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Introduction. SARS-CoV2 2019 infection represent global problem. At this moment there is no vaccine or efficient treatment of infected patients. Treatment with blood plasma rich with anti SARS-CoV2 specific antibodies represents rare safe and effective treatment of Covid19 patients. Patients and Material. A total number of 950 patients were analyzed in this study, whose samples were collected in time interval from 01.05. till 15.08.2020. Patients were enrolled in study from Covid-19 hospitals, out-clinics and as family members of SARS-CoV2 infected patients. Original ELISA tests were developed to measure the concentration of anti-S1S2 Spike and anti-Nucleoprotein (IgG, IgA, IgM) SARS-CoV2 antibodies. Blood convalescent plasma was selectively collected from recovered patients according to specific antibodies concentration. Results. The highest concentrations of anti-S1S2 Spike or anti-Nucleoprotein specific IgG antibodies were detected in patients with the moderate/heavy clinical form of infection as well as in group of family members of SARS-CoV2 infected patients. Extremely high concentration of anti S1S2 Spike IgG and anti-Nucleoprotein IgG was demonstrated in 3% and 6% (respectively) of patients recovered from severe Covid19. From our hospitalized patients 63% and 51% had modest antibody levels (anti S1S2 Spike and anti-Nucleoprotein, respectively). After 60 days, in our selected donors’ concentration of anti S1S2 Spike IgG antibodies increased in 67%, paralleled with increase of anti-Nucleoprotein IgG antibodies in 58% of donors. Conclusion. Originally developed ELISA tests enable novel protocol for selection of convalescent blood plasma donor.

Background/Aim. Oral squamous cell carcinoma (OSCC) is the most common tumor type of head and neck carcinomas, characterized by a high recurrence rate and patients? poor survival. Further elucidation of the function and regulation of the TP53, a pivotal tumor suppressor gene, would provide advances in predicting the clinical behavior, prognosis, and chemotherapy response of OSCC patients. Thus, we investigated the association of TP53 gene mutations with survival and response to cisplatin chemotherapy in human papilloma virus (HPV)-negative OSCC patients. Methods. The potential clinical relevance of TP53 mutations was analyzed in 82 patients with HPV-negative OSCC. All patients underwent radiotherapy, and 25 patients received cisplatin chemotherapy. A negative HPV status was deter-mined by type-specific polymerase chain reaction (PCR) for high-risk HPV 16, 18, 31, and 33. Targeted sequencing of TP53 exons 4?8 was assessed by Sanger sequencing. Results. Of 82 HPV-negative OSCC patients, 49 (59.79%) had TP53 mutations, and 26 patients (31.7%) carried pathogenic TP53 mutations. Patients with pathogenic TP53 mutations had significantly reduced overall survival (p = 0.009). Recurrence status, but not TP53 mutations, was an independent marker of poor survival in our cohort [hazard ratio (HR) = 4.733, 95% confidence interval (95% CI): 2.027?11.053, p = 0.0001]. In the sub-cohort of patients who underwent cisplatin-based chemotherapy, pathogenic TP53 mutations were predictors of poor response to chemotherapy (p = 0.026). Conclusion. Our findings indicate that pathogenic TP53 mutations in HPV-negative OSCC tumors could be a prognostic marker of patients? reduced overall survival. In addition, pathogenic TP53 mutations in HPV-negative OSCC could be a marker of poor chemotherapy response of OSCC patients.

Melanoma is one of the most aggressive tumors, and in the setting of rising incidence and mortality, there is an urgent need to identify new prognostic markers. Toll-like receptors (TLRs), are aberrantly expressed in numerous cancers, including melanoma. TLR signaling provides a microenvironment that is involved in antitumor immune response, chronic inflammation, cancer cell proliferation and evasion of immune destruction. In the present study, we investigated whether single nucleotide polymorphisms (SNPs) in TLR3 and TLR4 genes are associated with clinicopathologic features, progression and survival of melanoma patients. The study was conducted on 120 melanoma patients. DNA extracted from peripheral blood was genotyped for TLR3 polymorphisms rs5743312 and rs3775291 (L412F) and TLR4 polymorphisms rs4986790 (D299G) and rs4986791 (T399I), by TaqMan Real-Time PCR Assays. Kaplan-Meier survival curves were compared by the log-rank test. TLR3 polymorphism L412F was associated with a higher mitotic index ( P = 0.035). TLR4 D299G and T399I polymorphisms were associated with indicators of melanoma severity, nodal metastases ( P = 0.005 and P = 0.007, respectively) and advanced stage III ( P = 0.005 and P = 0.004, respectively). Cox regression analysis showed that the presence of tumor-infiltrating lymphocytes (TILs) predicted better overall survival (HR = 0.318; P = 0.004). TLR4 T399I polymorphism was significantly associated with worse survival, P = 0.025. The overall survival rates were significantly lower for patients carrying variant allele T of TLR4 T399I SNP (TC and TT genotypes combined) ( P = 0.008, log-rank test), compared to wild-type genotype CC. Our findings indicate that TLR4 polymorphisms T399I (rs4986791) and D299G (rs4986790) could be potential prognostic and survival markers for melanoma patients.

