Evaluation of core Biomarkers of Alzheimer's disease in saliva and plasma measured by chemiluminescent enzyme immunoassays on a fully automated platform.

Cerebrospinal fluid (CSF) core biomarkers of Alzheimer's disease (AD), including amyloid peptide beta-42 (Aß42), Aß42/40 ratio, and phosphorylated tau (pTau), are precious tools for supporting AD diagnosis. However, their use in clinical practice is limited due to the invasiveness of CSF collection. Thus, there is intensive research to find alternative, noninvasive, and widely accessible biological matrices to measure AD core biomarkers. In this study, we measured AD core biomarkers in saliva and plasma by a fully automated platform. We enrolled all consecutive patients with cognitive decline. For each patient, we measured Aß42, Aß40, and pTau levels in CSF, saliva, and plasma by Lumipulse G1200 (Fujirebio). We included forty-two patients, of whom 27 had AD. Levels of all biomarkers significantly differed in the three biofluids, with saliva having the lowest and CSF the highest levels of Aß42, Aß40, and pTau. A positive correlation of pTau, Aß42/40 ratio, and pTau/Aß42 ratio levels in CSF and plasma was detected, while no correlation between any biomarker in CSF and saliva was found. Our findings suggest that plasma but not saliva could represent a surrogate biofluid for measuring core AD biomarkers. Specifically, plasma Aß42/40 ratio, pTau/Aß42 ratio, and pTau could serve as surrogates of the corresponding CSF biomarkers.

Monitoring levetiracetam concentration in saliva during pregnancy is stable and feasible.

AIMS: This multicenter prospective cohort study (registration no. ChiCTR2000032089) aimed to investigate the relationship between saliva and plasma levetiracetam concentrations to determine whether saliva could be used for routine monitoring of levetiracetam during pregnancy. METHODS: The slot concentrations of levetiracetam in simultaneously obtained saliva and plasma samples were measured using UPLC-MS/MS. The correlations between saliva and plasma levetiracetam concentrations and the dose-normalized concentrations were compared among pregnant women in different stages and nonpregnant control participants with epilepsy. RESULTS: In total, 231 patients with 407 plasma and saliva sample pairs were enrolled from 39 centers. Linear relationships between salivary and plasma levetiracetam concentrations were reported in the enrolled population (r = 0.898, p < 0.001), including pregnant (r = 0.935, p < 0.001) and nonpregnant participants (r = 0.882, p < 0.001). Plasma concentrations were moderately higher than saliva concentrations, with ratios of saliva to plasma concentrations of 0.98 for nonpregnant women, 0.98, 1, and 1.12 for pregnant women during the first trimester, the second trimester, the and third trimester, respectively. The effective range of saliva levetiracetam concentration was found to be 9.98 µg/mL (lower limit) with an area under the curve (AUC) of 0.937 (95% confidence intervals, 0.915-0.959), sensitivity of 88.9%, specificity of 86.8%, and p < 0.001, to 24.05 µg/mL (upper limit) with an AUC of 0.952 (0.914-0.99), sensitivity of 100%, specificity of 92.3%, and p = 0.007. CONCLUSION: The saliva/plasma concentration ratio of levetiracetam remains constant during pregnancy and is similar to that in non-pregnant individuals. Monitoring levetiracetam concentration in saliva during pregnancy should be widely promoted.

Translocation of Antimicrobial Peptides across Model Membranes: The Role of Peptide Chain Length.

Cushioned lipid bilayers are structures consisting of a lipid bilayer supported on a solid substrate with an intervening layer of soft material. They offer possibilities for studying the behavior and interactions of biological membranes more accurately under physiological conditions. In this work, we continue our studies of cushion formation induced by histatin 5 (24Hst5), focusing on the effect of the length of the peptide chain. 24Hst5 is a short, positively charged, intrinsically disordered saliva peptide, and here, both a shorter (14Hst5) and a longer (48Hst5) peptide variant were evaluated. Experimental surface active techniques were combined with coarse-grained Monte Carlo simulations to obtain information about these peptides. Results show that at 10 mM NaCl, both the shorter and the longer peptide variants behave like 24Hst5 and a cushion below the bilayer is formed. At 150 mM NaCl, however, no interaction is observed for 24Hst5. On the contrary, a cushion is formed both in the case of 14Hst5 and 48Hst5, and in the latter, an additional thick, diffuse, and highly hydrated layer of peptide and lipid molecules is formed, on top of the bilayer. Similar trends were observed from the simulations, which allowed us to hypothesize that positively charged patches of the amino acids lysine and arginine in all three peptides are essential for them to interact with and translocate over the bilayer. We therefore hypothesize that electrostatic interactions are important for the interaction between the solid-supported lipid bilayers and the peptide depending on the linear charge density through the primary sequence and the positively charged patches in the sequence. The understanding of how, why, and when the cushion is formed opens up the possibility for this system to be used in the research and development of new drugs and pharmaceuticals.

