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The brown planthopper (BPH) is the most destructive insect pest in rice. Through a stylet, BPH secretes a plethora of salivary proteins into rice phloem cells as a crucial step of infestation. However, how various salivary proteins function in rice cells to promote insect infestation is poorly understood. Among them, one of the salivary proteins is predicted to be a carbonic anhydrase (Nilaparvata lugens carbonic anhydrase [NlCA]). The survival rate of the NlCA- RNA interference (RNAi) BPH insects was extremely low on rice, indicating a vital role of this salivary protein in BPH infestation. We generated NlCA transgenic rice plants and found that NlCA expressed in rice plants could restore the ability of NlCA- RNAi BPH to survive on rice. Next, we produced rice plants expressing the ratiometric pH sensor pHusion and found that NlCA- RNAi BPH induced rapid intracellular acidification of rice cells during feeding. Further analysis revealed that both NlCA- RNAi BPH feeding and artificial lowering of intracellular pH activated plant defense responses and that NlCA-mediated intracellular pH stabilization is linked to diminished defense responses, including reduced callose deposition at the phloem sieve plates and suppressed defense gene expression. Given the importance of pH homeostasis across the kingdoms of life, discovery of NlCA-mediated intracellular pH modulation uncovered a new dimension in the interaction between plants and piercing/sucking insect pests. The crucial role of NlCA for BPH infestation of rice suggests that NlCA is a promising target for chemical or trans-kingdom RNAi-based inactivation for BPH control strategies in plants. • Rapid intracellular acidification is a novel plant defense response • Brown planthopper (BPH) delivers carbonic anhydrase (NlCA) into rice for survival • NlCA counters intracellular acidification to suppress plant defense • pH-responsive regulators of defense responses are predicted to exist in the rice-BPH interaction The brown planthopper (BPH) is the most devastating insect pest in rice. Jiang et al. discover that BPH secretes a salivary carbonic anhydrase (NICA) to regulate the intracellular pH of the rice cell to facilitate its feeding and survival on rice plants and that NICA counters host intracellular pH acidification to diminished defense responses. [ABSTRACT FROM AUTHOR]

Objectives: The impacts of stress on inflammation, although hypothesized, have not been thoroughly examined, especially in relation to social and environmental factors and particularly within Black populations. This study aims to explore the biological mechanisms of embodiment linking stress and health to understand physiological changes in the body's response to psychological stress in a Nigerian population. Through a multidisciplinary approach, this study queries the relationship between stress, cortisol, and salivary C‐reactive protein (sCRP), a biomarker of inflammation, while also validating the use of sCRP as a potential and accurate stress indicator in the field. Methods: In this cross‐sectional study, 138 passive drool saliva samples (nfemale = 89 nmale = 49) were collected and assessed for sCRP and cortisol levels in adults. Participants also completed a short demographic survey and, to measure psychological stress, the General Health Questionnaire 12 (GHQ‐12). Relationships between sCRP and stress‐related variables (i.e., cortisol, GHQ‐12, and demographic data) were assessed using Spearman's correlations, simple regression, multivariable linear regression, and exploratory factor analysis. Results: sCRP levels ranged from 20.57 to 6879.41 pg/mL across all samples, with significant differences between female and male participants. The GHQ‐12 was not a significant predictor of sCRP variability. However, socio‐demographic factors such as body mass index (BMI), age, self‐reported sex, ethnic identity, and cortisol were significant predictors, collectively explaining 24%–27% of the variation in sCRP. Conclusion: Socio‐demographic predictors like BMI, age, sex, and particularly ethnic group experience in Nigeria encapsulate aspects of embodied stress, that significantly affect sCRP variability. [ABSTRACT FROM AUTHOR]

• This may be the first report on the salivary CRP pattern in active duty personnel. • We introduce new parameters to describe the salivary CRP morning pattern. • A sharp decline in salivary CRP after awakening may be associated with health. • Results have implications for using salivary CRP as a marker of dysfunction. C-reactive protein is a systemic inflammatory biomarker that is positively associated with the development of disease. Salivary C-reactive protein (sCRP) has previously been reported to have a diurnal rhythm with higher levels upon awakening and lower levels thereafter. The aims of this study were to evaluate the stability of sCRP across two days, characterize the daily sCRP pattern, compute morning sCRP parameters, and evaluate associations with biobehavioral health in US Navy Explosive Ordnance Disposal (EOD) technicians. Seventy male EOD technicians (age = 34.9 ± 6.5 years) participated in this study, which included a tablet-based survey, measures of health and fitness, and saliva collection. In a free-living setting, participants self-collected saliva on 2 consecutive days at WAKE, WAKE+30, WAKE+60, 4p.m., and 9p.m., for a total of 10 samples. Parameters (e.g., area under the curve) were computed to characterize the morning sCRP magnitude and pattern. Pearson product-moment correlation analyses were used to assess the stability of sCRP samples and parameters across the study period and to examine associations with biobehavioral health. Average sCRP concentrations for the 2-day period were evaluated using an analysis of variance with repeated measures. The stabilities between corresponding time points on Days 1 and 2 were very high (r s = 0.87–.94, all p s ≤ 0.001). sCRP concentrations were highest at WAKE, decreased by 73.6 % at WAKE+30, and then plateaued for the rest of the day. Parameter stabilities were good to excellent (r s = 0.77–.98, all p s ≤ 0.001). We also observed associations between sCRP parameters, self-reported health behaviors, and objective measures of health and fitness. In this study of a military population, we characterized sCRP as diurnal with robust stability across 2 consecutive days, which demonstrates the feasibility of sCRP as a biomarker. These results have significant implications for study methodology and for using sCRP as a marker of dysfunction or disease. [ABSTRACT FROM AUTHOR]

Background: Aedes aegypti presence, human-vector contact rates, and Aedes-borne virus transmission are highly variable through time and space. The Lower Rio Grande Valley (LRGV), Texas, is one of the few regions in the U.S. where local transmission of Aedes-borne viruses occurs, presenting an opportunity to evaluate social, urbanistic, entomological, and mobility-based factors that modulate human exposure to Ae. aegypti. Methodology & Principal findings: Mosquitoes were collected using BG-Sentinel 2 traps during November 2021 as part of an intervention trial, with knowledge, attitudes, and practices (KAP) and housing quality surveys to gather environmental and demographic data. Human blood samples were taken from individuals and a Bitemark Assay (ELISA) was conducted to quantify human antibodies to the Ae. aegypti Nterm-34kDa salivary peptide as a measure of human exposure to bites. In total, 64 houses were surveyed with 142 blood samples collected. More than 80% of participants had knowledge of mosquito-borne diseases and believed mosquitoes to be a health risk in their community. Our best fit generalized linear mixed effects model found four fixed effects contributed significantly to explaining the variation in exposure to Ae. aegypti bites: higher annual household income, younger age, larger lot area, and higher female Ae. aegypti abundance per trap night averaged over 5 weeks prior to human blood sampling. Conclusions: Most surveyed residents recognized mosquitoes and the threat they pose to individual and public health. Urbanistic (i.e., lot size), social (i.e., income within a low-income community and age), and entomological (i.e., adult female Ae. aegypti abundance) factors modulate the risk of human exposure to Ae. aegypti bites. The use of serological biomarker assays, such as the Bitemark Assay, are valuable tools for surveillance and risk assessment of mosquito-borne disease, especially in areas like the LRGV where the transmission of target pathogens is low or intermittent. Author summary: Aedes aegypti is a mosquito vector with public health importance on the global scale as it transmits viruses such as dengue, chikungunya, and Zika. Although transmission rates of dengue and Zika are low in the U.S., there are a few regions, including south Texas, where local transmission has occurred. Our study aimed to evaluate the factors associated with risk of exposure to these viruses using a serological bioassay that measured antibody response to an Ae. aegypti salivary protein to assess human-vector contact. We collected mosquitoes, took human-blood samples, and conducted urbanistic and demographic surveys in November 2021 in eight communities in the Lower Rio Grande Valley, Texas. Our knowledge, attitude, and practices survey found that most residents recognized adult mosquitoes, though few individuals knew someone personally who been sick with a mosquito-borne disease. Outdoor adult female Ae. aegypti abundance was positively associated with exposure to mosquito bites. Household income, individual age, and lot area also significantly affected exposure levels. The Bitemark Assay we used in this study can be utilized as a tool for entomological risk assessment and could be used as an alternative to infection exposure in areas where mosquito-borne disease levels are low. [ABSTRACT FROM AUTHOR]

Oral cancer, representing 2–4% of all cancer cases, predominantly consists of Oral Squamous Cell Carcinoma (OSCC), which makes up 90% of oral malignancies. Early detection of OSCC is crucial, and identifying specific proteins in saliva as biomarkers could greatly improve early diagnosis. Here, we proposed a strategy to pinpoint candidate biomarkers. Starting from a list of salivary proteins detected in 10 OSCC patients and 20 healthy controls, we combined a univariate approach and a multivariate approach to select candidates. To reduce the number of proteins selected, a Protein–Protein Interaction network was built to consider only connected proteins. Then, an over-representation analysis (ORA) determined the enriched pathways. The network from 172 differentially abundant proteins highlighted 50 physically connected proteins, selecting relevant candidates for targeted experimental validations. Notably, proteins like Heat shock 70 kDa protein 1A/1B, Pyruvate kinase PKM, and Phosphoglycerate kinase 1 were suggested to be differentially regulated in OSCC patients, with implications for oral carcinogenesis and tumor growth. Additionally, the ORA revealed enrichment in immune system, complement, and coagulation pathways, all known to play roles in tumorigenesis and cancer progression. The employed method has successfully identified potential biomarkers for early diagnosis of OSCC using an accessible body fluid. [ABSTRACT FROM AUTHOR]