Vitamin D receptor (VDR) gene polymorphisms could play a significant role in the susceptibility and pathogenesis of osteoarthritis (OA), the most common degenerative joint disorder in humans. The current study involved 94 OA patients and 100 healthy, asymptomatic controls. VDR variants FokI (rs2228570), TaqI (rs731236), ApaI (rs7975232) and EcoRV (rs4516035) were genotyped using TaqMan-based real-time PCR. Adjusted odds ratio (OR) analysis showed that VDR TaqI and FokI polymorphisms are significantly associated with susceptibility to OA (OR=1.986, P=0.001 and OR=1.561, P=0.017, respectively). Joint-specific analysis showed that the VDR TaqI polymorphism was associated with risk of hip OA (OR=1.930, P=0.005) and knee OA (OR=1.916, P=0.028), while the VDR FokI polymorphism was associated with higher risk of knee OA (OR=2.117, P=0.012). VDR TaqI and FokI polymorphisms are associated with the occurrence of persistent pain (P=0.005 and P=0.027, respectively), while ApaI was associated with a family history of OA (p=0.004). The VDR FokI and TaqI genetic variants significantly contribute to osteoarthritis susceptibility, the occurrence of persistent pain, and potentially to joint-specific OA risk.

Cyclin-dependent kinase inhibitor p21 (encoded by the CDKN1A gene) and cyclin D1 (encoded by the CCND1 gene) are important regulators of cell cycle progression and could have important effects in the complex process of neoplastic transformation. The current study aimed to investigate variants in CDKN1A (rs1801270, rs1059234) and CCND1 (rs9344) genes as potential risk and prognostic factors in oral squamous cell carcinoma (OSCC) patients. The study included 104 OSCC patients and 107 healthy individuals without a history of cancer. Genotypes were assessed by real-time PCR and TaqMan SNP genotyping. Significant differences in genotype distribution between OSCC cases and the control group were observed for the CCND1 rs9344 variant (p=0.017). According to the odds ratio (OR), adjusted for age and sex, the rs9344 heterozygous GA and homozygous mutated AA genotypes were associated with an increased OSCC susceptibility (OR=2.295, p=0.007; OR=2.029, p=0.037, respectively). Variants rs1801270 and rs105923 in CDKN1A were not associated with OSCC risk. There were no differences in overall survival among OSCC patients stratified by genotypes of the analyzed variants in CDKN1A and CCND1. Variant rs9344 in the CCND1 gene might be considered as a potential molecular risk factor for OSCC susceptibility but not for disease prognosis.

Background/Aim. The inflammatory response is of utmost importance in bone healing, but the precise role of cells and cytokines remains unclear. In our study, we examined the association between interleukin-1? (IL-1?), tumor necrosis factors alpha (TNF-?), monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) concentrations, fracture configuration, and callus formation. Methods. Serum cytokine concentrations were determined in 78 non-obese children with long bone fractures (group 1), 10 children with finger fractures (group 2), and 10 healthy controls (group 3). Blood samples were taken immediately after fracture upon hospital admission for groups 1 and 2. Differences in cytokine concentrations were analyzed among groups and catego-rized according to fracture configuration and callus formation. Results. IL-1? and TNF-? levels were lower in patients that went on to produce incomplete callus compared with patients that formed complete callus. Surprisingly, the average IL-1? concentration was highest in the healthy control group. The only significant correlation between IL-1? and TNF-? was in the group with moderate callus formation. MCP-1 level was slightly increased in all patient groups compared to controls, with no mutual difference. An average IL-8 level showed a clear decrease tendency from the group with incompletely formed callus toward the group with completely formed callus compared to controls, without significant difference. Children with epiphysiolysis had the lowest concentrations of cytokines compared with all other fracture types including transverse, oblique, and spiral. There were significantly lower concentrations of IL-1? and MCP-1 in patients with less fragment displacement compared with patients with greater fragment displacement. Conclusion. The systemic inflammatory response is important in physiological bone healing. High early production of IL-1?, TNF-?, and MCP-1 is associated with greater callus formation and better healing outcome, while increased IL-8 level is associated with poor callus formation and worse healing outcome. Our results indicate that epiphysiolysis and larger fragment displacement are associated with delayed fracture healing.

