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Objective: The objective was to develop a manually operatable, non-electric device to measure salivary spinnbarkeit for research, clinical and educational purposes., Materials and Methods: A newly developed device, named the Kamranmeter, was built and evaluated in a pilot study, comparing the spinnbarkeit of unstimulated and chew-stimulated saliva samples from healthy volunteers. The Neva Meter, which operates on electrical resistance, was used as a reference standard., Results: The study found no statistically significant differences between the two devices when measuring the spinnbarkeit of both unstimulated and stimulated saliva samples from healthy volunteers., Conclusions: We developed a hand-operated device to analyse salivary spinnbarkeit. The Kamranmeter performs comparably to the Neva Meter in measuring spinnbarkeit in saliva obtained from healthy volunteers., Clinical Relevance: The Kamranmeter has potential applications in diagnostic tests involving saliva, such as assessing spinnbarkeit to evaluate conditions like caries and dry mouth. Moreover, it holds promise for broader medical uses, including fertility assessments, where spinnbarkeit is a key indicator of cervical mucus quality. Its ease of handling is expected to make it well-suited for research, clinical, and educational applications., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Porcine epidemic diarrhea virus (PEDV) is a harmful coronavirus infecting pigs, which is resulting in substantial financial losses in the global pig industry. The lack of effective vaccines or treatments underscores the pressing need for new antiviral strategies. Antimicrobial peptides (AMPs), specifically cathelicidins such as LL-37, have demonstrated promising activity against a range of viruses. This study aims to elucidate the antiviral mechanisms of cathelicidins by examining their inhibitory capabilities against PEDV in vitro. Four pig-derived antimicrobial peptides (PMAP-36, PMAP-23, PR-39, and PG-1), together with chicken-derived CATH-B1 and human-derived LL-37 were analyzed for their anti-PEDV activity. Flow cytometry and fluorescent microscopy confirmed that LL-37 and CATH-B1 had strong inhibitory effects at non-toxic concentrations of 5 and 10 µM, significantly reducing GFP-PEDV infection of Vero cells both in co- and pre-incubation setups. In contrast, none of the porcine peptides exhibited any inhibitory effects, even at higher doses. Fluorogenic LL-37 was shown to enter VERO cells, indicative of a possible immunomodulatory antiviral mode of action. However, transmission electron microscopy clearly indicated that both LL-37 and CATH-B1 affected virus morphology and caused aggregation of viral particles, showing that peptide-virus interaction caused reduced virus infectivity. In conclusion, this analysis highlights the potential of LL-37 and CATH-B1 as inhibitors against PEDV, suggesting promising directions for innovative therapeutic antiviral strategies., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024. Published by Elsevier B.V.)

Objective: Until now, the clinically relevant improvement for the Xerostomia Inventory (XI) has not been defined. Therefore, our aim was to determine the Minimally Important Difference (MID) of the XI for improvement in dry-mouth symptoms in SjD patients., Method: The study recruited 34 SjD patients who underwent sialendoscopy of major salivary glands and 15 SjD patients in a nonintervention control group. XI scores were assessed at several time points. The MID was determined from the mean difference in XI scores between the groups with and without improvement., Results: In the control group, no significant XI score changes were seen. In the sialendoscopy group, a clinically relevant XI score change of four scale points was identified after 1 week. For a prolonged duration (≥16 weeks), a minimum reduction of seven scale points in the XI score was required to indicate clinically relevant improvement., Conclusion: In SjD patients, a minimum change of four points in the XI score indicates a clinically relevant improvement for evaluating short-term effects. For prolonged effects, a clinically relevant improvement requires a MID of seven points. The determination of the MID in XI could assist in future studies that evaluate changes in xerostomia., (© 2023 The Authors. Oral Diseases published by Wiley Periodicals LLC.)

Background: Olfactory stimulation with mastic resin, derived from the Pistacia lentiscus tree, demonstrated a bona fide sialagogic effect in healthy volunteers [1]. Its main volatile compound, α-pinene, also showed this effect. The current study aimed to validate the effect of mastic resin volatiles in chronic dry mouth patients with confirmed decreased saliva secretion., Methods: 41 chronic dry mouth patients with decreased unstimulated saliva secretion (<0.25 mL/min) were exposed to mastic resin volatiles as part of the diagnostic routine at the Saliva Clinic of Academic Centre for Dentistry Amsterdam. During their visit, dry-mouth questionnaires were conducted and samples of unstimulated whole saliva, chew-stimulated saliva, acid-stimulated saliva and mastic resin stimulated saliva were collected. Saliva flow rate, spinnbarkeit, pH, ion composition, MUC5B and MUC7 levels in all samples were analyzed., Results: Salivary flow rates increased by all stimuli when compared to the baseline unstimulated saliva (P<0.001). During olfactory mastic resin stimulation, the salivary spinnbarkeit (P<0.001) and sodium concentration (P<0.01) were increased compared to unstimulated saliva. MUC5B and MUC7 levels were increased during olfactory mastic resin stimulation compared to chew-stimulated saliva (P=0.016 and P<0.001, respectively). Spinnbarkeit correlated positively with MUC5B (R=0.399, P=0.002) and MUC7 levels (R=0.375, P=0.004). Results of dry-mouth questionnaires indicated reduced posterior palate dryness shortly after olfactory mastic resin stimulation (P=0.04)., Conclusions: Olfactory mastic resin stimulation increased mucous saliva secretion and reduced posterior palate dryness in a group of chronic dry mouth patients. These findings, validated in patients, underscore mastic resin scent as a beneficial and non-invasive sialagogic treatment for clinical applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Background: Gingivitis in response to biofilm formation may exhibit different trajectories. The purposes of the present study were to characterize the composition of the supragingival microbiota and salivary cytokine and protein levels in healthy individuals with different gingivitis patterns, to test the hypothesis that manifestations of gingivitis associate with specific profiles in terms of supragingival microbiota, salivary cytokines, and proteins., Methods: Forty orally and systemically healthy individuals refrained from all oral hygiene procedures for a period of 14 days, followed by a resolution period of 14 days with regular oral care. Supragingival plaque level and bleeding on probing (BOP) were recorded, and supragingival plaque as well as saliva samples were collected at baseline, day 14, and day 28. Based on change in BOP% from baseline to day 14, rapid ( n  = 15), moderate ( n  = 10), and slow ( n  = 15) responders were identified. Supragingival microbiota composition, salivary cytokine, and protein levels were compared between groups at baseline, day 14, and day 28., Results: A significantly higher baseline abundance of Capnocytophaga, Eikenella , and Campylobacter species were recorded in rapid responders, whereas a significantly higher baseline abundance of Streptococcus species were detected in slow responders. Slow responders expressed a high degree of resilience, with minimal difference in microbial composition at baseline and after 14 days of resolution (day 28). On the contrary, significant differences in relative abundance of members of the core microbiota, Streptococcus , Actinomyces , and Rothia species, was noted in baseline samples versus day 28 samples in rapid responders. Comparable baseline cytokine and protein levels were recorded in all groups., Conclusion: Supragingival microbiota composition, but not saliva cytokine and protein profiles, seems to influence the extent of the inflammatory response during development of gingivitis in systemically healthy individuals., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Salivary stones are hardened, stony calcifications that primarily develop in the drainage duct of a salivary gland. They can lead to obstruction of the saliva flow, resulting in swelling and pain. Since the aetiology of salivary stones remains largely unclear, this was further investigated in this PhD study. A case-control review of patient records showed that systemic diseases and lifestyle factors most likely do not play a role in their occurrence. The biochemical composition of salivary stones removed by oral-maxillofacial surgeons was examined, revealing that large salivary stones have a different inorganic composition than small salivary stones. Several salivary proteins were detected in submandibular salivary stones, including lysozyme, s-IgA, and -amylase. Clumping together of these proteins may play a role in the initial formation of salivary stones.

