Assessment of salivary miRNA, clinical, and demographic characterization in colorectal cancer diagnosis.

AIM: Colorectal cancer (CRC), as the third most frequent malignancy in the world, is the fourth major cause of cancer-related mortality. Its early detection contributes significantly to a reduction in mortality. The objective of this case-control research was to analyze the salivary expression of microRNA-29a (miR-29a) and microRNA-92a (miR-92a), and also to consider demographic, clinical, and nutritional habits for differentiation between CRC patients and healthy controls, especially in the early stages. METHOD: A standard checklist was used to obtain the demographic information, clinical features, and dietary habits of the case and control groups. Samplings of whole unstimulated saliva samples were obtained from 33 healthy persons and 42 CRC patients. Through real-time PCR, statistical analyses, and machine learning analyses, miR-29a and miR-92a salivary expression levels were evaluated. RESULTS: The mean salivary expression of miR-92a and miR-29a in CRC patients was significantly higher than in healthy controls (p < 0.001). The area under the receiver operating characteristic curve for miR-92a and miR-29a salivary biomarkers was 0.947 and 0.978, respectively. The sensitivity and specificity values for miR-92a were 95.24 % and 84.85 %, respectively, whereas sensitivity and specificity for miR-29a were equal to 95.20 % and 87.88 %, respectively. Multiple logistic regressions considering demographics, clinical features, and nutritional habits led to values of 95.35 % and 96.88 % as sensitivity and specificity, respectively, and machine learning analysis led to values of 88.89 % and 86.67 % as sensitivity and specificity, respectively. CONCLUSION: CRC could be accurately diagnosed based on miR-92a and miR-29a levels in saliva. Statistical analysis and machine learning might develop cost-effective models for the distinction of CRC using a noninvasive technique.

Effect of synbiotic mouthwash on oral mucositis induced by radiotherapy in oral cancer patients: a double-blind randomized clinical trial.

PURPOSE: Considering the complex pathobiology of oral mucositis, especially in oral cancer patients, the prevention and treatment of oral mucositis in patients undergoing radiotherapy remains an essential and clinically crucial unmet need. The present study aims to investigate and compare the effects of synbiotic mouthwash with normal saline mouthwash on the prevention and control of radiotherapy-induced oral mucositis in oral cancer patients. METHODS: Double-blind, randomized clinical trial (RCT) performed on 64 oral cancer patients who underwent radiotherapy (IRCT20201106049288N1, registration date: 2020-12-23). Patients were divided randomly into the case (32 subjects) and control (32 subjects) groups. All patients underwent intensity-modulated radiotherapy and received 6000 cGY of radiotherapy in 34 fractions. All patients received the usual treatment for mucositis, but in the case group, synbiotic mouthwash was prescribed and in the control group, normal saline mouthwash was prescribed from a day before the start to the end of radiotherapy treatment. Patients were monitored every session for 6 weeks to check the progression, oral involvement severity, and mucositis grade. RESULTS: The case group showed a significant reduction in the oral mucositis severity. The mucositis grade in the case group from the 7th session of oral examination was significantly lower than the control (p < 0.05), and this significant difference persisted until the last session of oral examination. Incidence rates of severe oral mucositis (grade 3) during the treatment period were 11.59% in the case and 36.45% in control (p < 0.001). CONCLUSION: Synbiotic mouthwash significantly reduces and prevents oral mucositis intensity in oral cancer patients undergoing radiotherapy.

Salivary noncoding RNA in the diagnosis of pancreatic cancer: Systematic review and meta-analysis.