Background/Aim. Lung cancer (LC) is one of the leading causes of mortality. Disease progression and advanced disease are characterized by the unprotective immune response due to M2 macrophage polarization, myeloid-derived suppressor cells (MDSC) activity, cytokine imbalance, and regulatory T lymphocyte activity. The aim of this study was to investigate the association between Th1/Th2 cytokines and CD14+B7H4+ monocyte (Mo) number in LC patients. Methods. We investigated principal Th1/Th2 cytokines and CD14+B7H4+ Mo number in blood and tumor microcirculation samples of 41 LC patients (III and IV clinical stage) and 30 healthy participants (control group). Results. The serum concentrations of investigated cytokines in all patients vs. heathy controls did not differ significantly. Stratification in groups according to tumor histology, disease extent, and tumor size revealed significant differences. LC patients with different histology types demonstrated significant differences, both in serum and tumor microcirculation samples. The presence of metastasis was associated with increased IFN-?/IL-4 in blood and increased IL-13 in tumor microcirculation samples. Tumor microcirculation samples of the largest tumors were characterized by the Th2 cytokine profile. Investigation of CD14+B7H4+ Mo in blood samples demonstrated a significant association of extreme value of this cell population with elevated IL-2/IL-13. Patients with the highest CD14+B7H4+ Mo number in tumor microcirculation samples demonstrated significant increments of IL-4, IL-13, IL-10, and TGF-?1. Conclusion. LC patients demonstrated polarization of cytokine response associated with microenvironment origin, tumor histology type, tumor size, and disease extent. The highest number of CD14+ B7H4+ monocytes is significantly associated with the Th2 cytokine profile.

Background/Aim. Recent data imply the significance of certain cytokines in the appearance and development of prostate cancer (PC), as well as their association with pathohistological and/or clinical characteristics of PC. The aim of this study was to examine the relationship between the IL-10 concentration with histopathological and clinical characteristics of PC patients. Methods. IL-10 concentration was determined in serum, urine and prostate massage secret (pms) samples of 88 CP patients (initially and after therapy), 20 benign prostatic hyperplasia (BPH) patients and 15 healthy controls. Results. Compared to BPH and control groups, PC patients had the highest average serum IL-10 concentration. Interestingly, BPH patients demonstrated the highest concentration of IL-6 in urine and pms samples. Also, patients with G3 gradus and with the highest Gleason score (4 + 4) demonstrated the highest IL-10 serum level. PC patients without any histopathological sign of tumor invasion had a significantly increased serum IL-10, either before or after the therapy, compared to the patient group with evident invasion of tumor cells. The therapy induced different IL-10 profile in serum and urine samples. After the therapy, there was a clear significant IL-10 increase in serum of patients with unfavorable Gleason score (4 + 4), with present infiltration of tumor cells in peritumoral tissue (lymphatic, vascular and combined) and in patients with high tumor volume. Conclusion. PC patients without any histopathological signs of tumor invasion before the therapy have significantly increased serum IL-10 concentration compared to those with the signs of tumor invasion. There is a clear dissociation of IL-10 values between a serum sample and local, urine and pms samples from a particular patient. After the therapy, high IL-10 serum concentration is present only in patients with high Gleason score, pre-sent infiltration of peritumoral tissue and high tumor volume.

