Saliva Samples Research Articles

Alterations in the Salivary Microbiome and Metabolism in Patients With Carotid Atherosclerosis from Rural Northeast China.

Periodontitis and atherosclerosis are both chronic inflammatory diseases with a high prevalence. Increasing evidence supports the independent association between severe periodontitis and atherosclerotic cardiovascular disease, in which oral microorganisms may play an important role. We aimed to evaluate the characteristic changes of salivary microbiome and metabolome in patients with carotid atherosclerosis (CAS) and periodontitis. The subjects were obtained from a cross-sectional study that included 1933 participants aged 40 years or older from rural northeast China. The study enrolled 48 subjects with CAS and 48 controls without CAS matched by sex, age, body mass index, and prevalence of hypertension, diabetes, and dyslipidemia. We performed full-length 16S rDNA gene sequencing and untargeted metabolomics of saliva samples from 96 subjects. We found that CAS was closely associated with an increased abundance of Streptococcus, Lactobacillus, and Cutibacterium. Furthermore, patients with CAS had higher prevalence of severe periodontitis than the control group. Notably, periodontal pathogens such as Tannerella and Anaeroglobus were not only associated with periodontitis but also enriched in patients with CAS, whereas periodontal health-associated Neisseria was more abundant in those without CAS. We also identified 2 lipid metabolism pathways, including glycerophospholipid and sphingolipid metabolism, as associated with CAS. The levels of trimethylamine N-oxide and inflammatory mediator leukotriene D4 were significantly higher in patients with CAS, whereas the levels of carnosine were significantly lower, than those in controls. Additionally, serum levels of inflammatory marker high-sensitivity C-reactive protein were significantly increased in CAS and positively correlated with the abundance of Anaeroglobus and leukotriene D4 in saliva. Our study suggests that characteristic changes in salivary microbiota and metabolites are closely related to CAS, and periodontitis and associated microorganisms may be involved in the initiation and progression of CAS.

Mixed-mode cationic exchange paper combined with direct infusion mass spectrometry, a sustainable approach to determine opioids in biosamples

Direct infusion mass spectrometry (DI-MS/MS) allows the rapid analysis of samples and minimizes solvent usage by avoiding chromatographic separation. However, it is prone to ion suppression due to matrix effects, especially when biosamples are processed. In this context, sample preparation appears as the perfect complement to get proper analytical results. In this article, a novel and sustainable planar sorptive phase containing mixed-mode cationic exchange (MCX) particles is presented to overcome the limitations of DI-MS/MS in bioanalysis. The new phase uses cellulose paper as a natural substrate over which the particles can be retained using polyacrylonitrile (PAN) as the chemical binder. The high porosity of the cellulose substrate simplifies the synthesis since a straightforward dip-coating technique can be applied. For this purpose, MCX particles are initially dispersed in a PAN solution prepared in N,N-Dimethylformamide. A segment of paper is later introduced into the slurry, and the evaporation of the solvent leaves a film of PAN-MCX composite over the paper, where the particles are entrapped but accessible for the analyte's isolation. The resulting MCX-paper has been evaluated for the extraction of selected opioids (codeine, methadone, morphine, naloxone, oxycodone, and tramadol) from saliva and urine samples and their determination by direct infusion tandem mass spectrometry. Working at the optimized conditions, the limits of detection were in the low μg L−1 range. The precision, expressed as relative standard deviation, was better than 12 % under intra-day and inter-day conditions. The accuracy, expressed as relative recoveries, provides values in the ranges 89–113 % and 74–121% for saliva and urine, respectively. The method was finally applied to analyze saliva and urine samples collected from patients under codeine treatment.

Comparative proteomic analysis of saliva from chewing tobacco users and oral cancer patients reveals shared biomarkers: A case control observational study

