Search Results (156)

This study compared the remineralization potential of three commercially available fluoride-free biomimetic formulations containing nanohydroxyapatite (nHAp), casein phosphopeptide–amorphous calcium phosphate (CPP-ACP), and bioactive glass (BAG) with a conventional 1450 ppm fluoride-containing dentifrice using cross-sectional microhardness and SEM analyses. Seventy extracted human mandibular molars were randomly allocated to five groups (n = 14). Artificial carious lesions were created by 72 h of demineralization followed by a 14-day pH-cycling regimen with twice-daily treatment applications. Cross-sectional microhardness was measured at depths of 30, 50, 100, and 150 µm, and SEM was used to evaluate morphological changes. Data were analyzed using one-way ANOVA with appropriate post hoc tests (Tukey’s HSD or Tamhane’s T2, depending on variance homogeneity) and repeated-measures ANOVA to assess depth-dependent differences among groups (α = 0.05). Demineralization significantly reduced microhardness at all depths (p < 0.05). At 30 µm, the nHAp, CPP-ACP, and fluoride formulations showed similar remineralization, whereas the BAG-containing formulation exhibited lower superficial recovery. At 100–150 µm, the nHAp formulation demonstrated the greatest recovery, followed by the CPP-ACP and BAG formulations, while the fluoride dentifrice showed progressively lower recovery. A significant depth-by-group interaction confirmed distinct depth-dependent remineralization profiles. Within the limitations of this in vitro study, all tested formulations promoted enamel remineralization but exhibited different depth-dependent recovery patterns. Full article

Background: Patients with White spot lesions (WSLs) are seeking minimally invasive remineralizing treatment. While fluoride is the gold standard of non-invasive caries therapy, its remineralizing impact is predominantly superficial. MI Paste One (MIP-1) combines casein phosphopeptide–amorphous calcium phosphate (CPP–ACP) with 1100 ppm fluoride. This in vitro study aimed to assess the effect of MIP-1 compared with other agents. Methods: Enamel slabs were demineralized to create a caries-like lesion and randomized into five groups (n = 10): G1, WSL/artificial saliva (AS) (negative control); G2, deionized water brushing; G3, fluoridated toothpaste brushing; G4, MIP-1 brushing; G5, topical MI Paste application. Color change (ΔE), gloss (GU), microhardness (VHN), roughness (Ra), and surface topography (SEM) were assessed at four time points: baseline, first demineralization, treatment, and second demineralization. Results: All active agents improved microhardness after treatment. After secondary demineralization, G4 achieved the highest hardness, significantly superior to G3 (p = 0.016). G5 occupied a statistically intermediate position. CPP–ACP-containing agents (G4, G5) achieved color improvement that was significantly superior to controls. Conclusions: MIP-1 outperformed fluoride toothpaste in resisting secondary demineralization of permanent enamel WSLs. CPP–ACP made a measurable independent contribution to demineralization resistance in permanent enamel. These findings highlight the need for further clinical investigation of MIP-1. Full article

Dental caries remains the most prevalent chronic disease worldwide, yet early enamel lesions are reversible if managed with appropriate remineralizing agents. This narrative review synthesizes current evidence on remineralizing agents, their mechanisms of action, and clinical applications, with a focus on dental materials used in preventive and minimally invasive dentistry. Traditional fluoride-based approaches enhance remineralization through fluorapatite formation; however, their effectiveness is limited when calcium and phosphate bioavailability is insufficient. Biomimetic agents, including casein phosphopeptide–amorphous calcium phosphate (CPP-ACP), bioactive glasses, tricalcium phosphate, and nano-hydroxyapatite, provide these bioavailable ions and demonstrate superior performance under challenging clinical conditions. Emerging therapies such as probiotics, photodynamic therapy, and laser-assisted mineralization show promise but require further clinical validation. Based on the primary mechanism of action, an original classification of remineralizing agents is proposed, grouping them into fluoride-based agents, calcium-phosphate systems, nanotechnology-based systems, biofilm modifiers, biomimetic and emerging systems, and adjunctive antimicrobial therapies. The review concludes that bioavailable calcium represents a critical limiting factor in remineralization under certain conditions, and that combination protocols incorporating multiple remineralizing agents, tailored to individual patient risk profiles, achieve superior outcomes compared to single-agent approaches. Clinicians are encouraged to adopt minimally invasive, patient-tailored remineralization strategies that arrest lesions before cavitation, preserving natural tooth structure and reducing the lifelong restorative burden. Full article