Triple-negative breast cancer (TNBC) is characterized by aggressive phenotype and a poorer prognosis compared to the estrogen and progesterone receptor positive, Her2 negative (ER + PR + Her2-) breast cancer. Increasing evidence suggests that sirtuins, a family of histone deacetylases, could have an important role in aggressiveness of TNBC's. The current study evaluated the potential clinical relevance of SIRT1, SIRT3 and SIRT6 gene expressions in two prognostically distinctive subtypes of breast cancer, the most aggressive TNBC and the least aggressive ER + PR + Her2- tumors. Total RNAs were isolated from 48 TNBC and 63 ER + PR + Her2- tumor samples. Relative gene expression was determined by SYBR Green RT-PCR and delta-delta Ct method, normalized to GAPDH. Mean gene expression of both SIRT1 and SIRT3 was significantly lower in the TNBC compared to ER + PR + Her2- tumors (p = 0.0001). Low SIRT1 and SIRT6 expressions associated with worse overall survival in ER + PR + Her2- patients (p = 0.039, p = 0.006, respectively), while TNBC patients with high SIRT1 tend to have a poor prognosis (p = 0.057). In contrast, high expression of SIRT3 in TNBC patients associated with higher histological grade (p = 0.027) and worse overall survival (p = 0.039). The Cox regression analysis revealed that low SIRT1 expression could be an independent prognostic marker of poor survival in ER + PR + Her2- breast cancers (HR = 11.765, 95% CI:1.234-100, p = 0.033). Observed differential expression of SIRT1, SIRT3 and SIRT6 genes in TNBC and ER + PR + Her2- subtypes, with opposite effects on patients' survival, suggests context-dependent mechanisms underlying aggressiveness of breast cancer. Further investigations are necessary to evaluate sirtuins as potential biomarkers and therapeutic targets in breast cancer.

Background/Aim. The group of genes, known as PAX (paired box), has a great role in organogenesis, as well as in maintaining the normal function of certain cells after the birth. In addition to these genes, the impact on the organogenesis, at the cellular level, has a transmembrane tetratricopeptid group of genes (TMTC). The term polymorphism in the human genome implies variations in the hereditary basis that occur in human populations, the presence of two or more different alleles of one genome in the population. The aim of the work was to determine whether there is an association of PAX3 and TMTC2 genes polymorphism with changes of the face morphology after skin tumor excision and direct suture closure. Methods. The study included 130 patients of both sexes, older than 50 years, with the medical indication for the elliptical surgical excision of the skin tumor. DNA was isolated from 5 mL of peripheral blood. Gene polymorphisms were analyzed with pre-designed single nucleotide polymorphisms (SNP) assays, by allelic discrimination method on REAL-TIME apparatus. The patients were subjected to a laser scanning preoperatively, and 7 and 90 days postoperatively, in order to obtain x, y and z coordinates of 5 cephalometric points on the face, which determined the shape of the medial cheek region. The shape of the medial cheek region, as well as the coordinates of 5 cepahlometric points, were compared among genotypes of both genes preoperatively, as well as 7 days and 90 days postoperatively. Results. A statistically significant difference in the shape of the medial cheek region between wildtype and mutant of PAX3 gene was found preoperatively, while the statistically significant difference in the shape of the medial cheek region was not found between wild-type and heterozygote, nor between wild-type and heterozygote and mutant of PAX3 gene, nor among genotypes of TMTC2 gene. Seven days and 90 days postoperatively, there were no statistically significant differences in the shape of the examined region among genotypes of both genes. Conclusion. Polymorphisms of PAX3 and TMTC2 genes are not associated with the change in the face morphology after the skin tumor excision and direct suture closure of the defect.

