cell micronucleus test was carried out. Results: Histological evaluation of the material was made according to the classification of leukoplakia WHO (2005). They are established 10 (14%) sites unmodified mucosa, 10 (14%) of the samples hyperkeratosis without atypia, 14 (19%) biopsies hyperkeratosis SIN1, 15 (21%) of hyperkeratosis SIN2, 10 (14%) -- SIN3 and 13 (18 %) of the cases of squamous cell carcinoma. Micronuclei, whose appearance is caused by violation of differentiation of epithelial cells, were found in patients with leukoplakia, the detection of micronuclei almost equally high as in patients with leukoplakia SIN2, and with SIN3 (a difference of 1.3 times (p <0,05, rxy = + 0.271)), and consequently the probability of occurrence of tumoral diseases of the oral mucosa or malignancy existing large. Conclusions: Thus, on the background of the general increase in proliferative activity of epithelial cells with increasing SIN, for each treatment group revealed the appearance of micronuclei in buccal cells. And the frequency of micronuclei and the fourth type of increases with hyperplasia, indicating an increase in the likelihood of malignancy and cancer of the oral mucosa in patients with leukoplakia SIN3.

Orcadian rhythm of melatonin receptor expression in the buccal epithelium of pregnant women and their newborn children: development of molecular markers for prognosis of delayed postnatal pathology

Orcadian rhythm of melatonin receptor expression in the buccal epithelium of pregnant women and their newborn children: development of molecular markers for prognosis of delayed postnatal pathology

Реакции мембраны клеток буккального эпителия человека на внешние микромеханические раздражители

The reaction of the shell of an adult living cell of human buccal epithelium to an external micromechanical stimulus was studied. The dependence of the magnitude of the reaction on the level of passive or active stimulation - the level of force exposure was established. It is shown that the cell can show both active and passive reactions to an external micromechanical stimulus, as well as control the level of exposure to adhesive forces.

In Vitro Culture Expansion and Characterization of Buccal Mucosal Epithelial Cells for Tissue Engineering Applications in Urethral Stricture After Transportation Using a Thermoreversible Gelation Polymer.

Introduction: The transportation of tissues from hospitals to clinical laboratories for cell therapy is an essential component of regenerative medicine. Previously, we used laboratory-cultured mucosal cells from buccal epithelium expanded and encapsulated using a scaffold-hybrid approach to the urethral stricture (BEES-HAUS) procedure. In this study, to improve the outcomes, we compared the thermoreversible gelation polymer (TGP) transportation procedure with conventional culture methods, and reported its advantages. Methods: Human buccal mucosal tissues in Phase I of the study were transported in Euro-Collins solution (ECS) and the cells obtained were cultured in two-dimensional (2D) Dulbecco's modified Eagle's medium (DMEM), CnT-Prime epithelial 2D differentiation medium (CnT-PR), and a three-dimensional (3D)-TGP scaffold. In Phase II, tissues were transported in a TGP cocktail and the ECS. The cells were cultured in 2D-DMEM and 3D-TGP, quantified, and characterized by immunohistochemistry. Results: The cells in 3D-TGP culture maintained epithelial morphology in a better manner compared with 2D-DMEM, in which they developed fibroblast-like morphology. The TGP-transported cells grew rapidly. Immunohistochemical analysis results for AE1/AE3, EGFR, integrin-β1, p63, and p75 were intensely positive in 3D-TGP. Conclusion: The TGP-based cocktail used in human buccal tissue transportation yielded cells with better morphology maintenance. The TGP scaffold provides an optimal in vitro environment wherein epithelial cells better maintain their native phenotype compared to those cultured through conventional methods. These results suggest using TGP for the transportation and culture of human buccal tissues for clinical applications. In addition, the use of a TGP-based cocktail for the transport of other tissues for regenerative medicine applications is worth further analysis.

“Rapid DNA” ― new horizons of forensic DNA profiling using full cycle genetic analyzers