Novel buffer for long-term preservation of DNA in biological material at room temperature.

The collection and preservation of biological material before DNA analysis is critical for inter alia biomedical research, medical diagnostics, forensics and biodiversity conservation. In this study, we evaluate an in-house formulated buffer called the Forensic DNA Laboratory-buffer (FDL-buffer) for preservation of biological material for long term at room temperature. Human saliva stored in the buffer for 8 years, human blood stored for 3 years and delicate animal tissues from the jellyfish Pelagia noctiluca comb jelly Beroe sp., stored for 4 and 6 years respectively consistently produced high-quality DNA. FDL-buffer exhibited compatibility with standard organic, salting out and spin-column extraction methods, making it versatile and applicable to a wide range of applications, including automation.

[Box: see text].

Influence of bottled salad dressings on the development of enamel erosion in the presence or absence of salivary pellicle - An in vitro study.

Background: Acidic beverages are believed to elevate the risk of enamel surface erosion. In addition to the intake of soft drinks, the increased consumption of salad dressings has been linked to a higher prevalence of dental erosion. Therefore, the current study aimed to investigate the influence of bottled salad dressings on the development of enamel erosion in the presence or absence of pellicle through in vitro experiment. Methods: Preliminary pH and calcium analyses of solutions were performed. Highest pH and calcium content was found for sandwich spread i.e., 4.69 and 55.4 mg/100 g grams, respectively. Eighty tooth specimens (measuring 4 × 4 × 3 mm) were prepared from extracted human premolars and randomly assigned to four groups (group 1: orange juice; group 2: eggless plain mayonnaise; group 3: sandwich spread; and group 4: thousand island dressing) with 20 samples in each group. Ten tooth specimens from each group were immersed in 20 ml of the respective solutions for 5 min (control group). The remaining ten tooth specimens from each group were submerged in 5 mL saliva vials for 3 min to facilitate salivary pellicle formation before being immersed in their respective solutions for 5 min (saliva-covered group). Pre and post-experimental assessments of enamel roughness and hardness were conducted using a surface roughness tester and Knoop Hardness indenter, respectively. Results: Overall, enamel roughness was notably elevated in the control group, with the eggless plain mayonnaise (0.52 ± 0.38) and thousand island dressing groups (0.57 ± 0.29) showing a significant increase in surface roughness post-test (p = 0.05). Nevertheless, there was no significant difference in the enamel roughness between the groups. On the other hand, regardless of the presence/absence of the salivary pellicle, a marked decrease in enamel hardness was observed among all groups except for group 3 (sandwich spread) with a mean score of 311.5 ± 82.6 (p < 0.05). Conclusion: A significant increase in surface roughness and reduction in enamel hardness was observed with salad dressings. However, in vitro formed salivary pellicle showed a protective effect against tooth erosion.

Brushing Methods for Patients Unable to Use a Mouthwash: A Preliminary Study of the Effects of Moisturizing Gel and Povidone-Iodine in Healthy Volunteers.

