Oral Fluid Research Articles

COMPARATIVE CHARACTERISTICS OF METHODS FOR THE PREVENTION OF DENTAL CARIES IN CHILDREN OF EARLY CHILDHOOD

The high prevalence and intensity of early childhood caries in Ukraine and in the world indicate insuffi cient attention of practical dentistry to the prevention of dental caries in young children. The development of adapted prevention programs considering the risk factors for the development of the disease remains relevant.Aim of the study. To compare the eff ectiveness of methods of pre- and postnatal prevention of early childhood dental caries based on correction of the microbial environment of the oral cavity and macro- and microelemental nutrition.Material and methods. The work was carried out within the framework of the research work of the Department of Pediatric Dentistry of the Bukovinian State Medical University «Development of methods of prevention and treatment of major dental diseases in children, taking into account the risk factors of their development» (DR # 0121U110122). The prenatal method of dental caries prevention was applied to a group of pregnant women (30 persons) whose average age was (25,17±1,05) years. It included the use of Elevit Pronatal and Iodomarin 200, one tablet daily, starting from the second trimester ofpregnancy, and a course of local probiotic therapy with BioGaya ProDentis chewable tablets, used for 20 days in the second trimester of pregnancy and 20 days before delivery. As a method of postnatal prevention of dental caries in children, a preventive complex was used, including the use of the probiotic BioGaya ProDentis and the vitamin and mineral complex Supervit. The course lasted three weeks twice a year. The preventive complex was applied to a group of children aged 2-3 years. The number of children in the control group was 30. The mean age of the group was (2.51±1.30) years. The eff ectiveness of preventive measures was evaluated by the prevalence and intensity of caries of primary teeth («kpv» and «kpppv»), the state of oral hygiene (M. E. Kuzmina index), the content of phosphate ions, inorganic and ionized calcium in the oral fluid of children, the content of mutans streptococci in the oral cavity and the concentration of sIgA in the oral fl uid of children. Statistical processing of the study results was performed using standard methods of variation. The study complied with all bioethical norms (conclusion of the Bioethics Commission of the Bukovinian State Medical University No. 4 dated 15.12.2022).Results of the study. The use of preventive complexes for early childhood caries, based on the supplementation of macro- and microelements and the infl uence on the microbial landscape of the oral cavity, are eff ective methods of prevention of caries of temporary teeth in children, which is confi rmed by a decrease in the prevalence and intensity of dental caries in children in the dynamics of their observation. Prenatal measures are more eff ective in the prevention of early childhood caries in children, as they optimize the processes of primary mineralization of hard dental tissues and thus strengthen them, reducing the risk of dental caries. Postnatal use of the drugs has a slightly lower preventive eff ect on early childhood caries, while it has a positive eff ect on the processes of secondary mineralization of tooth enamel, as it was found that the level of phosphate ions in the oral fl uid of children under these conditions increased by 23.72 % (p<0.05), and the level of total calcium decreased by 15.84 % and its free fraction by 25.86 % (p<0.05).Conclusions. Our results suggest that both methods of prevention of early childhood caries are eff ective, but prenatal prophylaxis is clinically more eff ective with respect to the hard tissues of temporary teeth in children, due to its eff ect on the processes of primary mineralization of enamel. Postnatal prophylactic measures have a positive eff ect on the secondary mineralization of the hard tissues of temporary teeth and the primary mineralization of permanent teeth and will have a clinical result in the long term.

The effect of oral cavity condition on expression of saliva biomarkers in patients with oral mucosa pathology

Introduction. Despite visual accessibility of the elements of tumor lesions in the oral mucosa, patients with this pathology rarely seek medical help. Regeneration of the mucosa is a long process taking varying time depending on the location of the lesions. Asymptomatic tumor process leads to its diagnosis at late stages.Aim. To develop clinical and proteomic approach to diagnosis of malignancies of the oral mucosa at the stage of out-patient dentist appointment.Clinical observations. Examples of extended clinical, dental and immunoenzymatic monitoring in patients with elements of tumor lesions of the oral mucosa with long-term course are presented.Conclusion. The use of extended clinical, dental and immunoenzymatic analysis of oral fluid in patients with elements of tumor lesions of the oral mucosa allows to diagnose this pathology early.