Introduction: Acquired pellicle (AP) acts as a membrane preventing acids from coming into direct contact with the tooth. Possibly, individuals with different dental health status present changes in its composition that could disrupt this function. Thus, the aim of this study was to compare the protein composition of the AP in adolescents with erosive tooth wear (ETW), caries, or sound. Methods: Calibrated examiners in BEWE index and ICDAS-merged Epi criteria assessed ETW and caries in a sample of 454 systemically healthy adolescents aged 12–15 years old. Thirty subjects from that sample were selected for this study: ETW group (n = 10; total BEWE ≥9 and absence of dentinal caries lesions); caries group (n = 10; total BEWE <9 and with at least one dentinal caries lesion), and sound group (n = 10; total BEWE <9 and absence of dentinal caries lesions). Two-hour-formation AP samples were taken from buccal, occlusal/incisal, palatal/lingual tooth surfaces. Protein composition was analysed by liquid chromatography-tandem mass spectrometry. Using mean reporter ion values, relative abundances of proteins were compared among the three groups to calculate for fold changes. Twofold protein increases or decreases were reported (t test, p < 0.05). Gene Ontology (GO) of included proteins was assigned. Results: Mean age of participants was 13.1 ± 1.14 years and 56.6% were females. The prevalence of ETW was of 66.6% and of dentinal caries of 33.3%. The GO analyses showed that the majority of detected proteins were stress response related. The ETW group disclosed upregulated relative abundance of antileukoprotease (2.85-fold in ETW vs. sound and 2.34-fold in ETW group vs. caries group); histatin (2.42-fold in ETW group vs. sound group and 2.20-fold in ETW group vs. caries group), and prolactin-induced protein (2.30-fold in ETW group vs. sound group and 2.06-fold in ETW group vs. caries group) (p < 0.05). Hemoglobin subunits alpha (HBA) and beta (HBB) showed decreased relative abundances in the ETW and caries groups when compared to the sound group (HBA: 0.42-fold in ETW group and 0.40-fold in caries group; HBB: 0.45-fold in ETW group and 0.38-fold in caries group; p < 0.05). Conclusion: AP from individuals with ETW showed differences when compared to other dental conditions, with relative abundance increasing of some stress response-associated proteins in ETW and a decrease in proteins related to salivary protection against acid challenges. [ABSTRACT FROM AUTHOR]

Energy‐dispersive Laue diffraction (EDLD) is a powerful method to obtain position‐resolved texture information in inhomogeneous biological samples without the need for sample rotation. This study employs EDLD texture scanning to investigate the impact of two salivary peptides, statherin (STN) and histatin‐1 (HTN) 21 N‐terminal peptides (STN21 and HTN21), on the crystallographic structure of dental enamel. These proteins are known to play crucial roles in dental caries progression. Three healthy incisors were randomly assigned to three groups: artificially demineralized, demineralized after HTN21 peptide pre‐treatment and demineralized after STN21 peptide pre‐treatment. To understand the micro‐scale structure of the enamel, each specimen was scanned from the enamel surface to a depth of 250 µm using microbeam EDLD. Via the use of a white beam and a pixelated detector, where each pixel functions as a spectrometer, pole figures were obtained in a single exposure at each measurement point. The results revealed distinct orientations of hydroxyapatite crystallites and notable texture variation in the peptide‐treated demineralized samples compared with the demineralized control. Specifically, the peptide‐treated demineralized samples exhibited up to three orientation populations, in contrast to the demineralized control which displayed only a single orientation population. The texture index of the demineralized control (2.00 ± 0.21) was found to be lower than that of either the STN21 (2.32 ± 0.20) or the HTN21 (2.90 ± 0.46) treated samples. Hence, texture scanning with EDLD gives new insights into dental enamel crystallite orientation and links the present understanding of enamel demineralization to the underlying crystalline texture. For the first time, the feasibility of EDLD texture measurements for quantitative texture evaluation in demineralized dental enamel samples is demonstrated. [ABSTRACT FROM AUTHOR]

[Display omitted] • Lipidomics identified 23 lipid classes low in glycolipids in tick salivary glands. • Phosphatidylcholine (PC; 26.66%) was the lipid class with the highest representation. • Tick salivary glycolipids were mostly represented by diacylglycerol (DG; 0.48%). • Individuals bitten by ticks produced more phosphatidylcholine-IgG antibodies. • DG levels were higher in tick salivary glands than in tick saliva. Ticks, as hematophagous ectoparasites, can manipulate host immune and metabolic processes, causing tick-borne allergies such as α-Gal syndrome (AGS). Glycolipids with bound galactose-alpha-1–3-galactose (α-Gal) are potential allergenic molecules associated with AGS. Nevertheless, proteins and lipids lacking α-Gal modifications may contribute to tick salivary allergies and be linked to AGS. In this study, we characterized the effect of deglycosylated tick salivary proteins without lipids on treated zebrafish fed with dog food formulated with mammalian (beef, lamb, pork) meat by quantitative proteomics analysis of intestinal samples. The characterization and functional annotations of tick salivary lipids with low representation of glycolipids was conducted using a lipidomics approach. Results showed a significant effect of treatment with saliva and saliva deglycosylated protein fraction on zebrafish abnormal or no feeding (p < 0.005). Treatment with this fraction affected multiple metabolic pathways, defense responses to pathogens and protein metabolism, which correlated with abnormal or no feeding. Lipidomics analysis identified 23 lipid classes with low representation of glycolipids (0.70% of identified lipids). The lipid class with highest representation was phosphatidylcholine (PC; 26.66%) and for glycolipids it corresponded to diacylglycerol (DG; 0.48%). Qualitative analysis of PC antibodies revealed that individuals bitten by ticks were more likely to produce PC-IgG antibodies (p < 0.001). DG levels were significantly higher in tick salivary glands (p < 0.05) compared with tick saliva and salivary fractions. The α-Gal content was higher in tick saliva than in deglycosylated saliva and lipid fractions. These results support a possible role for tick salivary proteins and lipids without α-Gal modifications in AGS. [ABSTRACT FROM AUTHOR]

Background: A high-performance sport like soccer requires training strategies that aim to reach peak performance at the right time for the desired competitions. Thus, the investigation of biochemical markers in saliva is a tool that is beginning to be used in athletes within the physical training process. There is still no evidence on universal saliva collection and analysis protocols in soccer. This review aims to map the use of saliva as a tool for analyzing athletic performance in soccer, from the biomarkers used to the validated protocols for these analyses. Methods: A broad systematic literature search was carried out in the electronic databases Web of Science, Livivo, Scopus, PubMed, LILACS and gray literature (Google Scholar and ProQuest). Two reviewers selected the studies and extracted data on the type of salivary collection used, the salivary biomarker evaluated and monitored. Results: Ninety-three articles were included. The most frequently analyzed salivary biomarkers were cortisol (n = 53), testosterone (n = 35), secretory immunoglobulin A (SIgA) (n = 33), salivary alpha amylase (n = 7), genetic polymorphisms (n = 4) and miRNAs (n = 2). The results of the studies indicated beneficial effects in monitoring salivary biomarkers in the assessment of sports performance, although most studies did not include a control group capable of comparison. Salivary collection and analysis protocols were varied and commonly not reported. Conclusions: This scoping review provides a comprehensive overview of the current landscape of salivary biomarker research in soccer. The findings underscore the importance of these biomarkers in assessing athletes' physiological responses and overall well-being. Future research should focus on refining methodologies, exploring additional biomarkers, and investigating the practical implications of salivary biomarker monitoring in soccer and other sports. [ABSTRACT FROM AUTHOR]

Anti-wear performance of human enamel in the mouth is closely related to the lubrication of salivary pellicle. It is well known that the inorganic hydroxyapatite (HA) of the enamel plays an important role in the adsorption and pellicle-forming of salivary proteins on the enamel, but the role of enamel matrix proteins remains unclear. In this study, the adsorption and lubrication behavior of salivary proteins on original, heated, and deproteinated enamel surfaces was comparatively investigated using an atomic force microscopy and nano-indentation/scratch techniques. Compared with that on the original enamel surface, the adsorption and lubrication behavior of salivary proteins remains almost unchanged on the heated enamel surface (where the enamel matrix proteins are denatured but the size of HA crystalline nanoparticles keeps constant) but exhibits an obvious compromise on the deproteinated enamel surface (where the enamel matrix proteins are removed and agglomeration of HA crystallites occurs). The HA agglomeration weakens the electrostatic interaction of enamel surfaces with salivary proteins to cause a distinct negative influence on the adsorption and pellicle-forming of salivary proteins. Further, the negative effect is confirmed with a quartz crystal microbalance with dissipation. In summary, by regulating enamel nanostructure for appropriate electrostatic interactions between salivary proteins and enamel surfaces, the enamel matrix proteins play an essential role in the adsorption and pellicle-forming of salivary proteins on human enamel, and then contribute to saliva lubrication, which provides the enamel with an anti-wear mechanism. The findings will promote and assist the design of enamel-inspired anti-wear materials. [ABSTRACT FROM AUTHOR]