Background: The management of white spot caries lesions (WSCLs) presents a clinical challenge due to a lack of standardized protocols. This study aimed to evaluate the knowledge, attitudes, and practices of dental practitioners in Russia regarding the diagnosis and treatment of WSCLs. Methods: A cross-sectional online survey was conducted from October 2024 to July 2025 among 397 dental professionals in Russia. A validated questionnaire assessed four domains: demographics, knowledge, attitudes, and clinical practices concerning WSCL management. Data were analyzed using descriptive statistics, Kruskal–Wallis tests, and Spearman’s correlation. Results: The majority of respondents were females (83%), with over half having less than five years of experience (55%). A majority of the practitioners (62.5%) demonstrated a good level of knowledge (mean score 12.8 ± 2.2). Knowledge was significantly better among female practitioners (p < 0.001) and inversely correlated with years of clinical experience (p < 0.001). Attitudes towards minimally invasive treatment were mainly positive (mean score 13.5 ± 2.6), with 86% of respondents showing a positive score (>10 points). While awareness of minimally invasive techniques like resin infiltration (84%) and remineralization (79.1%) was high, clinical practice varied. Remineralization was the most frequently used treatment (68% used casein phosphopeptide–amorphous calcium phosphate and 62% used fluoride), whereas resin infiltration was used by 46% and microabrasion by only 5% of the respondents. A majority of dentists (52%) preferred monitoring lesions rather than immediate intervention. Conclusions: Russian dental practitioners possess good theoretical knowledge and positive attitudes toward contemporary, minimally invasive management of WSCLs. However, a significant gap exists between knowledge and clinical practice, particularly concerning the use of resin infiltration and microabrasion. These findings underscore the need for continuous education on the topic. Full article

Background: Pathogenic germline BRCA1 and BRCA2 variants cause most hereditary breast and ovarian cancers. Widespread genetic testing has revealed thousands of variants with unknown effects on disease risk, known as variants of uncertain significance (VUS). BRCA VUS, the majority of which are missense, complicate genetic test interpretation and clinical decision-making. This study aimed to evaluate BRCA1 VUS pathogenicity with enhanced accuracy through computational, functional and structural methods. Methods: We characterized the structural distribution of BRCA1 variants. In silico tools scored known consequence variants within a specific region of BRCA1. The Molecular Feature Selection Tool (MolecularFeaST; Renwick Lab at Queen’s University; Kingston, ON, Canada) performed feature selection of the most discriminative tools. MATLAB (MATLAB R2024a; Mathworks; Natick, MA, USA) Classification Learner Application trained supervised machine learning models using combinations of the most accurate tools; the best model assigned pathogenicity prediction scores to VUS. Select VUS were functionally assessed through phosphopeptide binding pull-down assays and structurally analyzed on PyMOL (v2.4.1; Schrödinger Inc.; New York, NY, USA). Results: The RING and BRCT domains were identified as hotspots for missense pathogenic variants and VUS; BRCT was selected as the focus of the computational classifier. Nine in silico tools (CADD hg19, MetaRNN, ClinPred, VEST4, BayesDel AD, EVE, Eigen PC, gMVP and PolyPhen2) defined the BRCT-specific missense variant classifier. Twenty-two VUS (R1699P, F1704S, W1837L, W1712G, F1734S, V1804A, I1674V, V1804L, V1804I, I1807V, T1675S, I1764L, N1774I, E1698K, Q1848K, P1749S, A1669T, N1774H, L1839V, T1658I, L1705I, V1654L) demonstrated varying phosphopeptide binding ability and protein levels relative to the wildtype. Computational structural modeling contextualized VUS phosphopeptide interactions and structural implications. Conclusions: We provide in silico and functional evidence for the classification of BRCA1 BRCT VUS and highlight the utility of domain-specific computational approaches for characterizing missense variants in multi-domain genes. Full article

