A study on β\beta-defensin-2 and histatin-5 as a diagnostic marker of early childhood caries progression

Background: Recently, a continuous growth of interest has been observed in antimicrobial peptides (AMPs) in the light of an alarming increase in resistance of bacteria and fungi against antibiotics. AMPs are used as biomarkers in diagnosis and monitoring of oral cavity pathologies. Therefore, the determination of specific protein profiles in children diagnosed with early childhood caries (ECC) might be a basis for effective screening tests and specialized examinations which may enable progression of disease. Methods: The objective of the studies was to determine the role of histatin-5 and $\beta$-defensing-2 as a diagnostic marker of early childhood caries progression. In this work, results of concentration determination of two salivary proteins (histatin-5 and $\beta$-defensin-2) were presented. In addition, bacterial profiles from dental plaque in various stages of ECC and control were marked. The assessment of alteration in the concentration of these two proteins in a study group of children with various stages of ECC and a control group consisting of children with no symptoms was performed by enzyme-linked immunosorbent assays. Results: The statistical analysis showed a significant increase in the concentration of histatin-5 and $\beta$-defensin-2 in the study group compared to the control group and correlated with the progression of the disease. Conclusions: The confirmation of concentration changes in these proteins during the progression of dental caries may discover valuable disease progression biomarkers

Methods of biotyping of Streptococcus mutans : species with the routine test as a prognostic value in early childhood caries

Purpose. In order to investigate the suitability of Streptococcus mutans species biotyping by measuring the activity of selected enzymes from a commercial test, criteria were established for biotyping clinical strains from children with dental caries. In addition, the relationships between the selected biotypes, sensitivity to commonly used antibiotics, and early childhood caries were determined. Methods. A total of 142 S. mutans isolates from dental plaque of children with caries were divided into different biotypes. Patients were divided into two groups: noncavitated (1-2 in ICDAS) and cavitated (5-6 in ICDAS) lesions. Biotyping criteria were determined based on both the arbitrary method and the clusterization method. The susceptibility of the strains to amoxicillin, cefazolin, erythromycin, and teicoplanin was studied by diluting a solid medium. Results. Biotype I was the most common. Mean MIC values showed that the strains belonging to biotypes II and IV were the most sensitive to amoxicillin. For predetermined biotypes, observed differences were dependent on the severity of dental caries. Conclusions. The proposed method of S. mutans strains biotyping is relatively quick and simple to use, provided the application of suitable biotyping criteria, and may contribute to the effective prevention of dental caries induced by S. mutans

Relationship between Pyruvate Kinase Activity and Cariogenic Biofilm Formation in Streptococcus mutans Biotypes in Caries Patients

Streptococcus mutans (MS) and its biotype I are the strains most frequently found in dental plaque of young children. Our results indicate that in children pyruvate kinase (PK) activity increases significantly in dental plaque, and this corresponds with caries progression. The MS strains isolated in this study or their main glycolytic metabolism connected with PK enzymes might be useful risk factors for studying the pathogenesis and target points of novel therapies for dental caries. The relationship between PK activity, cariogenic biofilm formation and selected biotypes occurrence was studied. S. mutans dental plaque samples were collected from supragingival plaque of individual deciduous molars in 143 subjects. PK activity was measured at different time points during biofilm formation. Patients were divided into two groups: initial stage decay, and extensive decay. Non-parametric analysis of variance and analysis of covariance were used to determine the connections between S. mutans levels, PK activity and dental caries biotypes. A total of 143 strains were derived from subjects with caries. Biotyping data showed that 62, 23, 50, and 8 strains were classified as biotypes I, II, III, IV, respectively. PK activity in biotypes I, II, and IV was significantly higher in comparison to that in biotype III. The correlation between the level of S. mutans in dental plaque and PK activity was both statistically significant (p < 0.05) and positive. The greater the level of S. mutans in the biofilm (colony count and total biomass), the higher the PK activity; similarly, a low bacterial count correlated with low PK activity

Effect of a Lactobacillus salivarius probiotic on a double-species Streptococcus mutans and Candida albicans caries biofilm

