A small RNA, microRNA as a potential biomolecular marker to estimate post mortem interval in forensic science: a systematic review.

BACKGROUND: Post-mortem interval (PMI) is the cornerstone of the forensic field to investigate. The examination technique by seeing the changes in the body such as algor mortis, rigor mortis, and livor mortis is a traditional technique in which accuracy is influenced by many factors. A biomolecular technique that uses microRNA (miRNA) biomarkers is developing because miRNA has good stability than other RNA, so it meets the requirements to be used for PMI estimation. METHOD: Following the PRISMA guidelines, journals were taken from 5 databases: Scopus, Science Direct, PubMed, Embase, and Springer. The review was carried out by two people. Inclusion criteria in this review are original research, published in the last 10 years, discussing miRNA as a biomarker for PMI estimation, and free full access. While exclusion criteria are not original research and not using English. RESULT: Eighteen journals were reviewed in this study. The study was conducted using test animals (rats) and human samples with tissue sources taken from the liver, skeletal muscle, blood, bone, heart, skin, saliva, semen, brain, lung, vitreous humor, spleen, and kidney. miRNA expression levels after death showed different results based on miRNA target, tissue source, and others. DISCUSSION: The results of each study are different due to the use of different types of miRNA targets and tissue sources. miRNA has great potential to estimate PMI in forensic science, but it is necessary to control the influencing factors to obtain an accurate conclusion.

Accuracy of forensic age estimation using cementum annulation and dentin translucency in adult: a systematic review and meta-analysis.

Identification of the living and the dead individual is essential in routine forensic dental examinations. Age determination can be of great value in forensic odontology, not only in identifying bodies but also in relation to crime. When subjects have extensive changes that external features provide no information, teeth are often the only means of identification. Several procedures for age-at-death estimation in adults have been introduced. Two of them, cementum annulation and dentin translucency, are frequently used as a single dental indicator. Cementum annulation refers to an alternating dark and light band; each pair of it represents 1 year. Meanwhile, dentin translucency is the other dental physiological process that begins in the second or third decade of life and progresses with age. There are still few studies that compared both methods and their accuracy in estimating adult age at death. Therefore, this study aims to test and compare cementum annulation and dentin translucency accuracy by performing a systematic search on five online databases (Pubmed, Scopus, Ebsco, ScienceDirect, and Wiley). All the research articles must be published in the last 10 years, and the full paper must be available in English. Out of the total 1178 literature, 28 studies were recruited for qualitative analysis and 23 studies for meta-analysis. The results show that dentin translucency age estimation is more accurate than the cementum annulation method in the entire population. It is recommended to use the cementum annulation method for younger adults (15-44 years) and the dentin translucency method for the older ones (≥ 45 years).

Interferon-Gamma (IFNg) +874A/T Polymorphism Does Not Significantly Affect the Severity of Periodontitis.

OBJECTIVES: Interferon-gamma (IFNg) is an immune-regulatory cytokine with a role in host responses to periodontitis. Genetic factors have been reported to modify the corresponding protein expression. The objective of this study was to evaluate the association and role of IFNg polymorphisms, such as IFNg +874 A/T, and the susceptibility to periodontitis. MATERIALS AND METHODS: A total of 100 unrelated subjects were included in the present study. Genomic deoxyribonucleic acid (DNA) was obtained from peripheral blood of 43 patients with mild periodontitis and 57 patients with severe periodontitis. The determined clinical parameters of periodontitis included probing depth, clinical attachment loss, and papilla bleeding index. The oral hygiene indicators were also assessed. The level of IFNg was determined from the gingival crevicular fluid by enzyme-linked immunosorbent assay technique. The IFNg +874 A/T polymorphisms were analyzed from peripheral blood by the method of restriction fragment length polymorphism-polymerase chain reaction. STATISTICAL ANALYSIS: Statistical analysis of the results was conducted using chi-squared testing for categorical data. Independent t-tests and Mann-Whitney U tests were used for numeric data. Kruskal-Wallis testing was used to compare genotypes concerning for IFNg +874 A/T polymorphism. A p-value < 0.05 was assumed for statistical significance. RESULTS: Analysis of the IFNg +874 A/T polymorphism showed no significant differences with the level of IFNg. No significant differences were observed either in IFNg +874 A/T polymorphism between the subjects with mild periodontitis and those with severe periodontitis (p > 0.05). The subjects with severe periodontitis showed marginally but not significantly higher levels of IFNg compared with subjects with mild periodontitis (p > 0.05). CONCLUSION: The polymorphism of IFNg +874 A/T was not associated with the level of IFNg nor with the risk of periodontitis in this study.

