Buccal Cells Research Articles

Sequence Variants in PSMB8/PSMB9 Immunoproteasome Genes and Risk of Urothelial Bladder Carcinoma.

ThePSMB8andPSMB9immunoproteasomegenes are essential in cell processes, such as decisions on cell survival or death, the cell cycle, and cellular differentiation. Because recent evidence has demonstrated an immunological role for proteasomes in various malignancies, including urothelial bladder carcinoma (UBC), we evaluated single nucleotide polymorphisms (SNPs) inPSMB9andPSMB8. We determined any associations between these SNPs and susceptibility to UBC in the Saudi community. Samples of genomic DNA were taken from buccal cells of 111 patients with UBC and 78 healthy controls. TaqMan Real-Time PCR was used to determine genotype distributions and allele frequencies for thePSMB9rs17587 G>A andPSMB8rs2071543 G>T SNPs. We used SNPStats (https://www.snpstats.net) to choose each SNP's best interactive inheritance model. ThePSMB9rs17587 SNP was associated with the risk of UBC (odds ratio [OR] = 5.21,P< 0.0001). In contrast, thePSMB8rs2071543 SNP showed no association with UBC risk (OR = 1.13,P= 0.7871). In terms of genotypic distribution, the rs17587 G>A SNP was more frequent in UBC cases than controls in both the dominant (OR = 7.5; 95% confidence interval, 3.7-15.1;P= 0.0051) and recessive (OR = 17.11, 95% confidence interval 5.1-57.4;P= 0.0026) models. Genotypic distribution of thePSMB8rs2071543 G>T SNP was not significantly different between cases and controls in any interactive inheritance models (P> 0.05). These results suggest a potential role forPSMB9as a biomarker for increased UBC risk. Discovering more genetic variants within immunoproteasome genes related to antigen presentation could help further our understanding of this risk.

Deep learning methods for oral cancer detection using Raman spectroscopy

The Raman spectroscopy analysis has been applied to the detection and research of oral cancer. One of the essential works in this technique is the Raman spectral data analysis method, which is mainly divided into two categories: traditional machine learning and deep learning. Especially, the deep learning method is proved that it could obtain higher accuracy in oral cancer identification than the traditional machine learning method. The purpose of this study is to test, compare, and analyze the performance of existing classical deep neural network models (AlexNet, VGGNet, ResNet50, MobileNetV2, Transformer) that recognize multiple types of oral cancer tissues. To achieve this goal, 16,200 Raman spectra are first collected from 180 tissue samples of 90 patients who have undergone a surgical resection due to tongue squamous cell carcinoma, gingival squamous cell carcinoma, and buccal squamous cell carcinoma. Then, the models are trained and predicted at the patient level. The experimental results demonstrate that the ResNet50 has the best performance in the identification of oral cancer tissue and normal tissue with an overall accuracy rate of 92.81%, an overall precision rate of 92.93%, and an overall recall rate of 92.86%. With this foundation, we further develop a prototype intelligent detection system with above five classical deep neural network models to realize multi-types of oral cancer tissue detection. Hopefully, our work can provide a guide for oral cancer detection using the deep learning method with Raman spectroscopy analysis and promote the development of clinical diagnosis system for oral cancer. The code is available at https://github.com/ISCLab-Bistu/deep-learning-for-OSCC.

The Cultivation Modality and Barrier Maturity Modulate the Toxicity of Industrial Zinc Oxide and Titanium Dioxide Nanoparticles on Nasal, Buccal, Bronchial, and Alveolar Mucosa Cell-Derived Barrier Models.