BACKGROUND: Body‐color polymorphisms in insects are often explained by environmental selective advantages. Differential fitness related to body coloration has been demonstrated in Myzus persicae (Sulzer): performance of the red morph is in general better than that of the green morph on tobacco plants. However, the molecular mechanism involved is largely unclear. RESULTS: Here we showed that the red morph of M. persicae had higher expression of a carotenoid desaturase CarD763 in the whole body, salivary gland and saliva relative to the green morph. Also, 18% individuals displayed faded red body color 5 days post dsCarD763 treatment. Furthermore, knockdown of CarD763 in the red morph significantly prolonged the time needed to locate phloem and shortened the duration of phloem feeding. Honeydew production and survival rate decreased as well. In contrast, overexpression of CarD763 in tobacco leaves facilitated aphid feeding, enhanced honeydew production and improved the survival rate of aphids. Compared with those fed by dsGFP aphids, plants infested by dsCarD763‐treated aphids had higher ROS accumulation, lower lycopene content and photosynthetic rate, and maximum photon quantum yield. The reverse was true when plants overexpressed CarD763. CONCLUSION: These findings demonstrated that CarD763, a red morph‐specific salivary protein, could enhance aphid feeding and early colonization by promoting plant photosynthesis. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The evolution of hematophagy involves a series of adaptations that allow blood-feeding insects to access and consume blood efficiently while managing and circumventing the host's hemostatic and immune responses. Mosquito, and other insects, utilize salivary proteins to regulate these responses at the bite site during and after blood feeding. We investigated the function of Anopheles gambiae salivary apyrase (AgApyrase) in regulating hemostasis in the mosquito blood meal and in Plasmodium transmission. Our results demonstrate that salivary apyrase, a known inhibitor of platelet aggregation, interacts with and activates tissue plasminogen activator, facilitating the conversion of plasminogen to plasmin, a human protease that degrades fibrin and facilitates Plasmodium transmission. We show that mosquitoes ingest a substantial amount of apyrase during blood feeding, which reduces coagulation in the blood meal by enhancing fibrin degradation and inhibiting platelet aggregation. AgApyrase significantly enhanced Plasmodium infection in the mosquito midgut, whereas AgApyrase immunization inhibited Plasmodium mosquito infection and sporozoite transmission. This study highlights a pivotal role for mosquito salivary apyrase for regulation of hemostasis in the mosquito blood meal and for Plasmodium transmission to mosquitoes and to the mammalian host, underscoring the potential for strategies to prevent malaria transmission. Here, Pala et al. show that mosquito salivary apyrase regulates hemostasis in the mosquito blood meal, boosting malaria transmission. They demonstrate that immunizing against apyrase inhibits parasite transmission. [ABSTRACT FROM AUTHOR]

Simple Summary: Aphids are phloem feeders and are one of the most destructive pests of crop plants. Lipaphis erysimi is a specialist aphid that infests members of the Brassicaceae family and has detrimental impacts on the yield of oilseed and vegetable Brassicas in the Indian subcontinent. Recognising the critical need for developing aphid control strategies, we aimed to enhance the pool of gene targets for RNAi for developing resistance in Brassica crop plants against L. erysimi infestation. To understand the variations in expression profiles of genes while the aphid is feeding upon the host plant, as well as between two developmental stages (adults and nymphs), we conducted a two-way transcriptome-based differential expression analysis between feeding and non-feeding adults and between adults and nymphs. This led to the identification of important transcripts involved in detoxification, feeding, metabolism and aphid development. Our study provides valuable insights into the molecular mechanisms underlying feeding and development in aphids. It also generated an important resource for further research on aphid biology and for developing effective RNAi-based strategies for pest management and control. Lipaphis erysimi is a specialist aphid of the Indian subcontinent that causes significant yield losses in oilseed Brassicas. Several aphid genes have been used as preferred targets in RNAi-based transgenic plants for aphid resistance. In order to enhance the repertoire of potential target genes for aphid control and to identify the genes associated with aphid feeding and development, we performed a two-way comparative study of differential gene expression profiles between (i) feeding and non-feeding adults and (ii) adult and nymph developmental stages of L. erysimi. De novo RNA-seq of aphids using Illumina technology generated a final transcriptome comprising 52,652 transcripts. Potential transcripts for host selection, detoxification, salivary proteins and effectors, molecular chaperones and developmental genes were identified. Differential gene expression studies identified variations in the expression of 1502 transcripts between feeding and non-feeding adults and 906 transcripts between nymphs and adults. These data were used to identify novel target genes for RNAi-based aphid control and facilitate further studies on the molecular basis of aphid feeding and development. [ABSTRACT FROM AUTHOR]

Golgi-resident enzymes remain in place while their substrates flow through from the endoplasmic reticulum to elsewhere in the cell. COPI-coated vesicles bud from the Golgi to recycle Golgi residents to earlier cisternae. Different enzymes are present in different parts of the stack, and one COPI adaptor protein, GOLPH3, acts to recruit enzymes into vesicles in part of the stack. Here, we used proximity biotinylation to identify further components of intra-Golgi vesicles and found FAM114A2, a cytosolic protein. Affinity chromatography with FAM114A2, and its paralogue FAM114A1, showed that they bind to Golgi-resident membrane proteins, with membrane-proximal basic residues in the cytoplasmic tail being sufficient for the interaction. Deletion of both proteins from U2OS cells did not cause substantial defects in Golgi function. However, a Drosophila orthologue of these proteins (CG9590/FAM114A) is also localised to the Golgi and binds directly to COPI. Drosophila mutants lacking FAM114A have defects in glycosylation of glue proteins in the salivary gland. Thus, the FAM114A proteins bind Golgi enzymes and are candidate adaptors to contribute specificity to COPI vesicle recycling in the Golgi stack. [ABSTRACT FROM AUTHOR]

BACKGROUND: Saliva has a crucial role in determining the compatibility between piercing–sucking insects and their hosts. The brown planthopper (BPH) Nilaparvata lugens, a notorious pest of rice in East and Southeast Asia, secretes gelling and watery saliva when feeding on rice sap. Nlsalivap‐5 (NlSP5) and Nlsalivap‐7 (NlSP7) were identified as potential planthopper‐specific gelling saliva components, but their biological functions remain unknown. RESULTS: Here, we showed by transcriptomic analyses that NlSP5 and NlSP7 were biasedly expressed in the salivary glands of BPHs. Using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9‐mediated genome‐editing system, we constructed NlSP5 and NlSP7 homozygous mutants (NlSP5−/− and NlSP7−/−). Electrical penetration graph assay showed that NlSP5−/− and NlSP7−/− mutants exhibited abnormal probing and feeding behaviors. Bioassays revealed that the loss‐of‐function of NlSP5 and NlSP7 significantly reduced the fitness of BPHs, with extended developmental duration, shortened lifespan, reduced weight, and impaired fecundity and hatching rates. CONCLUSION: These findings deepen our understanding of the BPH–host interaction and may provide potential targets for the management of rice planthoppers. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Fermented food consumption is recommended for health and environmental purposes. While it is known to impact gut microbiota, further investigation is needed to establish connections with the oral microbiota. For this purpose, we investigated the effect of daily consumption of a model cheese containing 3 Lactic Acid Bacteria (LAB) species on the oral microbiota of rats following a 3-week diet. Cheese consumption transiently modifies the oral microbiota and leads to a transient persistence of LAB in the oral cavity of 1/3 of the animals. The origin of this variability was partly explained by an overrepresentation of salivary proteins involved in the response to oxidative stress in animals without LAB persistence. These findings highlight the significance of fermented foods in shaping the diversity of the oral microbiota. Additionally, they suggest that variations in the salivary proteome among individuals may influence the permissiveness of the oral microbiota towards exogenous microorganisms. [ABSTRACT FROM AUTHOR]

Salivary gland tumors are highly variable in clinical presentation and histology. The World Health Organization (WHO) classifies 22 types of malignant and 11 types of benign tumors of the salivary glands. Diagnosis of salivary gland tumors is based on imaging (ultrasound, magnetic resonance imaging) and fine-needle aspiration biopsy, but the final diagnosis is based on histopathological examination of the removed tumor tissue. In this pilot study, we are testing a new approach to identifying peptide biomarkers in saliva that can be used to diagnose salivary gland tumors. The research material for the peptidomic studies was extracts from washings of neoplastic tissues and healthy tissues (control samples). At the same time, saliva samples from patients and healthy individuals were analyzed. The comparison of the peptidome composition of tissue extracts and saliva samples may allow the identification of potential peptide markers of salivary gland tumors in patients' saliva. The peptidome compositions extracted from 18 tumor and 18 healthy tissue samples, patients' saliva samples (11 samples), and healthy saliva samples (8 samples) were analyzed by LC-MS tandem mass spectrometry. A group of 109 peptides was identified that were present only in the tumor tissue extracts and in the patients' saliva samples. Some of the identified peptides were derived from proteins previously suggested as potential biomarkers of salivary gland tumors (ANXA1, BPIFA2, FGB, GAPDH, HSPB1, IGHG1, VIM) or tumors of other tissues or organs (SERPINA1, APOA2, CSTB, GSTP1, S100A8, S100A9, TPI1). Unfortunately, none of the identified peptides were present in all samples analyzed. This may be due to the high heterogeneity of this type of cancer. The surprising result was that extracts from tumor tissue did not contain peptides derived from salivary gland-specific proteins (STATH, SMR3B, HTN1, HTN3). These results could suggest that the developing tumor suppresses the production of proteins that are essential components of saliva. [ABSTRACT FROM AUTHOR]