Colonoscopy is the gold standard for diagnosing inflammatory bowel disease (IBD). However, this invasive procedure has a high burden for pediatric patients. Previous research has shown elevated fecal amino acid concentrations in children with IBD versus controls. We hypothesized that this finding could result from increased proteolytic activity. Therefore, the aim of this study was to investigate whether fecal protease-based profiling was able to discriminate between IBD and controls. Protease activity was measured in fecal samples from patients with IBD (Crohn's disease (CD) n = 19; ulcerative colitis (UC) n = 19) and non-IBD controls (n = 19) using a fluorescence resonance energy transfer (FRET)-peptide library. Receiver operating characteristic (ROC) curve analysis was used to determine the diagnostic value of each FRET-peptide substrate. Screening the FRET-peptide library revealed an increased total proteolytic activity (TPA), as well as degradation of specific FRET-peptides specifically in fecal samples from IBD patients. Based on level of significance (p < .001) and ROC curve analysis (AUC > 0.85), the fluorogenic substrates W-W, A-A, a-a, F-h, and H-y showed diagnostic potential for CD. The substrates W-W, a-a, T-t, G-v, and H-y showed diagnostic potential for UC based on significance (p < .001) and ROC analysis (AUC > 0.90). None of the FRET-peptide substrates used was able to differentiate between protease activity in fecal samples from CD versus UC. This study showed an increased fecal proteolytic activity in children with newly diagnosed, treatment-naïve, IBD. This could lead to the development of novel, noninvasive biomarkers for screening and diagnostic purposes., (© 2024 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)

Objectives: This scoping review aims to provide an overview of the research and potential applications of carbon dots (CDs) for oral health purposes., Design: Systematic literature searches were performed on PubMed and Web of Science databases (up to February 2023). Two co-authors selected the published works independently and extracted the data in accordance with the PRISMA statement. Studies with the application of CDs for oral health purposes were included., Results: Among 152 articles, 19 articles were finally selected. Eight studies investigated the anti-microbial effects of CDs against, for example, oral pathogens, eight studies explored the applicability of CDs in relation to oral cancer, and three studies investigated CDs in relation to cell differentiation and tissue regeneration in oral health. The studies showed the promising potential of CDs in oral health, particularly for inducing bacterial killing by increasing reactive oxygen species, killing oral cancer cells via photodynamic therapeutic effects, and inducing dental pulp and periodontal bone regeneration., Conclusion: The findings show that CDs have the potential to be utilized in the future for various oral health purposes. Besides, these results underline the broad-spectrum applicability of CDs, crossing the borders of oral health., (© 2023 The Authors. Oral Diseases published by Wiley Periodicals LLC.)

Objectives: To study the effects of carbon dots (CDs), in combination with phytosphingosine (PHS), against acid-induced demineralization of hydroxyapatite in vitro., Methods: CDs were generated from citric acid and urea by microwave heating. Transmission electron microscope (TEM), FT-IR, and fluorescence intensity were used to characterize the CDs. A hydroxyapatite (HAp) model was used to investigate the protective effects of CDs, PHS, and their combinations with and without a salivary pellicle against acid-induced demineralization in vitro. Ca 2+ release as a parameter to evaluate the inhibition of demineralization was measured by capillary electrophoresis. The interactions between CDs, PHS, and HAp discs were investigated using a fluorescence detector., Results: Uniform-sized CDs were synthesized, showing typical optical characteristics. CDs exhibited no inhibition of acid-induced demineralization in vitro, in contrast to PHS. Notably, a pre-coating of CDs increased the protective effects of PHS against acid-induced demineralization, which was not disturbed by the presence of a salivary pellicle and Tween 20. Scanning electron microscope (SEM) confirmed the binding and layers formed of both CDs and PHS to the HAp surfaces. Based on fluorescence spectra CDs binding to HAp seemed to be dependent on Ca 2+ and PO 4 3- interactions., Conclusions: CDs combined with PHS showed protective effects against acid-induced demineralization of HAp discs in vitro., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: The objective of the study was to evaluate the therapeutic effects of carbon quantum dots (CQDs) in immunomodulation on non-obese diabetic (NOD) mice, as the model for Sjögren's syndrome (SS)., Methods: Carbon quantum dots were generated from Setaria viridis via a hydrothermal process. Their toxic effects were tested by cell viability and blood chemistry analysis, meanwhile therapeutic effects were investigated in NOD mice in the aspects of saliva flow, histology, and immune cell distribution., Results: Carbon quantum dots, with rich surface chemistry and unique optical properties, showed non-cytotoxicity in vitro or no damage in vivo. Intravenously applied CQDs alleviated inflammation in the submandibular glands in NOD mice after 6-week treatments. The inflammatory area index and focus score were significantly decreased in CQD-treated mice. Besides, the levels of anti-SSA and anti-SSB were decreased in the presence of CQDs. The stimulated saliva flow rates and weight of submandibular glands were significantly increased in CQD-treated mice by reducing the apoptosis of cells. The CD3 + and CD4 + T cells distributed around the ducts of submandibular glands were significantly decreased, while the percentage of Foxp3 + cells was higher in CQD-treated mice than that in the control group., Conclusions: Our findings suggest that CQDs may ameliorate the dysregulated immune processes in NOD mice., (© 2023 The Authors. Oral Diseases published by Wiley Periodicals LLC.)

Despite the availability of a wide range of (fluoridated) oral care products, there is a constant search under way for new substances that contribute to a healthy mouth. Laboratory research shows that the lipid phytosphingosine forms a molecular layer on hydroxyapatite and protects it against acid-induced demineralization and bacterial adhesion. In the future, phytosphingosine may be used in the future as a new ingredient in oral care products for the prevention of tooth erosion and biofilm-related disorders, such as caries, gingivitis and periodontitis.

