Oral Fluid Research Articles

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.

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.

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.

Salivary biomarkers: a promising approach for predicting immunotherapy response in head and neck cancers.

Head and neck cancers, including cancers of the mouth, throat, voice box, salivary glands, and nose, are a significant global health issue. Radiotherapy and surgery are commonly used treatments. However, due to treatment resistance and disease recurrence, new approaches such as immunotherapy are being explored. Immune checkpoint inhibitors (ICIs) have shown promise, but patient responses vary, necessitating predictive markers to guide appropriate treatment selection. This study investigates the potential of non-invasive biomarkers found in saliva, oral rinses, and tumor-derived exosomes to predict ICI response in head and neck cancer patients. The tumor microenvironment significantly impacts immunotherapy efficacy. Oral biomarkers can provide valuable information on composition, such as immune cell presence and checkpoint expression. Elevated tumor mutation load is also associated with heightened immunogenicity and ICI responsiveness. Furthermore, the oral microbiota may influence treatment outcomes. Current research aims to identify predictive salivary biomarkers. Initial studies indicate that tumor-derived exosomes and miRNAs present in saliva could identify immunosuppressive pathways and predict ICI response. While tissue-based markers like PD-L1 have limitations, combining multiple oral fluid biomarkers could create a robust panel to guide treatment decisions and advance personalized immunotherapy for head and neck cancer patients.

Monitoring of Respiratory Disease Patterns in a Multimicrobially Infected Pig Population Using Artificial Intelligence and Aggregate Samples

A 24/7 AI sound-based coughing monitoring system was applied in combination with oral fluids (OFs) and bioaerosol (AS)-based screening for respiratory pathogens in a conventional pig nursery. The objective was to assess the additional value of the AI to identify disease patterns in association with molecular diagnostics to gain information on the etiology of respiratory distress in a multimicrobially infected pig population. Respiratory distress was measured 24/7 by the AI and compared to human observations. Screening for swine influenza A virus (swIAV), porcine reproductive and respiratory disease virus (PRRSV), Mycoplasma (M.) hyopneumoniae, Actinobacillus (A.) pleuropneumoniae, and porcine circovirus 2 (PCV2) was conducted using qPCR. Except for M. hyopneumoniae, all of the investigated pathogens were detected within the study period. High swIAV-RNA loads in OFs and AS were significantly associated with a decrease in respiratory health, expressed by a respiratory health score calculated by the AI The odds of detecting PRRSV or A. pleuropneumoniae were significantly higher for OFs compared to AS. qPCR examinations of OFs revealed significantly lower Ct-values for swIAV and A. pleuropneumoniae compared to AS. In addition to acting as an early warning system, AI gained respiratory health data combined with laboratory diagnostics, can indicate the etiology of respiratory distress.

MiRNAs as Epigenetic Biomarkers in the Study of the Bidirectional Relationship between Type 2 Diabetes Mellitus and Periodontitis: A Systematic Review.

The objective of this study is to analyze the miRNA expression of oral fluids such as gingival crevicular fluid (GCF) in patients with periodontitis and Type 2 diabetes mellitus, and how these epigenetic biomarkers can influence the bidirectional relationship of these two inflammatory diseases. This review was conducted following the PRISMA criteria. PubMed, Scopus, Cochrane Library, Embase, and Web of Science databases were searched for clinical studies conducted on humans investigating, through GCF miRNA expression, the relationship between periodontal diseases and type 2 diabetes mellitus. In addition, the etiopathogenic pathways of the studied miRNAs were analyzed using the DIANA MIR path tool. A total of 1436 references were identified in the initial literature search, and seven articles were finally included in this review. Most of the articles included in this review were case-control studies and examined the expression of miRNAs in patients with periodontitis with or without diabetes. Due to their characteristics, miRNAs appear to be the ideal biomarkers for improving the understanding and knowledge of the etiopathogenic pathways that link both diseases. Among all the studied miRNAs, miR-146a, miR-155, miR-200b, miR-223, and miR-203 showed strong involvement in inflammatory and metabolic pathways, making them potential good diagnostic and prognostic biomarkers.

Cytokine profile of oral fluid in case of traumatic fractures of the lower jaw and the presence of third molars in patients` oral cavity

Objective. To analyze the level of cytokines in the oral fluid of patients with traumatic fractures of the mandible in the presence of third molars, to establish the influence of the latter on the oral cavity homeostasis, to determine the most informative indicators. Materials and methods. 82 people aged 18–60 years took part in the study, they were divided into 3 groups. Group 1 contained 43 patients with mandibular fracture and having more than two third molars. Group 2 consisted of 18 patients with mandibular fracture, without third molars. Group 3 included 21 people without a fracture of the mandible, who had more than two-thirds of the molars. Group 4 (control) contained the data of somatically and dentally healthy individuals, borrowed from special literature. The levels of IL-1β, IL-6, IL-8, and TNF-a in OF were determined using standard enzyme immunoassay kits. The data were processed statistically. Results. A comparison of the indicators of groups 1 and 3 revealed a significant increase in the level of IL-6 z = 4.15 (H = 20.43; p = 0.0001) and the level of IL-8 z = 3.08 (H = 12.29; p = 0.0064) in patients with a jaw fracture. A considerable increase in IL-8 levels was determined in the individuals from group 2 compared with group 3 – z = 2.74 (H = 12.29; p = 0.0064). The comparison of groups 1 and 4 in terms of TNF-a showed a tendency to a significant difference of z = 1.60 (H = 4.65; p = 0.19), and the data of group 2 in comparison with group 4 were significantly higher than z = 1.97 (H = 4.65; p = 0.19). Comparing the results of groups 3 and 4 with each other we revealed a considerable difference of z = 1.75 (H = 4.65; p = 0.19). Conclusion. The profile of cytokines level in the OF indicates that the presence of third molars in the oral cavity in mandibular fractures affects the homeostasis of the oral cavity negatively, which is the reason for expanding the indications for their removal in order to prevent infectious and inflammatory complications.