As common industrial by-products, airborne engineered nanomaterials are considered important environmental toxins to monitor due to their potential health risks to humans and animals. The main uptake routes of airborne nanoparticles are nasal and/or oral inhalation, which are known to enable the transfer of nanomaterials into the bloodstream resulting in the rapid distribution throughout the human body. Consequently, mucosal barriers present in the nose, buccal, and lung have been identified and intensively studied as the key tissue barrier to nanoparticle translocation. Despite decades of research, surprisingly little is known about the differences among various mucosa tissue types to tolerate nanoparticle exposures. One limitation in comparing nanotoxicological data sets can be linked to a lack of harmonization and standardization of cell-based assays, where (a) different cultivation conditions such as an air-liquid interface or submerged cultures, (b) varying barrier maturity, and (c) diverse media substitutes have been used. The current comparative nanotoxicological study, therefore, aims at analyzing the toxic effects of nanomaterials on four human mucosa barrier models including nasal (RPMI2650), buccal (TR146), alveolar (A549), and bronchial (Calu-3) mucosal cell lines to better understand the modulating effects of tissue maturity, cultivation conditions, and tissue type using standard transwell cultivations at liquid-liquid and air-liquid interfaces. Overall, cell size, confluency, tight junction localization, and cell viability as well as barrier formation using 50% and 100% confluency was monitored using trans-epithelial-electrical resistance (TEER) measurements and resazurin-based Presto Blue assays of immature (e.g., 5 days) and mature (e.g., 22 days) cultures in the presence and absence of corticosteroids such as hydrocortisone. Results of our study show that cellular viability in response to increasing nanoparticle exposure scenarios is highly compound and cell-type specific (TR146 6 ± 0.7% at 2 mM ZnO (ZnO) vs. ~90% at 2 mM TiO2 (TiO2) for 24 h; Calu3 93.9 ± 4.21% at 2 mM ZnO vs. ~100% at 2 mM TiO2). Nanoparticle-induced cytotoxic effects under air-liquid cultivation conditions declined in RPMI2650, A549, TR146, and Calu-3 cells (~0.7 to ~0.2-fold), with increasing 50 to 100% barrier maturity under the influence of ZnO (2 mM). Cell viability in early and late mucosa barriers where hardly influenced by TiO2 as well as most cell types did not fall below 77% viability when added to Individual ALI cultures. Fully maturated bronchial mucosal cell barrier models cultivated under ALI conditions showed less tolerance to acute ZnO nanoparticle exposures (~50% remaining viability at 2 mM ZnO for 24 h) than the similarly treated but more robust nasal (~74%), buccal (~73%), and alveolar (~82%) cell-based models.

Analysis of the cytotoxic and genotoxic effects in a population chronically exposed to coal mining residues

During coal mining activities, many compounds are released into the environment that can negatively impact human health. Particulate matter, polycyclic aromatic hydrocarbons (PAHs), metals, and oxides are part of the complex mixture that can affect nearby populations. Therefore, we designed this study to evaluate the potential cytotoxic and genotoxic effects in individuals chronically exposed to coal residues from peripheral blood lymphocytes and buccal cells. We recruited 150 individuals who lived more than 20 years in La Loma-Colombia and 120 control individuals from the city of Barranquilla without a history of exposure to coal mining. In the cytokinesis-block micronucleus cytome (CBMN-Cyt) assay, significant differences in the frequency of micronucleus (MN), nucleoplasmic bridge (NPB), nuclear bud (NBUD), and apoptotic cells (APOP) were observed between the two groups. In the buccal micronucleus cytome (BM-Cyt) assay, a significant formation of NBUD, karyorrhexis (KRX), karyolysis (KRL), condensed chromatin (CC), and binucleated (BN) cells was observed in the exposed group. Considering the characteristics of the study group, a significant correlation for CBMN-Cyt was found between NBUD and vitamin consumption, between MN or APOP and meat consumption, and between MN and age. Moreover, a significant correlation for BM-Cyt was found between KRL and vitamin consumption or age, and BN versus alcohol consumption. Using Raman spectroscopy, a significant increase in the concentration of DNA/RNA bases, creatinine, polysaccharides, and fatty acids was detected in the urine of individuals exposed to coal mining compared to the control group. These results contribute to the discussion on the effects of coal mining on nearby populations and the development of diseases due to chronic exposure to these residues.

Assessment of the quantity and purity of DNA obtained from buccal cells under different storage methods

Aim: This study aimed to verify the quantity and purity of DNA obtained from buccal cells under differentstorage conditions. Methods: Thirty students, between 18 and 23 years of age participated in the study. Three samples of genetic material were collected from each student (samples A, B, and C) through a mouth rinse with 5 mL of 3% glucose. The first phase of DNA extraction from sample A was carried out on the same day of sample collection, whereas samples B and C were stored in a refrigerator and a freezer, respectively, for 1 month before the first extraction phase. DNA extraction was performed with 10 M ammonium acetate and 1 mM EDTA. Sample purity was assessed by spectrophotometry. Statistical analyses were performed through descriptive analysis and analysis of variance ANOVA using the SPSS software, version 21.0. Results: the samples presented no statistically significant differences between the DNA quantity (p = 0.37) or quality (p = 0.16). Conclusion: the quantity and purity of DNA extraction from the three samples were satisfactory, and no differences in storage conditions were found. Uniterms: DNA. Mouth. Mucosa.