BACKGROUND: Pancreatic cancer is considered one of the most deadly malignancies, primarily because of its diagnostic challenges. We performed a systematic review and diagnostic meta-analysis to evaluate the diagnostic value of noncoding salivary RNAs in pancreatic cancer diagnosis. METHODS: Our investigation involved pertinent studies published in PubMed, Scopus, Web of Science, LIVIVO, Ovid and also the Google Scholar search engine. Specificity and sensitivity were calculated, as were positive and negative likelihood ratios (PLR and NLR), and the diagnostic odds ratio (DOR). The summary receiver-operating characteristics and area under the curve were plotted and assessed. RESULTS: This meta-analysis and systematic review involved and examined five studies that contained 145 study units with a total of 2731 subjects (1465 pancreatic cancer patients versus 1266 noncancer controls). The pooled specificity, sensitivity, NLR, PLR and DOR were 0.783 (95% CI: 0.759-0.805), 0.829 (95% CI: 0.809-0.848), 0.309 (95% CI: 0.279-0.343), 3.386 (95% CI: 2.956-3.879) and 18.403 (95% CI: 14.753-22.954), respectively, with the area under the curve (AUC) equal to 0.882. Subgroup analyses were conducted based on the saliva type (unstimulated and stimulated), mean age of patients, sample size, type of control, serum carbohydrate antigen 19-9 (CA19-9) level and type of salivary noncoding RNA (microRNA (miRNA) and long noncoding RNA (lncRNA)). CONCLUSIONS: The results of our systematic review and meta-analysis suggest that noncoding RNA biomarkers in the stimulated saliva could be a promising approach for accurate pancreatic cancer diagnosis in the early stages.

Assessment of the methylene blue mediated photodynamic therapy on BCL2 and BAX genes expression at mRNA level and apoptosis of head and neck squamous cell carcinoma cell line.

AIM: This study aimed to assess the effect of photodynamic therapy (PDT) on apoptosis of head and neck squamous cell carcinoma (HNSCC) cells by flow cytometry and evaluating BAX and BCL2 genes expression.

Evaluation of CSTB and DMBT1 expression in saliva of gastric cancer patients and controls.

BACKGROUND: Gastric cancer (GC) is the fifth most common cancer and the third cause of cancer deaths globally, with late diagnosis, low survival rate, and poor prognosis. This case-control study aimed to evaluate the expression of cystatin B (CSTB) and deleted in malignant brain tumor 1 (DMBT1) in the saliva of GC patients with healthy individuals to construct diagnostic algorithms using statistical analysis and machine learning methods. METHODS: Demographic data, clinical characteristics, and food intake habits of the case and control group were gathered through a standard checklist. Unstimulated whole saliva samples were taken from 31 healthy individuals and 31 GC patients. Through ELISA test and statistical analysis, the expression of salivary CSTB and DMBT1 proteins was evaluated. To construct diagnostic algorithms, we used the machine learning method. RESULTS: The mean salivary expression of CSTB in GC patients was significantly lower (115.55 ± 7.06, p = 0.001), and the mean salivary expression of DMBT1 in GC patients was significantly higher (171.88 ± 39.67, p = 0.002) than the control. Multiple linear regression analysis demonstrated that GC was significantly correlated with high levels of DMBT1 after controlling the effects of age of participants (R2 = 0.20, p < 0.001). Considering salivary CSTB greater than 119.06 ng/mL as an optimal cut-off value, the sensitivity and specificity of CSTB in the diagnosis of GC were 83.87 and 70.97%, respectively. The area under the ROC curve was calculated as 0.728. The optimal cut-off value of DMBT1 for differentiating GC patients from controls was greater than 146.33 ng/mL (sensitivity = 80.65% and specificity = 64.52%). The area under the ROC curve was up to 0.741. As a result of the machine learning method, the area under the receiver-operating characteristic curve for the diagnostic ability of CSTB, DMBT1, demographic data, clinical characteristics, and food intake habits was 0.95. The machine learning model's sensitivity, specificity, and accuracy were 100, 70.8, and 80.5%, respectively. CONCLUSION: Salivary levels of DMBT1 and CSTB may be accurate in diagnosing GCs. Machine learning analyses using salivary biomarkers, demographic, clinical, and nutrition habits data simultaneously could provide affordability models with acceptable accuracy for differentiation of GC by a cost-effective and non-invasive method.

Salivary biomarkers in breast cancer diagnosis: A systematic review and diagnostic meta-analysis.