INTRODUCTION:  Brushing older adults or intubated patients who are unable to rinse can transmit bacteria from dental plaque into the oral cavity and increase the risk of aspiration pneumonia. Therefore, this study examined brushing methods to prevent the spread of bacteria in the oral cavity.  Methods: Three types of brushing methods were performed on five volunteers by dental hygienists (water group: brushing with toothbrush bristles soaked in water; gel group: brushing with a moisturizing gel placed on the toothbrush; PV-I group: brushing with toothbrush bristles dipped in povidone-iodine). Neither group spat out the saliva or gargled during brushing but brushed while wiping the water/gel/PV-I solution with a sponge brush. The same five volunteers served as subjects for the three methods. Saliva was collected before and after brushing, and the number of colonies was determined using bacterial culture.  Results: The water group demonstrated a significantly increased number of bacteria in the saliva owing to the spread of bacteria from the dental plaque. The gel group prevented the spread of the bacteria. The PV-I group showed a significant decrease in the number of bacteria in the saliva after brushing. CONCLUSIONS:  Brushing with toothbrush bristles dipped in a povidone-iodine solution is recommended for intubated or older adult patients who cannot gargle.

Oral Mucosa and Saliva Alterations Related to Vape.

OBJECTIVES: Electronic nicotine delivery systems (e-cigarette, pod, and vape) are currently among the tobacco consumption of adolescents and young adults. The aim is to show oral mucosa and saliva alterations related to vape. MATERIAL AND METHODS: A vape-user patient, presenting a white plaque in the posterior region of the hard palate, underwent clinical examination, sialometry, pH evaluation, and excisional biopsy of the white lesion. Molecular changes in saliva and vape liquid were analyzed by vibrational spectroscopy. RESULTS: The histopathological analyses showed hyperparakeratosis without dysplasia. Formaldehyde, ketones, and aromatic hydrocarbon species were identified in e-cig liquid by the FTIR. CONCLUSIONS: The use of vape may be related to the development of hyperkeratotic lesions in the oral mucosa as well as significantly modify the patient's salivary patterns as the vape liquid presents carcinogenic and cytotoxic components in its composition.

Salivary oxytocin changes and effect of the season in sows kept in different farrowing systems: Farrowing crate and farrowing pen with temporary crating.

Alternative farrowing systems that have been developed in recent years could have a positive effect on the welfare of sows during farrowing and lactation. Oxytocin measurements in saliva may provide information about positive animal welfare status. The purpose of this study was to evaluate the changes in salivary oxytocin concentrations in sows during the lactation period in three different farrowing systems and in two different seasons. Crossbred Duroc sows (n = 34, average parity = 3.6 ± 1.80) were housed in conventional farrowing crates (FC) (n = 10) or in farrowing pens with temporary crating (TC), including SWAP (n = 12) and JFL15 (n = 12) in two different seasons: summer and winter. Saliva samples were collected for six days during lactation: days 2, 4, 12, 23, 25 (i.e., 1-day post-weaning) and 26 (i.e., 2-day post-weaning) after farrowing. Moreover, behavioral data from sows was recorded on days 2, 4, 12 and 23 after farrowing, using a 30-s scan sampling method for 3 min per pen to record the behaviors which were assessed by the same observer. The results showed that the salivary oxytocin concentrations were 472.5 pg/mL and 399.4 pg/mL higher in both TC (SWAP and JLF15, respectively) than in the FC in early-lactation period, and these differences were more pronounced in summer and at the end of lactation in winter. In terms of behavior, higher number of mother-young interactions were observed in TC than FC in early- and mid-lactation period. In conclusion, TC is associated to a higher salivary oxytocin concentration that could indicated an increased mother-young interaction, although oxytocin concentration can be influenced by other factors, such as season or day of lactation.

Impact of saliva incorporation on the rheological properties of in vitro gastric contents formulated from sour cream.

Rheological properties of gastric contents depend on the food ingested, and on the volume and composition of secretions from the host, which may vary. This study investigates the impact of saliva regular incorporation in the stomach after a meal on the rheological properties of gastric contents, considering two levels of salivary flow (low = 0.5 and high = 1.5 mL/min). In vitro chymes were obtained by mixing sour cream, simulated gastric fluid, two different volumes of oral fluid (at-rest human saliva, SSF for Simulated Salivary Fluid or water) and adjusting pH at 3. Chymes samples were characterized at 37°C for their particle size and rheological properties. Overall, particle size distribution was not different between samples: incorporating a larger volume of saliva resulted in more heterogeneity, but the surface area moment D[3,2] and volume moment D[4,3] did not differ significantly with the oral fluid type. Shear viscosity of chyme samples was higher when saliva was incorporated, in comparison with water or SSF. In addition, as shown from data extracted at γ ̇ $$ \dot{\gamma} $$ = 20 s-1 the higher the fluid volume the lower the shear viscosity, which is attributed to a dilution effect. However, this dilution effect was attenuated in the case of saliva, most likely due to its composition in organic compounds (e.g., mucins) contributing to the rheological properties of this biological fluid. In these in vitro conditions, both saliva and the salivation rate had a significant but slight impact on the rheological properties of gastric contents (of the order of 1-5 mPa s at γ ̇ $$ \dot{\gamma} $$ = 20 s-1).