Sex-dimorphic pathways in the associations between maternal trait anxiety, infant BDNF methylation, and negative emotionality.

Maternal antenatal anxiety is an emerging risk factor for child emotional development. Both sex and epigenetic mechanisms, such as DNA methylation, may contribute to the embedding of maternal distress into emotional outcomes. Here, we investigated sex-dependent patterns in the association between antenatal maternal trait anxiety, methylation of the brain-derived neurotrophic factor gene (BDNF DNAm), and infant negative emotionality (NE). Mother-infant dyads (N = 276) were recruited at delivery. Maternal trait anxiety, as a marker of antenatal chronic stress exposure, was assessed soon after delivery using the Stait-Trait Anxiety Inventory (STAI-Y). Infants' BDNF DNAm at birth was assessed in 11 CpG sites in buccal cells whereas infants' NE was assessed at 3 (N = 225) and 6 months (N = 189) using the Infant Behavior Questionnaire-Revised (IBQ-R). Hierarchical linear analyses showed that higher maternal antenatal anxiety was associated with greater 6-month-olds' NE. Furthermore, maternal antenatal anxiety predicted greater infants' BDNF DNAm in five CpG sites in males but not in females. Higher methylation at these sites was associated with greater 3-to-6-month NE increase, independently of infants' sex. Maternal antenatal anxiety emerged as a risk factor for infant's NE. BDNF DNAm might mediate this effect in males. These results may inform the development of strategies to promote mothers and infants' emotional well-being.

Mosaic 45,X/46, XX at amniocentesis with high-level mosaicism for 45,X in a pregnancy with a favorable fetal outcome and postnatal decrease of the 45,X cell line

We present mosaic 45,X/46, XX at amniocentesis with high-level mosaicism for 45,X in a pregnancy with a favorable fetal outcome and postnatal decrease of the 45,X cell line. A 20-year-old, primigravid woman underwent amniocentesis at 17 weeks of gestation because of the non-invasive prenatal testing (NIPT) result of-4.82 Z score in sex chromosome at 12 weeks of gestation suggestive of Turner syndrome in the fetus. Amniocentesis revealed a karyotype of 45,X [18]/46,XX [15], and simultaneous multiplex ligation-dependent probe amplification (MLPA) on the DNA extracted from uncultured amniocytes showed mosaic Turner syndrome. Prenatal ultrasound and parental karyotypes were normal. She was referred for genetic counseling at 24 weeks of gestation, and continuing pregnancy was encouraged. At 39 weeks of gestation, a 2550-g phenotypically normal female baby was delivered. The karyotypes of cord blood, umbilical cord and placenta were 45,X [24]/46,XX [16], 45,X [23]/46,XX [17] and 45,X [28]/46,X,del(X) (q23)[12], respectively. When follow-up at age two months, the neonate was phenotypically normal in development. The peripheral blood had a karyotypes of 45,X [16]/46,XX [24]. Interphase fluorescence in situ hybridization (FISH) analysis on 103 buccal mucosal cells showed normal disomy X signals in all cells. High-level mosaicism for 45,X in 45,X/46, XX at amniocentesis can be associated with a favorable fetal outcome, cytogenetic discrepancy in various tissues, and postnatal decrease of the 45,X cell line.