ObjectiveThe aim of this study was to compare the salivary proteomic profile of smokeless tobacco users with that of non-users and oral cancer patients using Liquid Chromatography-Mass Spectrometry/ Mass Spectrometry (LC-MS/MS). MethodsSaliva samples from 65 participants were collected in three groups: control (25 participants), smokeless tobacco users (25 participants), and oral cancer (15 participants). ResultsThe analysis revealed 343 protein groups with significantly altered abundance in the saliva samples (P < 0.05). Among these, 43 out of 51 dysregulated proteins in the smokeless tobacco group were also dysregulated in the oral cancer group. Notably, Apolipoprotein A1 (ApoA1) and Pon1 were found to be significantly increased in both smokeless tobacco users and oral cancer patients (p < 0.05). Furthermore, six out of the 20 most significantly altered proteins were mitochondrial proteins, and all of these were decreased relative to controls in both smokeless tobacco users and cancer samples. ConclusionThe proteomic profile of users of chewing (smokeless) tobacco (SLT) shows substantial overlap in the altered pathways and dysregulated proteins with those altered in oral cancer samples, suggesting that SLT use induces a shift toward an oncogenic state. Specifically indicated pathways included blood microparticles, platelet α-granules and protease inhibitors as well as indicators of oxidative stress and exogenous compound processing. What differentiates oral cancer samples from SLT users is enrichment of alterations related to cytoskeletal organisation and tissue remodelling. Clinical SignificanceThe findings emphasize the importance of salivary proteomic profiles because changes in certain proteins may be indicators for early oral cancer identification and risk assessment in smokeless tobacco users.

The associations of spirituality and Hispanic ethnicity with neuroendocrine biomarkers among patients with colorectal cancer

ObjectiveDealing with cancer evokes not only physical and emotional distress, but may also promote resilience through spirituality. Patients with cancer are vulnerable to neuroendocrine dysregulation. This longitudinal observational study examined the degree to which spirituality was associated with neuroendocrine biomarkers and the moderating role of Hispanic ethnicity. MethodsParticipants were adults who were recently diagnosed with colorectal cancer (n = 81, 55 years old, 66% male, 63% Hispanic, 72% advanced cancer, 7 months post-diagnosis). The domains of spirituality (faith, meaning, and peace) and ethnicity (Hispanic vs. non-Hispanic) were self-reported. Cortisol and alpha amylase (sAA) were assayed from saliva samples collected at waking and bedtime on seven consecutive days. Mean levels at waking and bedtime, and diurnal slopes over seven days were calculated. Age and cancer stage were covariates. Results: Overall, patients reported moderate to high levels of spirituality. General linear modeling revealed that greater faith was associated with higher levels of sAA at waking and bedtime as well as more blunted diurnal pattern of sAA only among Hispanic patients (p ≤ .045). Greater peace was associated with steeper diurnal pattern of sAA, regardless of ethnicity (B = 0.021, p = .005). Meaning and cortisol were not significantly associated with study variables. ConclusionsFindings indicate that presence of peace facing a cancer diagnosis associated with neuroendocrine regulation, whereas drawing on one's faith, particularly among Hispanic patients, associated with neuroendocrine dysregulation during the first months after the diagnosis. Further investigations of psychobiobehavioral moderators and mediators for healthy neuroendocrine functioning among patients with cancer are warranted.

Antimicrobial Efficacy of Mangifera indica, Mentha arvensis, and Chlorhexidine Mouthwashes on Streptococcus mutans and Candida albicans in Children: A Comparative In Vivo Study.

Herbal medicines are both promotive and preventive in inhibiting dental caries. Most commonly used natural plant extracts like mango (Mangifera indica) and pudina (Mentha arvensis) as an entire or together are scientifically well proven to be safe and effective against various oral health problems, especially in preventing dental caries and periodontal diseases. The present study aims to comparatively evaluate the effect of Mangifera indica, Mentha arvensis, and Chlorhexidine (CHX) (0.2%) mouthwashes on Streptococcus mutans (S. mutans) and Candida albicans (C. albicans) in children. By random allocation, 45 healthy children were selected between the age-group of 7 and 10 years of both genders from a residential school. They were divided into three study groups of 15 in each; group I-CHX (control group), group II-Mangifera indica mouthwash, and group III-Mentha arvensis mouthwash. Stimulated whole saliva sample collected for the amount of colony forming units and cultured for S. mutans and C. albicans. The data was tabulated, and statistical analysis was done with one-way variance analysis of the variance (ANOVA), Tukey's post hoc honestly significant difference (HSD) test, and Bonferroni and Kruskal-Wallis test. Mangifera indica (group II) and Mentha arvensis (group III) showed a significant reduction of salivary counts at baseline, 14th day, and 21st day period and minimal effect on C. albicans for Mangifera indica. Among the three types of mouthwash, the CHX was the most superior; it showed a statistically significant reduction overall. When compared to Mangifera indica with Mentha arvensis, Mentha arvensis showed a statistically significant reduction in S. mutans and C. albicans and had an almost equivocal effect like CHX. Both herbal mouthwashes had a significant reduction in the S. mutans and C. albicans in children. So, it can be considered an active ingredient in mouthwashes. It may serve as a natural antimicrobial mouthwash with no adverse effects. Kajjari S, K VB, Janardhanan S, et al. Antimicrobial Efficacy of Mangifera indica, Mentha arvensis, and Chlorhexidine Mouthwashes on Streptococcus mutans and Candida albicans in Children: A Comparative In Vivo Study. Int J Clin Pediatr Dent 2024;17(S-1):S78-S83.