Alteration of salivary LPO, MDA, LDH, glutathione, GPx, SOD and vitamins in oral submucous fibrosis: A three-level meta-analysis study.

This study aims to investigate the alteration of salivary biomarker profiling in the development of oral submucous fibrosis (OSMF) and to explore the influence of saliva in the diagnosis of OSMF. A systematic search of published articles using the PRISMA guidelines was conducted to identify relevant studies on OSMF and saliva. All eligible studies, including case-control, cross-sectional studies, cohort, and pilot studies, contained the evaluation of salivary biomarker profiling in patients with OSMF. Salivary biomarker data from 28 selected articles were categorized into nine groups, and their mean values were determined. A three-step meta-analysis was performed by grouping salivary biomarker profiling into more heterogeneous categories based on OSMF classification, considering functional, histological, and clinical grading. The salivary biomarker profiling analysis revealed significant alterations in all markers, indicating their efficacy in OSMF diagnosis. Subgroup analyses highlighted significant associations in oxidative stress and protein with increased mean values, particularly emphasizing lipid peroxidase (LPO), malondialdehyde (MDA), and lactate dehydrogenase (LDH). Conversely, decreased mean values were observed in glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and vitamins. Notably, OSMF grading analysis demonstrated a significant difference in weighted effect sizes for histological grading, particularly in stage IV. The study underscores the alteration of specific salivary biomarkers, particularly those associated with LPO, MDA, LDH, glutathione, GPx, SOD, and vitamins, in diagnosing and grading OSMF.

Saliva-derived secondary DNA transfer on fabric: The impact of varying conditions.

This study explored secondary DNA transfer involving saliva, a body fluid often encountered in forensic investigations. Various factors were examined to investigate their potential impact on the transfer of DNA from saliva stains deposited onto common types of fabric (cotton, nylon, and towel). We examined varying types of saliva moisture (wet, dry, and rehydrated) and different types of contact (controlled pressure and active/friction pressure) to quantitatively evaluate how such variables could impact transfer and possible conclusions surrounding saliva-derived DNA deposits. The transfer of DNA was generally least pronounced with more absorbent primary fabrics (cotton and towel materials) while a less absorbent primary fabric (nylon) exhibited a greater propensity for DNA transfer. There were significantly higher amounts of transferred DNA (p < 0.05) observed in wet saliva samples compared to dry and rehydrated saliva samples. Further, the use of active pressure (friction) appeared to result in more DNA transfer overall as compared to controlled pressure contact. Experiments conducted with wet saliva and active pressure (friction) demonstrated the highest likelihood of transfer, with the primary nylon and secondary towel fabric combination demonstrating the greatest average transfer percentage of 94.74 %. The variables explored in this study presented multiple combinations wherein a sufficient amount of DNA (≥ 240 pg total) was transferred to the secondary fabric, making it potentially suitable for STR-PCR amplification in our laboratory. The findings from this study indicate that the type of primary fabric receiving the saliva deposit, the type of saliva moisture, the type of secondary fabric and its moisture type, and the type of contact all have the potential to affect the quantity of DNA transferred and recovered. This study provides empirical data on the ease, and to what extent, DNA from saliva transfers between fabrics and aids DNA activity level evaluations. The significance of this research lies in its contribution to expanding our current understanding of DNA transfer involving saliva within forensic science and criminal investigations.

Relationship between oral hypofunction and salivary biomarkers in older adults: a cross-sectional study.