Effect of stabilizers on the detection of swine influenza A virus (swIAV) in spiked oral fluids over time.

Aggregated samples such as oral fluids (OFs) display an animal friendly and time and cost-efficient sample type for swine Influenza A virus (swIAV) monitoring. However, further molecular and biological characterization of swIAV is of particular significance. The reportedly inferior suitability of aggregated samples for subtyping of swIAV presents a major drawback compared to nasal swabs, still considered the most appropriate sample type for this purpose (Garrido-Mantilla et al. BMC Vet Res 15(1):61, 2019). In addition, the viral load in the original sample, storage conditions and characteristics of different swIAV strains might further compromise the eligibility of aggregated samples for molecular detection and subtyping. Therefore, the present study aimed to evaluate the suitability of stabilizing media to minimize the degradation of viral RNA and thus increase the detection and subtyping rate of swIAV by RT-qPCR in spiked OFs under different conditions (virus strain, storage temperature and viral load in the original sample) over a time span of 14days. The use of stabilizing media in spiked OFs resulted in a significant higher probability to detect swIAV RNA compared to OFs without stabilizers (OR = 46.1, p < 0.001). In addition,swIAV degradation over time was significantly reduced in samples suspended with stabilizer (OR = 5.80, p < 0.001), in samples stored at 4°C (OR = 2.53, p < 0.001) and in samples spiked with the avian derived H1N2 subtype (OR = 2.26, p < 0.01). No significant differences in swIAV RNA detection and degradation of swIAV RNA in spiked OFs over time were observed between the three different stabilizing media. Addition of stabilizers and storage of samples under cooled conditions significantly improved detection and subtyping of swIAV in spiked OFs.

INDICATORS OF FREE RADICAL OXIDATION AND ANTIOXIDANT PROTECTION IN THE ORAL FLUID OF CHILDREN WITH CHRONIC CATARRHAL GINGIVITIS AND DIABETIS MELLITUS

Failure of antioxidant mechanisms leads to an increase in lipid peroxidation products in the body, resulting in a non-specific cascade of cellular membrane damage. Proteins play a crucial role in metabolic processes, but when lipid peroxidation intensifies, protein modifications occur, leading to fragmentation, denaturation, and loss of biological activity. This disruption impairs tissue regenerative processes. The objective of this study was to analyze indicators of free radical oxidation and antioxidant protection in the oral fluid of children with chronic catarrhal gingivitis and type I diabetes mellitus. Materials and methods. A total of 170 children aged 12 to 16 were observed, including 130 who were examined and treated at the Endocrinology Department of the Regional Municipal Non-Profit Institution “Chernivtsi Regional Pediatric Clinical Hospital” for type 1 diabetes mellitus. Among them, 74 children had a disease duration of less than 5 years, including 65 with chronic catarrhal gingivitis. Sixty five children had diabetes for more than 5 years, with 44 of them diagnosed to have chronic catarrhal gingivitis. The children with chronic catarrhal gingivitis were further divided into groups based on their level of glycemic control: 1 child with optimal glycemic control, 66 with suboptimal glycemic control, and 42 with glycemic control at a high risk to life. The control group included 40 practically healthy children: 22 of them had clinically healthy periodontal tissues, and 18 children were diagnosed to suffer from chronic catarrhal gingivitis. Results. The study performed found out that children with chronic catarrhal gingivitis present increased indicators of lipid peroxide oxidation (protein oxidative modification, diene conjugates, malondialdehyde) and decreased activity of the enzyme in the antioxidant protective system of the oral fluid (whole protein, НS-groups, ceruloplasmin, activity of supermutase and catalase) in comparison with indicators of children without dental pathology. The most considerable changes were found in patients with chronic catarrhal gingivitis and type 1 diabetes mellitus, especially when the disease lasted more than 5 years (р&lt;0,05).