BACKGROUND: Salivary diagnostics and their utility as a nonaggressive approach for breast cancer diagnosis have been extensively studied in recent years. This meta-analysis assesses the diagnostic value of salivary biomarkers in differentiating between patients with breast cancer and controls. METHODS: We conducted a meta-analysis and systematic review of studies related to salivary diagnostics published in PubMed, EMBASE, Scopus, Ovid, Science Direct, Web of Science (WOS), and Google Scholar. The articles were chosen utilizing inclusion and exclusion criteria, as well as assessing their quality. Specificity and sensitivity, along with negative and positive likelihood ratios (NLR and PLR) and diagnostic odds ratio (DOR), were calculated based on random- or fixed-effects model. Area under the curve (AUC) and summary receiver-operating characteristic (SROC) were plotted and evaluated, and Fagan's Nomogram was evaluated for clinical utility. RESULTS: Our systematic review and meta-analysis included 14 papers containing 121 study units with 8639 adult subjects (4149 breast cancer patients and 4490 controls without cancer). The pooled specificity and sensitivity were 0.727 (95% CI: 0.713-0.740) and 0.717 (95% CI: 0.703-0.730), respectively. The pooled NLR and PLR were 0.396 (95% CI: 0.364-0.432) and 2.597 (95% CI: 2.389-2.824), respectively. The pooled DOR was 7.837 (95% CI: 6.624-9.277), with the AUC equal to 0.801. The Fagan's nomogram showed post-test probabilities of 28% and 72% for negative and positive outcomes, respectively. We also conducted subgroup analyses to determine specificity, sensitivity, DOR, PLR, and NLR based on the mean age of patients (≤52 or >52 years old), saliva type (stimulated and unstimulated saliva), biomarker measurement method (mass spectrometry [MS] and non-MS measurement methods), sample size (≤55 or >55), biomarker type (proteomics, metabolomics, transcriptomics and proteomics, and reagent-free biophotonic), and nations. CONCLUSION: Saliva, as a noninvasive biomarker, has the potential to accurately differentiate breast cancer patients from healthy controls.

Salivary cystatin S levels in children with early childhood caries in comparison with caries-free children; statistical analysis and machine learning.

BACKGROUND: Early childhood caries is the most common infectious disease in childhood, with a high prevalence in developing countries. The assessment of the variables that influence early childhood caries as well as its pathophysiology leads to improved control of this disease. Cystatin S, as one of the salivary proteins, has an essential role in pellicle formation, tooth re-mineralization, and protection. The present study aims to assess salivary cystatin S levels and demographic data in early childhood caries in comparison with caries-free ones using statistical analysis and machine learning methods. METHODS: A cross-sectional, case-control study was undertaken on 20 cases of early childhood caries and 20 caries-free children as a control. Unstimulated whole saliva samples were collected by suction. Cystatin S concentrations in samples were determined using human cystatin S ELISA kit. The checklist was collected from participants about demographic characteristics, oral health status, and dietary habits by interviewing parents. Regression and receiver operating characteristic (ROC) curve analysis were done to evaluate the potential role of cystatin S salivary level and demographic using statistical analysis and machine learning. RESULTS: The mean value of salivary cystatin S concentration in the early childhood caries group was 191.55 ± 81.90 (ng/ml) and in the caries-free group was 370.06 ± 128.87 (ng/ml). T-test analysis showed a statistically significant difference between early childhood caries and caries-free groups in salivary cystatin S levels (p = 0.032). Investigation of the area under the curve (AUC) and accuracy of the ROC curve revealed that the logistic regression model based on salivary cystatin S levels and birth weight had the most and acceptable potential for discriminating of early childhood caries from caries-free controls. Furthermore, using salivary cystatin S levels enhanced the capability of machine learning methods to differentiate early childhood caries from caries-free controls. CONCLUSION: Salivary cystatin S levels in caries-free children were higher than the children with early childhood caries. Results of the present study suggest that considering clinical examination, demographic and socioeconomic factors, along with the salivary cystatin S levels, could be usefull for early diagnosis ofearly childhood caries in high-risk children; furthermore, cystatin S is a protective factor against dental caries.