Association between Epstein-Barr virus and periodontitis: A meta-analysis.

PURPOSE: Previous studies have found that Epstein-Barr virus (EBV) is associated with periodontitis, though some controversy remains. This meta-analysis aimed to clarify and update the relationship between EBV and periodontitis as well as clinical parameters. METHODS: A comprehensive search was conducted in the PubMed and Scopus databases in December 2020. Original data were extracted according to defined inclusion and exclusion criteria. Outcomes were analyzed, including overall odds ratios (ORs) and 95% confidence intervals (CIs). A random-effects model was used, and publication bias was assessed by Egger's and Begg's tests. Sensitivity analysis was used to evaluate the stability of the outcome. RESULTS: Twenty-six studies were included in the present meta-analysis, involving 1354 periodontitis patients and 819 healthy controls. The included studies mostly showed high quality. The overall quantitative synthesis for the association between EBV and periodontitis was an increased odds ratio when subgingival EBV was detected OR = 7.069, 95% CI = 4.197-11.905, P<0.001). The results of subgroup analysis suggested that the association of EBV with periodontitis was significant in Asian, European, and American populations (P<0.001; P = 0.04; P = 0.003, respectively) but not in African populations (P = 0.29). Subgroup analysis by sample type showed that subgingival plaque (SgP), tissue and gingival crevicular fluid GCF were useful for EBV detection (P<0.001). EBV detection amplification methods included nested PCR, multiplex PCR and PCR (P<0.001; P = 0.05, P<0.001, respectively), but EBV detection by real-time PCR and loop-mediated isothermal amplification presented no significant result (P = 0.06; P = 0.3, respectively). For the clinical parameters of periodontitis, pocket depth (PD) and bleeding of probing (BOP) percentages were higher in the EBV-positive sites than in the EBV-negative sites (MD 0.47 [0.08, 0.85], P = 0.02; MD 19.45 [4.47, 34.43], P = 0.01). CONCLUSIONS: A high frequency of EBV detection is associated with an increased risk of periodontitis. The EBV association was particularly significant in all populations except in African populations. Subgigival plaque (SgP), tissue and GCF were not significantly different useful material for detecting EBV in periodontitis. Nested PCR and multiplex PCR are reliable methods for this purpose. In the presence of EBV, PD and BOP are reliable clinical parameters for gingival inflammation. However, some caution in such interpretation is justified due to heterogeneity among studies. A suggested extension could assess the parallel influence of other human herpesviruses.

Association of subgingival Epstein-Barr virus and periodontitis.

Background: The Epstein-Barr virus (EBV) is gaining interest as a possible agent in the etiology of periodontitis. Previous studies have shown controversy on whether EBV DNA in the subgingival periodontal pockets is associated with periodontitis. The aim of the present study was to seek the potential relationship between EBV and periodontitis. Methods: Data on socio-demographics, oral health, and periodontal health were recorded, and samples were collected from gingival crevicular fluid, using sterile paper point. This case-control study of 118 participants included 59 subjects with severe periodontitis and 59 control subjects with mild periodontitis. The EBV load was determined by quantitative real-time PCR. Results: EBV DNA was detected in 37.3% of the case samples and in 18.6% of the control samples. There was no significant difference in the load of EBV DNA between severe and mild periodontitis (p>0.05). The observed load of EBV DNA was up to 4.55x10 5 copies/mL. The detected EBV DNA was significantly associated with the plaque index and the oral hygiene index (all p<0.05). Conclusions: A significant association was not found, but EBV might contribute to periodontitis. Gingival crevicular fluid is useful for monitoring the EBV load by the real-time PCR technique.