The aim of the study was to evaluate the anti-cariogenic effects of Lactobacillus salivarius by reducing pathogenic species and biofilm mass in a double-species biofilm model. Coexistence of S. mutans with C. albicans can cause dental caries progression or recurrence of the disease in the future. Fifty-nine children with diagnosed early childhood caries (ECC) were recruited onto the study. The condition of the children’s dentition was defined according to the World Health Organization guidelines. The participants were divided into children with initial enamel demineralization and children showing dentin damage. The study was performed on the S. mutans and C. albicans clinical strains, isolated from dental plaque of patients with ECC. The effect of a probiotic containing Lactobacillus salivarius on the ability of S. mutans and C. albicans to produce a double-species biofilm was investigated in an in vitro model. The biomass of the formed/non-degraded biofilm was analyzed on the basis of its crystal violet staining. The number of colonies of S. mutans and C. albicans (CFU/mL, colony forming units/mL) forming the biofilm was determined. Microorganism morphology in the biofilm was evaluated using a scanning electron microscope (SEM). In vitro analysis demonstrated that the presence of S. mutans increased the number of C. albicans colonies (CFU/mL); the double-species biofilm mass and hyphal forms produced in it by the yeast. L. salivarius inhibited the cariogenic biofilm formation of C. albicans and S. mutans. Under the influence of the probiotic; the biofilm mass and the number of S. mutans; C. albicans and S. mutans with C. albicans colonies in the biofilm was decreased. Moreover; it can be noted that after the addition of the probiotic; fungi did not form hyphae or germ tubes of pathogenic potential. These results suggest that L. salivarius can secrete intermediates capable of inhibiting the formation of cariogenic S. mutans and C. albicans biofilm; and may inhibit fungal morphological transformation and thereby reduce the pathogenicity of C. albicans; weakening its pathogenic potential. Further research is required to prove or disprove the long-term effects of the preparation and to achieve preventive methods

Sperm antioxidant biomarkers and their correlation with clinical condition and lifestyle with regard to male reproductive potential

Measurement of sperm oxidative-antioxidant indicators is widely used in the assessment and detection of biochemical causes of male infertility. The main purpose of this study was to identify biomarkers that assist in diagnostics and monitoring of male reproductive potential. We performed the assessment of oxidative-antioxidant malondialdehyde (MDA), glutathione (GSH), and total redox antioxidant potential (TRAP) indicators in seminal plasma, seminogram, clinical condition, and lifestyle of people with reproductive problems. The combined assessment of GSH and TRAP as potential biomarkers of male infertility in semen plasma was characterized by the highest total sensitivity and specificity. Furthermore, we provide evidence that male reproductive potential is significantly correlated with basic sperm parameters, sperm cell membrane integrity, their morphology, lifestyle, eating habits, occupation, and mental health. Our results provide evidence on the importance of oxidative stress and defense against free radicals in diagnosing and monitoring men with infertility that are consistent with previously conducted research. We provide an alternative approach on the possibility of interpreting the combination of the biomarkers that can bring benefits to a multi-threaded approach to the diagnosis and treatment of male infertility

How much of the predisposition to Hashimoto's thyroiditis can be explained based on previously reported associations?

Purpose Our insight in the genetics of Hashimoto’s thyroiditis (HT) has become clearer through information provided by genome-wide association studies and candidate gene studies, but remains still not fully understood. Our aim was to assess how many different genetic risk variants contribute to the development of HT. Methods 147 HT cases (10.2% men) and 147 controls (13.6% men) were qualified for the analysis. Intrinsic and environmental factors were controlled for. Polymorphisms (SNP) were chosen based on the literature and included markers of the genes PTPN22, CTLA4, TG, TPO among others, and of genomic regions pointed by GWAS studies. SNP were typed on a microarray. Variants in the HLA-DRB1 gene were identified by Sanger sequencing. Results Multivariate predisposition to HT was modeled. Based on the investigated group, a model of seven variables was obtained. The variability explained by this model was assessed at only 5.4821% (p = 2 × 10-6), which indicates that many dozens of factors are required simultaneously to explain HT predisposition. Conclusions We analyzed genetic regions commonly and most significantly associated with autoimmune thyroid disorders in the literature, on a carefully selected cohort. Our results indicated a lack of possibility to predict the risk of HT development, even with a multivariate model. We therefore conclude that strong associations of single genetic regions with HT should be interpreted with great caution. We believe that a change in the attitude towards genetic association analyses of HT predisposition is necessary. Studies including multiple factors simultaneously are needed to unravel the intricacies of genetic associations with HT

A study on β-defensin-2 and histatin-5 as a diagnostic marker of early childhood caries progression