The Influence of Probiotics in Halitosis and Cariogenic Bacteria: A Systematic Review and Meta-Analysis

Background: The aim of this systematic review was to evaluate the therapeutic potential of probiotics in patients with halitosis and to assess whether probiotics can also be implemented as a preventative tool in oral health. Secondary objectives included the effect of probiotics on oral-health-related quality of life, as well as their safety. Materials and methods: An electronic literature search in Medline (PubMed), Scopus, Web of Science, and the Cochrane library was carried out for the identification and selection of relevant randomized controlled trials. Eligibility was based on inclusion criteria, which included RCTs published after 2013, and the outcome variables were volatile sulfur compound (VSC) levels, organoleptic scores, plaque, or saliva samples to assess cariogenic bacteria counts and/or pH levels. Results: Out of 192 identified records, 16 randomized controlled trials were included. Ten of those studied the effects of probiotics on halitosis and the other six analyzed the effect of probiotics on oral health parameters, such as cariogenic bacteria counts, pH levels, and salivary flow and quality. A total of 921 patients were evaluated. The risk of bias was assessed using the Cochrane risk-of-bias assessment tool version 2. Conclusions: Probiotics exhibit the potential for oral health management by reducing VSC levels, improving saliva quality, and enhancing oral-health-related quality of life. Combining probiotics with tongue scraping may sustain VSC reduction, while symbiotics show potential in reducing tongue coating. However, different bacterial strains have been used in the included studies; hence, the conclusions cannot be generalized, being one of the main limitations of this review. Future research should explore the probiotics’ potential to persist in the oral cavity post-treatment and employ standardized methodologies for conclusive efficacy assessment.

Differential Patterns of Gut and Oral Microbiomes in Hispanic Individuals with Cognitive Impairment.

Alzheimer's disease and related dementias (ADRD) have been associated with alterations in both oral and gut microbiomes. While extensive research has focused on the role of gut dysbiosis in ADRD, the contribution of the oral microbiome remains relatively understudied. Furthermore, the potential synergistic interactions between oral and gut microbiomes in ADRD pathology are largely unexplored. This study aims to evaluate distinct patterns and potential synergistic effects of oral and gut microbiomes in a cohort of predominantly Hispanic individuals with cognitive impairment (CI) and without cognitive impairment (NC). We conducted 16S rRNA gene sequencing on stool and saliva samples from 32 participants (17 CI, 15 NC; 62.5% female, mean age = 70.4 ± 6.2 years) recruited in San Antonio, Texas, USA. Correlation analysis through MaAslin2 assessed the relationship between participants' clinical measurements (e.g., fasting glucose and blood cholesterol) and their gut and saliva microbial contents. Differential abundance analysis evaluated taxa with significant differences between CI and NC groups, and alpha and beta diversity metrics assessed within-sample and group compositional differences. Our analyses revealed no significant differences between NC and CI groups in fasting glucose or blood cholesterol levels. However, a clear association was observed between gut microbiome composition and levels of fasting glucose and blood cholesterol. While alpha and beta diversity metrics showed no significant differences between CI and NC groups, differential abundance analysis revealed an increased presence of oral genera such as Dialister , Fretibacterium , and Mycoplasma in CI participants. Conversely, CI individuals exhibited a decreased abundance of gut genera, including Shuttleworthia , Holdemania , and Subdoligranulum , which are known for their anti-inflammatory properties. No evidence was found for synergistic contributions between oral and gut microbiomes in the context of ADRD. Our findings suggest that similar to the gut microbiome, the oral microbiome undergoes significant modifications as individuals transition from NC to CI. Notably, the identified oral microbes have been previously associated with periodontal diseases and gingivitis. These results underscore the necessity for further investigations with larger sample sizes to validate our findings and elucidate the complex interplay between oral and gut microbiomes in ADRD pathogenesis.