The mucosal pellicle (MP) is a biological film protecting the oral mucosa. It is composed of bounded salivary proteins and transmembrane mucin MUC1 expressed by oral epithelial cells. Previous research indicates that MUC1 expression enhances the binding of the main salivary protein forming the MP, MUC5B. This study investigated the influence of MUC1 structure on MP formation. A TR146 cell line, which does not express MUC1 natively, was stably transfected with genes coding for three MUC1 isoforms differing in the structure of the two main extracellular domains: the VNTR domain, exhibiting a variable number of tandem repeats, and the SEA domain, maintaining the two bound subunits of MUC1. Semi-quantification of MUC1 using dot blot chemiluminescence showed comparable expression levels in all transfected cell lines. Semi-quantification of MUC5B by immunostaining after incubation with saliva revealed that MUC1 expression significantly increased MUC5B adsorption. Neither the VNTR domain nor the SEA domain was influenced MUC5B anchoring, suggesting the key role of the MUC1 N-terminal domain. AFM-IR nanospectroscopy revealed discernible shifts indicative of changes in the chemical properties at the cell surface due to the expression of the MUC1 isoform. Furthermore, the observed chemical shifts suggest the involvement of hydrophobic effects in the interaction between MUC1 and salivary proteins.

Saliva diagnostics have become increasingly popular due to their non-invasive nature and patient-friendly collection process. Various collection methods are available, yet these are not always well standardized for either quantitative or qualitative analysis. In line, the objective of this study was to evaluate if measured levels of various biomarkers in the saliva of healthy individuals were affected by three distinct saliva collection methods: 1) unstimulated saliva, 2) chew stimulated saliva, and 3) oral rinse. Saliva samples from 30 healthy individuals were obtained by the three collection methods. Then, the levels of various salivary biomarkers such as proteins and ions were determined. It was found that levels of various biomarkers obtained from unstimulated saliva were comparable to those in chew stimulated saliva. The levels of potassium, sodium, and amylase activity differed significantly among the three collection methods. Levels of all biomarkers measured using the oral rinse method significantly differed from those obtained from unstimulated and chew-stimulated saliva. In conclusion, both unstimulated and chew-stimulated saliva provided comparable levels for a diverse group of biomarkers. However, the results obtained from the oral rinse method significantly differed from those of unstimulated and chew-stimulated saliva, due to the diluted nature of the saliva extract., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

In the oral cavity Candida albicans interacts with many oral bacteria, including Porphyromonas gingivalis , both physically and metabolically. The aim of this in vitro study was to characterize these interactions and study their effects on the survival of P. gingivalis . First, metabolic interactions were evaluated by counting the colony forming units (CFU) after co-culturing. The results indicated that the anaerobic bacterium P. gingivalis survives under aerobic conditions when co-cultured with C. albicans . This is due to the oxygen consumption by C. albicans as determined by a reduction in survival upon the addition of Antimycin A. By measuring the protease activity, it was found that the presence of C. albicans induced gingipain activity by P. gingivalis , which is an important virulence factor. Adherence of P. gingivalis to hyphae of C. albicans was observed with a dynamic flow system. Using various C. albicans mutants, it was shown that the mechanism of adhesion was mediated by the cell wall adhesins, members of the agglutinin-like sequence (Als) family: Als3 and Als1. Furthermore, the two microorganisms could be co-cultured into forming a biofilm in which P. gingivalis can survive under aerobic culturing conditions, which was imaged using scanning electron microscopy. This study has further elucidated mechanisms of interaction, virulence acquisition and survival of P. gingivalis when co-cultured with C. albicans . Such survival could be essential for the pathogenicity of P. gingivalis in the oxygen-rich niches of the oral cavity. This study has emphasized the importance of interaction between different microbes in promoting survival, virulence and attachment of pathogens, which could be essential in facilitating penetration into the environment of the host., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 The Authors.)

Background: Xerostomia, often associated with decreased saliva quality, poses challenges due to limited treatment efficacy. This study aimed to investigate alternative approaches to enhance saliva secretion through olfactory volatile stimulation with mastic resin and its main compound α-pinene, known for inhibiting acetylcholinesterase in vitro., Methods: The inhibitory effects of freshly prepared mastic resin extract oil and α-pinene oil on acetylcholinesterase (AChE) activity were measured in vitro. Eighty healthy participants were recruited and divided into two groups: exposed to mastic resin volatiles (n = 40) or α-pinene volatiles (n = 40). Saliva samples were collected pre, during and post exposure to analyze saliva flow rate, spinnbarkeit, ion composition and MUC5B levels., Results: Mastic resin extract oil and α-pinene oil inhibited AChE activity by 207 % and 22 %, respectively. Olfactory stimulation with these volatiles significantly increased saliva secretion rate without altering spinnbarkeit and ion composition. Salivary MUC5B concentration rose after exposure to mastic resin volatiles., Conclusions: Olfactory stimulation with mastic resin and α-pinene volatiles demonstrated a bona fide in vivo effect on saliva secretion, confirming their sialagogic capability, potentially as a result of local glandular AChE inhibition. These findings highlight the therapeutic potential of both volatile compounds in treating patients with xerostomia and hyposalivation through olfactory exposure., Competing Interests: Declaration of Competing Interest All authors declare that they have no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Objectives: To perform a systematic review and meta-analysis to determine the power of salivary electrolytes for the diagnosis of Sjögren's disease (SjD)., Methods: A literature search was conducted (last search March 2023) using PubMed and Web of Science and completed with a manual search. Articles were screened for reports of human salivary ion concentrations, comparing SjD patients with healthy controls and/or sicca patients. Articles not using the SjD classification criteria or performing the classification as part of the experimental design were excluded. Forest plots were used to present the meta-analyses results for each ion, distinguishing between salivary type (unstimulated and stimulated whole saliva, submandibular/sublingual and parotid saliva)., Results: A total of 21 out of 722 articles were eligible for inclusion. For SjD patients a significant increase in salivary ion concentration was observed for sodium, chloride and calcium when comparing to healthy controls. Significant differences between SjD and sicca patients were noted for sodium, chloride, phosphate, calcium, phosphate, nitrite and nitrate. Stimulated whole saliva showed larger variability in results between studies in comparison to other types of saliva (unstimulated whole saliva, submandibular/sublingual saliva and parotid saliva)., Conclusions: Despite differences in saliva type, salivary ion levels could be utilised for the screening for SjD. Making use of chloride in combination with sodium would be most promising for distinguishing SjD patients from healthy controls and adding phosphate to potentially make a distinguishment with sicca patients. Unstimulated whole saliva should be the first choice when testing salivary ion concentrations.