The aberrant DNA methylation plays a critical role in a number of different malignancies, including melanoma. DNA methylation is catalyzed by DNA methyltransferases (DNMTs), involved in methylation maintenance (DNMT1) and de novo DNA methylation (DNMT3A and DNMT3B). The current study investigated the association of genetic variants in the DNMT1 and DNMT3B with the clinicopathologic features and the clinical course of melanoma patients. In the present study, DNMT1 (rs2228612, rs2228611, and rs2114724) and DNMT3B (rs406193 and rs2424932) polymorphisms were examined in 123 melanoma patients. Single nucleotide polymorphisms were assessed using TaqMan SNPs Genotyping Assays according to the manufacturer's protocols. The carriers of the variant genotype of DNMT1 rs2228612 had poorer overall survival and recurrence-free survival, (P = 0.000 and 0.000, respectively), and an increased risk for adverse outcome [hazard ratio (HR) = 6.620, 95% confidence interval (CI): 2.214-19.791, P = 0.001]. DNMT1 rs2228612 was also associated with ulceration (P = 0.045), nodal status (P = 0.030), progression (P = 0. 007), and stage of disease (P = 0.003). Univariate analysis indicated that tumor-infiltrating lymphocytes could be a marker of good prognosis in melanoma patients (HR = 0.323, 95% CI: 0.127-0.855, P = 0.025), whereas the genotype distribution of the DNMT3B rs406193 polymorphism correlated significantly with the presence of tumor-infiltrating lymphocytes (P = 0.012). The multivariate analysis showed that the DNMT1 rs2228612 polymorphism (HR = 12.126, 95% CI: 2.345-62.715, P = 0.003) is an independent predictor of poor overall survival in melanoma patients. As expected, disease progression was also found to be an independent prognostic factor in melanoma patients (HR = 37.888, 95% CI: 3.615-397.062, P = 0.002). DNMT1 rs2228612 was found to be an independent predictor of poor overall survival in melanoma patients. DNMTs polymorphisms could serve as a potential target for novel therapeutic approaches.

A transplanted stem cell homing is a directed migration from the application site to the targeted tissue. Intrathecal application of stem cells is their direct delivery to cerebrospinal fluid, which defines the homing path from the point of injection to the brain. In the case of neurodegenerative diseases, this application method has the advantage of no blood–brain barrier restriction. However, the homing efficiency still needs improvement and homing mechanisms elucidation. Analysis of current research results on homing mechanisms in the light of intrathecal administration revealed a discrepancy between in vivo and in vitro results and a gap between preclinical and clinical research. Combining the existing research with novel insights from cutting-edge biochips, nano, and other technologies and computational models may bridge this gap faster.

Summary Association of vitamin D receptor (VDR) gene polymorphisms with sepsis risk and mortality was studied. VDR FokI CC genotype was associated with increased sepsis risk (OR = 13.396, p = .000009) compared to the TT genotype. Results suggest possible role of VDR FokI (rs2228570) as a molecular biomarker of increased sepsis risk.

Summary Background Vascular endothelial growth factor ( VEGF ), a key mediator of angiogenesis, is overexpressed in a wide variety of human cancers, including oral squamous cell carcinoma ( OSCC ). In this study, we examined whether individual polymorphisms within VEGF-A gene, rs699947 (−2578C/A), rs1570360 (−1154G/A), rs2010963 (−634G/C), rs3025039 (+936C/T) or their haplotypes are associated with an oral cancer risk and survival. Methods A case-control study was conducted on 114 OSCC patients and control group of 126 individuals without a previous cancer history, all the Caucasian race and the same ethnicity, matched by age and gender. VEGF-A genotypes were analyzed using TaqMan SNP Genotyping Assays, Applied Biosystems. Results The −1154 GG genotype was significantly associated with the decreased overall survival in OSCC patients ( p = 0.010, log rank test). Stratified analysis revealed that in patients with nodal metastases and stage III, −1154 GG genotype was related to worse survival, p = 0.009, p = 0.013, respectively. The multivariate analysis revealed that −1154 GG genotype is an independent adverse factor for survival in the OSCC (HR = 1.899, [1.138–3.168], p = 0.014). The +936 CC genotype was associated with advanced staged OSCC ( p = 0.050). The three polymorphisms, −2578, −1154 and −634 were in linkage disequilibrium (LD). The CAG haplotype could be associated with an increased oral cancer risk, OR = 7.967, [1.730–36.689], p = 0.008, while CGG haplotype could be associated with a decreased oral cancer risk, OR = 0.561, [0.326–0.964], p = 0.036. Conclusions VEGF-A −1154 GG genotype could be considered as a prognostic marker of poor survival in advanced-stage OSCC patients. Haplotypes of VEGF-A gene may be associated with susceptibility to OSCC .