Autistic Children/Adolescents Have Lower Adherence to the Mediterranean Diet and Higher Salivary IL-6 Concentration: Potential Diet-Inflammation Links?

Autism spectrum disorder (ASD) is one of the most prevalent neurodevelopmental disorders. Many patients with ASD often show behavioral problems at mealtimes, including food selectivity and atypical feeding behaviors. The Mediterranean diet (MD) has a beneficial effect on mental health for the general population across different ages. There is evidence that good adherence to the MD is effective in reducing peripheral inflammatory markers, such as the cytokine interleukin-6 (IL-6). The present study was designed to evaluate adherence to the MD in children with ASD using age- and sex-matched, typically developing individuals (TDs) as a control group and to determine whether differences in adherence to the MD are associated with salivary IL-6 and IL-6 receptor concentration. Twenty children and adolescents with ASD (mean age 9.95 ± 0.65 years) and twenty TDs (mean age: 9.85 ± 0.59 years) participated in this study (N = 16 males and N = 4 females in each group). Participants with ASD were enrolled in a psychiatric consultation in Valencia (Spain), and TDs were recruited from two public schools in Valencia. The parents of both ASD and TD groups answered the items in a validated Mediterranean Diet Quality Index for children and adolescents (KIDMED) questionnaire on their children's adherence to the MD. The mean adherence to MD score was significantly lower in the ASD group (9.10 ± 0.42) (range 6-12) than in the TD group (10.35 ± 0.31) (range 8-12) (p = 0.02, Mann-Whitney U test). There was no statistically significant association between adherence to the MD and age or sex in both groups, but there was a significant correlation between the total KIDMED score and body mass index (BMI) in the ASD group. Regarding the concentration of Il-6 and the Il-6 receptor in saliva samples, there were no significant differences between the two groups; however, linear regression analysis by group revealed significant associations between the adherence to MD score and the concentration of IL-6 and its receptor in saliva in the ASD group (p = 0.003, OR = 0.68, 95% CI 0.007 to -0.02; p = 0.009, OR = -0.64, 95% CI -0.01 to -0.00). In contrast, no significant associations were observed between the adherence to MD score and the concentration of IL-6 and its receptor in saliva in the TD group. Children and adolescents with ASD showed significantly lower adherence to the MD, which can contribute to nutritional deficits described in ASD, and the role of BMI composition (fat versus lean mass) needs to be further investigated in this group. The concentration of IL-6 and its receptor in saliva is associated with adherence to the MD, suggesting a possible link between IL-6 and diet in ASD. Further studies to clarify the associations between IL-6, psychiatric alterations, and diet in ASD are needed.

Difficulties in diagnosing rabies using the example of a clinical case in the Omsk region

Rabies is the deadliest zoonotic disease caused by the rabies virus, a member of the genus Lyssavirus. The disease occurs after a bite from a wild or domestic animal. The paper presents a clinical case of rabies with early diagnostic difficulties, particularlyclinical, epidemiological, and laboratory diagnostic features, in a patient suffering from alcoholism. The victim sought medical help 20 days after the dog bite. On day 29, a hydrophobia clinic was observed. The patient exhibited signs of visual and auditory hallucinations, aggressiveness, and disorientation in place and time, which resembled the development of alcoholic psychosis. Saliva samples were collected on day 3 of the disease, and its polymerase chain reaction test was negative for rabies virus. On day 29, the patient died. The diagnosis of rabies was confirmed antemortem and posthumously. The virus was detected in the saliva on day 9 of the disease and posthumously in the cerebral cortex, medulla oblongata, cerebellum, and salivary glands. Histologically, Babesh–Negri bodies were found in the brain tissue. Necessary preventive measures were implemented. The unfavorable outcome in the described case of rabies emphasizes the lack of awareness of the population (especially rural) about the risk of contracting rabies infection through contact with animals and the importance of timely post-exposure prophylaxis. Timely anti-rabies care remains the only effective means of preventing related deaths.

Effects of Mint Oils on the Human Oral Microbiome: A Pilot Study.