The Thermomechanical, Functional and Biocompatibility Properties of a Au–Pt–Ge Alloy for PFM Dental Restorations

A high-noble Au–Pt–Ge porcelain-fused-to-metal (PFM) dental alloy without the known adverse metallic elements and with the addition of germanium (Ge) was produced as a more cost-effective alternative to other precious alloying metals, with investigations for determining the functionality and clinical use of this alloy. The thermomechanical, biocompatibility, durability, workability and economic characteristics of the produced dental alloy were investigated. These properties were investigated with in vitro biocompatibility testing on human gingival fibroblasts (HGFs); static immersion testing for metal ion release; DSC analysis; hardness, tensile testing, density and coefficient of thermal expansion (CTE) measurements; metallographic and SEM/EDX microstructure investigations; and finally with the production of a test PFM dental bridge. The results of the thermomechanical testing showed alloy properties suitable for dental restorations and clinical use, with somewhat lower mechanical properties, making the alloy not suitable for extensive multiunit fixed restorations. The microstructure investigations showed segregations of Ge in the homogeneous alloy matrix, which reduce the alloy’s mechanical properties. The produced PFM dental bridge showed excellent workability of the alloy in a dental laboratory setting, as well as a high standard of the final dental restoration. The ion release was negligible, well below any harmful quantities, while the cell viability examination showed significantly higher viability ratings on polished alloy samples as compared to as-cast samples. The results showed that a dental substructure in direct contact with oral tissue and fluids should be highly polished. The performed investigations showed that the produced PFM dental alloy is suitable for clinical use in producing high-quality dental restorations with high biocompatibility for patients prone to metal allergies.

Validation of an LC-MS/MS Method for Analysis of 58 Drugs of Abuse in Oral Fluid and Method Comparison with an Established LC-HRMS Method.

Liquid chromatography-mass spectrometry (LC-MS) methods for detection of multiple drugs of abuse (DoA) in oral fluid (OF) samples are being implemented in many clinical routine laboratories. Therefore, there is a need to develop new multi-analyte methods with simple sample pre-treatment and short analysis times. The purpose of this work was to validate a method detecting 58 DoA to be used with two different OF sampling kits, the saliva collection system (SCS) from Greiner Bio-One and Quantisal from Immunalysis, using the same sample pretreatment and analytical method. A set of 110 samples collected with the SCS kit was further compared to an LC-HRMS (high resolution mass spectrometry) method in another laboratory. The method was successfully validated, with precision and accuracy of ≤15% and z-scores of <2 for external controls. Using a sensitive LC-MS/MS instrument, the detection limits were <1 µg/L in neat oral fluid. In the comparative study between the LC-MS/MS and LC-HRMS methods using SCS samples, a good agreement was observed. Discrepancies were limited to lower concentration ranges, attributable to differences in cut-off thresholds between the methods. This work contributes to the development of LC-MS multi-analyte methods for OF samples, which are suitable for clinical routine laboratories.

Efficiency of chewing load deficiency correction in clinical dental practice

Weakening of the chewing act, typical for modern humans due to consumption of soft food, causes a deficit of chewing load and has a negative effect on the dental system. To combat hypodynamia of the masticatory apparatus with the aim of preventing destructive processes in its structures, a gnathotraining system was developed, including electrical stimulation of the masticatory muscles themselves in physiological parameters, which develops the effect of “imaginary chewing”, and gnathotraining using chewing gum. In addition, functional training of the masticatory apparatus using chewing exercises was proposed. Considering that chewing gum practically does not compensate for the deficit of chewing load observed in modern society, the staff of the Kazan State Medical University and the Kazan National Research Technological University developed the “Denta Fit” dental trainer and a dental training system. It has been established that dental and jaw training stimulates secretion of saliva, its mineralizing potential, functional resistance of tooth enamel, allows achieving a good level of oral hygiene, reduces the rate of increase in the intensity of dental caries, as well as the severity of muscular-articular disorders of the temporomandibular joints, enhances neuromuscular activity of the masticatory muscles and regional blood flow. In children with permanent bite, dental and jaw training maintains the hydrogen index (pH) of oral fluid at a neutral level and provides, in combination with a toothbrush, optimal hygienic dental care, contributes to the formation of a correct dental occlusion. Thus, the effects of increased neuromuscular activity of the masticatory muscles, as well as regional vascular tone, arising during dental and jaw training, indirectly indicate that the dental and jaw system receives adequate mechanical loads necessary for the normal functioning of all its links. The results of the conducted studies indicate the effectiveness and feasibility of using dental training to improve dental status, which serves as a new approach to the prevention and treatment of dental diseases.