BACKGROUND: The study is a part of a comprehensive research conducted at the Russian Center of Forensic Medical Expertise of the Ministry of Health of the Russian Federation with the ultimate goal of development and implementation of new effective technologies for molecular genetic identity testing and kinship analysis in the realm of national forensic expertise.&#x0D; AIMS: The objective of the study is to evaluate the possibilities of the Rapid DNA technology in application to forensic DNA analysis. One may expect that this technology in the future will ensure the creation of an operational and highly effective procedure for forensic DNA profiling.&#x0D; MATERIAL AND METHODS: The methodical basis of the work is the genotyping of polymorphic STR-loci of chromosomal DNA in biological material using high-tech fully automated (full cycle) analytical platform RapidHIT: namely, RapidHIT 200 (Integenx, USA) and RapidHIT-ID (Life Technologies/Thermo Fisher, IntegenX (USA) instruments.&#x0D; RESULTS: Buccal epithelium and blood samples, while being the most common in a forensic DNA testing, have been examined as a validation test objects. Then experiments were carried out with real expert material such as cigarette butts, hair, nails, bones, chewing gum; swabs from different surfaces were examined as test samples as well.&#x0D; CONCLUSIONS: Based on the data obtained, it can be stated that the Rapid DNA technology as it is implemented in the full-cycle genetic analyzers IntegenX RapidHIT 200 (Integenx, USA) and RapidHIT ID (Thermo Fisher, IntegenX, USA), according to the criteria studied, in general supports the idea of to be introduced into the practice of forensic DNA analysis.

Histopathological Features and Protein Markers of Arrhythmogenic Cardiomyopathy.

Arrhythmogenic cardiomyopathy (ACM) is a heritable heart muscle disease characterized by syncope, palpitations, ventricular arrhythmias and sudden cardiac death (SCD) especially in young individuals. It is estimated to affect 1:5,000 individuals in the general population, with >60% of patients bearing one or more mutations in genes coding for desmosomal proteins. Desmosomes are intercellular adhesion junctions, which in cardiac myocytes reside within the intercalated disks (IDs), the areas of mechanical and electrical cell-cell coupling. Histologically, ACM is characterized by fibrofatty replacement of cardiac myocytes predominantly in the right ventricular free wall though left ventricular and biventricular forms have also been described. The disease is characterized by age-related progression, vast phenotypic manifestation and incomplete penetrance, making proband diagnosis and risk stratification of family members particularly challenging. Key protein redistribution at the IDs may represent a specific diagnostic marker but its applicability is still limited by the need for a myocardial sample. Specific markers of ACM in surrogate tissues, such as the blood and the buccal epithelium, may represent a non-invasive, safe and inexpensive alternative for diagnosis and cascade screening. In this review, we shall cover the most relevant biomarkers so far reported and discuss their potential impact on the diagnosis, prognosis and management of ACM.

ASSESSMENT OF FS-1 GENOTOXIC EFFECT ON THE BUCCAL EPITHELIUM OF RABBITS

Antimicrobial activity of the drug FS-1 is due to the content of iodine halogen, which is quite active. Currently, potassium triiodide-based preparations, such as Lugol's solution, Iodopyron, Betadine and others, are already used as antiseptics. Studies have already been conducted confirming the ability of FS-1 to increase the permeability of the bacterial cell membrane, which allows antibiotics to act more efficiently, and in vitro and in vivo studies have been conducted to evaluate the cytotoxicity and genotoxicity of FS-1 in laboratory animals with systemic exposure. In this experiment, effect of FS-1 on the site of primary contact, the buccal epithelium, was studied. Based on the experiment results, we could conclude about the possible use of FS-1 as an oral antiseptic. Three rabbits were used in the experiment, buccal epithelium smears of which were taken before the administration of 4 mg/kg FS-1 (control samples) and after the administration of FS-1 (experimental samples). Total period of FS-1 daily administration was 14 days, which is a period for self-renewal of the epithelium. Obtained smears of the cells were examined under a microscope for the presence of cytogenetic aberration. As a result, there was no significant change after administration of FS-1, since the level of cells with karyolysis, karyorrhexis, micronuclei, and nuclear protrusions remained at about the same level. Therefore, it was concluded that FS-1 does not cause cytogenotoxic effect upon prolonged exposure to rabbit buccal epithelium.

Preparation, Characterization and Evaluation of Guar Films Impregnated with Relaxing Peptide Loaded into Chitosan Microparticles

Background: Biological molecules such as therapeutic proteins and peptides have provided ample opportunities for further improvements in health and wellbeing. However, such biomacromolecules face stability challenges regarding premature processing and digestion, whenever any enteric delivery route is considered. Hence, oral guar films entrapping peptide-containing chitosan microparticles have been developed as a new delivery system to carry and protect therapeutic relaxing peptides. Results: The composite films were characterized regarding physicochemical properties using FTIR as well as peptide encapsulation efficiency, cytotoxicity, and peptide permeability through buccal and intestinal cell-based models. The composite films with 0.18% (w/w) of peptide demonstrated ca. 0.63% of peptide loading efficiency into the chitosan microparticles and to be safe for buccal and intestinal epithelial cells up to 24 h of contact. The permeability assays through TR146 cells to mimic buccal mucosa epithelium and Caco-2/HT29-MTX to mimic duodenum conditions showed that the incorporation of peptide chitosan microparticles into guar-gum oral films offered a faster permeability, when compared with the free peptide and peptide-chitosan-microparticles-loaded chitosan. Conclusion: This study suggests that peptide-loaded chitosan microparticles into guar-gum oral films might be a promising carrier for enhancement of the buccal and intestinal absorption of a relaxing peptide.