The oral microbiome is a diverse and complex ecosystem essential for maintaining oral and systemic health. Our study is the first to define the oral microbial community in Egyptian young adults and investigate the effects of natural antimicrobials on the oral microbiome. SuperMint (SM) is a proprietary blend of peppermint, Japanese mint, bergamot mint, and spearmint essential oils encapsulated in a tiny soft beadlet. This work aimed to evaluate the effects of SM beadlets on the oral microbiome. This study recruited twenty healthy participants. A baseline investigation of the oral microbiome of the selected participants was performed by collecting saliva and swab samples before treatment. Treatment included chewing four SM beadlets twice a day for 7 days, and then, post-administration saliva and swab samples were collected at the end of treatment. The oral microbiome samples were analyzed by the high-throughput amplicon sequencing of 16S rRNA gene fragments, and the community composition was determined. The results showed that the abundance of some microbial genera and families decreased after using SM, including Prevotella, Streptococcus, Neisseria, and Haemophilus. However, some genera showed inconsistent patterns. We also found that the subject's gender and SM usage were significantly associated with diverse microbial composition. The results suggest that SM treatment decreased the abundance of several bacteria associated with halitosis and periodontal diseases, such as Actinomyces and Streptococcus. Furthermore, Corynebacterium species increased and Streptococcus decreased after SM usage. More research is needed to fully understand the antimicrobial effects of mint oils and their potential applications in maintaining good oral health.

Analysis of the immune response using FTIR spectroscopy in mothers and their newborns with different vaccination schemes for COVID-19

The SARS-CoV-2 outbreak has provoked more than 6 million deaths worldwide. The scarcity of effective treatments and its virulence converted the vaccines into an essential tool to face it. The most used vaccines were the mRNA, adenovirus vector, and inactivated whole-virus. However, nowadays, infants aged < 6 months are not eligible for any vaccines against COVID-19, and their immunization relies on passive immunity. In this research, we investigated the humoral and cellular immune response generated on newborns of SARS-CoV-2 vaccinated mothers with mRNA or viral vector (VV) vaccine employing Fourier transformed infrared (FTIR) spectroscopy in saliva samples. For this purpose, saliva samples of newborns and their mothers were collected; the population was divided into two groups, VV and mRNA, which were subdivided into three subgroups: before pregnancy (BP), at the first (FTP) and second (STP) trimesters of pregnancy. The samples were analyzed using FTIR spectroscopy, and the bands associated with the humoral and cellular immune responses, such as IgG, IgA, and IFN-γ were analyzed. The integrated areas were calculated and compared to elucidate the quantity of those immunoglobins and the cytokine. Likewise, the correlation of the humoral and cellular immune response between the newborns and their mothers and the correlation between cellular and humoral immune response was also evaluated. The VV vaccine produced a significant humoral and cellular immune response in newborns and their mothers when they received it at the STP compared with the mRNA vaccine, evidencing statistical significance. However, no correlation was observed between newborns and their mothers when the vaccine was applied in this trimester of pregnancy. When administered BP, the mRNA vaccine generated more humoral immunity in newborns and their mothers. Nevertheless, compared with the VV vaccine, it only showed statistical significance in the mothers, highlighting that IgG showed a moderate positive correlation between the newborns and their mothers.

Surface plasmon resonance (EC-SPR) sensor based on molecularly imprinted strategy for the determination of stimulant modafinil in saliva samples

A highly sensitive and selective molecularly imprinted polymer-based electrochemical-surface plasmon resonance (MIP-based EC-SPR) sensor was developed for the determination of modafinil in saliva samples. The optimal MIPpy film was synthesized using a modafinil concentration of 1 mM and a constant potential electropolymerization method. The fabricated sensor exhibited a wide linear range (0.1 μM to 10 μM), low detection limits (LOD: 50 nM, LOQ: 170 nM), and high selectivity towards modafinil, with selectivity coefficients of 8.5, 6.2, and 12.7 for adrafinil, armodafinil, and caffeine, respectively. The sensor also demonstrated excellent reproducibility (RSD: 4.2 %) and long-term stability (92 % response after one month). Furthermore, the applicability of the sensor in real samples was validated using spiked saliva samples, with recovery values ranging from 94.7 % to 103.5 %. These results highlight the potential of the MIP-based EC-SPR sensor as a powerful tool for the rapid, sensitive, and selective detection of modafinil in complex biological matrices.