Salivary proteins of insect herbivores can suppress plant defenses, but the roles of many remain elusive. One such protein is glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the saliva of the Recilia dorsalis (RdGAPDH) leafhopper, which is known to transmit rice gall dwarf virus (RGDV). Here we show that RdGAPDH was loaded into exosomes and released from salivary glands into the rice phloem through an exosomal pathway as R. dorsalis fed. In infected salivary glands of R. dorsalis, the virus upregulated the accumulation and subsequent release of exosomal RdGAPDH into the phloem. Once released, RdGAPDH consumed H2O2 in rice plants owing to its –SH groups reacting with H2O2. This reduction in H2O2 of rice plant facilitated R. dorsalis feeding and consequently promoted RGDV transmission. However, overoxidation of RdGAPDH could cause potential irreversible cytotoxicity to rice plants. In response, rice launched emergency defense by utilizing glutathione to S-glutathionylate the oxidization products of RdGAPDH. This process counteracts the potential cellular damage from RdGAPDH overoxidation, helping plant to maintain a normal phenotype. Additionally, salivary GAPDHs from other hemipterans vectors similarly suppressed H2O2 burst in plants. We propose a strategy by which plant viruses exploit insect salivary proteins to modulate plant defenses, thus enabling sustainable insect feeding and facilitating viral transmission. Salivary proteins of insect herbivores can suppress plant defenses. Here, Wang et al show that exosomal GAPDH in leafhopper saliva reduces the hydrogen peroxide in rice plants to supress resistance to leafhopper feeding and viral transmission. [ABSTRACT FROM AUTHOR]

Introduction: Damage Associated Molecular Patterns (DAMPs) can initiate and amplify immune responses and can play an important role in the sustenance and progression of periodontal diseases. S100A8, S100A9 and their heterodimer, S100A8/9, are DAMPs or alarmins currently being evaluated for their potential as biomarkers in periodontal diseases. Aim: To quantify and compare salivary levels of S100A8, S100A9 and S100A8/9 in periodontal health, gingivitis or stage 1 periodontitis and stages 3 or 4 periodontitis. Materials and Methods: A cross-sectional analytical study was carried out in the Department of Periodontics, Saveetha Dental College and Hospital, Chennai, India, from December 2022 to February 2023. Periodontal examination and saliva sample collection were done for sixty-eight consecutively enrolled subjects who met the inclusion and exclusion criteria. Study subjects were categorised as group 1, which comprised participants with clinical periodontal health (n=20), group 2, subjects with gingivitis or stage 1 periodontitis (n=20), and group 3, which included patients with stages 3 or 4 periodontitis (n=28). Detailed clinical examination and periodontal charting were done in all study subjects. Saliva samples were processed and stored at -80°C, and enzyme-linked immunosorbent assay was done to quantify S100A8, S100A9 and S100A8/9. Data were analysed using International Business Machines (IBM) Statistical Package for the Social Sciences (SPSS) software version 25.0. Salivary levels of S100A8, S100A9 and S100A8/9 were expressed as mean and standard deviation values. Mean values of the three salivary proteins were compared using the non parametric Kruskal-Wallis test. Bonferroni adjusted pairwise comparisons were also done. The p-values less than 0.05 were considered statistically significant. Results: The mean age of study participants was 38.97±7.62 years. Salivary levels of S100A8 were 41.34±14.34, 141.95± 185.25 and 901.37±65.60 pg/mL in groups 1, 2 and 3, respectively. A statistically significant difference was present among the three groups for mean salivary levels of S100A8 (p-value<0.001). Pairwise comparisons showed that salivary S100A8 values in group 3 were significantly higher than the values observed in clinical health (group 1) (p-value<0.001) and those seen in the gingivitis or stage 1 periodontitis group (group 2) (p-value<0.001). Mean salivary levels of S100A9 and S100A8/9 did not show any statistically significant differences between the groups. Conclusion: There is an altered expression of S100A8, S100A9 and S100A8/9 in periodontal diseases compared to clinical health. Salivary levels of S100A8 are markedly different in cases of advanced periodontal destruction than in periodontal health and early stages of periodontal disease. Salivary S100A8 merits potential as a biomarker for periodontal diseases. [ABSTRACT FROM AUTHOR]

The aim of this in vivo study was to compare total protein present in the salivary films (F) and acquired enamel pellicle (AEP) on eroded and non-eroded surfaces in patients suffering from GORD symptoms with and without GORD diagnosis (GORD, No-GORD). Thirty-nine patients suffering from GORD symptoms and erosive tooth wear on lower first molars and an unaffected posterior occlusal surface in the same quadrant were recruited from Guy's hospital, London. Salivary film and AEP were collected from the eroded and uneroded occlusal surfaces, using 0.5% sodium dodecyl sulphate (SDS)-soaked filter papers. Total protein concentration was analysed using bicinchoninic acid assay (BCA). Statistical analysis was conducted using Shapiro–Wilk, ANOVA, and Tukey's tests (p < 0.05), comparing four GDS sample types and GORD vs. No-GORD groups. The level of significance was set as p < 0.05. Data were compared between eroded and uneroded surfaces in the same patient with GORD symptoms, as well as between those with or without a GORD diagnosis (GORD, No-GORD). The AEP total protein concentration from the eroded [2.17 (0.49) mg/mL] and uneroded surfaces [2.24 (0.66) mg/mL] of the GORD group were statistically significantly lower than those on eroded [3.27 (1.01) mg/mL] and uneroded [3.33 (1.57) mg/mL] surfaces in the No-GORD group (p = 0.007) (p = 0.008), respectively. No statistically significant differences were observed for film and AEP between eroded and uneroded surfaces (p > 0.05). [ABSTRACT FROM AUTHOR]

The Aedes aegypti mosquito is a vector of many infectious agents, including flaviviruses such as Zika virus. Components of mosquito saliva have pleomorphic effects on the vertebrate host to enhance blood feeding, and these changes also create a favorable niche for pathogen replication and dissemination. Here, we demonstrate that human CD47, which is known to be involved in various immune processes, interacts with a 34-kilodalton mosquito salivary protein named Nest1. Nest1 is up-regulated in blood-fed female A. aegypti and facilitates Zika virus dissemination in human skin explants. Nest1 has a stronger affinity for CD47 than its natural ligand, signal regulatory protein α, competing for binding at the same interface. The interaction between Nest1 with CD47 suppresses phagocytosis by human macrophages and inhibits proinflammatory responses by white blood cells, thereby suppressing antiviral responses in the skin. This interaction elucidates how an arthropod protein alters the human response to promote arbovirus infectivity. Editor's summary: Aedes aegypti mosquitos can transmit arboviruses during blood feeding, and components of mosquito saliva may enhance both blood feeding and virus transmission. Marin-Lopez et al. identified a 34-kDa mosquito salivary protein called Nest1, highly expressed in the saliva of female A. aegypti mosquitos, as a high-affinity binder to human CD47. In human skin explants infected with Zika virus, Nest1 increased virus burden and inhibited some CD47-governed immune functions like phagocytosis and proinflammatory responses in immune cells, thus inhibiting antiviral responses. These findings provide insight into how mosquito salivary proteins can potentially facilitate arbovirus infection. —Christiana Fogg [ABSTRACT FROM AUTHOR]

BACKGROUND: Citrus huanglongbing (HLB) is a devastating disease caused by Candidatus Liberibacter asiaticus (CLas) that affects the citrus industry. In nature, CLas relies primarily on Diaphorina citri Kuwayama as its vector for dissemination. After D. citri ingests CLas‐infected citrus, the pathogen infiltrates the insect's body, where it thrives, reproduces, and exerts regulatory control over the growth and metabolism of D. citri. Previous studies have shown that CLas alters the composition of proteins in the saliva of D. citri, but the functions of these proteins remain largely unknown. RESULTS: In this study, we detected two proteins (DcitSGP1 and DcitSGP3) with high expression levels in CLas‐infected D. citri. Quantitative PCR and Western blotting analysis showed that the two proteins were highly expressed in the salivary glands and delivered into the host plant during feeding. Silencing the two genes significantly decreased the survival rate for D. citri, reduced phloem nutrition sucking and promoted jasmonic acid (JA) defenses in citrus. By contrast, after overexpressing the two genes in citrus, the expression levels of JA pathway‐associated genes decreased. CONCLUSION: Our results suggest that CLas can indirectly suppress the defenses of citrus and support feeding by D. citri via increasing the levels of effectors in the insect's saliva. This discovery facilitates further research into the interaction between insect vectors and pathogens. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Ticks can transmit many pathogens, including arboviruses, to their vertebrate hosts. Arboviruses must overcome or evade defense mechanisms during their passage from the tick gut to the hemolymph, salivary glands, and the feeding site in the host skin. This review summarizes current knowledge of defense mechanisms in specific tick tissues and at the feeding site in the host skin. We discuss the possible roles of these defense mechanisms in viral infection and transmission. The responses of tick salivary proteins to arbovirus infection are also discussed. This review provides information that may help accelerate research on virus–tick interactions. [ABSTRACT FROM AUTHOR]

Dental caries is a multifactorial chronic disease resulting from the intricate interplay among acid‐generating bacteria, fermentable carbohydrates, and several host factors such as saliva. Saliva comprises several proteins which could be utilized as biomarkers for caries prevention, diagnosis, and prognosis. Mass spectrometry‐based salivary proteomics approaches, owing to their sensitivity, provide the opportunity to investigate and unveil crucial cariogenic pathogen activity and host indicators and may demonstrate clinically relevant biomarkers to improve caries diagnosis and management. The present review outlines the published literature of human clinical proteomics investigations on caries and extensively elucidates frequently reported salivary proteins as biomarkers. This review also discusses important aspects while designing an experimental proteomics workflow. The protein–protein interactions and the clinical relevance of salivary proteins as biomarkers for caries, together with uninvestigated domains of the discipline are also discussed critically. [ABSTRACT FROM AUTHOR]