On the markers for schizophrenia

Summary. The study of various markers indicating an imbalance in both the neurotransmitter and immune systems observed in patients with schizophrenia is currently of great scientific and practical interest. These studies allow us to deepen knowledge about the etiology and pathogenesis of schizophrenic spectrum disorders, to facili-tate the classification and conduct of differential diagnosis in complex diagnostic cases. The article presents the results of laboratory studies of DMT-1, CD4, CD20, CD34 and dopamine in 24 people suffering from paranoid schizophrenia, and 14 healthy individuals. The aim of the study was to determine DMT-1, CD4, CD20, CD34 and dopamine as possible markers for the diagnosis of schizophrenia using the minimally invasive method.Materials and methods: Buccal epithelium was taken in 2 groups of patients suffering from paranoid schizophrenia receiving and not receiving therapy with psychotropic drugs. The third group consisted of healthy volunteers. The samples were studied by the immunocytochemical method followed by visualization using scanning laser confocal microscopy and morphometric analysis.Results and discussion: : It was revealed that the markers studied were found to be associated with the nature of mental disorder, but not with the influence of psychotropic drugs used in it. Conclusion: The results of this study may indicate the possibility of minimally invasive diagnosis of schizophrenia using this method; however, the authors note that the results of the study should be considered preliminary. It is necessary to expand the number of patients and compare them with groups of patients suffering from other mental disorders, which is certainly important for assessing the adequacy of the proposed method and the possibility of its use in differential diagnosis.

Electron Spin Resonance Evaluation of Buccal Membrane Fluidity Alterations by Sodium Caprylate and L-Menthol.

The ability of sodium caprylate and l-menthol to fluidize phospholipid bilayers composed of lipids simulating the buccal epithelium was investigated using electron spin resonance (ESR) to evaluate the action of these agents as permeation enhancers. 5-Doxyl stearic acid (5-DSA) and 16-doxyl stearic acid (16-DSA) were used as spin labels to identify alterations in membrane fluidity near the polar head groups or inner acyl regions of the lipid bilayer, respectively. The molecular motion of both 5-DSA and 16-DSA showed increased disorder near the polar and inner hydrophobic regions of the bilayer in the presence of sodium caprylate suggesting fluidization in both the regions, which contributes to its permeation enhancing effects. L-menthol decreased the order parameter for 16-DSA, showing membrane fluidization only in the inner acyl regions of the bilayer, which also corresponded to its weaker permeation enhancing effects. The rapid evaluation of changes in fluidity of the bilayer in the presence of potential permeation enhancers using ESR enables improved selection of effective permeation enhancers and enhancer combinations based on their effect on membrane fluidization.

Вплив поліморфізму гена інтерлейкіну-10 на перебіг муковісцидозу в дітей

Актуальність. Муковісцидоз (МВ) — генетичне захворювання, перебіг якого залежить не тільки від мутації гена трансмембранного регулятора МВ, але й від інших генів-модифікаторів. Ген інтерлейкіну-10 (ІЛ-10) входить до переліку таких модифікуючих факторів. Мета. Удосконалення медичної допомоги хворим на МВ шляхом уточнення прогнозу перебігу на підставі визначення патогенетичної ролі поліморфізму гена-модифікатора запалення ІЛ-10. Матеріали та методи. 42 дитини з діагнозом МВ обстежені за стандартною методикою. Визначення поліморфізму G1082A гена IL-10 проводили за допомогою полімеразної ланцюгової реакції в реальному часі. ДНК виділяли з клітин букального епітелію. Результати. У групі дітей, хворих на МВ, із мутацією G1082A гена IL-10 встановлена більш рання маніфестація при кишкових симптомах і більш пізня — при легеневих ознаках, тяжкий перебіг захворювання (зниження об’єму форсованого видоху за 1 секунду — 70,0 (65,0; 72,0) %), ранній розвиток бронхоектазів легенів (72 %) та пневмофіброзу (95,4 %), цирозу печінки (36,4 %), переважання серед респіраторних збудників Pseudomonas aeruginosa (59,1 %) у бактеріальному пейзажі мокротиння, зміни імунного статусу (підвищення циркулюючих імунних комплексів — 8,65 (7,6; 10,55) од. та зниження індексу активності нейтрофілів спонтанного — 0,31 (0,24; 0,78) од. Висновки. Охарактеризовано фенотип МВ, який асоційований з мутацією гена IL-10 G1082A.