Biophysical parameters of oral fluid, periodontal tissues and buccal epithelial cells in adolescent athletes in the process of complex dental treatment

Annotation. The aim of the work was to evaluate the impact of the developed complex for the dental treatment of adolescent athletes on the biophysical parameters of their oral fluid, periodontal tissues and buccal epithelial cells. The study involved 42 boys aged 12-14, professionally involved in sports, with major dental diseases. The studies included spectrocolorimetric assessment of functional hyperemia of periodontal tissues, spectrocolorimetric assessment of the degree of inflammation in periodontal tissues using the Schiller-Pysarev test solution, assessment of oral fluid pH stability as one of the indicators of the level of non-specific resistance in the body and in the oral cavity in particular, and assessment of the charge state of cells buccal epithelium, the percentage of mobile nuclei and plasmalemma cells in the electric field and the ratio of the amplitudes of their movement. The computer application STATISTICA 6.1 was used for statistical analysis of the results and assessment of their reliability and errors. A statistically significant difference between alternative quantitative traits with a distribution corresponding to the normal law was evaluated using the Student's t-test (the difference was considered statistically significant at p&lt;0.01). The use of the proposed therapeutic complex, which included “Albumin Egg Plus”, “Chlorophyll Spray”, “Active Sulfur”, “Active Fluoride” and toothpastes “Biodentis” and “Meridol”, in adolescent athletes of the main group helped to eliminate capillary spasms and increase blood flow in them under the action of masticatory load, which was already a normal physiological response, disrupted in the initial state. At the same time, the staining of the gums in Schiller-Pisarev solution decreased in children-athletes, which indicates a decrease in the degree of inflammation. Complex prophylaxis allowed to reduce the interval of fluctuations in the oral fluid pH value in individual samples for 1 year more than 4.67 times, which indicates a certain normalization of functional adaptive-compensatory reactions responsible for its stability. Under the influence of the developed therapy, the charge state of the buccal epithelial cells was also normalized. The obtained results testify to the positive influence of the developed complex on the biophysical parameters of oral fluid, periodontal tissues and buccal epithelial cells in adolescent athletes.

A closer look at the resection margins of buccal mucosa cancer: Their influence on local recurrence free survival.

The adequate surgical margin for local control of buccal mucosa squamous cell carcinoma (BMSCC) is under debate. This study investigates surgical margins and other factors associated with local recurrence free survival (LRFS) in a large cohort of BMSCC patients. Multiple factors were evaluated retrospectively in 97 patients with BMSCC. Cox-regression and Kaplan-Meier curves were used for analysis. The local recurrence rate was 23%. The tumor-free margin was <5.0 mm in 89% of the patients and the deep margin was significantly more often inadequate. Multivariate analysis associated pT3-classification, former smokers, tumor-free margin status, and postoperative (chemo)radiation (PO(ch)RT) with local recurrence. Re-resections did not improve LRFS in patients with <5.0 mm tumor-free margins. Adequate tumor-free margins are pivotal for LRFS of BMSCC. PO(ch)RT, not re-resection, can improve LRFS in patients with <5.0 mm tumor-free margins.

Stressful life events and accelerated biological aging over time in youths

Experiencing adversity in childhood and adolescence, including stressful life events (SLEs), may accelerate the pace of development, leading to adverse mental and physical health. However, most research on adverse early experiences and biological aging (BA) in youths relies on cross-sectional designs. In 171 youths followed for approximately 2 years, we examined if SLEs over follow-up predicted rate of change in two BA metrics: epigenetic age and Tanner stage. We also investigated if rate of change in BA was associated with changes in depressive symptoms over time. Youths aged 8–16 years at baseline self-reported Tanner stage and depressive symptoms at baseline and follow-up and provided saliva samples for DNA at both assessments. Horvath epigenetic age estimates were derived from DNA methylation data measured with the Illumina EPIC array. At follow-up, contextual threat interviews were administered to youths and caregivers to assess youths’ experiences of past-year SLEs. Interviews were objectively coded by an independent rating team to generate a SLE impact score, reflecting the severity of all SLEs occurring over the prior year. Rate of change in BA metrics was operationalized as change in epigenetic age or Tanner stage as a function of time between assessments. Higher objective SLE impact scores over follow-up were related to a greater rate of change in epigenetic age (β = 0.21, p = .043). Additionally, among youths with lower—but not higher—Tanner stage at baseline, there was a positive association of SLE impact scores with rate of change in Tanner stage (Baseline Tanner Stage × SLE Impact Score interaction: β = − 0.21, p = .011). A greater rate of change in epigenetic age was also associated with higher depressive symptom levels at follow-up, adjusting for baseline symptoms (β = 0.15, p = .043). Associations with epigenetic age were similar, although slightly attenuated, when adjusting for epithelial (buccal) cell proportions. Whereas much research in youths has focused on severe experiences of early adversity, we demonstrate that more commonly experienced SLEs during adolescence may also contribute to accelerated BA. Further research is needed to understand the long-term consequences of changes in BA metrics for health.