Evaluation of Periodontal Clinicoradiographic Status and Whol Salivary Prostaglandin E2 Levels among Users of Water Pipe and Cigarettes.

The objective was to evaluate the periodontal clinicoradiographic status and whole salivary prostaglandin E2 (PgE2) levels among users of water pipe and cigarettes. Demographic data, duration of smoking (pack years), and familial history of smoking were recorded using a questionnaire. Participants were allocated into three groups based on their smoking status: group 1: self-reported cigarette smokers (CS); group 2: self-reported water-pipe-users; and group 3: non-smokers. The assessment included measurements of full-mouth plaque and gingival indices (PI and GI), as well as probing depth (PD), clinical attachment loss (CAL), and marginal bone loss (MBL). Unstimulated whole saliva samples were collected and PgE2 levels were measured. Group comparisons were done and p<0.05 was considered statistically significant. Thirty-three, 34 and 33 individuals were included in groups 1, 2 and 3, respectively. Full mouth PI (p<0.05), GI (p<0.05), PD (p<0.05) and mesial (p<0.05) and distal (p<0.05) MBL were statistically significantly higher among patients in groups 1 and 2 than group 3. The scores of CAL in groups 1 and 2 were 3.45 ± 0.97 and 3.62 ± 1.2 mm, respectively. None of the individuals in the control group displayed CAL. PgE2 levels were statistically significantly higher among patients in groups 1 (231.5 ± 66.3 pg/ml) (p<0.05) and 2 (231.5 ± 66.3 pg/ml) (p<0.05) compared with group 3 (76.6 ± 10.6 pg/ml). In groups 1 and 2, a statistically significant relationship was observed between pack-years, the duration of water-pipe smoking, and the levels of PgE2 and PD. There is no difference in periodontal clinicoradiographic status and whole salivary PgE2 levels between CS and waterpipe-users; however, these parameters are worse in CS and water-pipe users than in non-smokers.

Enhanced modified poly-tyrosine voltammetric sensor for the quantification detection of salivary pepsin

Pepsin as an aspartic acid protease member and one of the three foremost proteolytic enzymes in the digestive system is essential to be detected. An electrochemically polymerized tyrosine film on carbon paste electrode (pTyr/CPE) has been synthesized by electro-polymerization donating an affordable electrochemical sensor to sense salivary pepsin as a diagnostic technique for gastroesophageal reflux disease (GRD) due to saliva collection is non-invasive and relatively comfortable. The pTyr/CPE was applied for Voltammetric sensing of pepsin and its quantification in phosphate buffer solution of pH 2.0 (PBS). Scanning electron microscopy (SEM) was conducted to learn the surface morphology. Cyclic voltammetry (CV), differential pulse voltammetry (DPVs), chronoamperometry (CA), and electrochemical impedance spectroscopy (EIS) were developed to realize the electrocatalytic activity of the sensor. The pTyr/CPE proceeded as a sensitive detector to pepsin with two linear ranges from 1 to 20 & 20 to 100 ng/mL donating two limits of detection as 0.5 & 0.09 ng/mL, respectively, and high selectivity toward pepsin, as well as stability and fast response of 1.5 s. Consequently, it is guessed that the pTyr/CPE sensor could be supportive for the initial diagnosis of GRD through the detection of pepsin in saliva. Finally, we quantified the pepsin levels in saliva samples of LPR patients (n = 2), showing that the results were agreeable with those from the electrochemical sensor.

Molecular microbiological profiling of bottled unsweetened tea beverages: A screening experiment

To explore the potential storage and safety of drinking leftover bottled tea beverages from various manufacturers after direct drinking from bottles, we conducted a screening experiment on the growth of salivary bacteria in plastic bottles of tea. The diluted saliva samples from 10 participants were inoculated into the test bottled beverages, which resulted in bacteria, particularly former members of the genus Lactobacillus, growing in some green tea beverages with a neutral pH. In contrast, tea beverages with less bacterial growth contained Streptococcus spp., and the leftovers may be safe to store and drink again.

Association of opium use and tobacco smoking with α-, β-, and γ-human papillomavirus oral infection.