Concentrations of Delta 9-tetrahydrocannabinol (THC) in oral fluid at different time points after use: An individual participant meta-analysis

BackgroundDelta 9-tetrahydrocannabinol (THC) concentrations in oral fluid (OF) at different time points after cannabis administration and factors related to these concentrations have not been previously described in a meta-analysis. This information is critical for better understanding of these tests for detection of prior cannabis use and cannabis impairment. Objectives1: To describe the summary statistics of THC concentrations at different time points after cannabis administration. 2: To describe the relationship between the variables of dose of THC, frequency of using cannabis, route of administration (i.e., inhaled or ingested), OF collection device and sex, with THC concentrations in OF, based on bivariate analyses. 3: To describe the independent contribution of each of the variables in Objective 2, based on a multivariate analysis of THC concentrations. MethodsA meta-analysis of studies from two databases (PubMed and Scopus) was conducted. Our inclusion criteria included published empirical articles that administered natural cannabis to subjects in a controlled setting, with OF drug tests showing the exact THC concentrations in OF for each subject (i.e., raw data) for at least two time points after cannabis administration using confirmatory methods. Seven studies of tests with published raw data for OF THC after cannabis administration met these criteria (n observations=1157). ResultsSummary statistics showed OF THC concentrations by time after use were highly dispersed at every time point, positively skewed, and declined over time. Many positive OF THC concentrations were found after 24-hours in one study, but most studies did not conduct observations past 24 hours. In a multivariate analysis, we found that increased dose, increased frequency of cannabis use, and inhaled (versus ingested) cannabis were statistically related to higher OF THC concentrations. OF collection with the intercept DOA device was significantly higher than expectorant (i.e. saliva) and being male (versus female) were only significant in a bivariate analysis. Too little data existed to reliably analyze the possible influence of other variables of age, race and body mass index (BMI) on OF THC concentrations. DiscussionFalse negatives exist when the tests are used to detect prior use. OF test results are related to confounders of frequency of cannabis use and inhaled (versus ingested) cannabis. OF tests can produce positives at a cut-off 1.0 ng/mL well beyond 24 hours. The tests are not valid to detect cannabis impairment. More information is needed on the influence of potential confounders for OF concentrations. We do not have a good idea of the degree to which the subjects in these studies are representative of persons who use cannabis. Overall, more research is needed for these tests to be used in workplace settings.

Pipette-tip microextraction using carboxymethylated wooden-based sawdust sorbent for the determination of antidepressants in oral fluid by direct infusion mass spectrometry

Matrix complexity is a challenging scenario in sample preparation and the scientific community is in the search of novel materials capable of replacing the classical sorbents to overcome the sensitivity and selectivity issues it presents. In this article, the use of a carboxymethylated wooden-based sawdust sorbent pipette-tip microextraction is introduced for the determination of four antidepressants (imipramine, amitriptyline, fluoxetine, and clomipramine) in human oral fluids. The different variables affecting the extraction and elution procedures, including sample pH, ionic strength, number of pushing cycles, dilution of the sample, eluent, and volume of the eluent have been evaluated by liquid chromatography – ultraviolet detection. Moreover, the proposed method was validated by direct infusion coupled to tandem mass spectrometry with limits of detection of 0.2 μg/L, limits of quantification of 0.6 μg/L, linear range spanning from 0.6 to 320 μg/L and intra-day precision (expressed as relative standard deviation) better than 14.2 %. Intra-day accuracy (expressed as relative recovery) ranged from 85.4 to 102.9 %, while inter-day accuracy was between 97.0 and 103.9 %. Finally, the analysis of two oral fluids from a patient under amitriptyline treatment were analyzed, providing values within the linear range of the proposed method.

Extraction of synthetic cathinones from biological samples: A systematic review

Analysis of biological samples containing synthetic cathinones has become a significant area of interest for researchers in recent years, because synthetic cathinones can cause adverse reactions such as delusions and hallucinations. Generally, the intricate nature of the sample matrix and exceedingly low drug concentration in biological samples present significant challenges for the determination of target substances. Accordingly, scientists were dedicated to the advancement of effective sample preparation techniques to address these difficulties. However, the current reviews involved in synthetic cathinones all focused on detection methods. A systematic review focusing on sample preparation for the quantification of synthetic cathinones is still lacking. In light of that, this paper reviewed the most commonly used sample preparation techniques for synthetic cathinones. Recent advances in sample preparation techniques for synthetic cathinones in different biological samples such as blood, urine, oral fluid and hair samples were discussed. In addition, we prospected the challenges and the future perspectives of biological sample preparation techniques for synthetic cathinones determination.

"Optical Properties and Translucency of ImplantSupported PEEK Crowns: Spectrophotometric Evaluation"

AIM- To evaluate the difference in optical properties and translucency parameters of implant supported PEEK crowns at different time intervals. MATERIAL AND METHODS: Optical properties and translucency parameters of implant supported PEEK crowns were assessed in 35 patients using spectrophotometer RESULT: Results revealed noticeable changes in translucency parameters and optical properties of implant supported PEEK crowns at 3 months. CONCLUSION- Our study on 35 patients on implant-supported PEEK crowns revealed that glazing improves surface roughness and opalescence, enhancing aesthetic appeal. Although, oral fluids containing intrinsic and extrinsic factors still adversely affect PEEK's color stability and surface quality