Background: Maize is a vital crop, supporting 19.5% of global calorie intake. However, maize is vulnerable to even brief periods of drought which substantially reduces seed set and therefore yield. Methods: To identify proteins involved in responses of maize to drought, soluble proteins were extracted from leaf and silk tissues of Zea mays and protein abundance and phosphorylation status were quantified relative to well-watered controls. Label-free quantification and phosphopeptide enrichment were applied to the same biological samples and over 300 proteins were identified with significantly different changes. Results: Proteins known to be involved in drought responses were identified, such as the abscisic acid pathway and transcription factors. Of particular interest is a group of dehydrins quantified at both total protein and phosphopeptide levels, permitting insight into stoichiometry. The biological function of dehydrins in the model plant Arabidopsis thaliana is known to be regulated by phosphorylation. Conclusions: Translation of protein function from model plant to crops remains highly challenging because genome duplication has created complex sets of orthologous and homologous proteins. By focusing on proteomic changes during crop stress responses, this work enables the identification of known and novel proteins, substantially aiding the transfer of knowledge from model plants to crops. Full article

Muscle fibers exhibit high plasticity: both the fast-twitch fiber type and slow-twitch fiber type can mutually transform under the regulation of phosphorylation. In this study, we characterized the muscle fiber profiles and phosphoproteomes of the m. extensor digitorum longus (EDL) and m. soleus (SOL) in slow-growing Xueshan chickens and fast-growing Ross 308 broilers. Fiber-type distribution was quantified by immunohistochemistry and RT-qPCR of MYH7B, MYH1A and MYH1B. TMT-based phosphoproteomics, combined with bioinformatic analysis, was used to identify differentially expressed phosphopeptides (DEPs) in two comparisons: Ross 308 SOL vs. Ross 308 EDL, and Xueshan EDL vs. Ross 308 EDL. The proportion of type I (slow-twitch) fibers in EDL was significantly higher in Xueshan chickens than in Ross 308 broilers (15.53% vs. 6.14%, p < 0.05), with no significant differences in fiber distribution or diameter between the SOL and EDL in Xueshan chickens (p > 0.05). A total of 3226 phosphopeptides corresponding to 1762 phosphoproteins were identified, with serine as the most abundant phosphorylated amino acid (73.92%). PDHA1, PHKB and PGAM1 were identified as the key DEPs common to both comparison groups. Bioinformatic analyses revealed that reversible site-specific phosphorylation regulates avian muscle fiber-type transformation mainly via the glycolysis/gluconeogenesis pathway. Full article

Abscisic acid (ABA) is a major phytohormone regulating plant growth and stress responses. Subclass III SnRK2 kinases and clade A type 2C protein phosphatases (PP2Cs) are core components of ABA signaling. Despite advances from phosphoproteomics, major gaps remain, particularly in mapping PP2C dephosphorylation targets and SnRK2-dependent phosphorylation dynamics under non-stress conditions. Here, we performed large-scale LC–MS/MS phosphoproteomic analyses using the subclass III SnRK2 triple mutant srk2dei and the constitutively active PP2C mutant abi1–1C, with and without ABA treatment in Arabidopsis thaliana. We identified 2757 and 2886 differentially regulated phosphopeptides in srk2dei and abi1–1C, respectively. Beyond known ABA signaling components, these datasets revealed numerous previously uncharacterized candidate proteins involved in metabolism, membrane transport, transcription, and cytoskeletal regulation. Integrative analysis uncovered a core set of candidate proteins oppositely regulated by SnRK2-mediated phosphorylation and ABI1-mediated dephosphorylation, defining a coordinated hierarchical network. These results indicate that the SnRK2–PP2C module functions not only in stress-induced ABA responses but also as a central regulator of phosphorylation homeostasis under basal conditions. This study provides a systematic framework for the global SnRK2–PP2C phosphorylation network and reframes ABA signaling as a dynamic homeostatic system. Full article

Energy gels are widely used by athletes to maintain performance during endurance activities; however, their acidic composition may pose a risk to dental health. This study aimed to evaluate the pH of four commercial energy gels at different dilutions with artificial saliva and to assess the potential neutralizing effect of casein phosphopeptide–amorphous calcium phosphate (CPP–ACP). An in vitro experimental design was conducted using four commercial gels (FullGas Hydrogel, ENA Energy Gel, UltraTech Gel, and Maverick Race Gel). Serial dilutions with artificial saliva (1:2, 1:5, 1:7, and 1:10) were prepared, and pH was measured using indicator strips at 0, 15, and 30 min at 37 °C. The effect of CPP-ACP was evaluated in the 1:2 dilution. Undiluted gels showed highly acidic pH values ranging from 2.0 to 3.0. Dilutions of 1:2 and 1:5 remained significantly more acidic than artificial saliva (p < 0.001). From 1:7 dilution onward, pH values increased and approached salivary levels (approximately 7.0), with no significant differences compared with artificial saliva. The addition of CPP-ACP significantly increased pH in the 1:2 dilution (p < 0.05), although the effect was limited in more diluted conditions. These findings suggest that commercial energy gels may represent a source of acidic exposure under in vitro conditions, which could be relevant for dental health. Adequate dilution, particularly ≥1:7, was associated with a reduction in acidity under the experimental conditions tested, although its clinical relevance cannot be directly inferred, while CPP-ACP may provide a limited buffering effect under concentrated exposure conditions. Full article