Gingival enlargement is a common clinical sign in the gingival diseases associated with orthodontic treatment. Its biological mechanisms are not completely understood; nevertheless, the biochemical changes associated with these inflammatory and overgrowth processes could alter the post-translational protein modifications occurring in various locations within the mouth. Here, changes in the profiles of the carbonylated and phosphorylated proteins in saliva were examined in donors with gingival enlargement (seven men and seven women) and healthy donors (six men and eight women). The sociodemographic characteristics of both groups did not present significant differences. Carbonylation was measured by a quantitative immunoassay (Dot Blot), whereas the profiles of the phosphorylated proteins were visualized by SDS-PAGE with quercetin staining. Some phosphopeptides were also identified using a typical LC-MS-MS approach. Our results showed that gingival enlargement induced a significant increase in oxidative damage in salivary proteins. While a significant reduction in phosphorylation was observed at the stain level in SDS-PAGE, there was a slight increase in the number of phosphorylated proteins identified by MS in samples with gingival enlargement. [ABSTRACT FROM AUTHOR]

Phlebotomus argentipes is the predominant sandfly vector of leishmaniasis in the Indian subcontinent. India and Sri Lanka primarily report visceral and cutaneous leishmaniasis caused by Leishmania donovani. We compared Ph. argentipes from two locations, focusing on its morphological, molecular, and salivary protein characteristics. Sandflies were captured using CDC light traps and cattle-baited net traps. Species identification and morphological comparisons were carried out using standard taxonomic keys. DNA extracted from 12 Sri Lankan sandfly samples was PCR-amplified and sequenced for the variable region of Cytochrome oxidase subunit I. Existing DNA sequences of India from GenBank were utilized for a phylogenetic analysis between Sri Lanka and India. Salivary protein profiles were studied using SDS-PAGE, Western blot, and electrospray ionization/LC/MS/MS. The morphological similarities observed between female Ph. argentipes from India and Sri Lanka suggest the presence of Ph. argentipes var. glaucus. A phylogenetic analysis showed genetic divergence between Ph. argentipes populations, but both shared a similar salivary protein profile. A common, strong 30 kDa immunogenic band comprised PagSP05, PagSP06, and PagSP17 proteins of Ph. argentipes. The similarity between the immunogenic salivary proteins suggests their potential use as common markers for vector exposure or immune response stimulants across regions. The use of multiple samples for each category of serum would improve the comprehensiveness of the immunogenic profiles obtained. [ABSTRACT FROM AUTHOR]

Background and Objectives: An oral lichen planus (OLP) chronic lesion refers to a group of oral potentially malignant disorders (OPMDs) that still lack a proper understanding from the point of view of relevant biomarkers for diagnostics and prognosis. The aim of the study was to assess the salivary histamine levels in patients with oral lichen planus lesions. Materials and Methods: The study included a group of 76 patients with oral lichen planus. General diseases and medication taken, smoking habits, severity of pain assessed using a visual analogue scale (VAS), oral hygiene status, and duration of OLP were evaluated. ELISA diagnostics for histamines in saliva levels were assessed. Results: The histamine levels in the OLP group were higher (0.468) in comparison with the control group (0.056), without a statistically significant value p = 0.090 (Mann–Whitney U Test). The median age of 76 OLP patients was 63 years (min 22.0–max. 81), with the biological sex being 80.3% females and 15 19.7% males. The average duration of OLP lesion presence was 29.4 months (SD 37.1) and the median value was 14.5 months. The median of the VAS was 3.0. OLP assessment in accordance with the Malhotra methodology showed the highest frequency—30.3% for only two of the point areas involved and 17.1% for three points. Clinical assessment of the different OLP grades, severity, and oral site involvement and the VAS in correlation with histamine salivary levels showed a lack of statistical significance in the investigated population. Conclusions: Undertaking further research could provide further possibilities for searching for general factors in OLP development. [ABSTRACT FROM AUTHOR]

Featured Application: Monitoring salivary biomarkers demonstrates a potential application to assess the internal training load and recovery status of elite futsal players during congested competition periods, especially total protein and salivary immunoglobulin A. Tracking biomarker changes provides a practical tool for coaching staff to guide adjustments to training loads, nutrition, and recovery protocols to mitigate risks and optimize athlete performance and health. A congested match and training schedule could alter internal load, and this could be reflected in biomarkers of stress and immunity in elite futsal players. The aim of this study was to analyze the effects of a congested match and training schedule on internal load and levels of total protein, total oxidant status (TOS), total antioxidant capacity (TAC), oxidative stress index, and the concentrations of salivary immunoglobulin A (SIgA) in 17 professional players from the same Portuguese elite futsal club (age: 23.07 ± 6.76 years old; height: 1.75 ± 0.06 m; body mass: 75.47 ± 7.47 kg; experience in playing in elite: 5.38 ± 2.03 years) who performed 5 matches and 16 training sessions in a period of 27 days. The salivary content of total protein, TOS, TAC, oxidative stress index, and SIgA were calculated before and after the training sessions and the unofficial matches under study. Saliva sampling was conducted 10 min before each match or training session and 40 min after (post-match and post-training). The MANOVA of repeated measures showed a significant difference for total protein and SIgA (p < 0.01). Total protein (sphericity = 0.007; statistical power = 0.818) and SIgA (sphericity = 0.018; statistical power = 0.693) are highly correlated with the time factor. The main findings revealed several key points: (a) There was a significant increase in total protein, SIgA, and TAC during acute load (pre- vs. post-session) in both training and match contexts. Specifically, total protein and SIgA displayed notable increments in both training and match settings, while TAC exhibited significant increases exclusively during matches. (b) No changes in TOS and oxidative stress index were observed during acute load in either training or match contexts. (c) A positive trend was noted between the chronic load during a congested week of the precompetitive season and the decrease in total protein and SIgA levels. (d) Additionally, a positive correlation between internal training loads and oxidative/antioxidant responses was found, as expressed by the oxidative stress index, without significant differences (p-value > 0.05) in acute and chronic loads during congested matches and training schedules. [ABSTRACT FROM AUTHOR]

Mosquito‐borne viruses (MBVs) are a large class of viruses transmitted mainly through mosquito bites, including dengue virus, Zika virus, Japanese encephalitis virus, West Nile virus, and chikungunya virus, which pose a major threat to the health of people around the world. With global warming and extended human activities, the incidence of many MBVs has increased significantly. Mosquito saliva contains a variety of bioactive protein components. These not only enable blood feeding but also play a crucial role in regulating local infection at the bite site and the remote dissemination of MBVs as well as in remodeling the innate and adaptive immune responses of host vertebrates. Here, we review the physiological functions of mosquito salivary proteins (MSPs) in detail, the influence and the underlying mechanism of MSPs on the transmission of MBVs, and the current progress and issues that urgently need to be addressed in the research and development of MSP‐based MBV transmission blocking vaccines. [ABSTRACT FROM AUTHOR]

Objective: To determine which salivary proteins adhere onto sport mouthguards, and to evaluate the effectiveness of different cleaning strategies in removing deposited protein. Methods: Fifteen healthy volunteers used a mouthguard for 1 h. The deposited salivary proteins were analyzed using gel electrophoresis and Western blotting techniques and compared with the protein composition of unstimulated saliva. In addition, the effectiveness of two different cleaning strategies to remove proteins from the mouthguards were compared: rinsing the mouthguards after use with cold tap water and cleaning the mouthguard with a soluble effervescent tablet. Results: Gel electrophoresis showed deposition of proteins of 50–60 kDa and 14 kDa on the mouthguards used in the mouth for 1 h. Western blotting identified these bands as amylase and lysozyme, respectively. Rinsing the mouthguard with cold tap water after use removed 91% of the total amount of deposited proteins, while cleaning with an effervescent tablet removed 99%. Conclusions: During the use of mouthguards, salivary proteins are deposited on their surface. Because salivary proteins can potentially affect bacterial adhesion to mouthguards, proper cleaning after use is recommended. Cleaning the mouthguard with cold tap water or using an effervescent tablet both seem to be effective strategies to remove proteins deposited on sport mouthguards. [ABSTRACT FROM AUTHOR]

Simple Summary: Piglets raised at a farm for meat production experience stress that affects their welfare. Recently, it has been demonstrated that several salivary proteins are up- or down-regulated in stressful situations. These could, therefore, be used as biomarkers for stress. This review revisits the definition of stress and defines the protein composition of porcine saliva to finally propose a panel of six salivary stress biomarkers. Identifying the potential presence of stress at the pig farm is fundamental since it affects pig welfare. As a result, a reliable and straightforward tool to monitor stress could record the welfare status of the animals. Although numerous methods to assess the welfare of pigs have been developed in the past, no gold standard has been established yet. Recently, the value of saliva as a tool to identify chronic stress in piglets was explored, as it can be collected fast and non-invasively. Since the protein composition, i.e., the proteome of porcine saliva, responds to stress, the affected proteins could be used as salivary stress biomarkers. The present review first defines stress and its relationship with welfare. Next, the porcine gland-specific salivary proteome is characterized. Finally, six potential salivary biomarkers for stress are proposed, i.e., odorant-binding protein, vomeromodulin-like protein, chitinase, lipocalin-1, long palate lung and nasal epithelium protein, and alpha-2-HS-glycoprotein. [ABSTRACT FROM AUTHOR]