The aim was to test if probiotics counteract oral dysbiosis during 14 days of sugar stress and subsequently help restore oral homeostasis. Eighty healthy individuals received either probiotics ( n = 40) or placebo lozenges ( n = 40) for 28 days and rinsed with a 10% sucrose solution 6-8 times during the initial 14 days of the trial. Saliva and supragingival samples were collected at baseline, day 14, and day 28. Saliva samples were analyzed for levels of pro-inflammatory cytokines, albumin, and salivary enzyme activity. The supragingival microbiota was characterized according to the Human Oral Microbiome Database. After 14 days of sugar stress, the relative abundance of Porphyromonas species was significantly higher ( p = 0.03) and remained significantly elevated at day 28 in the probiotic group compared to the placebo group ( p = 0.004). At day 28, the relative abundance of Kingella species was significantly higher in the probiotic group ( p = 0.03). Streptococcus gordinii and Neisseria elongata were associated with the probiotic group on day 28, while Streptococcus sobrinus was associated with the placebo group on day 14 and day 28. On day 28, the salivary albumin level was significantly lower in the probiotic group. The present study demonstrates a potential stabilizing effect on the supragingival microbiota mediated by consumption of probiotics during short-term sugar stress.

The present study aims to test whether probiotics protect against experimental gingivitis incited by 14 days of oral hygiene neglect and/or subsequently support the restoration of oral homeostasis. Eighty systemically and orally healthy participants refrained from oral hygiene procedures for 14 days, followed by 14 days with regular oral hygiene procedures. Additionally, participants consumed either probiotics ( n = 40) or placebo ( n = 40) throughout the trial. At baseline, day 14, and day 28, supragingival plaque score and bleeding-on-probing percentage (BOP %) were registered, and supragingival plaque and saliva samples were collected. The supragingival microbiota was characterized using 16S sequencing, and saliva samples were analyzed for levels of pro-inflammatory cytokines and proteases. At day 28, the relative abundance of Lautropia ( p = 0.014), Prevotella ( p = 0.046), Fusobacterium ( p = 0.033), and Selenomonas ( p = 0.0078) genera were significantly higher in the placebo group compared to the probiotics group, while the relative abundance of Rothia ( p = 0.047) species was associated with the probiotics group. Streptococcus sanguinis was associated with the probiotics group, while Campylobacter gracilis was associated with the placebo group. No difference was observed in salivary cytokines, albumin, or any enzyme activity. The present study suggests that probiotics support the resilience of the oral microbiota in the resolution period after gingivitis.

Microbiome modulation, aiming to restore a health-compatible microbiota, is a novel strategy to treat periodontitis. This study evaluated the modulation effects of antimicrobial peptide LL-31 and its D-enantiomer (D-LL-31) on saliva-derived microcosm biofilms, spiked with or without Porphyromonas gingivalis . To this end, one-day-old biofilms were incubated for 24 h with biofilm medium alone, or medium containing 40 µM LL-31 or D-LL-31, after which biofilms were grown for 5 days. Biofilms were assessed at 1 day and 5 days after intervention for the total viable cell counts, dipeptidyl peptidase IV (DPP4) activity, P. gingivalis amount (by qPCR) and microbial composition (by sequencing). The results showed that D-LL-31, not LL-31, significantly reduced the total viable cell counts, the P. gingivalis amount, and the DPP4 activity of the biofilms spiked with P. gingivalis , but only at 1 day after intervention. In the biofilms spiked with P. gingivalis , D-LL-31 tended to reduce the α-diversity and the compositional shift of the biofilms in time as compared to the control and LL-31 groups. In conclusion, D-LL-31 showed a better performance than LL-31 in biofilm modulation. The biofilm modulation function of the peptides could be impaired when the biofilms were in a severely dysbiotic state.

Plant products have been employed in medicine for centuries. As the world becomes more health-conscious, there is a growing interest in natural and minimally processed products for oral health care. This has led to an increase in research into the bioactive compounds found in plant products, particularly monoterpenes. Monoterpenes are known to have beneficial biological properties, but the specific mechanisms by which they exert their effects are not yet fully understood. Despite this, some monoterpenes are already being used in oral health care. For example, thymol, which has antibacterial properties, is an ingredient in varnish used for caries prevention. In addition to this, monoterpenes have also demonstrated antifungal, antiviral, and anti-inflammatory properties, making them versatile for various applications. As research continues, there is potential for even more discoveries regarding the benefits of monoterpenes in oral health care. This narrative literature review gives an overview of the biological properties and current and potential applications of selected monoterpenes and their derivatives in oral health care. These compounds demonstrate promising potential for future medical development, and their applications in future research are expected to expand.

The binding of Host Defense Peptides (HDPs) to the endotoxin of Gram-negative bacteria has important unsolved aspects. For most HDPs, it is unclear if binding is part of the antibacterial mechanism or whether LPS actually provides a protective layer against HDP killing. In addition, HDP binding to LPS can block the subsequent TLR4-mediated activation of the immune system. This dual activity is important, considering that HDPs are thought of as an alternative to conventional antibiotics, which do not provide this dual activity. In this study, we systematically determine, for the first time, the influence of the O-antigen and Lipid A composition on both the antibacterial and anti-endotoxin activity of four HDPs (CATH-2, PR-39, PMAP-23, and PMAP36). The presence of the O-antigen did not affect the antibacterial activity of any of the tested HDPs. Similarly, modification of the lipid A phosphate (MCR-1 phenotype) also did not affect the activity of the HDPs. Furthermore, assessment of inner and outer membrane damage revealed that CATH-2 and PMAP-36 are profoundly membrane-active and disrupt the inner and outer membrane of Escherichia coli simultaneously, suggesting that crossing the outer membrane is the rate-limiting step in the bactericidal activity of these HDPs but is independent of the presence of an O-antigen. In contrast to killing, larger differences were observed for the anti-endotoxin properties of HDPs. CATH-2 and PMAP-36 were much stronger at suppressing LPS-induced activation of macrophages compared to PR-39 and PMAP-23. In addition, the presence of only one phosphate group in the lipid A moiety reduced the immunomodulating activity of these HDPs. Overall, the data strongly suggest that LPS composition has little effect on bacterial killing but that Lipid A modification can affect the immunomodulatory role of HDPs. This dual activity should be considered when HDPs are considered for application purposes in the treatment of infectious diseases.