J Oral Pathol Med (2012) 41: 779–787 Background: Genetic polymorphisms of vitamin D receptor gene (VDR) and genes involved in vitamin D metabolism pathway, CYP27B1 and CYP24B1, may affect individual susceptibility to oral squamous cell carcinoma. The purpose of this study was to evaluate the associations between VDR, CYP27B1 and CYP24A1 gene polymorphisms with oral cancer risk and survival. Methods: Study cohort consisted of 110 patients with oral cancer and 122 healthy controls. The genotypes of the analysed genes were determined by PCR–RFLP or real-time PCR method. Results: The significant decrease of oral cancer risk was observed in individuals with heterozygote genotype of CYP24A1 gene (rs2296241) (odds ratio 0.281, P = 0.000) in comparison with wild type. Patients with VDR FokI ff wild type genotype had significantly worse overall survival (P = 0.012, log rank) compared with heterozygous and mutated genotype combined. A stratified analysis by the lymph node involvement and tumour stage showed that ff is associated with poor survival in groups with and without lymph node involvement (P = 0.025, P = 0.040, respectively) and in stage III tumours (P = 0.026). Multivariate Cox’s regression analysis revealed that VDR FokI could be considered an independent prognostic factor. Conclusion: Our findings indicate that CYP24A1 gene polymorphism might have an influence on the susceptibility to oral cancer. VDR FokI polymorphism was associated with worse survival and could be considered as an independent prognostic marker.

Oral Diseases (2011) 17, 794–800 Objectives: Recent studies indicate various molecular abnormalities in oral squamous cell carcinomas (OSCC), including DNA methylation of tumor-associated genes. Although promoter hypermethylation of Wnt pathway antagonists RUNX3 (Runt-related transcription factor 3) and Wnt inhibitory factor 1 (WIF1) has been identified as a common event in a number of carcinomas, methylation status and the role of RUNX3 as a possible tumor suppressor in oral and head and neck cancer are yet controversial. The aim of our study is to determine the occurrence of RUNX3 and WIF1 genes hypermethylation and correlation with tumor and host-related factors and prognosis in tongue carcinomas. Material and Methods: In 76 patients with tongue carcinoma, RUNX3 and WIF1 genes promoter hypermethylation analysis was assessed by nested methylation-specific PCR method. Results: Hypermethylation of WIF1 and RUNX3 genes promoters was observed in 35% and 25% of carcinomas, respectively. RUNX3 gene promoter hypermethylation was significantly associated with lymph node involvement (P = 0.013) and tumor stage (P = 0.006), but not with the overall survival. Occurrence of RUNX3 and WIF1 genes comethylation was associated with nodal status (P = 0.058) and tumor stage (P = 0.055). Conclusions: Our findings indicate that RUNX3 and WIF1 are frequently aberrantly methylated and that RUNX3 promoter methylation could be considered as a potential prognostic marker in tongue carcinoma.

Adult mesenchymal stem cells (MSCs) have recently become a potent tool in regenerative medicine. Due to certain shortcomings of obtaining bone marrow MSCs, alternate sources of MSCs have been sought. In this work, we studied MSCs from dental pulp (DP-MSCs) and dental follicle (DF-MSCs), isolated from the same tooth/donor, to define differences in their phenotypic properties, differentiation potential, and immunomodulatory activities. Both cell types showed colony-forming ability and expressed typical MSCs markers, but differed in the levels of their expression. DF-MSCs proliferated faster, contained cells larger in diameter, exhibited a higher potential to form adipocytes and a lower potential to form chondrocytes and osteoblasts, compared with DP-MSCs. In contrast to DF-MSCs, DP-MSCs produced the transforming growth factor (TGF)-β and suppressed proliferation of peripheral blood mononuclear cells, which could be neutralized with anti-TGF-β antibody. The treatment with toll-like receptor 3 (TLR3) agonist augmented the suppressive potential of both cell types and potentiated TGF-β and interleukin-6 secretions by these cells. TLR4 agonist augmented the suppressive potential of DF-MSCs and increased TGF-β production, but abrogated the immunosuppressive activity of DP-MSCs by inhibiting TGF-β production and the expression of indolamine-2,3-dioxygenase-1. Some of these effects correlated with the higher expression of TLR3 and TLR4 by DP-MSCs compared with DF-MSCs. When transplanted in imunocompetent xenogenic host, both cell types induced formation of granulomatous tissue. In conclusion, our results suggest that dental MSCs are functionally different and each of these functions should be further explored in vivo before their specific biomedical applications.