BACKGROUND: Oral health problems have increased among older adults. Oral hypofunction is characterized by seven signs and symptoms: oral uncleanness, oral dryness, decline in occlusal force, decline in the movement function of the tongue and lips, decline in tongue pressure, decline in masticatory function, and decline in swallowing function, the latter being a significant risk factors for oral frailty. Recent research has suggested that salivary biomarkers can be used to assess not only oral diseases, including dental caries and periodontitis, but also systemic diseases, such as cancer and diabetes mellitus. This cross-sectional study investigated the relationship between oral hypofunction and the levels of salivary biomarkers. METHODS: In total, 116 patients, aged 65 years or older, were included in this cross-sectional study. If three or more signs or symptoms in seven kinds of tests met the criteria of each test, oral hypofunction was diagnosed. The levels of biomarkers in the saliva collected from the patients were analyzed using an enzyme-linked immunosorbent assay. RESULTS: In total, 63.8% of patients were diagnosed with oral hypofunction. Multivariable linear regression analysis showed that calprotectin levels in the saliva were significantly related to oral moisture and masticatory function. Furthermore, 8-OHdG levels in saliva were associated with the movement function of the tongue and lips and oral hygiene level, and salivary AGE correlated only with the movement function of the tongue and lips. Multiple logistic regression analysis revealed that calprotectin levels in the saliva were significantly correlated with the prevalence of oral hypofunction, even after adjusting for age, sex, and periodontal status. However, none of the biomarker levels in the saliva had a significant relationship with the number of examinations outside the reference range. CONCLUSIONS: Calprotectin, 8-OHdG, and AGE levels are associated with oral hypofunction in older adults.

Saliva stress biomarkers in ERCP trainees before and after familiarisation with ERCP on a virtual simulator.

Background: Stress during the early ERCP learning curve may interfere with acquisition of skills during training. The purpose of this study was to compare stress biomarkers in the saliva of trainees before and after familiarisation with ERCP exercises on a virtual simulator. Methods: Altogether 26 endoscopists under training, 14 women and 12 men, completed the three phases of this study: Phase 1. Three different ERCP procedures were performed on the simulator. Saliva for α-amylase (sAA), Chromogranin A (sCgA), and Cortisol (sC) were collected before (baseline), halfway through the exercise (ex.), and 10 min after completion of the exercise (comp.); Phase 2. A three-week familiarisation period where at least 30 different cases were performed on the virtual ERCP simulator; and Phase 3. Identical to Phase 1 where saliva samples were once again collected at baseline, during, and after the exercise. Percentage differences in biomarker levels between baseline and exercise (Diffex) and between baseline and completion (Diffcomp) during Phase 1 and Phase 3 were calculated for each stress marker. Results: Mean % changes, Diffex and Diffcomp, were significantly positive (p < 0.05) for all markers in both Phase 1 and Phase 3. Diffex in Phase 1 was significantly greater than Diffex in Phase 3 (p < 0.05) for sAA and sCgA. Diffcomp for sAA in Phase 1 was significantly greater than Diffcomp in Phase 3 (p < 0.05). No significant differences were found in sC concentration between Phases 1 and 3. Conclusion: This study shows that familiarisation with the ERCP simulator greatly reduced stress as measured by the three saliva stress biomarkers used with sAA being the best. It also suggests that familiarisation with an ERCP simulator might reduce stress in the clinical setting.

Dysrhythmic saliva microbiota in mobile phone addicts with sleep disorders and restored by acupuncture.

Background: Mobile phone addiction (MPA) greatly affects the biological clock and sleep quality and is emerging as a behavioral disorder. The saliva microbiota has been linked to circadian rhythms, and our previous research revealed dysrhythmic saliva metabolites in MPA subjects with sleep disorders (MPASD). In addition, acupuncture had positive effects. However, the dysbiotic saliva microbiota in MPASD patients and the restorative effects of acupuncture are unclear. Objectives: To probe the circadian dysrhythmic characteristics of the saliva microbiota and acupunctural restoration in MPASD patients. Methods: MPASD patients and healthy volunteers were recruited by the Mobile Phone Addiction Tendency Scale (MPATS) and the Pittsburgh Sleep Quality Index (PSQI). Saliva samples were collected every 4 h for 72 h. After saliva sampling, six MPDSD subjects (group M) were acupuncturally treated (group T), and subsequent saliva sampling was conducted posttreatment. Finally, all the samples were subjected to 16S rRNA gene sequencing and bioinformatic analysis. Results: Significantly increased MPATS and PSQI scores were observed in MPDSD patients (p< 0.01), but these scores decreased (p<0.001) after acupuncture intervention. Compared with those in healthy controls, the diversity and structure of the saliva microbiota in MPASD patients were markedly disrupted. Six genera with circadian rhythms were detected in all groups, including Sulfurovum, Peptostreptococcus, Porphyromonas and Prevotella. There were five genera with circadian rhythmicity in healthy people, of which the rhythmicities of the genera Rothia and Lautropia disappeared in MPASD patients but effectively resumed after acupuncture intervention. Conclusions: This work revealed dysrhythmic salivary microbes in MPASD patients, and acupuncture, as a potential intervention, could be effective in mitigating this ever-rising behavioral epidemic.