Determination of endogenous GHB in ante-mortem whole blood, urine, and oral fluid by LC–MS/MS: The effect of different additives and storage conditions on the stability of GHB in blood

Two challenges in detecting γ-hydroxybutyric acid (GHB) intake are its endogenous presence and in vitro production after sampling. This study developed an LC–MS/MS method for selective GHB determination in human antemortem blood, urine, and oral fluid at endogenous concentrations. Furthermore, the stability of GHB in blood samples and its endogenous concentrations in samples taken under controlled circumstances were investigated. Samples were extracted in methanol/acetonitrile and processed by anion exchange solid-phase extraction. GHB was separated from structural isomers using a reversed–phase LC column with anion properties. The validated limit of quantification was 0.005 µg/mL in blood and 0.010 µg/mL in urine and oral fluid, at which the relative reproducibility standard deviation and bias were <15 %. The mean extraction recovery was ≥90 %. The average GHB concentration increased by 1.2 µg/mL in fluoride/citrate- preserved blood after 28 days of storage at 4°C; however, in fluoride/oxalate (FX)-preserved blood, the mean concentration increased by only 0.055 µg/mL. No change was observed at −20°C. In 105 randomly selected samples of FX-preserved blood collected for forensic antemortem toxicological analysis, all concentrations were <0.066 µg/mL, even after long-term storage at −20°C. In blood, urine, and oral fluid samples from a clinical study of GHB intake, endogenous baseline levels from 30 participants ranged from 0.0069–0.050, 0.024–0.38, and 0.034–0.93 µg/mL, respectively. These results demonstrate that the current cut-off level of 5 µg/mL for discriminating between endogenous and exogenous GHB in antemortem blood could be considerably lower for FX-preserved blood stored at −20°C.

Effervescent-assisted Liquid-phase Microextraction Employing Switchable Hydrophilicity Solvent for the Determination of Cortisol and Testosterone in Oral Fluid by High-Performance Liquid Chromatography-Diode Array Detection.

Cortisol and testosterone are important biomarkers for diagnosing complex disorders, including polycystic ovary syndrome and Cushing's syndrome, where symptomatology usually overlaps with other prevalent disorders. This work proposes, for the first time, an analytical method based on a switchable hydrophilicity solvent as an extraction phase for the determination of cortisol and testosterone in oral fluid (OF) by high-performance liquid chromatography with diode-array detection. The optimized extraction conditions consisted of 1000µL of OF, 100µL of decanoic acid solution (65mg/mL), 170µL of Na2CO3, 900µL of H2SO4 and 150µL of acetonitrile for dilution. The method was validated, and coefficients of determination higher than 0.9926, the limit of detection of 4.55ng/mL and the limit of quantification of 15.00ng/mL were obtained. Intra-day precision varied from 5.6% to 11.9%, inter-day precision ranged from 6.1% to 13.5%, and relative recoveries ranged from 98.9% to 104.6% for cortisol, and 89.1% to 103.9% for testosterone. This methodology was successfully applied to five OF samples from volunteers. Moreover, the greenness of this methodology was evaluated based on the sample preparation metric of sustainability achieving a global score of 7.37 which can be considered sustainable.

Assessment of clinical and laboratory parameters of oral сavity in patients with orthodontic systems

equipment (braces) on the biochemical parameters of oral fluid. Materials and methods. The study involved patients aged 18 to 22 years who are undergoing orthodontic treatment with a bracket system. Thegroup of examined persons was 25 people. The control group included 15 practically healthy people without braces. All patients underwent dental status assessment (Green – Vermillion hygiene index – simplified IG-Y; PMA inflammation index), laboratory examination of oral fluid before and after controlled oral hygiene. The biochemical analysis included determination of the acid-base balance in the oral fluid using a pH meter, assessment of free radical oxidation by chemiluminogram parameters by biochemiluminescence and by levels of molecular products of lipid peroxidation (diene and triene conjugates, Schiff bases), activity of the enzyme alkaline phosphatase, concentrations of total protein and lactate. Results and discussions. In patients undergoing orthodontic treatment with a bracket system, an insufficient level of oral hygiene and inflammatory manifestations in the gums of moderate severity were revealed. The level of the acid-base balance of the oral fluid before controlled hygiene was equal to 6.8±0.19, that is, below the range of normal pH values in the oral cavity. After hygiene, the pH of the oral fluid returned to the norm of 7.1±0.2. In patients with braces, activation of free radical oxidation and an increase in lipoperoxidation parameters – triene conjugates and Schiff bases, increased activity of alkaline phosphatase, total protein and lactate levels took place.Cоnclusion. After hygienic treatment of the oral cavity, the biochemical parameters did not differ statistically significantly from the control data.