Saliva has shown considerable promise as a diagnostic medium for point-of-care (POC) and over-the-counter (OTC) diagnostic devices due to the non-invasive nature of its collection. However, a significant limitation of saliva-based detection is undesirable interference in a sensor's readout caused by interfering components in saliva. In this study, we develop standardized sample treatment procedures to eliminate bubbles and interfering molecules while preserving the sample's target molecules such as spike (S) protein and glucose. We then test the compatibility of the pretreatment system with our previously designed SARS-CoV-2 and glucose diagnostic biosensing systems for detecting S protein and glucose in subject saliva. Ultimately, the effectiveness of each filter in enhancing biomarker sensitivity is assessed. The results show that a 20 mg nylon wool (NW) filter shows an 80% change in viscosity reduction with only a 6% reduction in protein content, making it an appropriate filter for the salivary S protein diagnostic system. Meanwhile, a 30 mg cotton wool (CW) filter is identified as the optimal choice for salivary glucose detection, achieving a 90% change in viscosity reduction and a 60.7% reduction in protein content with a minimal 4.3% reduction in glucose content. The NW pretreatment filtration significantly improves the limit of detection (LOD) for salivary S protein detection by five times (from 0.5 nM to 0.1 nM) and it reduces the relative standard deviation (RSD) two times compared to unfiltered saliva. Conversely, the CW filter used for salivary glucose detection demonstrated improved linearity with an R2 of 0.99 and a sensitivity of 36.6 μA/mM·cm2, over twice as high as unfiltered saliva. This unique filtration process can be extended to any POC diagnostic system and optimized for any biomarker detection, making electrochemical POC diagnostics more viable in the current market. [ABSTRACT FROM AUTHOR]

The present study identified 151 proteins from the salivary gland of cowpea aphid Aphis craccivora Koch, using LC-MS/MS analysis. These included enzymes mainly involved in the digestion and detoxification of secondary metabolites and proteins related to cell development and molecular function. Enzymes like peroxidase, trehalase, cytochrome P450 monooxygenase, glutathione peroxidase, esterase, peptidase, carboxypeptidase, maltase, and beta-galactosidase were prevalent in the proteome. Additionally, several proteins were assigned to cellular and molecular functions of salivary gland. These proteins may be involved in host-plant interactions. Comprehensively, these results provide a database for elucidating aphid-plant interactions at the molecular level in the future. [ABSTRACT FROM AUTHOR]

Abstract Introduction This longitudinal study assessed the association between salivary protein composition and the clinical onset/severity of oral mucositis (OM) in patients with head and neck tumours treated with intensity-modulated-radiotherapy (IMRT). Methods Saliva samples/clinical data were obtained from 40 head and neck cancer patients treated at Guy’s Hospital before -IMRT(T0) and after-IMRT (T1 = 6 m, T2 = 12 m) (ethics approval/consent). Salivary flow rate, total protein concentration, and secretion rate were determined from saliva samples and compared with pre-treatment values. OM was assessed, total/specific salivary proteins, including mucin 5B and 7, IgA, cystatin-S, albumin, and α-amylase, were quantified. Results 95% patients experienced OM during IMRT, with 33 subjects reaching grade 2&3. At T1, there was a significant reduction in salivary flow rate, total protein secretion rate, α-amylase and cystatin-S compared to baseline. Remarkably IMRT did not significantly alter mucin 5B and 7, or the IgA secretion rate at any time point. At T1, all the analyzed proteins were associated with the OM outcomes. In addition, there was a significant inverse correlation between IgA concentration at T0 and the severity of OM during IMRT. Conclusion This study revealed significant associations between several salivary proteins and OM in patients with head and neck cancer undergoing IMRT. Further longitudinal studies are needed to confirm these results. Clinical significance The study contributes to the understanding of certain salivary proteins association with OM. This could be the first step towards identifying potential salivary markers that could offer perspectives for personalized medicine approaches to improve their quality of life (QoL). Research question What is the association between salivary proteins and the occurrence and severity of OM in head and neck cancer patients? Aim To assess the association between salivary protein composition with the clinical onset/severity of oral mucositis (OM) in head and neck cancer patients treated with intensity modulated radiotherapy. Null hypothesis There is no association between salivary proteins and onset/severity of OM in HNC patients.

Introduction: This longitudinal study assessed the association between salivary protein composition and the clinical onset/severity of oral mucositis (OM) in patients with head and neck tumours treated with intensity-modulated-radiotherapy (IMRT). Methods: Saliva samples/clinical data were obtained from 40 head and neck cancer patients treated at Guy's Hospital before -IMRT(T0) and after-IMRT (T1 = 6 m, T2 = 12 m) (ethics approval/consent). Salivary flow rate, total protein concentration, and secretion rate were determined from saliva samples and compared with pre-treatment values. OM was assessed, total/specific salivary proteins, including mucin 5B and 7, IgA, cystatin-S, albumin, and α-amylase, were quantified. Results: 95% patients experienced OM during IMRT, with 33 subjects reaching grade 2&3. At T1, there was a significant reduction in salivary flow rate, total protein secretion rate, α-amylase and cystatin-S compared to baseline. Remarkably IMRT did not significantly alter mucin 5B and 7, or the IgA secretion rate at any time point. At T1, all the analyzed proteins were associated with the OM outcomes. In addition, there was a significant inverse correlation between IgA concentration at T0 and the severity of OM during IMRT. Conclusion: This study revealed significant associations between several salivary proteins and OM in patients with head and neck cancer undergoing IMRT. Further longitudinal studies are needed to confirm these results. Clinical significance: The study contributes to the understanding of certain salivary proteins association with OM. This could be the first step towards identifying potential salivary markers that could offer perspectives for personalized medicine approaches to improve their quality of life (QoL). Research question: What is the association between salivary proteins and the occurrence and severity of OM in head and neck cancer patients? Aim: To assess the association between salivary protein composition with the clinical onset/severity of oral mucositis (OM) in head and neck cancer patients treated with intensity modulated radiotherapy. Null hypothesis: There is no association between salivary proteins and onset/severity of OM in HNC patients. [ABSTRACT FROM AUTHOR]

The infectious inoculum of a sand fly, apart from its metacyclic promastigotes, is composed of factors derived from both the parasite and the vector. Vector-derived factors, including salivary proteins and the gut microbiota, are essential for the establishment and enhancement of infection. However, the type and the number of bacteria egested during salivation is unclear. In the present study, sand flies of Phlebotomus papatasi were gathered from three locations in hyperendemic focus of zoonotic cutaneous leishmaniasis (ZCL) in Isfahan Province, Iran. By using the forced salivation assay and targeting the 16S rRNA barcode gene, egested bacteria were characterized in 99 (44%) out of 224 sand flies. Culture-dependent and culture-independent methods identified the members of Enterobacter cloacae and Spiroplasma species as dominant taxa, respectively. Ten top genera of Spiroplasma, Ralstonia, Acinetobacter, Reyranella, Undibacterium, Bryobacter, Corynebacterium, Cutibacterium, Psychrobacter, and Wolbachia constituted >80% of the saliva microbiome. Phylogenetic analysis displayed the presence of only one bacterial species for the Spiroplasma, Ralstonia, Reyranella, Bryobacter and Wolbachia, two distinct species for Cutibacterium, three for Undibacterium and Psychrobacter, 16 for Acinetobacter, and 27 for Corynebacterium, in the saliva. The abundance of microbes in P. papatasi saliva was determined by incorporating the data on the read counts and the copy number of 16S rRNA gene, about 9,000 bacterial cells, per sand fly. Both microbiological and metagenomic data indicate that bacteria are constant companions of Leishmania, from the intestine of the vector to the vertebrate host. This is the first forced salivation experiment in a sand fly, addressing key questions on infectious bite and competent vectors. Author summary: Female sand flies salivate during feeding on vertebrate blood and natural sugars. During salivation, they may release microorganisms associated with the salivary glands and digestive tract, i.e. viruses, bacteria, and other pathogens, into the feeding substrates. However, the type and the number of bacteria egested during salivation is unclear. Here, by sing the forced salivation experiment and targeting the 16S rRNA barcode gene, egested bacteria were characterized in sand flies. Culture-dependent and culture-independent methods identified the members of Enterobacter cloacae and Spiroplasma species as dominant taxa, respectively. The abundance of microbes in the saliva of each P. papatasi sand fly was determined to be around 9,000 bacterial cells. The lack of a vaccine and the failure of treatment in leishmaniasis provide many motivations to better understand the factors cause the inflammatory response. The findings of this study can improve our insight into measuring the effect of vector-derived bacteria on the improvement or deterioration of leishmaniasis. [ABSTRACT FROM AUTHOR]

Calcium binding protein, spermatid associated 1 (CABS1) is a protein most widely studied in spermatogenesis. However, mRNA for CABS1 has been found in numerous tissues, albeit with little information about the protein. Previously, we identified CABS1 mRNA and protein in human salivary glands and provided evidence that in humans CABS1 contains a heptapeptide near its carboxyl terminus that has anti-inflammatory activities. Moreover, levels of an immunoreactive form of CABS1 were elevated in psychological stress. To more fully characterize human CABS1 we developed additional polyclonal and monoclonal antibodies to different sections of the protein and used these antibodies to characterize CABS1 in an overexpression cell lysate, human salivary glands, saliva, serum and testes using western blot, immunohistochemistry and bioinformatics approaches exploiting the Gene Expression Omnibus (GEO) database. CABS1 appears to have multiple molecular weight forms, consistent with its recognition as a structurally disordered protein, a protein with structural plasticity. Interestingly, in human testes, its cellular distribution differs from that in rodents and pigs, and includes Leydig cells, primary spermatogonia, Sertoli cells and developing spermatocytes and spermatids, Geodata suggests that CABS1 is much more widely distributed than previously recognized, including in the urogenital, gastrointestinal and respiratory tracts, as well as in the nervous system, immune system and other tissues. Much remains to be learned about this intriguing protein. [ABSTRACT FROM AUTHOR]