The Acute Effects of Kinesio Taping on Drooling in Children with Cerebral Palsy: A Randomized Placebo-Controlled Trial.

INTRODUCTION: This study purpose of determining the short-term effects of kinesio taping on drooling in children with cerebral palsy (CP). METHODS: CP were randomly divided into 3 groups as the kinesio tape (n = 16), sham tape (n = 16), and control (n = 16) groups. The drooling severity and frequency were assessed with the Drooling Severity and Frequency Scale, and the amount of saliva was measured by the 5-minute drooling quotient. All outcome measurements were repeated for all children at the baseline, after 45 minutes, and after 2 days of application. RESULTS: There was a significant decrease found in drooling severity, frequency, and amount in the kinesio taping group (p < .05). There was no significant difference in the sham taping and control groups (p > .05). CONCLUSION: The use of kinesio tape in drooling reduced drooling severity, frequency, and amount.

Biomarkers in Alzheimer's disease.

Alzheimer's disease (AD) is among the most common neurodegenerative disorders. AD is characterized by deposition of neurofibrillary tangles and amyloid plaques, leading to associated secondary pathologies, progressive neurodegeneration, and eventually death. Currently used diagnostics are largely image-based, lack accuracy and do not detect early disease, ie, prior to onset of symptoms, thus limiting treatment options and outcomes. Although biomarkers such as amyloid-ß and tau protein in cerebrospinal fluid have gained much attention, these are generally limited to disease progression. Unfortunately, identification of biomarkers for early and accurate diagnosis remains a challenge. As such, body fluids such as sweat, serum, saliva, mucosa, tears, and urine are under investigation as alternative sources for biomarkers that can aid in early disease detection. This review focuses on biomarkers identified through proteomics in various biofluids and their potential for early and accurate diagnosis of AD.

Multikingdom oral microbiome interactions in early-onset cryptogenic ischemic stroke.

Although knowledge of the role of the oral microbiome in ischemic stroke is steadily increasing, little is known about the multikingdom microbiota interactions and their consequences. We enrolled participants from a prospective multicentre case-control study and investigated multikingdom microbiome differences using saliva metagenomic datasets (n = 308) from young patients diagnosed with cryptogenic ischemic stroke (CIS) and age- and sex-matched stroke-free controls. Differentially abundant taxa were identified using Analysis of Compositions of Microbiomes with Bias Correction (ANCOM-BC2). Functional potential was inferred using HUMANn3. Our findings revealed significant differences in the composition and functional capacity of the oral microbiota associated with CIS. We identified 51 microbial species, including 47 bacterial, 3 viral, and one fungal species associated with CIS in the adjusted model. Co-abundance network analysis highlighted a more intricate microbial network in CIS patients, indicating potential interactions and co-occurrence patterns among microbial species across kingdoms. The results of our metagenomic analysis reflect the complexity of the oral microbiome, with high diversity and multikingdom interactions, which may play a role in health and disease.

Revealing microbial community characteristics in healthy human, cat and canine salivas and looking for species-specific microbes.