Експресія мРНК галектину-9 при ожирінні у дітей з поліморфізмами гена лактази

Мета. Вивчення асоціації експресії мРНК галектину-9 (Gal-9) та мальабсорбції лактози при ожирінні у дітей з поліморфізмами (SNP) гена лактази (LCT) та дослідження ефективності лікування лактазної недостатності з використанням препаратів екзогенної лактази. Матеріали та методи. Обстежено 70 дітей з ожирінням (індекс маси тіла &gt; 95-го перцентиля), а також 16 дітей без ожиріння віком 6–18 років. Визначено SNP LCT (матеріал для дослідження — венозна кров) методом полімеразної ланцюгової реакції (ПЛР) у реальному часі, експресії мРНК Gal-9 (матеріал для дослідження — букальний епітелій) методом ПЛР у реальному часі зі зворотною транскрипцією, дослідження мальабсорбції лактози за допомогою водневого дихального тесту. Серед дітей з ожирінням першу групу спостереження становили 38 дітей з генотипом С/С 13910, другу групу — 32 дитини з фенотипово ідентичними генотипами С/Т 13910 та Т/Т 13910, р &gt; 0,05. У дітей з першої групи спостереження також визначався рівень експресії мРНК Gal-9 та мальабсорбції лактози після використання препаратів екзогенної лактази. Результати. Генотип С/С 13910 реєструвався у 38 (54,3 %), генотип С/Т 13910 — у 22 (31,4 %), генотип Т/Т — у 10 (14,3 %) пацієнтів. Мальабсорбція лактози у дітей з генотипом С/С 13910 становила в середньому 32,7 ± 10,4 рmm, у дітей з генотипами С/Т 13910 — 26,3 ± 4,9 рmm (р &gt; 0,05), при генотипі Т/Т 13910 та у дітей без ожиріння була відсутня (р &lt; 0,05). Середній рівень експресії мРНК Gal-9 у дітей з генотипом С/С 13910 дорівнював 564,3 ± 32,8 УО ∆mRNA Gal-9/mRNA actin, у дітей з генотипами С/Т і Т/Т 13910 — 61,04 ± 15,30 УО ∆mRNA Gal-9/mRNA actin, р &lt; 0,01. Важливим є той факт, що в підгрупі дітей з генотипом С/С 13910 і мальабсорбцією лактози (n = 20) зареєстрований найнижчий середній рівень експресії мРНК Gal-9 (42,47 ± 13,30 УО ∆mRNA Gal-9/mRNA actin), тоді як у підгрупі дітей з генотипом С/С 13910 без мальабсорбції лактози (n = 18) — найвищий (1086,73 ± 52,60 УО ∆mRNA Gal-9/mRNA actin), що перебільшив рівень експресії мРНК Gal-9 у дітей без ожиріння та мальабсорбції лактози (313,34 ± 19,70 УО ∆mRNA Gal-9/mRNA actin), р &lt; 0,01. Після використання препаратів екзогенної лактази у дітей з генотипом С/С 13910 та мальабсорбцією лактози протягом 1 місяця рівень експресії мРНК Gal-9 наблизився до рівня 246,21 ± 15,70 УО ∆mRNA Gal-9/mRNA actin, тоді як при використанні лише низьколактозної дієти збільшився незначно — до 58,72 ± 21,10 УО ∆mRNA Gal-9/mRNA actin, р &lt; 0,01. Цікаво, що у дітей з генотипом С/С 13910 без мальабсорбції лактози на фоні низьколактозної дієти рівень експресії мРНК Gal-9 знизився до рівня 388,38 ± 20,40 УО ∆mRNA Gal-9/mRNA actin, наблизившись до рівня здорових дітей без ожиріння. Висновки. У дітей з генотипом С/С 13910 рівень експресії мРНК Gal-9 залежить від мальабсорбції лактози, яка потребує замісної терапії, що поєднує низьколактозну дієту та використання препаратів екзогенної лактази. За відсутності мальабсорбції лактози використання лише низьколактозної дієти у дітей з генотипом С/С 13910 наближає рівень експресії мРНК Gal-9 до рівня здорових дітей без ожиріння.