Probe Electrospray Ionization coupled to a Quadrupole Time-Of-Flight: A feasibility study for the detection of cocaine and its derivatives in oral fluid

Probe Electrospray Ionization coupled to a Quadrupole Time-Of-Flight: A feasibility study for the detection of cocaine and its derivatives in oral fluid

Development of a concentration method for simple and sensitive detection of SARS-CoV-2 in saliva using a magnetic nanoparticle kit

BackgroundWhen diagnosing viral infections in humans and animals, the presence of virus in a sample in trace amounts that are below the analytical sensitivity of the detection system may cause false negative results and inaccurate diagnosis. We previously reported the development of a simple virion concentration technique using 12ml large-volume samples that can dramatically improve diagnostic sensitivity by increasing analytical sensitivity by 100-fold over conventional methods. The present study was conducted to further improve the simplicity and versatility of this method. We constructed a simple and highly sensitive method for the detection of SARS-CoV-2 in human saliva after concentration using a magnetic nanoparticle conjugated with polyethylene glycol (PEG). ResultsPerformance of the method was evaluated by comparing a combination of automated nucleic acid extraction and RT-qPCR or triplex RT-LAM detection in a spiked sample of 20ml saliva collected from healthy humans. The method theoretically achieved 300-fold concentration of spiked SARS-CoV-2 in saliva, enabling 10- to 1000-fold higher analytical sensitivity for detection compared to conventional RNA extraction methods. ConclusionsThis newly developed method allows for easy and reliable concentration of the virion in less than 60min, improving the analytical sensitivity of the SARS-CoV-2 test. Further, the method allows for easy and reliable enrichment of the virus in less than 60min, improving the analytical sensitivity of the SARS-CoV-2 test. This method is easily used for highly sensitive virus detection from a variety of human oral fluid samples and may also be applied to rapid and labor-saving screening tests by pooling a large number of samples.

The Role of Interleukin-8 in the Estimation of Responsiveness to Steps 1 and 2 of Periodontal Therapy.

To investigate the association between interleukin-8 (IL-8) levels in gingival crevicular fluid (GCF) and total oral fluid (TOF) and the responsiveness to steps 1 and 2 of periodontal therapy. One-hundred and fifty-nine patients affected by periodontitis received steps 1 and 2 of periodontal therapy. At baseline, TOF and GCF samples were collected and analysed for IL-8 (Il-8TOF/IL-8GCF) using flow cytometry. Therapy outcomes were relative proportions of residual periodontal pockets (PPD%), pocket closure (PC) rates and pocket probing depth (PPD) reductions; these were associated with IL-8TOF/IL-8GCF. High IL-8TOF was significantly associated with higher residual PPD% (p = 0.044) and lower PPD reduction compared to low IL-8TOF (high 0.79 ± 1.20 mm vs. low 1.20 ± 1.20 mm, p < 0.001) in non-smokers, while in smokers high IL-8GCF was related to lower PPD reduction (high 0.62 ± 1.22 mm vs. low 0.84 ± 1.12 mm, p = 0.009). Furthermore, high baseline IL-8TOF was significantly associated with poorer PC rates compared to medium and low concentrations in both non-smokers (high 41% vs. medium 55% vs. low 58%, p < 0.001) and smokers (high 34% vs. medium 44% vs. low 46%, p < 0.001). High IL-8 concentrations at baseline had a significant impact on residual PPD%, PC rates and PPD reduction. The findings suggest that, especially in non-smokers, baseline IL-8 levels collected from the TOF could serve as a component in the estimation of responsiveness to steps 1 and 2 of periodontal therapy.