As two kinds of increasingly popular pets, the saliva of cat or canine is most likely to be left at the crime scene compared with the common types of body fluids in forensics. Accurately identifying the species of saliva samples found at the crime scene involving pets will help the investigators find available testing materials, reduce the consumption of reagents and save the investigative time of the case. Therefore, it is necessary to explore the characteristics and differences of saliva microbiomes of cat, canine and human. In this study, 16S rRNA gene amplicon sequencing technology was used to reveal microbial communities of saliva samples of healthy human, cat, and canine. Alpha diversity analyses indicated that canine saliva demonstrated the highest microbial diversity, followed by cat saliva, whereas human saliva microbial diversity was the lowest. The saliva samples of the three species all had their own unique microbial community compositions, and the dominant phyla of canine and cat salivas were Proteobacteria and Bacteroidete, while the dominant phyla of human saliva were Firmicutes and Proteobacteria. There was no significant statistical difference in the salivary microbiota obtained by the two collection methods (cotton swab and liquid saliva). The gender of cats and canines might have no effect on the salivary microbiota, but the different breeds had an impact on their saliva microbiomes. Principal coordinates analysis, non-metric multidimensional scaling analysis and random forest analysis all indicated significant differences in microbial community structures among the three species, allowing inference on the species sources of saliva samples by microbiome method. Differential microbial biomarkers for the salivas of three species were screened out using a variety of bioinformatics analyses, and the results demonstrated that Prevotella melaninogenica, Veillonella parvula, and Haemophilus parainfluenzae could be used as species-specific microbial biomarkers of human saliva. The detections of human species-specific microbes provide a potential method for determining human saliva.

Salivary IgG and IgA in newborn calves and the possible role in the assessment of passive immunity transfer.

Introduction: The transfer of immunoglobulins from the mother to newborns is widely recognized as a critical event for safeguarding offspring against potentially life-threatening infectious diseases. Mainly for this reason, this study aimed to assess the concentrations of immunoglobulin G (IgG) and immunoglobulin A (IgA) in the saliva of newborn calves and explore its potential use for monitoring passive immunity transfer from cows to calves, as also to evaluate how colostrum intake affects serum and saliva IgG and IgA concentrations. Methods: The quality of colostrum samples was evaluated using an optical refractometer before administration to the calves. Saliva and blood samples from 24 calves were obtained at the day of birth (T0) and 2 days after (T2) for determination of serum concentrations of total protein by refractometer, IgG and IgA (both on serum and saliva) by ELISA test. Results: Positive correlations were observed between salivary IgA at T2 and salivary IgG at T2. A significant increase in both IgG and IgA levels in calf serum and saliva was noted. Salivary IgA levels can reflect salivary IgG levels. Discussion: These findings suggest the potential utility of IgA in monitoring passive immunity transfer, and do not exclude saliva as an alternative, practical, and non-invasive matrix for assessing passive immunity transfer.

Single-molecule characterization of salivary protein aggregates from Parkinson's disease patients: a pilot study.

Saliva is a convenient and accessible biofluid that has potential as a future diagnostic tool for Parkinson's disease. Candidate diagnostic tests for Parkinson's disease to date have predominantly focused on measurements of α-synuclein in CSF, but there is a need for accurate tests utilizing more easily accessible sample types. Prior studies utilizing saliva have used bulk measurements of salivary α-synuclein to provide diagnostic insight. Aggregate structure may influence the contribution of α-synuclein to disease pathology. Single-molecule approaches can characterize the structure of individual aggregates present in the biofluid and may, therefore, provide greater insight than bulk measurements. We have employed an antibody-based single-molecule pulldown assay to quantify salivary α-synuclein and amyloid-ß peptide aggregate numbers and subsequently super-resolved captured aggregates using direct Stochastic Optical Reconstruction Microscopy to describe their morphological features. We show that the salivary α-synuclein aggregate/amyloid-ß aggregate ratio is increased almost 2-fold in patients with Parkinson's disease (n = 20) compared with controls (n = 20, P < 0.05). Morphological information also provides insight, with saliva from patients with Parkinson's disease containing a greater proportion of larger and more fibrillar amyloid-ß aggregates than control saliva (P < 0.05). Furthermore, the combination of count and morphology data provides greater diagnostic value than either measure alone, distinguishing between patients with Parkinson's disease (n = 17) and controls (n = 18) with a high degree of accuracy (area under the curve = 0.87, P < 0.001) and a larger dynamic range. We, therefore, demonstrate for the first time the application of highly sensitive single-molecule imaging techniques to saliva. In addition, we show that aggregates present within saliva retain relevant structural information, further expanding the potential utility of saliva-based diagnostic methods.