Pyruvate kinase M2 (PKM2) influences meat quality through glycolysis and also exhibits its moonlighting function as a protein kinase that catalyzes protein phosphorylation. However, it remains unclear whether PKM2 phosphorylates myofibrillar proteins, thereby affecting postmortem myofibrillar protein stability. This study investigates PKM2’s non-canonical kinase function using quantitative phosphoproteomics and an in vitro myofibrillar protein incubation model to identify its phosphorylation substrates and functional impacts. The quantitative phosphoproteomics identified 441 phosphoproteins, 881 phosphopeptides, and 1040 phosphorylation sites. Notably, the myosin regulatory light chain (MRLC) was identified as a likely candidate phosphorylation substrate of PKM2 in vitro. The interaction between PKM2 and MRLC was confirmed using co-immunoprecipitation (Co-IP) and Western blotting. Furthermore, MRLC phosphorylation by PKM2 significantly inhibited its degradation and enhanced its stability. This work establishes an in vitro biochemical framework for the moonlighting role of glycolytic enzymes, suggesting a potential mechanistic pathway that might influence myofibrillar protein stability during meat aging. Full article

The management of dental caries has evolved from the traditional mechanical approach of “extension for prevention” to a biologically oriented philosophy centered on preserving natural tooth structures. Minimally invasive dentistry (MID) emphasizes early detection, risk assessment, prevention, and conservative intervention based on the lesion’s activity and depth. This review outlines current evidence on non-operative, micro-invasive, and minimally invasive strategies, including fluoride therapy, remineralizing agents such as casein phosphopeptide–amorphous calcium phosphate (CPP-ACP), self-assembling peptides that promote biomimetic enamel repair, sealants, and resin infiltration. Minimally invasive operative methods employ advanced technologies for selective tissue removal—chemomechanical systems (Carisolv, Papacarie, Brix3000), sono-and airabrasion, and new-generation polymeric and ceramic burs (SmartBur, Cerabur) designed to preserve sound dentin. Laser photoablation, particularly with erbium lasers (Er:YAG, Er,Cr:YSGG), enables precise cavity preparation with minimal thermal and mechanical stress. These approaches enhance patient comfort, reduce anesthesia requirements, and maintain tooth vitality. Despite limitations related to cost, equipment, and operator sensitivity, MID represents not only a set of refined clinical techniques but also a comprehensive, evidence-based treatment philosophy founded on biological principles, structural preservation, and the promotion of long-term oral health. Full article

Dental erosion has emerged as a significant modern oral health problem, characterized by the chemical dissolution of tooth structure resulting from frequent exposure to intrinsic or extrinsic acids. With a high global prevalence ranging from 30% to 50% in children and 20% to 40% in adults, its management is a clinical priority to prevent long-term complications like dentine hypersensitivity and functional impairment. This review outlines the multifactorial etiology of erosion, encompassing dietary acids, gastroesophageal reflux, and reduced salivary flow. The historical context of oral care is explored, leading to a discussion on contemporary management strategies centered on remineralization. Fluoride ions play a crucial role by inhibiting demineralization, facilitating the formation of acid-resistant fluorapatite, and exerting antibacterial effects. A major focus is placed on advanced biomimetic, calcium phosphate-based topical agents such as Casein Phosphopeptide–Amorphous Calcium Phosphate (CPP-ACP), functionalized Tricalcium Phosphate (fTCP), and Hydroxyapatite (HAP), which effectively replenish lost minerals. The review further explores innovative methods, such as laser-assisted and electrically enhanced remineralization. Finally, it outlines next-generation regenerative strategies, including self-assembling peptides (P11-4), stem cell therapies, 3D bioprinting, and gene-editing (CRISPR) technologies, which aim to biologically regenerate lost enamel and dentine. The field is rapidly evolving from a preventive to a restorative paradigm, with future directions focusing on biologically based, minimally invasive therapies to fully restore tooth structure and function. Full article