Biochemical parameters and oral fluid microcrystallization in young patients who use a vape device

The study aims s are researching the negative effect of nicotine-containing vapes on physical and chemical parameters of oral fluid and morphotyping of dried oral fluid drop among young somatically healthy individuals. Material and methods. The study included 20 people between the ages of 18 and 24 years old, divided into 2 groups: the «Vape» group (n=12) human only nicotine-containing vape for at least 2 years and the «Control» group (n=8) not human nicotine in any form. pH, thiocyanate (rhodonide) concentrations, surface tension, oral fluid volume, hygiene indices and periodontal indices were studied. The morphotype of dried oral fluid drop was analyzed. Results. In the «Control» group in 3 cases gingivitis chronic K05.1 was diagnosed, and in the patients of the «Vape» group in 9 cases. It was found that the rhodonide content in the «Vape» group before «vaping» was higher than in the «Control» group and was Me= 1.31 mM/L and Me=0.58 mM/L respectively p=0.012. After vape application, the concentration of rhodonides in oral fluid increased in 1.16 times (p=0.0077), nitrite ions content from Me=0.09 mg/L to Me=0.32 mg/L (p=0.07), pH increased to Me=7.64; (LQ=7.45; HQ=7.67) (p=0.07) (p=0.07). The volume of RW in the Vape group remains 2.8 times smaller than in the Control group (p=0.012). The microcrystallization pattern of the oral fluid drop in the «Vape» group before «steaming» more often corresponded to type II A, and after «steaming» in 33.4% cases type III was determined, in 25% – type II B. Conclusions. Thus, in «vapers» of 18–24 years old statistically reliable reduction of oral fluid volume, high values of hygiene index shift of acid-base balance in the direction of alkalosis creates conditions in the oral cavity for the development of periodontal diseases and the emergence of cariesogenic situation. There was an increase in the concentration of rhodanide and nitrite ions, which in combination with other components of vape aerosols can be considered as procarcinogens, syncarcinogens and cocarcinogens. The change in the morphology of facies pattern, type of microcrystallization of oral fluid drop in the dynamics of vape use corresponds to the vector of changes observed in other inflammatory diseases of the oral cavity

Comparative analysis of the parameters of local immunity of oral fluid and the duration of the course in children with systemic lupus erythematosus, juvenile scleroderma, juvenile dermatomyositis, juvenile idiopathic arthritis and systemic vasculitis

Immunoglobulins are considered to be the main factor of specific antimicrobial protection of oral tissues.The study was conducted on the basis of the University Children’s Clinical Hospital of the I.M. Sechenov First Moscow State Medical University of the N.F. Filatov Clinical Institute of Child Health, in the Department of Rheumatology. The study involved 316 children, with diseases: systemic lupus erythematosus (SLE), juvenile dermatomyositis (YDM), juvenile limited scleroderma (YOSD), juvenile systemic scleroderma (YSSD), juvenile idiopathic arthritis (JIA), systemic vasculitis (SV) and Behcet’s disease (BB) and were divided into age groups according to the duration of the course of the underlying disease. The analysis of the results revealed a decrease in the content of IgG, IgA, sIgA in saliva in children with rheumatic diseases in all age groups compared with the control group (children without somatic diseases). With the duration of the course of the disease up to 2 years, the most pronounced decrease in the content of IgG, IgA, sIgA in saliva was revealed, more than 4–10 times (p&lt;0.05), and with a duration of more than 2 years, a decrease of 2–3 times (p&lt;0.01), compared with the control group. The deficiency of these immunoglobulins shows that in children with rheumatic diseases, regardless of the nosology, there is a violation of the local immunity of the oral cavity, especially at the onset of the underlying disease (in groups up to 2 years of the duration of the course of the underlying disease), with pronounced autoimmune inflammation of tissues and blood vessels, and this is also facilitated by the reception, during this period of shock doses of glucocorticosteroids, cytostatics, immunosuppressors, which leads to severe inflammatory processes of the oral mucosa, periodontal and hard tissues of the teeth.