АССОЦИАЦИИ ПОЛИМОРФИЗМА ГЕНА РЕЦЕПТОРА ВИТАМИНА D (VDR) И РЕЦЕССИИ ДЕСНЫ В СМЕННОМ ПРИКУСЕ

Subject of study. The development of new approaches to early diagnosis and identification of predisposition to the formation of gingival recession in children is an urgent task of modern dentistry. It is known that vitamin D plays an important role, affecting, among other things, the immune system and calcium-phosphorus homeostasis through the vitamin D receptor (VDR). In connection with the above, a promising direction is the study of polymorphisms of the vitamin D receptor gene to determine markers of gingival recession in a removable bite.&#x0D; Purpose: to study the relationship of the polymorphic marker Bsm I (rs1544410 T/C) of the VDR gene with a predisposition to gingival recession in children in a removable bite living in the Republic of Tatarstan.&#x0D; Methodology. We examined 81 children with gingival recession in a removable bite (8 ± 1 years old) living in the Republic of Tatarstan. DNA was isolated from buccal epithelial cells. Genotyping of the rs1544410 polymorphism of the VDR gene was performed using a real-time PCR.&#x0D; Results. A positive relationship was found for the rs1544410 T allele of the VDR gene with class II gingival recession in a removable bite according to Miller's classification (p = 0.021, r = 0.258), and recession depth (p = 0.029, r = 0.24). &#x0D; Conclusions. 1. A positive correlation was found between the risk allele T of the BsmI (rs1544410) polymorphism of the VDR gene and clinical manifestations of gingival recession in a removable bite in a sample of the population of the Republic of Tatarstan. 2. The distribution of alleles and genotypes of the BsmI (rs1544410) polymorphism of the VDR gene did not differ between the observation and control groups.

DNA Damage on Buccal Epithelial Cells, Personal Working in the Rubber Industry Occupationally Exposed to Carbon Disulfide (CS2).

The most significant industrial utilization of carbon disulfide (CS2) has been in the manufacture of cellulose rayon, cellophane, and rubber industry. CS2 prompts expanded recurrence of chromosomal variations in laborers occupationally exposed to CS2. In the current study, the DNA analysis was carried out from exfoliated buccal epithelial cells from rubber industry workers exposed to CS2 and an equal number of healthy control subjects. Both the control and experimental subjects were categorized by their smoking habits such as smokers (S) and non-smokers (NS). Furthermore, experimental subjects were further separated based on their exposure period. Students t-test statistical tools were used to analyze the final results. The present analysis identified a high frequency of DNA damage in rubber industry workers (16.55±0.43) than control subjects (9.8±0.21). Also, maximum number of DNA damage detected in smoking experimental group (18.27±0.02) than non-smoking experimental (15.02±0.01) and smoking control groups (10.25±0.04 ). Smoking habits synergistically increased the DNA damage in the rubber industry workers exposed to CS2.

Comparison of Nucleus and Cytoplasm Diameter of Buccal Mucosa Cells in Cigarette Smokers and Nonsmokers: A Cytomorphometric Study using Feulgen and Papanicolaou Stains

Comparison of Nucleus and Cytoplasm Diameter of Buccal Mucosa Cells in Cigarette Smokers and Nonsmokers: A Cytomorphometric Study using Feulgen and Papanicolaou Stains

Artisanal Gem Mining in Brazil: A Source of Genotoxicity and Exposure to Toxic Elements.