Effect of photodynamic therapy on expression of HRAS, NRAS and caspase 3 genes at mRNA levels, apoptosis of head and neck squamous cell carcinoma cell line.

OBJECTIVES: This study aimed to assess the effect of photodynamic therapy (PDT) on expression of CASP3, NRAS and HRAS genes at mRNA levels, and apoptosis of head and neck squamous cell carcinoma (HNSCC) cell line. MATERIALS AND METHODS: In order to complete the present in vitro study, HNSCC cell line (NCBI C196 HN5) purchased from Pasteur Institute. Cells were divided into four groups; Group 1: photodynamic treatment (laser + methylene blue (MB) as photosensitizer), group 2: MB, group 3: laser (with 660 nm wavelength), and group 4: control (without any treatment). To determine the optimal concentration of MB, in a pilot study, toxicity of MB in different concentration was assessed using MTT assay. Cells in group 1, 2 and 3 was treated at optimal concentration of MB (1.6 µg/mL). Gene expression at mRNA levels was assessed after 24 h incubation, using real-time (qRT)-PCR. The expression of BAX and BCL2 genes at the mRNA levels was analyzed to evaluate apoptosis. 2-ΔΔCt values of BCL2, BAX, CASP3, NRAS, and HRAS in groups was analyzed using ANOVA. Tukey's HSD and Games Howell test was used to compare between two groups. RESULTS: Over-expression of BAX (p < 0.001), CASP3 (p < 0.001) and down-regulation of BCL2 (p = 0.004), HRAS (p = 0.023) and NRAS (p = 0.045) were noted in group 1 (PDT), compared with the control group. Treatment by laser alone induce down-regulation of CASP3 (p < 0.05), BAX (p < 0.05), BCL2 (p > 0.05), HRAS (p > 0.05) and NRAS (p > 0.05). CONCLUSION: PDT caused down-regulation of NRAS, HRAS and BCL2 and over-expression of CASP3 and BAX genes at mRNA levels in HNSCC cell line. The present study raises the possibility that the role of MB on BCL2 down-regulation and BAX and CASP3 over-expression was higher than laser alone while it seems that laser alone was more effective than MB in HRAS and NRAS down-regulation.

Er,Cr:YSGG laser surface treatment of gamma titanium aluminide: Scanning electron microscopy-energy-dispersive X-ray spectrometer analysis, wettability and Eikenella corrodens and Aggregatibacter actinomycetemcomitans bacteria count-in vitro study.

Dental implants play an important role in oral health. Titanium dental implants must endure the complex microflora environment of the oral cavity. Moreover, bacterial infections have been considered as one of the most important factors of implant failure. The issue of dental improvement through modification of chemical composition and surface treatment has received considerable critical attention. γ-TiAl as a novo biocompatible material revealed a slower corrosion rate in biological media rather Ti-6Al-4V. The objective of this study is to investigate the effect of Er,Cr:YSGG laser on γ-TiAl in comparison with sandblasted and acid-etched samples as the control groups and machined samples.Wettability, surface roughness, surface topography, scanning electron microscopy-energy dispersive X-ray spectrometer analysis of surface and subsurface of samples were investigated and bacteria counts of two periodontal bacterial strains (Aggregatibacter actinomycetemcomitans and Eikenella corrodens) were evaluated on the Er,Cr:YSGG laser surface-treated sandblasted and acid-etched and machined samples.The results of this investigation show that Er,Cr:YSGG laser surface treatment affects surface roughness, surface topography, wettability, chemical composition of the surface and bacteria count. Scanning electron microscopy-energy dispersive X-ray spectrometer analysis of the sample revealed the increment of titanium and oxygen content and reduction of aluminum content in the surface and subsurface layer. A. actinomycetemcomitans and E. corrodens count were found from the lowest level to highest in the sandblasted and acid-etched samples, laser samples and machined samples, respectively.Using controlled parameters of Er,Cr:YSGG laser ensured no significant adverse alteration. The findings to emerge from this study revealed the significant correlation between microbial count and wettability. Furthermore, the contact angle strongly correlated with surface roughness.