Obesity correlated to a higher risk of acquiring periodontitis: a cross-sectional study.

BACKGROUND: The present study aimed to investigate the correlation between obesity and periodontitis, among other risk factors for periodontitis.   Methods:  In total, 262 Indonesian male and female subjects were analysed for body mass index (BMI), oral hygiene, plaque index, and clinically evaluated periodontitis. Statistical analysis was performed using Spearman tests and Pearson chi-square tests to estimate the correlation between BMI and periodontitis. Multivariate binary logistic analysis was conducted between covariate and periodontitis. P<0.05 was considered as statistically significant.   Results: The prevalence of obesity was 48.47%. There were positive correlations between BMI and periodontal status for healthy-mild periodontitis, moderate, and severe periodontitis respectively. BMI and periodontitis crude odds ratio (OR) = 2.31 (95% CI 1.41-3.78); p < 0.05, adjusted OR of BMI among other variables, was 1.88 (95%CI 1.05-3.37); p < 0.05. Exploration of the ROC curve found a BMI cut off point of 24.785 kg/m2.  Conclusion: Obesity by BMI measurement of ≥ 25kg/m2 correlated to a higher risk of acquiring periodontitis compared to normal-weight individuals.

Vertical Mandibular and Trunk Symmetry in Indonesian Orthodontic Patients

Abstract Objective: To analyze differences in vertical mandibular and trunk symmetry in orthodontic patients. Material and Methods: This was a cross-sectional study of 129 growing orthodontic patients who sought orthodontic treatment at the Dental Hospital Universitas Sumatera Utara, Indonesia. Mandibular symmetry index was observed with pre-treatment panoramic radiography based on Kjellberg's technique and trunk symmetry was evaluated based on questionnaires and visual observation. Vertical mandibular asymmetry was decided if the index of asymmetry was lower than 93.7%. The bivariate analysis used the chi-squared and Fisher's exact tests, with a significance level of 5%. Results: There was a significant association between vertical mandibular and trunk symmetry (p<0.05). The prevalence odds ratio for the association with vertical mandibular asymmetry was 3.007 (95% CI = 1.016-8.905) for trunk asymmetry. Conclusion: The necessity to consider trunk symmetry could be included in orthodontics treatment of any malocclusion with vertical mandibular asymmetry that might require a multidisciplinary approach in the future.

Immunoglobulin G Levels in the Gingival Crevicular Fluid of Menopausal Patients with Periodontitis

Abstract Objective: To measure the level of immunoglobulin G (IgG) in the gingival crevicular fluid (GCF). Material and Methods: A total of 158 patients aged >45 years were examined for periodontitis and interviewed regarding their menopausal status. The non-menopause group entailed female patients with periodontitis without menopause (n=23). The menopause group included females who stopped menstruating since >1 year, had a pocket depth of 4-5 mm, and did not have other systemic conditions (n=40). Samples were selected based on periodontal and menopausal status. In total, 63 samples of GCF were collected from the participants and tested using an enzyme-linked immunosorbent assay kit for IgG. Results: The median level of IgG in the menopause group was 39.50 (g/mL, whereas that of the non-menopause group was 41.08 (g/mL. There was a positive correlation between the plaque index and IgG level in both groups. In contrast, there was a negative correlation between age and IgG level. However, there was no correlation between plaque index and age regarding the IgG level in both groups (p>0.05). Conclusion: The IgG levels in the menopause group were lower than those in the non-menopause group. As such, menopausal females should take great care of their overall health, including the periodontium.