Changes in heavy metal levels in the oral fluid of young patients with and without plaque-induced gingivitis undergoing treatment with braces

Relevance. Research into the effects of nickel-titanium alloys and heavy metal ions on the development of inflammatory oral diseases is becoming increasingly significant in orthodontics. The relationship between toxic elements and inflammatory responses remains under-researched and controversial. Data on the connection between heavy metals and inflammatory reactions could influence the choice of orthodontic appliance materials and guide the subsequent management and monitoring of patients with braces. The question of the safe use of nickel-titanium alloy braces continues to be a relevant concern.Purpose. To assess the levels of heavy metals in the oral fluid of young patients with braces, both in those who developed plaque-induced gingivitis and those who did not.Materials and Methods. The study involved 50 clinically healthy patients (Groups I and II health categories) with intact teeth, including patients with compensated dental caries (isolated carious lesions – Grade I) and no periodontal diseases (PMA index &lt;20), presenting with dental crowding and malocclusion (K07.2, K07.3). The average age of the patients was 22.1 ± 2.7 years, all of whom were undergoing treatment with metal braces. Oral fluid samples were analyzed for 12 heavy metal elements using inductively coupled plasma mass spectrometry (ICP-MS) in a laboratory setting. Additionally, lysozyme activity and oral fluid pH were measured before appliance placement and six months into orthodontic treatment.Results. Adaptation to braces in young patients is frequently complicated by the development of gingivitis and periodontitis due to impaired oral hygiene and the mechanical pressure exerted on teeth and gums, leading to elemental imbalances. Patients with gingivitis during orthodontic treatment are particularly at risk of developing such imbalances and therefore require timely correction of their mineral metabolism.

Immunological indicators of the risk of the transition from chronic gastritis to gastric ulcer in adolescents

As is known, gastric ulcer (GCC) and duodenal ulcer (duodenal ulcer) are chronic, complex-etiology diseases, the mechanism of which is based on complex transformations of nervous, hypothalamic-pituitary, hypothalamic-pituitary-adrenal and local gastroduodenal processes that lead to a change in trophism in the mucous membrane of the stomach and duodenum. In turn, the incipient ulcerative defect is a source of irritation of the enteric receptors, supporting disorders of neurohormonal regulation. Thus, the resulting “vicious circle” causes the chronic course of peptic ulcer disease. During the study of chronic diseases of gastroduodenal localization, including peptic ulcer disease, many doctrines of its occurrence have been proposed. However, the issue of the origin of peptic ulcer disease still remains insufficiently studied; approaches and tactics for managing patients with erosive and ulcerative lesions of the upper gastrointestinal tract are being developed. The aim of the study was to evaluate immunological indicators that determine the risk of chronic gastritis turning into gastric ulcer. The study included 120 children aged 14-17 years, permanently residing in the Bukhara region of the Republic of Uzbekistan. Taking into account the importance of H. Pylori in the development of HCG, patients were distributed depending on the association of HCG with H. Pylori. To study cytokines, blood tests of patients were performed to determine the titer of antibodies (IgG and IdM) to H. Pylori, immunoglobulins A, M, G in blood serum, secretory IgA in oral fluid, IL-1β, IL-10, TNFα, VEGF in the serum of patients. At the same time, blood sampling was carried out in the periods before and 3 weeks (21 days) after treatment. Thus, the study of the value of immunological blood parameters in the progression of HCG to gastric ulcer disease dictates the need to develop immunological indicators for predicting the progression of HCG to gastric ulcer in adolescents. An increase in TNFα levels indicates a high risk of developing gastric ulcer as a result of vascular damage and impaired local circulation. In the study, an established increase in the level of IL-1β indicates activation of the synthesis of proinflammatory cytokines in HCG, especially in HCG associated with H. Pylori. The use of these recommendations contributes to the monitoring of HCG, improving the effectiveness of diagnosis and the correct choice of treatment for patients with HCG, reducing mortality and disability at the same time.

Inter-laboratory harmonization of microsphere immunoassays for SARS-CoV-2 antibody detection in dried blood spots and oral fluids.

Dried blood spots (DBS) and oral fluids (OF) are easily attainable biospecimen types that have enabled population scale antibody monitoring for SARS-CoV-2 exposure and vaccination. However, the degree to which the two different biospecimen types can be used interchangeably remains unclear. To begin to address this question, we generated contrived DBS (cDBS) and OF (cOF) from serum panels from SARS-CoV-2 infected, vaccinated, and uninfected individuals. The contrived samples were evaluated using SARS-CoV-2 multiplexed microsphere immunoassays (MIAs) at two different institutions. Intra-laboratory tests revealed near perfect agreement between cDBS and cOF for N and S antigens, as evidenced by κ = 0.97-1 and 98%-100% agreement. Inter-laboratory comparisons were equally robust for both N (κ = 0.94-0.96; 97.5%-98 % agreement) and S (κ = 0.98 -1.0; 99.0%-100%). Furthermore, assays were transferred between labs, including methods and reagents, and a subset of cDBS and cOF samples (n = 52) were tested. Qualitative concordance remained high (κ = 0.94-1.0; 97.5%-100% agreement), confirming that integrity of the assays is retained upon transfer. In summary, our results provide evidence that DBS and OF can be used interchangeably across laboratories and institutions for the qualitative assessment of SARS-CoV-2 antibody determinations.