Introduction: Aedes spp. are the most prolific mosquito vectors in the world. Found on every continent, they can effectively transmit various arboviruses, including the dengue virus which continues to cause outbreaks worldwide and is spreading into previously non-endemic areas. The lack of widely available dengue vaccines accentuates the importance of targeted vector control strategies to reduce the dengue burden. High-throughput tools to estimate human-mosquito contact and evaluate vector control interventions are lacking. We propose a novel serological tool that allows rapid screening of human cohorts for exposure to potentially infectious mosquitoes. Methods: We tested 563 serum samples from a longitudinal pediatric cohort study previously conducted in Cambodia. Children enrolled in the study were dengue-naive at baseline and were followed biannually for dengue incidence for two years. We used Western blotting and enzyme-linked immunosorbent assays to identify immunogenic Aedes aegypti salivary proteins and measure total anti-Ae. aegypti IgG. Results: We found a correlation (rs=0.86) between IgG responses against AeD7L1 and AeD7L2 recombinant proteins and those to whole salivary gland homogenate. We observed seasonal fluctuations of AeD7L1+2 IgG responses and no cross-reactivity with Culex quinquefasciatus and Anopheles dirus mosquitoes. The baseline median AeD7L1+2 IgG responses for young children were higher in those who developed asymptomatic versus symptomatic dengue. Discussion: The IgG response against AeD7L1+2 recombinant proteins is a highly sensitive and Aedes specific marker of human exposure to Aedes bites that can facilitate standardization of future serosurveys and epidemiological studies by its ability to provide a robust estimation of human-mosquito contact in a highthroughput fashion. [ABSTRACT FROM AUTHOR]

Background: Mid- to late-stage Parkinson's disease (PD) is often linked with worsened and significant impairment of motor activities, but existing prognostic markers do not adequately capture the risk of loss of balance in PD patients. This study aims to develop a risk prognostic model for mid- to late-stage PD and identify prognostic factors that are indicative of impending loss of balance and falls. Methods: The study included 307 participants of which 75 were diagnosed with idiopathic PD and 232 were neurological or non-neurological controls. Among the PD group, 46 were early-stage (Hoehn and Yahr [H&Y] = 1,2) with no significant loss of balance while 29 were mid- to late-stage (H&Y = 3,4,5) which is characterized by loss of balance and falls. Multivariable logistic regression (MLR) was used to develop a prognostic model for mid- to late-stage PD. Model discrimination was assessed by ROC curves. The model was internally validated through bootstrapping and calibration plots. Results: The relevant factors identified and included in the final MLR model were shortness of breath, age, swollen joints, heme oxygenase-1 (HO-1) protein, and total salivary protein. The model had an AUC of 0.82 (95% CI = 0.71–0.92) and was well calibrated (calibration slope = 0.77, intercept = 0.03). The likelihood of shortness of breath (OR = 7.91, 95% CI = 1.63–45.12) was significantly higher among mid- to late-stage PD than early-stage. Age and total salivary protein were also significantly higher among mid- to late-stage PD. Conclusion: The MLR prognostic model for mid- to late-stage PD may assist physicians in identifying patients at high risk for loss of balance and falls. [ABSTRACT FROM AUTHOR]

Aphids are insect pests that suck phloem sap and introduce salivary proteins into plant tissues through saliva secretion. The effector of salivary proteins plays a key role in the modulation of host plant defense responses and enhancing aphid host adaptation. Based on previous transcriptome sequencing results, a candidate effector cyclin-dependent kinase-like (CDK) was identified from the grain aphid Sitobion avenae. In this study, the function of SaCDK in wheat defense response and the adaptation of S. avenae was investigated. Our results showed that the transient overexpression of SaCDK in tobacco Nicotiana benthamiana suppressed cell death triggered by mouse pro-apoptotic protein-BAX or Phytophthora infestans PAMP-INF1. SaCDK, delivered into wheat cells through a Pseudomonas fluorescens-mediated bacterial type III secretion system, suppressed callose deposition in wheat seedlings, and the overexpression of SaCDK in wheat significantly decreased the expression levels of salicylic acid and jasmonic acid signaling pathway-related genes phenylalanine ammonia lyase (PAL), pathogenesis-related 1 protein (PR1), lipoxygenase (LOX) and Ω-3 fatty acid desaturase (FAD). In addition, aphid bioassay results showed that the survival and fecundity of S. avenae were significantly increased while feeding on the wheat plants carrying SaCDK. Taken together, our findings demonstrate that the salivary protein SaCDK is involved in inhibiting host defense response and improving its host adaptation, which lays the foundation to uncover the mechanism of the interaction of cereal aphids and host plants. [ABSTRACT FROM AUTHOR]

Mosquitoes transmit many disease-relevant flaviviruses. Efficient viral transmission to mammalian hosts requires mosquito salivary factors. However, the specific salivary components facilitating viral transmission and their mechanisms of action remain largely unknown. Here, we show that a female mosquito salivary gland-specific protein, here named A. aegypti Neutrophil Recruitment Protein (AaNRP), facilitates the transmission of Zika and dengue viruses. AaNRP promotes a rapid influx of neutrophils, followed by virus-susceptible myeloid cells toward mosquito bite sites, which facilitates establishment of local infection and systemic dissemination. Mechanistically, AaNRP engages TLR1 and TLR4 of skin-resident macrophages and activates MyD88-dependent NF-κB signaling to induce the expression of neutrophil chemoattractants. Inhibition of MyD88-NF-κB signaling with the dietary phytochemical resveratrol reduces AaNRP-mediated enhancement of flavivirus transmission by mosquitoes. These findings exemplify how salivary components can aid viral transmission, and suggest a potential prophylactic target. Synopsis: Mosquito saliva contains factors that promote flavivirus infection in the animal host. This study reveals that the mosquito salivary protein AaNRP mediates cutaneous recruitment of flavivirus-susceptible myeloid cells to mosquito bite sites, thus aiding flavivirus transmission. AaNRP is a female-mosquito-specific salivary protein. AaNRP stimulates skin-resident macrophages via TLR1/4-MyD88-NF-κB signaling to release chemoattractants for neutrophils. The AaNRP-induced influx of myeloid cells promotes flaviviral cutaneous infection. Dietary supplementation with resveratrol suppresses AaNRP-promoted flavivirus transmission by mosquitoes. AaNRP is a female-specific mosquito salivary protein that promotes Zika and dengue virus infection by stimulating skin macrophage-dependent neutrophil recruitment. [ABSTRACT FROM AUTHOR]

Vitamin B12 (cobalamin or Cbl) functions as a cofactor in two important enzymatic processes in human cells, and life is not sustainable without it. B12 is obtained from food and travels from the stomach, through the intestine, and into the bloodstream by three B12-transporting proteins: salivary haptocorrin (HC), gastric intrinsic factor, and transcobalamin (TC), which all bind B12 with high affinity and require proteolytic degradation to liberate Cbl. After intracellular delivery of dietary B12, Cbl in the aquo/hydroxocobalamin form can coordinate various nucleophiles, for example, GSH, giving rise to glutathionylcobalamin (GSCbl), a naturally occurring form of vitamin B12. Currently, there is no data showing whether GSCbl is recognized and transported in the human body. Our crystallographic data shows for the first time the complex between a vitamin B12 transporter and GSCbl, which compared to aquo/hydroxocobalamin, binds TC equally well. Furthermore, sequence analysis and structural comparisons show that TC recognizes and transports GSCbl and that the residues involved are conserved among TCs from different organisms. Interestingly, haptocorrin and intrinsic factor are not structurally tailored to bind GSCbl. This study provides new insights into the interactions between TC and Cbl. [ABSTRACT FROM AUTHOR]

Objectives: The presence of prostate‐specific antigen (PSA) in saliva and salivary glands has been reported. Nevertheless, its release pathway in these glands remains to be elucidated. Here, we showed PSA subcellular distribution focusing on its plausible route in human salivary parenchyma. Materials and Methods: Sections of parotid and submandibular glands were subjected to the immunohistochemical demonstration of PSA by the streptavidin–biotin method revealed by alkaline phosphatase. Moreover, ultrathin sections were collected on nickel grids and processed for immunocytochemical analysis, to visualize the intracellular distribution pattern of PSA through the observation by transmission electron microscopy. Results: By immunohistochemistry, in both parotid and submandibular glands PSA expression was detected in serous secretory acini and striated ducts. By immunocytochemistry, immunoreactivity was retrieved in the cytoplasmic compartment of acinar and ductal cells, often associated with small cytoplasmic vesicles. PSA labeling appeared also on rough endoplasmic reticulum and in the acini's lumen. A negligible PSA labeling appeared in most of the secretory granules of both glands. Conclusions: Our findings clearly support that human parotid and submandibular glands are involved in PSA secretion. Moreover, based on the immunoreactivity pattern, its release in oral cavity would probably occur by minor regulated secretory or constitutive‐like secretory pathways. [ABSTRACT FROM AUTHOR]