Association of a Polymorphism in the Gene Encoding Methylenetetrahydrofolate Dehydrogenase 1 (MTHFD1) 1958G> A with Orofacial Cleft

ABSTRACT Objective: To evaluate the possible association of a polymorphism in the gene encoding methylenetetrahydrofolate dehydrogenase 1 (MTHFD1), 1958G>A, with the susceptibility to orofacial cleft in an Indonesian population. Material and Methods: A total of 200 stored secondary biological samples from 30 cases of orofacial cleft and 170 unaffected controls were analyzed to determine the polymorphism status at base 1958. The analysis was conducted using the PCR-restriction fragment length polymorphism technique after digestion with the Msp1 restriction enzyme. The samples were then subjected to agarose gel electrophoresis to investigate the presence or absence of the following fragments: genotype GG, 196, 86 and 40 base pairs (bp); genotype AA, 282 and 28 bp and genotype AG, 282, 196, 86, 40 and 28 bp. The test groups were compared using the Chi-square test. Results: The wild-type allele containing 1958G, as well as the genotype GG, were significantly more common in the control group than in the orofacial cleft group. Conclusion: The MTHFD1 1958G>A polymorphism was significantly associated with orofacial cleft susceptibility in the tested Indonesian population.

Detection of DNA adduct 8-hydroxy-2'-deoxyguanosine (8-OHdG) as a toxicity bioindicator to the effects of nickel on Ni-Cr alloy prosthesis users.

Previous studies have suggested that exposure to Ni from Ni-Cr alloys can affect the human body through oxidative stress. The present study discusses the effect of nickel from Ni-Cr alloy prostheses on the formation of DNA Adduct 8-Hydroxy-2'-Deoxyguanosine (8-OHdG), evaluated based on creatinine and 8-OHdG concentrations in urine, determined with LC-MS/MS, for a Ni-Cr alloy user group and a never-user control group. The mean creatinine and 8-OHdG concentrations were not significantly different between the test groups, although highest levels were observed for the in the Ni-Cr user group. It is suggested that samples with relatively high creatinine and/or 8-OHdG levels are further studied in more detail for stability of concentrations and for the effect of contributing factors.

Sex Identification Based on Tooth Crown Trait Analysis Among the Mongoloid Race

Abstract Objective: To determine whether anterior and posterior tooth crown traits exhibit sexual dimorphism and identify traits characteristic to the Mongoloid race, especially among the Indonesian population. Material and Methods: This study cross-sectional study analyzed 108 dental casts from 36 males and 72 females. The traits analyzed included winging, shoveling, double shoveling, canine mesial ridge, canine distal accessory ridge, hypocone, metaconule, Carabelli's cusp, protostylid, metaconulid, enteconulid, and hypoconulid. All tooth crown traits were scored based on the ASUDAS scoring system. Descriptive statistics were used to calculate the absolute and relative frequencies. The Chi-square tests was used to determine significant differences in anterior and posterior tooth crown traits between males and females. Level of significance was set at 5%. Results: None of the traits showed sexual dimorphism. Moreover, the most common traits among the Mongoloid race were hypocone (94.4%) and shoveling (86.1%). Conclusion: Although none of the traits exhibited sexual dimorphism, most of them had a higher incidence among females than males. Nonetheless, further research including adequate samples and a similar number of females and males, is needed, especially for population studies.

Early Detection of Bacillus anthracis From Saliva in Anticipation of a Bioterrorism Attack

Objective: To assess potential for early detection of oral infection by B. anthracis spores for preparedness of a bioterrorism attack. Material and Methods: The laboratory study used saliva with a range of initial anthrax concentrations, to compare detection by direct observation from conventional blood agar culture and by anthrax-specific PCR after a shorter culture in BHI broth. Three types of saliva were collected: stimulated saliva, unstimulated/whole saliva, and unstimulated/whole saliva with antibiotic treatment (for negative control). Using bivariate Kruskal-Wallis and Mann-Whitney tests for statistical analysis for factors that could affecting anthrax detection, significant differences between the test groups was assumed at p<0.05. Results: From unstimulated whole saliva heat shock treated at 62.50C, B. anthracis growth was detected with both methods. PCR detection from a BHI broth culture could shorten the time to diagnosis in comparison to conventional culture in blood agar. Conclusion: Saliva can provide useful samples for diagnosis of oropharyngeal anthrax. In comparison to conventional culture on blood agar, shorter-term culture in BHI broth provides potential for earlier detection and diagnosis.