Environmental and occupational exposure to toxic metals has led many people around the world to have serious health problems. Mining activities contribute to an increased risk of exposure to these elements. In this work, a study of environmental biomonitoring and routes of exposure to toxic metals in a region of artisanal mining was performed. This study was carried out in the district of Taquaral de Minas, located in the Jequitinhonha Valley in the state of Minas Gerais. The valley is one of the wealthiest and highest gem-producing areas in Brazil. Five artisanal mines were sampled (Bode, Pirineu, Pinheira, Lajedo, and Marmita). Several potentially toxic metals (Be, Zn, Mn, Ba Cd, Hg, and U) were investigated in the soils and dust over the rocks and the soils. Samples from 22 individuals occupationally exposed and 17 unexposed persons, who formed the reference group, were analyzed for trace elements by an inductively coupled plasma mass spectrometer. The genotoxicity was evaluated by the micronucleus test in buccal mucosa epithelial cells, where the following changes were scored: micronuclei (MN) binucleate (BN) cells and kariolytic (KL) cells. The MN test showed significantly increased frequencies in all alterations of exposed individuals compared to the controls (p < 0.05, Student's t-test). The urine analysis showed levels of Cr, Ni Ba, Pb, and As in the blood, which were higher than the ATSDR recommended levels. The association between the MN test and the trace element concentrations found in the blood and urine was significant (p < 0.05). The higher the number of years of working, the higher the concentrations in the blood were, due to chronic exposure. The results of the present study indicate environmental contamination and a potential risk to the health of miners, suggesting an intervention.

Quantitation by Liquid Chromatography-Nanoelectrospray Ionization-High-Resolution Tandem Mass Spectrometry of Multiple DNA Adducts Related to Cigarette Smoking in Oral Cells in the Shanghai Cohort Study.

We developed a liquid chromatography-nanoelectrospray ionization-high-resolution tandem mass spectrometry (LC-NSI-HRMS/MS) method for simultaneous quantitative analysis of 5 oral cell DNA adducts associated with cigarette smoking: (8R/S)-3-(2'-deoxyribos-1'-yl)-5,6,7,8-tetrahydro-8-hydroxypyrimido[1,2-a]purine-10(3H)-one (γ-OH-Acr-dGuo, 1) from acrolein; (6S,8S and 6R,8R)-3-(2'-deoxyribos-1'-yl)-5,6,7,8-tetrahydro-8-hydroxy-6-methylpyrimido[1,2-a]purine-10(3H)-one [(6S,8S)γ-OH-Cro-dGuo, 2; and (6R,8R)γ-OH-Cro-dGuo, 3] from crotonaldehyde; 1,N6-etheno-dAdo (4) from acrylonitrile, vinyl chloride, lipid peroxidation, and inflammation; and 8-oxo-dGuo (5) from oxidative damage. Oral cell DNA was isolated in the presence of glutathione to prevent artifact formation. Clear LC-NSI-HRMS/MS chromatograms were obtained allowing quantitation of each adduct using the appropriately labeled internal standards. The accuracy and precision of the method were validated, and the assay limit of quantitation was 5 fmol/μmol dGuo for adducts 1-4 and 20 fmol/μmol for adduct 5. The assay was applied to 80 buccal cell samples selected from those collected in the Shanghai Cohort Study: 40 from current smokers and 40 from never smokers. Significant differences were found in all adduct levels between smokers and nonsmokers. Levels of 8-oxo-dGuo (5) were at least 3000 times greater than those of the other adducts in both smokers and nonsmokers, and the difference between amounts of this adduct in smokers versus nonsmokers, while significant (P = 0.013), was not as great as the differences of the other DNA adducts between smokers and nonsmokers (P-values all less than 0.001). No significant relationship of adduct levels to risk of lung cancer incidence was found. This study provides a new LC-NSI-HRMS/MS methodology for the quantitation of diverse DNA adducts resulting from exposure to the α,β-unsaturated aldehydes acrolein and crotonaldehyde, inflammation, and oxidative damage which are all associated with carcinogenesis. We anticipate application of this assay in ongoing studies of the molecular epidemiology of cancers of the lung and oral cavity related to cigarette smoking.

Multiple-scattering simulator-trained neural network for intensity diffraction tomography.