Background and Purpose: High label uptake in 68 Ga-PSMA-11 PET/CT recently identified a bilateral nasopharyngeal structure as a salivary gland (SG)-like additional 'area of interest', to be considered in conditions affecting SGs. These structures were termed 'tubarial glands'. We aimed to further characterize their histological and immunohistochemical position compared to established SGs., Methods: Tubarial gland tissue was compared with parotid, submandibular, sublingual, palatal and labial SGs tissue using immunohistological techniques., Results: Expression of acinar cell-associated aquaporin-5 (AQP5) was detected in tubarial glands, in an apical location associated in control, established SGs with polarized, secretory acinar cells. Keratin14 (KRT14) expression in cells peripheral to AQP5 + clusters also suggested presence of myoepithelial cells. α-amylase, prolactin-induced protein, proline rich protein Haelll subfamily 2, and Muc5B expression suggests mucous acinar cell presence, and presence of muco-serous acinar cells peripheral to putative mucous acinar cells. Expression of adrenergic receptor-β1 by acinar-like cells of the tubarial gland suggests ability to transduce sympathetic neuronal signaling. In terms of ductal architecture, tubarial glands contained large excretory-like ducts (similar to all other SGs), and squamous ducts, comprised of intermingled KRT14 + and KRT7 + cells. These squamous ducts were also observed in palatal, sublingual and labial SGs. No striated or intercalated ducts were observed, similar to palatal SGs., Conclusion: Based on histological and immunohistochemical analyses, the tubarial glands resemble SGs. They most convincingly echo characteristics of the palatal SGs in terms of ductal cells, and both the palatal and labial SGs when considering acinar cells., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Dry mouth has a complex aetiology which makes proper diagnosis complicated. Until now, dry mouth diagnosis has mainly focused on the overall oral dryness, without taking into account regional differences within the mouth. This research showed, among other things, that there are unique patterns of oral dryness, each with its own cause. For example, patients suffering from Sjogren's syndrome mainly experienced dryness of the posterior of the palate. Patients with dry mouth due to the side effects of medication, in contrast, experienced the front part of the tongue as the driest. These findings suggest that mapping of intraoral dryness may be a useful diagnostic tool to differentiate between possible causes of dry mouth.

Objectives: Sjögren's disease (SjD) patients use various interventions to relief their oral dryness. However, the use and efficacy of these interventions have only partially been evaluated. The present study aims to investigate whether there is an association between the perceived oral dryness and discomfort of SjD patients and their use of specific interventions., Materials and Methods: A cross-sectional study was performed among SjD patients, who completed several questionnaires to assess the severity of their oral dryness and an inventory of dry-mouth interventions. The perceived efficacy of each intervention was reported on a 5-point Likert-scale., Results: The questionnaires were returned by 92 SjD patients. For relief of oral dryness, they mostly used "eating fruit", "drinking tea", "moistening the lips", "drinking water, and "drinking small volumes" (> 50%). Three interventions had a frequency of use ranging from 2-6 times/day, whereas, "drinking water" and "drinking small volumes" showed higher frequencies (> 14). The highest overall efficacy (≥ 3.5) was reported for "chewing gum" and "using a mouth gel". Furthermore, various dry-mouth interventions showed significant associations with oral dryness scores and/or patients' discomfort. For example, "drinking small volumes" and "using XyliMelts" were associated with the Bother Index score., Conclusion: Great variation was found in the use of dry-mouth interventions by the participants and the severity of the oral dryness and/or patients' discomfort seemed to affect their choice of intervention. Notably, the mostly used interventions did not show the highest reported efficacy., Clinical Relevance: These findings might help SjD patients and clinicians in their choice of effective dry-mouth interventions., (© 2023. The Author(s).)

A systematic review and meta-analysis were conducted to investigate the SARS-CoV-2 viral load in human saliva and compared it with the loads in oropharyngeal swabs, nasopharyngeal swabs, and sputum. In addition, the salivary viral loads of symptomatic and asymptomatic COVID-19 patients were compared. Searches were conducted using four electronic databases: PubMed, Embase, Scopus, and Web of Science, for studies published on SARS-CoV-2 loads expressed by C T values or copies/mL RNA. Three reviewers evaluated the included studies to confirm eligibility and assessed the risk of bias. A total of 37 studies were included. Mean C T values in saliva ranged from 21.5 to 39.6 and mean copies/mL RNA ranged from 1.91 × 10 1 to 6.98 × 10 11 . Meta-analysis revealed no significant differences in SARS-CoV-2 load in saliva compared to oropharyngeal swabs, nasopharyngeal swabs, and sputum. In addition, no significant differences were observed in the salivary viral load of symptomatic and asymptomatic COVID-19 patients. We conclude that saliva specimen can be used as an alternative for SARS-CoV-2 detection in oropharyngeal swabs, nasopharyngeal swabs, and sputum., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2023 Mouri R. J. Faruque et al.)

Salivary stones (sialoliths) are calcified structures located in the ductal system of the major salivary glands. Their exact cause is not clear but in general they are characterized by concentric inorganic (hydroxyapatite) layers. The formation is a slow intermittent process which may result in enlargement of the sialolith causing obstruction of saliva secretion resulting in mealtime related pain and swelling of the affected salivary gland. Various studies reported the presence of organic material such as proteins and lipids in the core of sialoliths. In the present study the protein composition of twenty submandibular sialoliths was analyzed. It was found that proteins contributed on average 5% to the dry weight of submandibular stones whereby small salivary stones contained more extractable proteins than large salivary stones. Using a combination of SDS-PAGE gel electrophoresis and Western blotting, we identified α-amylase (in all stones; 100%), lysozyme (95%), lactoferrin (85%), secretory-IgA (75%), MUC7 (60%), complement C4 (60%) and C-reactive protein (35%). The presence, and the combinations, of lactoferrin, lysozyme, s-IgA and α-amylase in sialoliths was confirmed by ELISA. The gradually increasing size of a sialolith might provoke a local inflammatory response in the duct of the submandibular gland whereby the relatively low concentrations of lactoferrin and lysozyme may originate from neutrophils. The interaction of lactoferrin with s-IgA could contribute to the accumulation of lactoferrin in sialoliths. In summary, these results suggest a new pathophysiological role for lactoferrin, in the formation of sialoliths., (© 2022. The Author(s).)

Objectives: The purposes of this study were to localize monocyte chemoattractant protein-1-induced protein-1 (MCPIP-1) and its suppressor mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT-1) in gingival tissues and to profile their protein expression levels in relation to the clinical inflammation, Porphyromonas gingivalis colonization, and interleukin (IL)-8 levels., Materials and Methods: Study samples were collected from two independent study populations: (1) Gingival tissues were collected from eight periodontally healthy individuals and eight periodontitis patients to localize MCPIP-1 and MALT-1 immunohistochemically, and (2) forty-one gingival tissue samples with marginal, mild, or moderate to severe inflammation were collected from 20 periodontitis patients to determine MCPIP-1 and MALT-1 levels using immunoblots, P. gingivalis levels with qPCR, P. gingivalis gingipain activities with fluorogenic substrates, and IL-8 levels with multiplex technique., Results: MCPIP-1 was detectable in the epithelium and in connective tissue, being especially prominent around the blood vessel walls in healthy periodontal tissues. MALT-1 was observed at all layers of gingival epithelium and especially around the accumulated inflammatory cells in connective tissue. No difference in gingival tissue MCPIP-1 and MALT-1 levels was observed in relation to the severity of gingival inflammation. MALT-1 levels were elevated (p = 0.023) with the increase in tissue P. gingivalis levels, and there was an association between MALT-1 and IL-8 levels (β = 0.054, p = 0.001)., Conclusions: Interactions of MALT-1 levels with gingival tissue P. gingivalis counts and IL-8 levels suggest that activation of MALT-1 can take part in P. gingivalis-regulated host immune responses., Clinical Relevance: Pharmacological targeting the crosstalk between immune response and MCPIP-1/MALT-1 may have benefits in periodontal treatment., (© 2023. The Author(s).)