Head and neck squamous cell carcinomas (HNSCCs) are linked to tobacco smoking, opium use, and human papillomavirus (HPV) infection. However, little is known about the association of HPV infection with risk factors of HNSCCs, including opium and tobacco use. This cross-sectional analysis of a national multi-center case-control study in Iran included 498 HNSCC cases and 242 controls. We investigated the association of opium and tobacco use with α- (n = 21), β- (n = 46), and γ-HPV (n = 52) types in saliva samples using type-specific bead-based multiplex genotyping assays (TS-MPG). We found that α-HPV positivity was significantly associated with tobacco smoking (OR = 10.35; 95% CI = 1.15, 93; p = .03), but not with opium use (OR = 1.06; 95% CI = 0.41, 2.76; p = .89). Additionally, tobacco smoking correlated with an elevated risk of β-species 2 HPV infection (OR = 1.28; 95% CI = 1.04, 1.58; p = .020). Conversely, opium use showed a positive association with γ-species 12 HPV infection (OR = 5.67; 95% CI = 1.43, 22.44; p = .013). These findings indicate that tobacco and opium use may influence the risk of HPV infection in different ways depending on the HPV genus and species. Further studies are needed to replicate these findings in other populations.

Safety and immunogenicity of the live-attenuated hRVFV-4s vaccine against Rift Valley fever in healthy adults: a dose-escalation, placebo-controlled, first-in-human, phase 1 randomised clinical trial

Rift Valley fever virus, a pathogen to ruminants, camelids, and humans, is an emerging mosquito-borne bunyavirus currently endemic to Africa and the Arabian Peninsula. Although animals are primarily infected via mosquito bites, humans mainly become infected following contact with infected tissues or fluids of infected animals. There is an urgent need for adequate countermeasures, especially for humans, because effective therapeutics or vaccines are not yet available. Here we assessed the safety, tolerability, and immunogenicity of a next-generation, four-segmented, live-attenuated vaccine candidate, referred to as hRVFV-4s, in humans. A first-in-human, single-centre, randomised, double-blind, placebo-controlled trial was done in Belgium in which a single dose of hRVFV-4s was administered to healthy volunteers aged 18-45 years. Participants were randomly assigned using an interactive web response system. The study population encompassed 75 participants naive to Rift Valley fever virus infection, divided over three dosage groups (cohorts) of 25 participants each. All participants were followed up until 6 months. Using a staggered dose escalating approach, 20 individuals of each cohort were injected in the deltoid muscle of the non-dominant arm with either 104 (low dose), 105 (medium dose), or 106 (high dose) of 50% tissue culture infectious dose of hRVFV-4s as based on animal data, and five individuals per cohort received formulation buffer as a placebo. Primary outcome measures in the intention-to-treat population were adverse events and tolerability. Secondary outcome measures were vaccine-induced viraemia, vaccine virus shedding, Rift Valley fever virus nucleocapsid antibody responses (with ELISA), and neutralising antibody titres. Furthermore, exploratory objectives included the assessment of cellular immune responses by ELISpot. The trial was registered with the EU Clinical Trials Register, 2022-501460-17-00. Between August and December, 2022, all 75 participants were vaccinated. No serious adverse events or vaccine-related severe adverse events were reported. Pain at the injection site (51 [85%] of 60 participants) was most frequently reported as solicited local adverse event, and headache (28 [47%] of 60) and fatigue (28 [47%] of 60) as solicited systemic adverse events in the active group. No vaccine virus RNA was detected in any of the blood, saliva, urine, or semen samples. Rift Valley fever virus nucleocapsid antibody responses were detected in most participants who were vaccinated with hRVFV-4s (43 [72%] of 60 on day 14) irrespective of the administered dose. In contrast, a clear dose-response relationship was observed for neutralising antibodies on day 28 with four (20%) of 20 participants responding in the low-dose group, 13 (65%) of 20 responding in the medium-dose group, and all participants (20 [100%] of 20) responding in the high-dose group. Consistent with the antibody responses, cellular immune responses against the nucleocapsid protein were detected in all dose groups, whereas a more dose-dependent response was observed for the Gn and Gc surface glycoproteins. Neutralising antibody titres declined over time, whereas nucleocapsid antibody responses remained relatively stable for at least 6 months. The hRVFV-4s vaccine showed a high safety profile and excellent tolerability across all tested dose regimens, eliciting robust immune responses, particularly with the high-dose administration. The findings strongly support further clinical development of this candidate vaccine for human use. The Coalition for Epidemic Preparedness Innovations with support from the EU Horizon 2020 programme.