Aim: To evaluate the salivary biomarkers and plaque index after a treatment with a propolis-contained toothpaste. Materials and Methods: This is a longitudinal, randomized, double-blind study where 76 participants were randomized into two groups: Group I: Fluoridated Red Propolis toothpaste; Group II: Fluoridated toothpaste. The participants were selected in a municipality without fluoridated public water. All participants received standardized oral hygiene instructions from the same instructor for 3 daily brushings (after breakfast, after lunch, and before bed) for a period of 2 min; Saliva samples were collected before (D0) and after 28 days (D28) of treatment for analysis of pH and total protein, amylase, and IL-10. Saliva was collected in the initial consultation and on return, totaling two collections. All samples were collected under the same conditions, by the same operator and between 9:00 am and 11:00 am in order to minimize the influence of circadian rhythm on salivary flow. Results: On D0 and D28, the various treatments had no effect on total salivary proteins (G1: P = 0.0746; G2: P = 0.2144), and the pH stayed about the same. Additionally, there was no change in the amylase activity in G1 (P = 0.1877) or G2 (P = 0.4674). Significant decreases in G1 (P < 0.0001) and G2 (P = 0.03) were observed with IL-10. There was no statistically significant difference in the salivary flow between the BRP toothpaste-treated group (P = 0.172) and the commercial fluoridated toothpaste-treated group (P = 0.329). Compared to G2 (P = 0.03), G1 showed a superior decline in the plaque index (P = <0.0001). Conclusions: After 28 days of using the toothpastes, there were no changes in the amylase, pH, or total protein indicators. After 28 days, there was a decrease in the propolis group’s IL-10 dose and plaque index. [ABSTRACT FROM AUTHOR]

Food consumption can alter the biochemistry and redox status of human saliva, and the serving temperature of food may also play a role. The study aimed to explore the immediate (3 min) and delayed (30 min) effects of hot tea (57 ± 0.5 °C) ingestion and cold tea (8 ± 0.5 °C) ingestion on the salivary flow rate and salivary redox-relevant attributes. The saliva was collected from 20 healthy adults before, 3-min after and 30-min after the tea ingestion. The hot or cold deionised water at the same temperatures were used as control. The salivary flow rate and redox markers in hot tea (HBT), cold tea (CBT), hot water (HW) and cold water (CW) group were analysed and compared. The results demonstrated that neither the black tea nor the water altered the salivary flow rate; the black tea immediately increased the salivary thiol (SH) and malondialdehyde (MDA) content while reduced salivary uric acid (UA) significantly. The tea ingestion showed a tendency to elevate the ferric reducing antioxidant power (FRAP) in saliva, although not significantly. The water ingestion decreased the MDA content immediately and increased the UA level significantly. Cold water was found to induce a greater delayed increase in total salivary total protein (TPC) than the hot water. In conclusion, the black tea ingestion affects the redox attributes of human saliva acutely and significantly, while the temperature of drink makes the secondary contribution. [ABSTRACT FROM AUTHOR]

Introduction: Cutaneous leishmaniasis is a neglected vector-borne parasitic disease prevalent in 92 countries with approximately one million new infections annually. Interactions between vector saliva and the human host alter the response to infection and outcome of disease. Methods: To characterize the human immunological responses developed against saliva of Phlebotomus duboscqi, a Leishmania major (L. major) vector, we repeatedly exposed the arms of 14 healthy U.S volunteers to uninfected P. duboscqi bites. Blood was collected a week after each exposure and used to assess total IgG antibodies against the proteins of P. duboscqi salivary gland homogenate (SGH) and the levels of IFN-gamma and IL-10 from peripheral blood mononuclear cells (PBMCs) stimulated with SGH or recombinant sand fly proteins. We analyzed skin punch biopsies of the human volunteer arms from the insect bite site and control skin site after multiple P. duboscqi exposures (four volunteers) using immunohistochemical staining. Results: A variety of immediate insect bite skin reactions were observed. Late skin reactions to insect bites were characterized by macular hyperpigmentation and/or erythematous papules. Hematoxylin and eosin staining showed moderate mononuclear skin infiltrate with eosinophils in those challenged recently (within 2 months), eosinophils were not seen in biopsies with recall challenge (6 month post bites). An increase in plasma antigen-specific IgG responses to SGH was observed over time. Western Blot results showed strong plasma reactivity to five P. duboscqi salivary proteins. Importantly, volunteers developed a cellular immunity characterized by the secretion of IFN-gamma upon PBMC stimulation with P. duboscqi SGH and recombinant antigens. Discussion: Our results demonstrate that humans mounted a local and systemic immune response against P. duboscqi salivary proteins. Specifically, PduM02/SP15-like and PduM73/adenosine deaminase recombinant salivary proteins triggered a Th1 type immune response that might be considered in future development of a potential Leishmania vaccine. [ABSTRACT FROM AUTHOR]

Astringency has an important impact on the taste quality of tea infusion, a process which occurs when polyphenols complex with salivary proteins to form an impermeable membrane. (-)-Epigallocatechin gallate (EGCG) is the main astringent compound found in green tea and mucin is the main protein present in saliva. Determining the turbidity of EGCG–mucin mixtures is an effective method to quantify the astringency intensity of EGCG solutions. In this study, the effects of taste-related, substances present during green tea infusion, on the turbidity of EGCG–mucin mixtures was investigated under the reacting conditions of a pH value of 5.0, at 37 °C, and for 30 min. The results showed that epicatechins, caffeic acid, chlorogenic acid, and gallic acid reduced the turbidity of EGCG–mucin mixtures, while rutin increased turbidity. Metal ions increased the turbidity of EGCG–mucin mixtures. These can be arranged by effectiveness as Al3+ > K+ > Mg2+ > Ca2+. Caffeine, theanine, and sodium glutamate all decreased the turbidity values of EGCG–mucin mixtures, but sucrose had a weak effect. Further experiments confirmed that the turbidity of green tea infusion–mucin mixture indicated the astringent intensity of green tea infusion, and that the turbidity was significantly correlated with the contents of tea polyphenols and EGCG. [ABSTRACT FROM AUTHOR]

Background: Insecticide resistance (IR) is one of the major threats to malaria vector control programs in endemic countries. However, the mechanisms underlying IR are poorly understood. Thus, investigating gene expression patterns related to IR can offer important insights into the molecular basis of IR in mosquitoes. In this study, RNA-Seq was used to characterize gene expression in Anopheles gambiae surviving exposure to pyrethroids (deltamethrin, alphacypermethrin) and an organophosphate (pirimiphos-methyl). Results: Larvae of An. gambiae s.s. collected from Bassila and Djougou in Benin were reared to adulthood and phenotyped for IR using a modified CDC intensity bottle bioassay. The results showed that mosquitoes from Djougou were more resistant to pyrethroids (5X deltamethrin: 51.7% mortality; 2X alphacypermethrin: 47.4%) than Bassila (1X deltamethrin: 70.7%; 1X alphacypermethrin: 77.7%), while the latter were more resistant to pirimiphos-methyl (1.5X: 48.3% in Bassila and 1X: 21.5% in Djougou). RNA-seq was then conducted on resistant mosquitoes, non-exposed mosquitoes from the same locations and the laboratory-susceptible An. gambiae s.s. Kisumu strain. The results showed overexpression of detoxification genes, including cytochrome P450s (CYP12F2, CYP12F3, CYP4H15, CYP4H17, CYP6Z3, CYP9K1, CYP4G16, and CYP4D17), carboxylesterase genes (COEJHE5E, COE22933) and glutathione S-transferases (GSTE2 and GSTMS3) in all three resistant mosquito groups analyzed. Genes encoding cuticular proteins (CPR130, CPR10, CPR15, CPR16, CPR127, CPAP3-C, CPAP3-B, and CPR76) were also overexpressed in all the resistant groups, indicating their potential role in cross resistance in An. gambiae. Salivary gland protein genes related to 'salivary cysteine-rich peptide' and 'salivary secreted mucin 3' were also over-expressed and shared across all resistant groups. Conclusion: Our results suggest that in addition to metabolic enzymes, cuticular and salivary gland proteins could play an important role in cross-resistance to multiple classes of insecticides in Benin. These genes warrant further investigation to validate their functional role in An. gambiae resistance to insecticides. [ABSTRACT FROM AUTHOR]

Background: The present study aimed to estimate the salivary immunoglobulin A and total salivary proteins in minor RAS and in healthy controls. 50 patients with known history of minor RAS were diagnosed based on the major criteria with the absence of any other systemic disease or conditions and presenting with single and multiple ulcers and 50 healthy control group were included in the study. Minor aphthous were selected as they are more common than the major and herpetiform ulcers. Unstimulated saliva was collected by spit method in a calibrated measuring cup from 100 participants (50 RAS and 50 healthy controls) to assess the salivary IgA levels and total salivary proteins. A detailed case history and informed consent was taken from all the participants prior to our study. As results we found the mean age of study group was 31.58 years where as in control group it was 30.16 years with a range of 20-50 years in study group and 19-50 years in the control group with a p value of 0.03 that's significant. Out of 50 participants 36 (72%) were males and 14 (28%) were females in the study group where as in control group 38 (76%) were males and 12 (24%) were females. When compare between group A and group B showed significantly higher in study group as compare control group and concluded in our study RAS showed a male predominance with maximum number of patients in the age group of 20-50 years. The salivary immunoglobulin A levels was significantly higher in minor RAS group than the healthy control group. [ABSTRACT FROM AUTHOR]