Impact of vitamin C supplementation on placental DNA methylation changes related to maternal smoking: association with gene expression and respiratory outcomes

BackgroundMaternal smoking during pregnancy (MSDP) affects development of multiple organ systems including the placenta, lung, brain, and vasculature. In particular, children exposed to MSDP show lifelong deficits in pulmonary function and increased risk of asthma and wheeze. Our laboratory has previously shown that vitamin C supplementation during pregnancy prevents some of the adverse effects of MSDP on offspring respiratory outcomes. Epigenetic modifications, including DNA methylation (DNAm), are a likely link between in utero exposures and adverse health outcomes, and MSDP has previously been associated with DNAm changes in blood, placenta, and buccal epithelium. Analysis of placental DNAm may reveal critical targets of MSDP and vitamin C relevant to respiratory health outcomes.ResultsDNAm was measured in placentas obtained from 72 smokers enrolled in the VCSIP RCT: NCT03203603 (37 supplemented with vitamin C, 35 with placebo) and 24 never-smokers for reference. Methylation at one CpG, cg20790161, reached Bonferroni significance and was hypomethylated in vitamin C supplemented smokers versus placebo. Analysis of spatially related CpGs identified 93 candidate differentially methylated regions (DMRs) between treatment groups, including loci known to be associated with lung function, oxidative stress, fetal development and growth, and angiogenesis. Overlap of nominally significant differentially methylated CpGs (DMCs) in never-smokers versus placebo with nominally significant DMCs in vitamin C versus placebo identified 9059 candidate “restored CpGs” for association with placental transcript expression and respiratory outcomes. Methylation at 274 restored candidate CpG sites was associated with expression of 259 genes (FDR < 0.05). We further identified candidate CpGs associated with infant lung function (34 CpGs) and composite wheeze (1 CpG) at 12 months of age (FDR < 0.05). Increased methylation in the DIP2C, APOH/PRKCA, and additional candidate gene regions was associated with improved lung function and decreased wheeze in offspring of vitamin C-treated smokers.ConclusionsVitamin C supplementation to pregnant smokers ameliorates changes associated with maternal smoking in placental DNA methylation and gene expression in pathways potentially linked to improved placental function and offspring respiratory health. Further work is necessary to validate candidate loci and elucidate the causal pathway between placental methylation changes and outcomes of offspring exposed to MSDP.Clinical trial registration ClinicalTrials.gov, NCT01723696. Registered November 6, 2012. https://clinicaltrials.gov/ct2/show/record/NCT01723696.

Combined toxic effects of aflatoxin B2 and the protective role of resveratrol in Swiss albino mice

In this study, the toxic effects of aflatoxin B2 (AFB2) on Swiss albino mice and the protective effects of resveratrol were investigated. Physiological (body weight, liver and kidney weight), biochemical (aspartate aminotransferase-AST, alanine transaminase-ALT, blood urea nitrogen-BUN, creatinine, malondialdehyde-MDA and glutathione-GSH) and cytogenetic parameters (micronucleus-MN in buccal epithelium, erythrocyte and leukocyte cells and chromosomal aberrations-CAs) were used to determine the toxic effects. Additionally, scavenging effects of resveratrol against superoxide, hydrogen peroxide (H2O2) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals were also investigated. In experimental period, mice were divided into six groups and the groups were treated with tap water, 10 mg/kg b.w resveratrol, 20 mg/kg b.w resveratrol, 20 µg/kg b.w. AFB2, 10 mg/kg b.w resveratrol + 20 µg/kg b.w AFB2, 20 mg/kg b.w resveratrol + 20 µg/kg b.w AFB2, respectively. As a result, the scavenging effects of resveratrol increased with increasing dose and the superoxide, H2O2 and DPPH radical scavenging activity of resveratrol were 74.9%, 79.1% and 49.2%, respectively. AFB2 administration caused a significant decrease in physiological parameters, and these decreases regressed in AFB2 + resveratrol treated groups. Serum ALT and AST activities, BUN and creatinine levels were higher in the AFB2 treated group compared to the control group and serious abnormalities were found in MDA and GSH levels in the kidney and liver. In the group treated with AFB2 + 20 mg/kg resveratrol, ALT, AST, BUN and creatinine levels decreased significantly and GSH levels increased compared to only-AFB2 treated group. AFB2 triggered MN formation in buccal epithelium, erythrocyte and leukocyte cells and CAs in bone marrow cells. The application of 20 mg/kg resveratrol together with AFB2 was decreased the MN and CAs frequency. Resveratrol exhibited a recovery effect in the range of 40.9–80.5% against AFB2 toxicity in all tested parameters. In this study, it was determined that AFB2 caused serious changes in selected physiological, biochemical and cytogenetic parameters while resveratrol displayed a protective role against these toxic effects.