Introduction: The oral pathogen Porphyromonas gingivalis is not only associated with periodontitis but also with systemic diseases elsewhere in the body. The mechanisms by which P. gingivalis travels from the oral cavity to other organs in the body are largely unknown. This review describes the four putative mechanisms supported by experimental evidence, which enable translocation of P. gingivalis over the oral mucosa, endothelial barriers and subsequent dissemination into the bloodstream., Mechanisms: The first mechanism: proteolytic enzymes secreted by P. gingivalis degrade adhesion molecules between tissue cells, and the extracellular matrix. This weakens the structural integrity of the mucosa and allows P . gingivalis to penetrate the tissue. The second is transcytosis: bacteria actively enter tissue cells and transfer to the next layer or the extracellular space. By travelling from cell to cell, P. gingivalis reaches deeper structures. Thirdly, professional phagocytes take up P. gingivalis and travel to the bloodstream where P. gingivalis is released. Lastly, P. gingivalis can adhere to the hyphae forming Candida albicans. These hyphae can penetrate the mucosal tissue, which may allow P. gingivalis to reach deeper structures., Conclusion: More research could elucidate targets to inhibit P. gingivalis dissemination and prevent the onset of various systemic diseases., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

This study evaluated the effect of phytosphingosine (PHS) and bioactive glass-ceramic (Biosilicate) on dental enamel in terms of color alteration (ΔE), microhardness, and surface roughness when submitted to erosive challenge (EC). Sixty specimens of bovine teeth (6×6×2mm) were obtained. Initial color (Easyshade, VITA), KHN (HMV-2, Shimadzu), and Ra (SJ-201P, Mitutoyo) measurements were performed. Specimens were separated into groups according to treatments: PHS, 10% Biosilicate, PHS+10% Biosilicate, and artificial saliva (control) and submitted to EC with Coca-Cola for 2 min. This cycle was repeated 4 times daily/15 days. Between cycles, specimens remained in artificial saliva (2 h/37°C). After daily cycles, they were also stored in artificial saliva at 37ºC. Final color, microhardness, and surface roughness measurements were done. Color and KHN data were analyzed by one-way ANOVA, Tukey's test; and Ra, by 2-way ANOVA, repeated measures, and Tukey's test (p<.05). The highest ΔE occurred in Saliva+EC (p<.05). Groups treated with PHS presented lower color change than Saliva+EC (p<.05). All the groups presented mean values above the 50:50% perceptibility (50:50%PT) and acceptability (50:50%AT) thresholds, except for control that showed mean value above 50:50%PT but below 50:50%AT. Biosilicate+EC showed higher relative microhardness than Saliva+EC (p<.05), but was similar to PHS+EC and PHS+Biosilicate+EC. Final enamel surface roughness increased for all the groups (p<.05), except for the control. The Biosilicate may prevent enamel mineral loss induced by erosion better than saliva. The PHS associated or not to Biosilicate demonstrated better color stability than saliva.

Objectives: MUC5B plays a multifactorial role in oral health. As a consequence, decreased MUC5B output leads to impaired salivary functions and xerostomia. Synthetic combinatorial technologies have been used to develop functional peptide libraries by phage display e.g. for therapeutic purposes. In this light, our primary aim was to identify peptide sequences with specific selectivity for salivary MUC5B in vitro using phage display. Our secondary aims were to analyze their effect on salivary spinnbarkeit in situ and their effect on acid-induced demineralization in vitro., Methods: MUC5B binding phages were selected by phage display. Peptide affinity to MUC5B was evaluated using MUC5B coated hydroxyapatite (HA) granules. The MUC5B binding peptides (MBPs) were then examined for their effects on salivary spinnbarkeit and protective effect on acid-induced demineralization in vitro. A competitive ELISA was performed to identify the binding epitope on MUC5B using F2, a MUC5B specific antibody., Results: MBP-12 and MBP-14 displayed the highest affinity to MUC5B. MBP-12 mildly stabilized the spinnbarkeit of serous saliva after overnight incubation and of mucous saliva at all timepoints tested. The addition of MBP-12 to a pellicle of unstimulated saliva on HA discs showed no additive protective effect against acid-induced demineralization. Epitope characterization suggested sulfo-Lewis a SO3-3Gal&#95;1-3GlcNAc (galactose residue) as MBP-12 binding site on MUC5B., Conclusions: The use of phage display in generating MBPs was successful. Characterization of the MBPs revealed a mild effect on spinnbarkeit in case of mucous saliva. Possibly, combinatorial peptide libraries might contribute to the development of novel formulations to treat xerostomia., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Food-derived carbon quantum dots (CQDs) can relatively easily be synthesized and chemically manipulated for a broad spectrum of biomedical applications. However, their toxicity may hinder their actual use. Here, Spinacia oleracea -derived CQDs i.e ., CQD-1 and CQD-2, were synthesized by means of different shredding methods and followed by a microwave-assisted hydrothermal approach. Subsequently, these CQDs were analyzed in vitro and in an in vivo mice model to test their biocompatibility and potential use as bioimaging agents and for activation of osteogenic differentiation. When comparing CQD-1 and CQD-2, it was found that CQD-1 exhibited 7.6 times higher photoluminescent (PL) emission intensity around 411 nm compared to CQD-2. Besides, it was found that the size distribution of CQD-1 was 2.05 ± 0.08 nm, compared with 2.14 ± 0.04 nm for CQD-2. Upon exposure to human bone marrow-derived mesenchymal stem cells (hBMSCs) in vitro , CQD-1 was endocytosed into the cytoplasm and significantly increased the differentiation of hBMSCs up to 10 μg mL -1 after 7 and 14 days. Apparently, the presence of relatively low doses of CQD-1 showed virtually no toxic or histological effects in the major organs in vivo . In contrast, high doses of CQD-1 (1 mg mL -1 ) caused cell death in vitro ranging from 35% on day 1 to 80% on day 3 post-exposure, and activated the apoptotic machinery and increased lymphocyte aggregates in the liver tissue. In conclusion, S. oleracea -derived CQDs have the potential for biomedical applications in bioimaging and activation of stem cells osteogenic differentiation. Therefore, it is postulated that CQD-1 from S. oleracea remains potential prospective material at appropriate doses and specifications., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors. Published by Elsevier Ltd.)