Background: Cleft palate (CP) is a prevalent craniofacial malformation, with the TGFβ pathway playing a critical role. Recent evidence links autophagy to regulating mouse embryonic palatal mesenchyme (MEPM) cells, but its interaction with TGFβ-activated phosphorylation cascades remains largely unknown. Here, we investigated the interplay between these pathways during palatogenesis. Methods: H&E and IHC analyses revealed increased expression of Beclin 1 and LC3 during the critical period of palatal shelf elevation and fusion (E13.5–E15.5). Bulk RNA sequencing (Bulk RNA-seq) further revealed enrichment of autophagy-related pathways and their interaction with TGFβ signaling. TMT-based phosphoproteomics was performed on TGFβ2-treated MEPM cells. Results: We identified 23,471 peptides and 3952 proteins, including 6339 phosphopeptides corresponding to 2195 phosphoproteins. Differential analysis found 477 phosphopeptides with increased abundance and 53 with decreased abundance, revealing the enrichment of seven serine (p-Ser) motifs (RxxS, SDxD, SDxE, SP, SxDE, SxEE, SxxxxD) and one threonine (p-Thr) motif (TP). Notably, kinase-substrate enrichment analysis identified CSNK2A as a previously unrecognized phosphorylation regulator, together with MAPKs and CDKs. Functional enrichment showed significant involvement of mTOR, MAPK, and autophagy/mitophagy pathways. Conclusions: Our findings revealed that TGFβ2 reshapes the MEPM phosphoproteome through Smad-independent pathway, expanding the palate-specific phospho-signaling atlas beyond the canonical Smad cascade. Full article

Background: Abscisic acid (ABA) is a key phytohormone that regulates plant growth and stress responses through protein phosphorylation. While ABA-induced phosphosignaling has been extensively studied in Arabidopsis thaliana, its conservation and divergence across plant species remain unclear. Methods: Here, we performed phosphoproteomic analysis using LC-MS/MS in Arabidopsis, rice (Oryza sativa), and soybean (Glycine max) to compare ABA-responsive phosphorylation profiles among monocots, dicots, and legumes. Results: ABA treatments on Arabidopsis, rice, and soybean seedlings yielded approximately 24,604, 18,865, and 24,930 phosphopeptides, respectively. Comparative analyses revealed both conserved and species-specific ABA-responsive phosphoproteins. Conclusions: This work provides insights into the evolutionary diversification of ABA signaling and its potential applications in improving crop stress tolerance. Full article

This study aimed to develop bioactive toothpaste and compare its remineralization potential on initial enamel lesions with toothpaste containing other active agents. Sixty extracted human maxillary incisors were randomly assigned to six groups: Group EXP (Experimental toothpaste), Group SRP (Sensodyne Repair & Protect), Group ZAC (Zubio Active Carbon Whitening), Group GTM (GC Tooth Mousse), Group CSP (Colgate Sensitive Pro-Relief), and Group ASS (Artificial saliva, control). Artificial caries were induced by immersion in a demineralization solution for three days. Specimens then underwent a seven-day pH-cycling protocol, during which toothpaste was applied twice daily for two minutes. Analyses were performed at baseline, post-demineralization, and post-remineralization using ATR-FTIR, SEM-EDS, and Vickers micro-hardness testing. Statistical analyses were conducted using SPSS (version 27.0, IBM Corp., Chicago, IL, USA). All treatment groups, except the control, showed significant microhardness recovery after remineralization, with the highest increase in group CSP followed by group EXP (p < 0.05). Granular surface deposits were observed, most pronounced in groups SRP and GTM (p < 0.05). Calcium and phosphorus contents increased in all groups (p < 0.05), with calcium highest in group GTM and phosphorus in group EXP. The mineral-to-matrix ratio increased in all groups, and a statistically significant difference was identified between the experimental toothpaste (EXP) and the other toothpaste formulations (p < 0.05). It is hypothesized that pomegranate seed essential oil may exhibit a remineralizing effect due to its content of anthocyanidins, anthocyanins, and various polyphenolic compounds. Therefore, the development of a toothpaste with enhanced remineralization potential was targeted by incorporating pomegranate seed essential oil into the experimental formulation in addition to bioactive agents such as bioactive glass, hydroxyapatite, and casein phosphopeptide. Full article