The purpose of this study was to evaluate the associations of variability in pulp sensitivity with sex, psychosocial variables, the gene that encodes for the enzyme catechol-O-methyltransferase (COMT), and chronic painful conditions (temporomandibular disorders [TMDs]).The study was composed of 97 subjects (68 women and 29 men aged 20-44 years). The electric (electric pulp tester) and cold (refrigerant spray) stimuli were performed on mandibular lateral incisors. The results were expressed as pain threshold values for electric pulp stimulation (0-80 units) and as pain intensity scores (visual numeric scale from 0-10) for cold stimulation. The Research Diagnostic Criteria for TMD were used to assess TMD, depression, and somatization. DNA extracted from peripheral blood was genotyped for 3 COMT polymorphisms (rs4680, rs6269, and rs165774) using the real-time TaqMan method. Multivariate linear regression was used to investigate the joint effect of the predictor variables (clinical and genetic) on pulp sensitivity (dependent variables).Threshold responses to electric stimuli were related to female sex (P .01) and the homozygous GG genotype for the rs165774 polymorphism (P .05). Pain intensity to cold stimuli was higher in TMD patients (P .01) and tended to be higher in women. Multivariate linear regression identified sex and the rs165774 COMT polymorphism as the determinants of electric pain sensitivity, whereas TMD accounts for the variability in the cold response.Our findings indicate that sex/a COMT gene variant and TMD as a chronic painful condition may contribute to individual variation in electric and cold pulp sensitivity, respectively.

Summary Oral squamous cell carcinoma (OSCC) is characterized by high mortality rate and rising incidence. The aim of this study was to examine the methylation of particular tumor suppressor genes promoters in OSCC and to correlate the methylation status with the tumor-host features and patients survival. The genes selected for our investigation are involved in key cellular processes of malignant transformation, including cell cycle control (p16), apoptosis (Death Associated Protein Kinase, DAPK), Wnt signaling (Adenomatous Polyposis Coli, APC), cell–cell adhesion (E-cadherin, E-cad), and DNA repair (O6-methylguanine-DNA methyltransferase, MGMT, Werner syndrome gene, WRN). In 77 patients with OSCC, hypermethylation was determined by nested methylation-specific PCR method. Methylation of p16 gene promoter was detected in 58.4% of samples, E-cad in 42.9%, DAPK in 36.8%, MGMT in 33.8%, WRN in 23.8%, and APC in 18.2% of OSCC samples. Patients with E-cad promoter methylation had significantly worse overall survival (p = 0.039, log-rank test), while hypermethylation of other genes did not have prognostic significance. Stratified analysis by the lymph node involvement and tumor stage showed that E-cad promoter methylation is associated to poor survival in N positive (p = 0.024), and stage III tumors (p = 0.027), as an opposite to N0 and stage II tumors, where E-cad methylation did not have prognostic significance (p = 0.849, p = 0.794, respectively). Our findings indicate that multiple genes are frequently aberrantly methylated in OSCC, and that E-cad promoter methylation should be considered as a potential molecular marker for the poor survival in advanced OSCC.

Emerging studies suggest that dietary components can affect gene expression through epigenetic mechanisms. Epigenetic modifications are heritable and potentially reversible changes in gene expression that do not require changes in the DNA sequence. The main mechanisms of epigenetic control in mammals are DNA methylation, histone modifications, and RNA silencing. The potential reversibility of epigenetic changes suggests that they could be modulated by nutrition and bioactive food compounds. Thus, epigenetic modifications could mediate environmental signals and provide a link between susceptibility genes and environmental factors in the etiology of cancer. Elucidating the impact of nutrition on epigenetic mechanisms may serve as a tool to predict an individuals' susceptibility to cancer, provide dietary recommendations, or provide therapeutic applications of natural compounds against cancer. The optimal duration and the dose necessary for a chemopreventive effect require further studies. There is limited information about tissue specificity and temporal aspects of dietary treatments. Species differences need to be considered when interpreting results from various models. Importantly, molecular mechanisms of bioactive dietary components should be investigated in greater detail in human intervention studies. Although some of these issues remain controversial, this review mainly focuses on promising data that support the developing field of Nutritional Epigenetics.