Development of a Diagnostic Microfluidic Chip for SARS-CoV-2 Detection in Saliva and Nasopharyngeal Samples.

The novel coronavirus SARS-CoV-2 was first isolated in late 2019; it has spread to all continents, infected over 700 million people, and caused over 7 million deaths worldwide to date. The high transmissibility of the virus and the emergence of novel strains with altered pathogenicity and potential resistance to therapeutics and vaccines are major challenges in the study and treatment of the virus. Ongoing screening efforts aim to identify new cases to monitor the spread of the virus and help determine the danger connected to the emergence of new variants. Given its sensitivity and specificity, nucleic acid amplification tests (NAATs) such as RT-qPCR are the gold standard for SARS-CoV-2 detection. However, due to high costs, complexity, and unavailability in low-resource and point-of-care (POC) settings, the available RT-qPCR assays cannot match global testing demands. An alternative NAAT, RT-LAMP-based SARS-CoV-2 detection offers scalable, low-cost, and rapid testing capabilities. We have developed an automated RT-LAMP-based microfluidic chip that combines the RNA isolation, purification, and amplification steps on the same device and enables the visual detection of SARS-CoV-2 within 40 min from saliva and nasopharyngeal samples. The entire assay is executed inside a uniquely designed, inexpensive disposable microfluidic chip, where assay components and reagents have been optimized to provide precise and qualitative results and can be effectively deployed in POC settings. Furthermore, this technology could be easily adapted for other novel emerging viruses.

Post-COVID-19 Pain Is Not Associated with DNA Methylation Levels of the ACE2 Promoter in COVID-19 Survivors Hospitalized Due to SARS-CoV-2 Infection.

One of theories explaining the development of long-lasting symptoms after an acute severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection include changes in the methylation pattern of the host. The current study aimed to investigate whether DNA methylation levels associated with the angiotensin-converting enzyme 2 (ACE2) promoter are different when comparing individuals previously hospitalized due to COVID-19 who then developed long-lasting post-COVID pain with those previously hospitalized due to COVID-19 who did not develop post-COVID-19 pain symptoms. Non-stimulated saliva samples were obtained from a cohort of 279 (mean age: 56.5, SD: 13.0 years old, 51.5% male) COVID-19 survivors who needed hospitalization. Clinical data were collected from hospital medical records. Participants were asked to disclose pain symptoms developed during the first three months after hospital admission due to COVID-19 and persisting at the time of the interview. Methylations of five CpG dinucleotides in the ACE2 promoter were quantified (as percentages). Participants were evaluated up to 17.8 (SD: 5.3) months after hospitalization. Thus, 39.1% of patients exhibited post-COVID-19 pain. Most patients (77.05%) in the cohort developed localized post-COVID-19 pain. Headache and pain in the lower extremity were experienced by 29.4% of the patients. Seven patients received a post-infection diagnosis of fibromyalgia based on the presence of widespread pain characteristics (11.6%) and other associated symptoms. No significant differences in methylation percentages at any CpG location of the ACE2 promoter were identified when comparing individuals with and without post-COVID-19 pain. The current study did not observe differences in methylation levels of the ACE2 promoter depending on the presence or absence of long-lasting post-COVID-19 pain symptoms in individuals who needed hospitalization due to COVID-19 during the first wave of the pandemic.

The Effects of Nicotine Pouches and E-Cigarettes on Oral Microbes: A Pilot Study.

It remains uncertain whether nicotine pouches and electronic cigarettes alter the oral environment and result in a high presence of periodontopathogenic bacteria in saliva, compared to that among cigarette users or non-tobacco users. In this study, saliva samples were collected from respondents using nicotine pouches, electronic cigarettes, and conventional cigarettes, alongside a control group of non-tobacco users. Polymerase chain reaction was used to identify clinical isolates of the following periodontal bacteria: Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, Fusobacterium nucleatum, Fusobacterium periodonticum, Porphyromonas endodontalis, and Rothia mucilaginosa. The presence of some periodontal pathogens was detected in the saliva samples from users of nicotine pouches, electronic cigarettes, and conventional cigarettes but not in samples taken from the control group. Therefore, the initial results of this pilot study suggest that the presence of periodontopathogenic bacteria in the saliva of nicotine pouch and electronic cigarette users could alter the oral microbiome, leading to periodontal diseases. However, further quantitative investigation is needed.