Objective: This study evaluated the in vitro and in situ effects of phytosphingosine (PHS) associated with tooth brushing on color stability, surface roughness, and microhardness of dental enamel., Methods: Sixty-four specimens of bovine teeth (6 × 6 × 2 mm) were separated into 8 groups (n = 8): S + TB: PHS (spray) + tooth brushing; TB + S: tooth brushing + PHS (spray); I + TB: PHS (immersion) + tooth brushing; TB + I: tooth brushing + PHS (immersion); TB: tooth brushing; S: PHS spray; I: immersion in PHS solution, and Saliva: immersion in saliva. Tooth brushing simulation (Mavtec, Brazil) was performed (356 rpm on 3.8 cm area by the toothbrush - Soft Tek) for 1, 7, 15, and 30 days. PHS remained in contact with specimens for 15 min. The specimens were evaluated before and after tooth brushing for color alteration (Easy Shade, VITA), and surface roughness (Model SJ-201P Mitutoyo), and Knoop microhardness (HMV-2, Shimadzu Corporation). For the in situ analyses, 8 participants were recruited and received an intraoral device with 6 fragments of bovine enamel (6 × 6 × 2 mm). The properties evaluated were the same as those of the in vitro study. Participants were randomized following best results of in vitro tested protocols, for 15 days: TB, TB + S, I + TB. Data obtained by in vitro (two-way ANOVA, Tukey, p < .05) and in situ (one-way ANOVA, Tukey, p < .05) studies were analyzed., Results: The in vitro study showed that greater color change was found after 30 days. The greatest differences in surface roughness occurred between the initial value and after 1 day. Regarding microhardness, the highest values occurred after 15 and 30 days, which showed similar results. The in situ study showed greater color changes for the TB and I + TB, and greater surface roughness changes for TB as well as a similar increase in microhardness for the PHS protocols, which were higher than TB., Conclusions: Phytosphingosine leads to an increase in performance regarding color stability, surface roughness, and microhardness when applied. In general, the application of PHS after brushing showed a positive impact on its performance., Clinical Relevance: Phytosphingosine proved to be interesting for compound prevention formulations in the dentistry field., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: In this study, we investigated the salivary film thickness and the MUC5B levels at various intra-oral locations in healthy volunteers, with a focus on the palate. Besides, measurements of the palatal surface area were included to explore the possible relationships between the palatal surface area and the palatal salivary film and MUC5B levels., Materials and Methods: The salivary film thickness was determined using filter strips, which were pressed to the mucosal surfaces of five different intra-oral locations; conductance was then analysed using a Periotron. After elution of the strips, the MUC5B levels at various intra-oral locations were determined using ELISA. The palatal surface area was measured using an intra-oral scanner. The surface area was subsequently calculated using the software., Results: The anterior tongue had the thickest salivary film and also the highest levels of MUC5B, while the anterior palate had the thinnest salivary film and lowest MUC5B levels. There was no association between the palatal surface area and the salivary film thickness of the palate., Conclusion: The salivary film and MUC5B levels are unequally distributed over the intra-oral regions of the soft tissues. The lack of association between the palatal surface area and the salivary film thickness indicates that a larger surface area is not associated with a relative thinner palatal salivary film., Clinical Relevance: The results of the current study increase our understanding of saliva distribution in the oral cavity and could be used as reference values for future studies., (© 2022. The Author(s).)

Background: The salivary glycoprotein MUC5B plays a versatile role in maintaining oral health. It contributes to lubrication, pellicle formation, antimicrobial defense, and water retention, and its glycans are an important nutrient for oral bacteria. This review aimed to describe the role of MUC5B in oral health and examine changes in its levels and composition in cases of hyposalivation and xerostomia., Highlight: In cases of hyposalivation, the reduction of total salivary MUC5B levels and MUC5B glycosylation patterns due to Sjögren's syndrome (SS) and medication intake appeared insignificantly limited. In patients with SS, xerostomia was related to reduced MUC5B levels at the anterior tongue. In cases of xerostomia, MUC5B glycosylation might be reduced, yet other factors such as total protein concentration, MUC7 levels and glycosylation, and salivary spinnbarkeit are involved. In contrast to SS- and medication-induced hyposalivation, radiotherapy in the head and neck region leads to a bona fide reduction in salivary MUC5B levels., Conclusion: Our findings suggest that MUC5B levels are clearly impaired in hyposalivation and xerostomia related to radiotherapy in the head and neck region versus those related to SS and medication intake. A reduction in glycosylation in the case of dry mouth appears associated with MUC5B and MUC7 as well as other factors., Competing Interests: Conflict of interest The authors declare no conflict of interest., (Copyright © 2022 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.)

Objectives: Sjögren's syndrome (SS) patients should be involved in the development of new saliva substitutes at an early stage. The purpose of the current study was to explore the preferences of these patients regarding various product characteristics of potential new saliva substitutes., Material and Method: A questionnaire was distributed among SS patients. They could anonymously indicate their preferences for saliva substitute characteristics using 5-point Likert scales., Results: Fifty-nine SS patients filled in the questionnaire. According to their opinion, the most ideal saliva substitute has a thin-watery consistency with a neutral flavour that should be applied as a spray. Patients demand a prolonged alleviation of dry mouth complaints and neutralization of harmful bacteria. The patients mainly object against the presence of artificial sweeteners and alcohol in saliva substitutes, but have limited objections against the presence of vegetable-based ingredients and natural enzymes. Major objections were against the potential side effects "bitter taste" and "discoloration of teeth". Age and severity of xerostomia affected desire of flavours. Younger patients preferred menthol flavour, while respondents with severe xerostomia preferred the use of "neutral flavours" significantly more., Conclusion: The most ideal saliva substitute has thin-watery consistency in spray form with a neutral flavour and providing long alleviation of dry mouth complaints. Besides, it should not contain artificial sweeteners or alcohol, and should not have a bitter taste or cause discoloration of the teeth., Clinical Relevance: Investigating the opinion of SS patients provides tailoured insights into their preference, which may contribute to the development of more effective saliva substitutes., (© 2022. The Author(s).)