Relationship of Age, Body Mass Index, Bone Density, and Menopause Duration with Alveolar Bone Resorption in Postmenopausal Women

Objective: To analyze the relationship between age, body mass index (BMI), bone mineral density (BMD), and alveolar bone resorption with menopause duration in postmenopausal women. Material and Methods: A cross-sectional study was developed involving 59 subjects, aged 45 to 80 years and categorized the duration of menopause as ≤5 years and >5 years. Body mass index measurement and menopause duration were collected. Bone loss seen on radiography was measured by drawing a vertical line from the cementoenamel in the distal part of the 36 teeth and the mesial portion of 46 teeth to the base of the bone marked by the lamina dura intact. Categorical determinations of age, BMI, BMD, and alveolar bone resorption were based on receiver operating characteristic (ROC) curves. Were used Pearson correlation and Spearman correlation tests with the significance level set at 5%. Results: The majority of subjects (54.2%) with menopause duration >5 years were aged >54.5 years, most had BMI >24.2 kg/m2 (39%), had bone resorption >2.95 mm (52.5%), and had bone density ≤73.89 (49.2%). Pearson and Spearman correlation tests showed no significant correlation between age, BMI, bone density, and alveolar bone resorption (p>0.05). Conclusion: The longer the duration of menopause showed a tendency for lower bone density and higher age, BMI, and bone resorption.

Positive Correlation Between the Level of Interferon-Gamma and the Severity of Periodontitis

Abstract Objective: To examine the IFN-γ levels in patients with periodontitis and determine the difference in the levels of IFN-γ with the severity of the disease. Material and Methods: The study design was cross-sectional, and the sample consisted of 31 patients, aged between 18 and 64 years. Plaque index (PlI), calculus index (CI), and papillary bleeding index (PBI) were measured. Pocket depth (PD), recession, and clinical attachment loss (CAL) (mm) were measured at six sites per teeth. For mild/moderate periodontitis, pocket depth ≥4 mm in 1-3 sites was required, while the essential criteria for severe periodontitis were pocket depth ≥5 mm, clinical attachment loss >3 mm in more than 3 sites ≥2 quadrants. The IFN-γ levels were measured by performing enzyme-linked immunosorbent assay with the gingival crevicular fluid (GCF) samples. The measurements were made in two different sites, and the severity of periodontitis was categorized based on the pocket depth, attachment loss, and the remaining natural teeth. Kruskal-Wallis test, Mann-Whitney test, and Spearman's rank correlation coefficient were Results: The levels of IFN-γ (pg/mL) were correlated with the severity of the periodontal status, with p <0.05. Clinical parameters of periodontitis also correlated with the level of IFN-γ (pg/mL) Conclusion: Subjects with periodontitis presented greater levels of IFN-γ (pg/mL) in GCF than the periodontal healthy individuals. This result showed the role of IFN-γ in the inflammation.