BACKGROUND: Recently, a continuous growth of interest has been observed in antimicrobial peptides (AMPs) in the light of an alarming increase in resistance of bacteria and fungi against antibiotics. AMPs are used as biomarkers in diagnosis and monitoring of oral cavity pathologies. Therefore, the determination of specific protein profiles in children diagnosed with early childhood caries (ECC) might be a basis for effective screening tests and specialized examinations which may enable progression of disease METHODS: The objective of the studies was to determine the role of histatin-5 and β-defensing-2 as a diagnostic marker of early childhood caries progression. In this work, results of concentration determination of two salivary proteins (histatin-5 and β-defensin-2) were presented. In addition, bacterial profiles from dental plaque in various stages of ECC and control were marked. The assessment of alteration in the concentration of these two proteins in a study group of children with various stages of ECC and a control group consisting of children with no symptoms was performed by enzyme-linked immunosorbent assays RESULTS: The statistical analysis showed a significant increase in the concentration of histatin-5 and β-defensin-2 in the study group compared to the control group and correlated with the progression of the disease CONCLUSIONS: The confirmation of concentration changes in these proteins during the progression of dental caries may discover valuable disease progression biomarker

Contribution of selected oxidant-antioxidant markers in oral lichen planus in adults

Liszaj płaski jamy ustnej (OLP, ang. oral lichen planus) jest przewlekłą chorobą zapalną błon śluzowych. Wśród czynników etiologicznych choroby wyróżnia się specyficzne i niespecyficzne mechanizmy immunologiczne. Niemniej jednak rozważane są również mechanizmy związane ze stresem oksydacyjnym. Praca przedstawia patomechanizm liszaja płaskiego jamy ustnej, ze szczególnym uwzględnieniem roli stresu oksydacyjnego, jego genezy, rozwoju i konsekwencji występowania w organizmie. U 92 pacjentów podzielonych na trzy grupy badane (kontrolna, postać siateczkowa, postać nadżerkowa) zostały zbadane poziomy całkowitej zdolności antyoksydacyjnej (TAC, ang. total antioxidant capacity), glutationu (GSH) oraz malonylodialdehydu (MDA). Oznaczenia stężeń wybranych parametrów przeprowadzono metodami spektrofotometrycznymi oraz, w przypadku MDA, fluorymetryczną. Powyższe wskaźniki zostały przedstawione jako stężenie w jednostce objętości śliny oraz w przeliczeniu na białko zawarte w ślinie.Wyniki: Stężenie malonylodialdehydu (MDA) było znacząco wyższe we wszystkich grupach z liszajem, w porównaniu do grupy kontrolnej zdrowych osób. Stężenia całkowitej zdolności antyoksydacyjnej (TAC) oraz glutationu (GSH) były znacząco niższe w grupach badanych, w porównaniu do grupy kontrolnej. Zmiany stężeń oznaczanych wskaźników istotnie różniły się w zależności od postaci choroby i jej nasilenia oraz nie zależały od wieku i płci pacjentów.Oral lichen planus (OLP) is a chronic inflammatory disease of the mucous membranes. Among the etiological factors of the disease, specific and non-specific immune mechanisms are distinguished. However, the mechanisms associated with oxidative stress are also contemplated. This work presents the patomechanism of oral lichen planus, with a particular focus on the role of oxidative stress, its origins, development and consequences of its occurrence in the organism.The total amount of 92 patients was divided into three groups (control, reticular form, erosive form), and have been undergone the examination of levels of total antioxidant capacity (TAC), glutathione (GSH) and malondialdehyde (MDA). The assessment of the selected parameters’ concentration was performed by spectrophotometric and, in the case of MDA, fluorimetric methods. Results were expressed as concentration per volume unit of saliva and per amount of saliva protein.Results: The concentration of malondialdehyde (MDA) was significantly higher in all groups with OLP in comparison to the control group of healthy individuals. Concentrations of total antioxidant capacity (TAC) and glutathione (GSH) were significantly lower in the examined groups compared to the control group. Changes in concentrations of those markers differed significantly depending on the form of the disease and its severity and didn’tt depend on age and gender of the patients

The role of the saliva antioxidant barrier to reactive oxygen species with regard to caries development

Objectives: The aim of this study was to evaluate the role of the antioxidant barrier in the saliva of children with caries, and its impact on the colonization of cariogenic bacteria. Methods: This is a cross-sectional study of 81 children aged 1–5 years. Antioxidant levels and salivary bacterial profiles were measured. Patients were divided into two groups as follows: initial stage decay, termed non-cavitated (1–2 in International Caries Detection and Assessment System (ICDAS)), and extensive decay, termed cavitated lesions (5–6 in ICDAS). The control group includes children without caries. Results: The linear regression model demonstrated that the GSH, GSSG, GSH/GSSG, and total antioxidant capacity levels are influenced (P < 0.05) by: the stage of caries and the dominant bacterial strain. Compared with the other groups (P < 0.001), the highest antioxidant parameters were recorded in the saliva of patients with cavitated lesions. Discussion: Our results indicate that the high levels of antioxidants in saliva increase significantly in children in line with the salivary cariogenic bacterial profiles and caries progression