MicroRNAs (miRNAs) are known to regulate most biological processes and have been found dysregulated in a variety of diseases, including multiple sclerosis (MS). In this study, we characterized miRNAs that associate with susceptibility to develop experimental autoimmune encephalomyelitis (EAE) in rats, a well-established animal model of MS. Using Illumina next-generation sequencing, we detected 544 miRNAs in the lymph nodes of EAE-susceptible Dark Agouti and EAE-resistant Piebald Virol Glaxo rats during immune activation. Forty-three miRNAs were found differentially expressed between the two strains, with 81% (35 out of 43) showing higher expression in the susceptible strain. Only 33% of tested miRNAs displayed differential expression in naive lymph nodes, suggesting that a majority of regulated miRNAs are EAE dependent. Further investigation of a selected six miRNAs indicates differences in cellular source and kinetics of expression. Several of the miRNAs, including miR-146a, miR-21, miR-181a, miR-223, and let-7, have previously been implicated in immune system regulation. Moreover, 77% (33 out of 43) of the miRNAs were associated with MS and other autoimmune diseases. Target genes likely regulated by the miRNAs were identified using computational predictions combined with whole-genome expression data. Differentially expressed miRNAs and their targets involve functions important for MS and EAE, such as immune cell migration through targeting genes like Cxcr3 and cellular maintenance and signaling by regulation of Prkcd and Stat1. In addition, we demonstrated that these three genes are direct targets of miR-181a. Our study highlights the impact of multiple miRNAs, displaying diverse kinetics and cellular sources, on development of pathogenic autoimmune inflammation.

Genetic polymorphisms of vitamin D receptor gene (VDR) and genes involved in vitamin D metabolism pathway, CYP27B1 and CYP24B1, may affect individual susceptibility to oral squamous cell carcinoma. The purpose of this study was to evaluate the associations between VDR, CYP27B1 and CYP24A1 gene polymorphisms with oral cancer risk and survival.Study cohort consisted of 110 patients with oral cancer and 122 healthy controls. The genotypes of the analysed genes were determined by PCR-RFLP or real-time PCR method.The significant decrease of oral cancer risk was observed in individuals with heterozygote genotype of CYP24A1 gene (rs2296241) (odds ratio 0.281, P = 0.000) in comparison with wild type. Patients with VDR FokI ff wild type genotype had significantly worse overall survival (P = 0.012, log rank) compared with heterozygous and mutated genotype combined. A stratified analysis by the lymph node involvement and tumour stage showed that ff is associated with poor survival in groups with and without lymph node involvement (P = 0.025, P = 0.040, respectively) and in stage III tumours (P = 0.026). Multivariate Cox's regression analysis revealed that VDR FokI could be considered an independent prognostic factor.Our findings indicate that CYP24A1 gene polymorphism might have an influence on the susceptibility to oral cancer. VDR FokI polymorphism was associated with worse survival and could be considered as an independent prognostic marker.

Objective. Several studies have suggested that the metabolism of alcohol is modulated by the polymorphisms in genes encoding ethanol-metabolizing enzymes, including alcohol dehydrogenase 1C, ADH1C, and cytochrome P450-dependent monooxygen- ase, CYP2E1. Genetic polymorphisms of ethanol-metabolizing enzymes may affect individual susceptibility to oral cancer. The purpose of this study was to investigate the associations between ADH1C and CYP2E1 gene polymorphisms with oral squamous cell carcinoma in an ethnically homogeneous Cau- casian population. Design. Case-control study. Setting. Serbian national general hospital. Subjects and Methods. The study was conducted on 123 oral cancer patients and a control group of 177 individuals of the Caucasian race and the same ethnicity, matched in age and gender, without previous cancer history. The control group consisted of 120 population-based and 57 hospital-based con- trols of heavy-drinking individuals. Genetic polymorphisms of ADH1C SspI, ADH1C HaeIII, CYP2E1 RsaI, and CYP2E1 Ins were determined by the polymerase chain reaction and restriction fragment length polymorphisms. Results. After adjustment by potential confounders, the significant increase of oral cancer risk, independent of alcohol drinking, was observed in individuals with the variant ADH1C SspI*2/*2 genotype (odds ratio, 3.029; P = .014) and in combined ADH1C SspI*1/*2 and ADH1C SspI*2/*2 genotypes (odds ratio, 2.605; P = .002), compared to the ADH1C*1/1* wild type. The associa- tion of other polymorphisms under study was not observed. Conclusion. This study suggested that the ADH1C SspI poly- morphism could play a significant role in the etiology of oral cancer, whereas ADH1C HaeIII, CYP2E1 RsaI, and CYP2E1 Ins could have minor influence.