Polymorphism of Serotonin Transporter SLC6A4 (5-HTTLPR) Gene in Cheilitis Angularis Patients

Abstract Objective: To determine the relationship between the Serotonin transporter SLC6A4 (5-HTTLPR) gene polymorphism in cheilitis angularis patients. Material and Methods: We conducted a descriptive analysis of 100 DNA samples extracted from the blood serum of 50 patients with cheilitis angularis and 50 patients without cheilitis angularis. Analysis of the Serotonin transporter SLC6A4 (5-HTTLPR) gene polymorphism was observed by carrying out PCR method followed by electrophoresis for the analysis, without the usage of restriction enzyme. The Chi-square test was used for statistical analysis Results: In the cheilitis angularis group there were 24 samples with SS genotype, 23 samples with LS genotype, and 3 samples with LL genotype. Whereas in the non-cheilitis angularis group, there were 5 samples with SS genotype, 18 samples with LS genotype, and 27 samples with LL genotype. In the cheilitis angularis group found 71 S alleles and 29 L alleles, and in the non-cheilitis angularis group 28 S alleles and 72 L alleles were found. A statistically significant difference was found between the groups (p<0.001) Conclusion: There were significant differences in the distribution of the Serotonin transporter SLC6A4 (5-HTTLPR) gene polymorphism between patients with and without cheilitis angularis.

Areca nut extract demonstrated apoptosis-inducing mechanism by increased caspase-3 activities on oral squamous cell carcinoma.

Background: Oral squamous cell carcinoma is a neoplasm of keratinocyte cells of oral mucosa epithelium that can potentially spread through lymphatic tissue or blood vessel. Although areca nut is one of the plants with a risk of inducing that cancer, areca nut is believed to have high antioxidant properties. Due to the current interest in the apoptosis effects from areca nut for oral cancer treatment, we investigated its ability to induce apoptosis and caspase-3 activity in oral cancer cell lines: HSC-2 and HSC-3. Methods: We examined the effect of areca nut on apoptosis and caspase-3 activity in HSC-2 and HSC-3 cells. Flow cytometry was conducted for the quantification of the cells that were apoptotic and expressing the caspase-3 enzyme for 24 and 48 hours. Results: Areca nut induced a significant increase (p<0.01) in late apoptosis of HSC-2 cells and mostly occurred over 48 hours. The study also found that in HSC-3, there were significant increases (p<0.01) the percentage of cells in early apoptosis after 24 hours and late apoptosis at 48 hours. Caspase-3 activity increased after 24 and 48 hours of areca nut exposure in both cells. Conclusions: The study showed that areca nut could be considered as a potential anticancer agent through its capability in inducing a caspase-dependent apoptosis.

Methylation of tumor suppressor genes p16(INK4a), p27(Kip1) and E-cadherin in carcinogenesis.

Not only genomic mutations but also abnormal epigenetic methylation can significantly contribute to gene silencing and carcinogenesis. Methylation is particularly often observed in the CpG islands of the promoter regions in the regulatory genes. However, there are considerable differences in the incidence of methylation e.g. in the tumor suppressor genes, so that aberrant methylation of p16(INK4a) is relatively frequently observed in tumors, p27(Kip1) methylation is rare, and the incidence of E-cadherin methylation occurs at an intermediate rate. Although true genomic defects are generally much less common than methylation, parallel tendencies for both are often observed, probably reflecting the different levels of evolutionary advantage for tumor cells from inactivation of different genes. This also suggests that loss of p27 expression could be more a consequence of carcinogenesis, while lost p16 expression is a true oncogenic event. Due to the role of p27 in maintaining cellular quiescence, however, loss of its expression can still be a useful partial indicator of the aggressiveness of cancer. Loss of E-cadherin or its catenin partners of cellular adhesion will result in increasing invasiveness and metastatic potential of neoplastic cells but, because of several alternative routes to the same effect, incidence of lost expression for one component gene like E-cadherin does not need to be very high. Similarly, there must be a relatively high number of genes with modest or low incidence of aberrant silencing by methylation, to reflect multiple alternatives for the multistep process of carcinogenesis. Nevertheless, methylation of different genes also shows characteristic differences between different cancer and tumor types, and the epigenetic methylation patterns therefore have considerable diagnostic and prognostic potential. Realising this potential requires efficient methods for profiling the status of methylation. Such profiling methods have only recently become available and are still under relatively rapid development.