Recovering 3D phase features of complex biological samples traditionally sacrifices computational efficiency and processing time for physical model accuracy and reconstruction quality. Here, we overcome this challenge using an approximant-guided deep learning framework in a high-speed intensity diffraction tomography system. Applying a physics model simulator-based learning strategy trained entirely on natural image datasets, we show our network can robustly reconstruct complex 3D biological samples. To achieve highly efficient training and prediction, we implement a lightweight 2D network structure that utilizes a multi-channel input for encoding the axial information. We demonstrate this framework on experimental measurements of weakly scattering epithelial buccal cells and strongly scattering C. elegans worms. We benchmark the network's performance against a state-of-the-art multiple-scattering model-based iterative reconstruction algorithm. We highlight the network's robustness by reconstructing dynamic samples from a living worm video. We further emphasize the network's generalization capabilities by recovering algae samples imaged from different experimental setups. To assess the prediction quality, we develop a quantitative evaluation metric to show that our predictions are consistent with both multiple-scattering physics and experimental measurements.

Characterization of the external exposome and its contribution to the clinical respiratory and early biological effects in children: The PROMESA cohort study protocol.

Air pollution contains a mixture of different pollutants from multiple sources. However, the interaction of these pollutants with other environmental exposures, as well as their harmful effects on children under five in tropical countries, is not well known. This study aims to characterize the external exposome (ambient and indoor exposures) and its contribution to clinical respiratory and early biological effects in children. A cohort study will be conducted on children under five (n = 500) with a one-year follow-up. Enrolled children will be followed monthly (phone call) and at months 6 and 12 (in person) post-enrolment with upper and lower Acute Respiratory Infections (ARI) examinations, asthma development, asthma control, and genotoxic damage. The asthma diagnosis will be pediatric pulmonologist-based and a standardized protocol will be used. Exposure, effect, and susceptibility biomarkers will be measured on buccal cells samples. For environmental exposures PM2.5 will be sampled, and questionnaires, geographic information, dispersion models and Land Use Regression models for PM2.5 and NO2 will be used. Different statistical methods that include Bayesian and machine learning techniques will be used for the ambient and indoor exposures-and outcomes. This study was approved by the ethics committee at Universidad Pontificia Bolivariana. To estimate i) The toxic effect of particulate matter transcending the approach based on pollutant concentration levels; ii) The risk of developing an upper and lower ARI, based on different exposure windows; iii) A baseline of early biological damage in children under five, and describe its progression after a one-year follow-up; and iv) How physical and chemical PM2.5 characteristics influence toxicity and children's health.

Study of Serial Exposures of an Astringent Green Tea Flavonoid Extract with Oral Cell-Based Models.

It is well known that repeated exposure to phenolic compounds (PCs) raises astringency perception. However, the link between this increase and the oral cavity's interactions with salivary proteins (SPs) and other oral constituents is unknown. To delve deeper into this connection, a flavonoid-rich green tea extract was tested in a series of exposures to two oral cell-based models using a tongue cell line (HSC3) and a buccal mucosa cell line (TR146). Serial exposures show cumulative PC binding to all oral models at all concentrations of the green tea extract; however, the contribution for the first and second exposures varies. The tongue mucosal pellicle (HSC3-Mu-SP) may contribute more to first-stage astringency (retaining 0.15 ± 0.01 mg mL-1 PCs at the first exposure), whereas the buccal mucosal pellicle (TR146-Mu-SP) retained significantly less (0.08 ± 0.02 mg mL-1). Additionally, increased salivary volume (SV+), which simulates the stimulation of salivary flow brought by a food stimulus, significantly enhances PC binding, particularly for TR146 cells: TR46-Mu-SP_SV+ bound significantly higher total PC concentration (0.17 ± 0.02 mg mL-1) than the model without increased salivary volume TR146-Mu-SP_SV- (0.09 ± 0.03 mg mL-1). This could be associated with a higher contribution of these oral cells for astringency perception during repeated exposures. Furthermore, PCs adsorbed in the first exposure to cell monolayer models (+TR146 and +HSC3) change the profile of PCs bound to these models in the second exposure. Regarding the structure binding activity, PCs with a total higher number of hydroxyl groups were more bound by the models containing SP. Regarding the SP, basic proline-rich proteins (bPRPs) may be involved in the increased perception of astringency upon repeated exposures. The extent of bPRP precipitation by PCs in mucosal pellicle models for both cell lines (HSC3 and TR146) in the second exposure (76 ± 13 and 83 ± 6%, respectively) was significantly higher than in the first one (25 ± 14 and 5 ± 6%, respectively).