The fluorescent carbon dots (CDs) have found their extensive applications in sensing, bioimaging, and photoelectronic devices. In general terms, the synthesis of CDs is straight-forward, though their subsequent purification can be laborious. Therefore, there is a need for easier ways to generate solid CDs with a high conversion yield. Herein, we used collagen waste as a carbon source in producing solid CDs through a calcination procedure without additional chemical decomposition treatment of the raw material. Considering a mass of acid has destroyed the original protein macromolecules into the assembled structure with amino acids and peptide chains in the commercial extraction procedure of collagen product. The residual tissues were assembled with weak intermolecular interactions, which would easily undergo dehydration, polymerization, and carbonization during the heat treatment to produce solid CDs directly. The calcination parameters were surveyed to give the highest conversion yield at 78%, which occurred at 300°C for 2 h. N and S atomic doping CDs (N-CDs and S-CDs) were synthesized at a similar process except for immersion of the collagen waste in sulfuric acid or nitric acid in advance. Further experiments suggested the prepared CDs can serve as an excellent sensor platform for Fe 3+ in an acid medium with high anti-interference. The cytotoxicity assays confirmed the biosafety and biocompatibility of the CDs, suggesting potential applications in bioimaging. This work provides a new avenue for preparing solid CDs with high conversion yield., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The handling editor HM declared a shared affiliation with the author DL at the time of review., (Copyright © 2022 Qin, Fu, Zhang, Shao, Qin, Gui, Wang, Guo, Chen, Jiang, Wu, Bikker and Luo.)

Purpose: Saliva distribution over the palatal surface plays an important role in the perception of dry mouth. It is envisaged that non-invasive estimation of the palatal surface area by anthropometric measurements of the head and face can be useful in the assessment of oral dryness. For this purpose, the relationship between the palatal surface area and anthropometric measurements of the head and face was investigated., Methods: The palatal surface was measured in 51 healthy volunteers using an intra-oral scanner. The distances between anthropometric landmarks of the head and face were determined using an anatomical sliding caliper. Correlations between the palatal surface area and the anthropometric landmarks were investigated., Results: The median palatal surface area for the total study population was found to be 2120.6 mm 2 . Virtually, all anthropometric measurements showed significant differences between females and males. Various head and face measurements had a significant correlation with the palatal surface area. However, these correlations disappeared when the participants were stratified based on their sex, with the exception of mandibular length and palatal width in females., Conclusion: The surface area of the palate correlates with nearly all anthropometric measurements of the head and face included in this study. Yet, the clinical applicability seems limited to females., (© 2022. The Author(s).)

Human salivary histatin 1 (Hst1) exhibits a series of cell-activating properties, such as promoting cell spreading, migration, and metabolic activity. We recently have shown that fluorescently labeled Hst1 (F-Hst1) targets and activates mitochondria, presenting an important molecular mechanism. However, its regulating signaling pathways remain to be elucidated. We investigated the influence of specific inhibitors of G protein-coupled receptors (GPCR), endocytosis pathways, extracellular signal-regulated kinases 1/2 (ERK1/2) signaling, p38 signaling, mitochondrial respiration and Na+/K+-ATPase activity on the uptake, mitochondria-targeting and -activating properties of F-Hst1. We performed a siRNA knockdown (KD) to assess the effect of Sigma-2 receptor (S2R) /Transmembrane Protein 97 (TMEM97)-a recently identified target protein of Hst1. We also adopted live cell imaging to monitor the whole intracellular trafficking process of F-Hst1. Our results showed that the inhibition of cellular respiration hindered the internalization of F-Hst1. The inhibitors of GPCR, ERK1/2, phagocytosis, and clathrin-mediated endocytosis (CME) as well as siRNA KD of S2R/TMEM97 significantly reduced the uptake, which was accompanied by the nullification of the promoting effect of F-Hst1 on cell metabolic activity. Only the inhibitor of CME and KD of S2R/TMEM97 significantly compromised the mitochondria-targeting of Hst1. We further showed the intracellular trafficking and targeting process of F-Hst1, in which early endosome plays an important role. Overall, phagocytosis, CME, GPCR, ERK signaling, and S2R/TMEM97 are involved in the internalization of Hst1, while only CME and S2R/TMEM97 are critical for its subcellular targeting. The inhibition of either internalization or mitochondria-targeting of Hst1 could significantly compromise its mitochondria-activating property., (© 2022. The Author(s).)

Acanthamoeba castellanii ( A. castellanii ) can cause Acanthamoeba keratitis, a sight-threatening infection, as well as a fatal brain infection termed granulomatous amoebic encephalitis, mostly in immunocompromised individuals. In contrast, Naegleria fowleri ( N. fowleri ) causes a deadly infection involving the central nervous system, recognized as primary amoebic encephalitis, mainly in individuals partaking in recreational water activities or those with nasal exposure to contaminated water. Worryingly, mortality rates due to these infections are more than 90%, suggesting the need to find alternative therapies. In this study, antiamoebic activity of a peptide based on the structure of the antibiotic tyrocidine was evaluated against A. castellanii and N. fowleri . The tyrocidine-derived peptide displayed significant amoebicidal efficacy against A. castellanii and N. fowleri . At 250 μg/mL, the peptide drastically reduced amoebae viability up to 13% and 21% after 2 h of incubation against N. fowleri and A. castellanii. , whereas, after 24 h of incubation, the peptide showed 86% and 94% amoebicidal activity against A. castellanii and N. fowleri . Furthermore, amoebae pretreated with 100 μg/mL peptide inhibited 35% and 53% A. castellanii and N. fowleri , while, at 250 μg/mL, 84% and 94% A. castellanii and N. fowleri failed to adhere to human cells. Amoeba-mediated cell cytopathogenicity assays revealed 31% and 42% inhibition at 100 μg/mL, while at 250 μg/mL 75% and 86% A. castellanii and N. fowleri were inhibited. Assays revealed inhibition of encystation in both A. castellanii (58% and 93%) and N. fowleri (73% and 97%) at concentrations of 100 and 250 μg/mL respectively. Importantly, tyrocidine-derived peptide depicted minimal cytotoxicity to human cells and, thus, may be a potential candidate in the rational development of a treatment regimen against free-living amoebae infections. Future studies are necessary to elucidate the in vivo effects of tyrocidine-derived peptide against these and other pathogenic amoebae of importance., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

Previously we described the discovery of a Bacillus spp. specific peptidase activity related to d-stereospecific peptidases (DSPs). The peptidase showed a strong preference for d-leucine and d-valine amino acids. These amino acids are present in the structure of the non-ribosomal peptide (NRP) antibiotics gramicidin A, B and C and polymyxin E. To examine if the Bacillus spp. DSP-related peptidase can hydrolyze these NRPs, the effect of gramicidin A and C and polymyxin E on peptidase activity in Bacillus anthracis culture supernatant was monitored. It was found that both gramicidins inhibited the DSP-related activity in a competitive manner. MALDI-TOF analysis revealed that upon incubation with B. anthracis culture supernatant gramicidin A hydrolyzation products appeared. This study shows that the Bacillus spp. specific DSP-like peptidase was potentially produced by the bacteria to gain intrinsic resistance against NRP antibiotics. These results are of utmost importance in research towards antimicrobial resistance, whereas transfer of DSP-related activity to other clinically relevant pathogens can be a serious threat to human health., (© 2022 The Authors. Environmental Microbiology Reports published by Society for Applied Microbiology and John Wiley & Sons Ltd.)