Environmentally friendly screen-printed electrodes for the selective detection of 4-bromo-2,5-dimethoxyphenethylamine (2C-B) in forensic analysis.

In response to the growing need for sustainable analytical methods, this study explores the repurposing of screen-printed electrodes (SPEs) that would otherwise be discarded. This involves recoating the working electrode surface with a graphite (Gr) and chitosan (CTS) dispersion, creating a reusable SPE (SPE-Gr/CTS). Demonstrating its utility, SPE-Gr/CTS was employed for the detection of 4-bromo-2,5-dimethoxyphenethylamine (2C-B), a phenylethylamine commonly used for recreational proposes. Identifying 2C-B in fluid oral and seized samples is of great interest for forensic and toxicological applications. The 2C-B detection using SPE-Gr/CTS was optimized in Britton-Robinson buffer solution (0.1 mol L-1) at pH 2.0, employing square-wave adsorptive stripping voltammetry. The electrochemical behavior of 2C-B on SPE-Gr/CTS exhibited one irreversible oxidation and a reversible redox process. The proposed method presented a dynamic linear range for 2C-B determination (0.05 to 7.5 μmol L-1) with a low LOD (0.015 μmol L-1). Moreover, the stability of 2C-B electrochemical responses on SPE-Gr/CTS was confirmed using the same or different electrodes (N = 3), with a relative standard deviation of less than 5.0%. Interference studies with seventeen other illicit drugs and adulterants demonstrated that the proposed method is selective for 2C-B detection even in the presence of these substances. Real seized and oral fluid samples containing 2C-B were analyzed using this method, and the results were confirmed by LC-MS. The proposed device demonstrates to be an environmentally friendly and selective sensor for 2C-B detection in forensic analysis, offering a rapid and straightforward screening method for seized and biological samples. In addition, a portable and sensitive determination of 2C-B in forensic samples is presented with minimal sample consumption (50 μL).

Oropharyngeal swab sampling for PRRSV detection in large-scale pig farms: a convenient and reliable method for mass sampling

BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) has significant productivity and economic impacts in swine herds. Accurately determining the PRRSV status at the herd level is crucial for producers and veterinarians to implement strategies to control and eliminate the virus from infected herds. This study collected oropharyngeal swabs (OSs), nasal swabs (NSs), oral fluid swabs (OFs), rectal swabs (RSs), and serum samples continuously from PRRSV challenged pigs under experimental conditions and growing pigs under field conditions. Additionally, OSs and serum samples were collected from individual sows from 50 large-scale breeding farms, and the collection of OSs does not require the sows to be restrained. Ct values of PRRSV were detected in all samples using real-time reverse transcriptase-polymerase chain reaction (RT-qPCR).ResultsIn PRRSV challenged pigs, OSs showed a higher PRRSV-positive rate until the end of the observation period. The Ct values of OSs were significantly lower than those of NSs, OFs, and RSs at 2, 8, 12, 14 and 20 days post-challenge (DPC) (P < 0.05). For growing pigs, the positivity rate of PRRSV in OSs was higher than that in other sample types at 30, 70, and 110 days of age. In sows, 24,718 OSs and 6259 serum samples were collected, with PRRSV-positive rate in OSs (9.4%) being significantly higher than in serum (4.1%) (P < 0.05). However, the Ct values of PRRSV RNA in serum were significantly lower than those in OSs (P < 0.001).ConclusionsThe OSs sample type yielded higher PRRSV-positive rates for longer periods compared to NSs, RSs, OFs and serum samples for PRRSV detection in infected pigs. Therefore, OSs has a good potential to be a convenient, practical, and reliable sample type for implementing mass sampling and testing of PRRSV in large-scale pig farms.