Effects of fixed orthodontic treatment with and without chlorhexidine mouthwash on vitality of oral mucosal cells reflected by cell nuclear indexes: A preliminary 3-phase before-after clinical trial

Effects of mouthwashes on the vitality of oral mucosal cells have not been determined in orthodontic patients. We aimed to assess, for the first time, the effects of fixed orthodontic treatment with and without chlorhexidine (CHX) mouthwash on the oral mucosal cell vitality. All patients meeting the eligibility criteria were consecutively included until the desired sample size was reached. Oral buccal mucosal cell samples were taken immediately before orthodontic treatment. For each patient, 20 metal brackets and 4 bands were installed on the teeth. Cell samples were recollected after one month of treatment. Then, the patients used an ethanol-free 0.12% CHX mouthwash two times a week for one month. Sampling was repeated at the end of the second month. Papanicolaou staining was used for micronucleus screening of the indexes: micronucleus (MIC), karyorrhexis (KR), karyolysis (KL), and broken eggs (BE). The repair index (RI) was calculated as RI=KR+KL/BE+MIC. Comparisons of nuclear changes over 3 intervals were done using the Friedman and Dunn-Bonferroni tests (α=0.05, β<0.05). This prospective before-after clinical trial was performed on 408 observations of 34 patients (14 males, 20 females, mean age: 16.68±3.75 years) at 3 intervals (×4 parameters each). The means of MIC, KR, KL, BE, and RI were respectively 1.312±1.219, 0.241±0.564, 0.426±0.657, 0.115±0.224, and 0.476±0.360 before treatment. They were 1.348±1.171, 0.215±0.236, 0.406±0.369, 0.124±0.187, and 0.511±0.310 at the first interval and 1.909±1.263, 0.368±0.174, 0.615±0.269, 0.253±0.150, and 0.529±0.195 at the second interval. Friedman showed significant time-dependent changes for all variables (P<0.0005) except RI. Dunn-Bonferroni showed that except MIC (P=0.017), KR/KL/BE changed insignificantly after orthodontic treatment (P≥0.974). MIC/KR/KL/BE increased significantly after the addition of CHX (P<0.0005). CHX mouthwash, together with orthodontic treatment, has a strong deteriorating effect on nuclear indexes associated with the vitality of buccal mucosal cells. Nuclear changes caused by orthodontic treatment alone might be negligible.

Successful Strategy of Pre-implantation Genetic Testing for Beta-Thalassemia (c.17A&gt;T Mutation)-Hb E Disease Using Multiplex Fluorescent PCR and Mini-Sequencing

Objectives: Hemoglobin E disease, c.26G&gt;A variant of beta-globin gene, is the most common hemoglobinopathy in Asia. Compound heterozygotes inheriting Hb E disease and beta-thalassemia generate beta-thalassemia-Hb E disease with severe anemia. This study aimed to develop a pre-implantation genetic testing for monogenic disorders (PGT-M) protocol for beta–thalassemia (c.17A&gt;T mutation)-Hb E disease (c.26G&gt;A mutation) using multiplex fluorescent polymerase chain reaction (PCR) and mini-sequencing. Materials and Methods: bthalw1 primers were used to amplify a beta-globin gene fragment covering both mutations, i.e. beta– thalassemia (c.17A&gt;T) and Hb E disease. D21S11 microsatellite marker was included for contamination detection. Novel mini-sequencing primers were designed and tested for detection of both mutations. Results: Pre-clinical work up of the optimized PGT-M protocol using 20 single buccal cells of a heterozygous subject showed 100% amplification efficiency and 0% allele drop out (ADO) rate for both primers. In clinical PGT-M cycle, 15 embryos were subjected to biopsy. The results showed two normal, one heterozygous for beta-thalassemia, six heterozygous for Hb E disease, one affected for beta-thalassemia-Hb E disease and five with ambiguous results. Two normally diagnosed embryos were chosen for transfer, one singleton pregnancy was obtained. A healthy baby boy was resulted. Postnatal testing confirmed PGT results. Conclusions: Novel PGT-M protocols for beta-thalassemia-Hb E disease using multiplex fluorescent PCR and mini-sequencing were developed and described here. The protocol was applied in a clinical PGT-M cycle and gave rise to one successful pregnancy and consequently a healthy baby boy. Mini-sequencing was proved to be rapid, accurate and cost-effective protocol for PGT-M.