Interleukin-33 and Osteoprotegerin Levels in Gingival Crevicular Fluid and Saliva in Chronic Periodontitis and Their Correlation to Diabetes Mellitus: A Cross-Sectional Study

Background: This study investigates the presence of interleukin (IL)-33 and osteoprotegerin (OPG) in saliva and gingival crevicular fluid (GCF) samples of patients with chronic periodontitis and whether or not they are correlated with diabetes mellitus. Methods: Sixty subjects were included in this study: 20 chronic periodontitis patients (CP), 20 diabetic chronic periodontitis patients (CPDM) and 20 systemically and periodontally healthy subjects. GCF and saliva samples were collected from all participants. Enzyme linked immunosorbent assay (ELISA) kits were used for assaying IL-33 and OPG levels. Analysis of variance was used to compare means of the CP and CP-DM groups with the control. Correlation analyses were performed to find the value of Spearman's correlation. A receiver operating characteristic (ROC) curve was constructed to determine the cut-off values of the markers to differentiate between the groups. Areas under the ROC curve (AUCs) were compared using z-statistic. Results: IL-33, in both GCF and saliva, was significantly higher in the CP and CP-DM groups compared to the control, and significantly higher in the CP-DM group compared to the CP group in saliva. On the other hand, the level of OPG in GCF and saliva was significantly lower in the CP and CP-DM groups compared to the control, but was of no significance when comparing the CP-DM and CP groups. Conclusion: IL-33 seems to play a role in the pathogenesis of periodontal disease, while OPG may have a protective function. Diabetes may affect and influence the expression of IL-33. Thus, they could be utilized as diagnostic biomarkers for chronic periodontitis either in saliva or GCF

Polymorphisms of IL-17A and IL-17F in Periodontal Disease: A Case-Control Study

Background: Increased interleukin-17 (IL-17) leads to the production of proinflammatory mediators and increases local inflammation. Interleukin-17 may also promote receptor activator of nuclear factor kappa-B ligand (RANKL) expression on gingival fibroblasts, T cells, and B cells, resulting in alveolar bone resorption. Interleukin-17A and IL-17F levels in saliva and gingival crevicular fluid (GCF), were found to be elevated in periodontitis patients. Thus, IL-17A and IL-17F polymorphisms were hypothesized to be associated with a risk of periodontitis. Methods: The present study was conducted on 60 subjects, including 20 stage II grade B periodontitis patients, 20 stage III grade C periodontitis patients, and 20 healthy controls. Blood samples were drawn from the subjects and analyzed for IL-17A G-197A and IL-17F 7488T/C genetic polymorphisms using the TaqMan assay. Results: There was a significant statistical difference between the distribution of the different genotypes and the different alleles in the three groups for IL-17A G-197A with the A allele presence indicating a risk of periodontitis. Conclusions: Interleukin-17A G-197A polymorphism is significantly associated with different clinical forms of periodontitis in the Egyptian population. The A allele could be considered a risk factor for periodontal diseases

Study of microRNAs-21/221 as potential breast cancer biomarkers in Egyptian women

microRNAs (miRNAs) play an important role in cancer prognosis. They are small molecules, approximately 17–25 nucleotides in length, and their high stability in human serum supports their use as novel diagnostic biomarkers of cancer and other pathological conditions. In this study, we analyzed the expression patterns of miR-21 and miR-221 in the serum from a total of 100 Egyptian female subjects with breast cancer, fibroadenoma, and healthy control subjects. Using microarray-based expression profiling followed by real-time polymerase chain reaction validation, we compared the levels of the two circulating miRNAs in the serum of patients with breast cancer (n = 50), fibroadenoma (n = 25), and healthy controls (n = 25). The miRNA SNORD68 was chosen as the housekeeping endogenous control. We found that the serum levels of miR-21 and miR-221 were significantly overexpressed in breast cancer patients compared to normal controls and fibroadenoma patients. Receiver Operating Characteristic (ROC) curve analysis revealed that miR-21 has greater potential in discriminating between breast cancer patients and the control group, while miR-221 has greater potential in discriminating between breast cancer and fibroadenoma patients. Classification models using k-Nearest Neighbor (kNN), Naïve Bayes (NB), and Random Forests (RF) were developed using expression levels of both miR-21 and miR-221. Best classification performance was achieved by NB Classification models, reaching 91% of correct classification. Furthermore, relative miR-221 expression was associated with histological tumor grades. Therefore, it may be concluded that both miR-21 and miR-221 can be used to differentiate between breast cancer patients and healthy controls, but that the diagnostic accuracy of serum miR-21 is superior to miR-221 for breast cancer prediction. miR-221 has more diagnostic power in discriminating between breast cancer and fibroadenoma patients. The overexpression of miR-221 has been associated with the breast cancer grade. We also demonstrated that the combined expression of miR-21 and miR-221can be successfully applied as breast cancer biomarkers

ACE gene polymorphism and serum ACE level with Progression of Nephropathy in Type 2 Diabetic Patients

Background. One of the most common complications of diabetes mellitus (DM) is diabetic nephropathy (DN).  Angiotensin- converting enzyme (ACE) gene was the first candidate gene of renin-angiotensin system (RAS) for predisposition to DN.Objective. Investigation whether the ACE insertion/deletion (I/D) polymorphism is associated with Egyptian type 2 diabetic patients (T2DM) with nephropathy. In addition, the study investigated the relationship between variants of ACE I/D gene polymorphism and serum ACE level and the progression of nephropathy in Egyptian T2DM patients.Methods.  A total of 85 T2DM patients (45 with nephropathy and 40 without nephropathy) besides 45 healthy (non-diabetic) age-matched subjects were recruited in this study for comparison. The (I/D) polymorphism of the ACE gene was investigated using PCR and serum ACE levels were determined using ELISA.Results. The frequency of ACE DD genotype and D allele was significantly higher in DN patients when compared to control healthy subjects and diabetic patients without nephropathy. In addition our results showed a significant association between DD genotype of ACE gene and elevated serum ACE level.Conclusion. The present study showing a strong association between the D allele and/or DD homozygous of ACE gene and diabetic patients developed nephropathy. In addition, individuals with D allele have higher levels of serum ACE compared to those having I allele. ACE gene polymorphism and serum ACE level may serve as a susceptibility biomarker for nephropathy in type 2 diabetic patients.Â

Galectin-1 (Gal-1) and Galectin-3 (Gal-3) levels in seminal plasma and serum in azoospermic patients versus fertile men: A cross-sectional study

Introduction: Galectin-1 (Gal-1) and galectin-3 (Gal-3) are expressed by many immune cells and receive considerable attention in the context of immunity. We aimed to compare between seminal plasma and serum levels of Gal-1 and Gal-3 in azoospermic patients and fertile men. Materials and methods: This cross-sectional study was conducted at the andrology outpatient clinic from January (2022) to September (2022). A total of 90 participants were enrolled and divided into two equal groups: azoospermic and normal group. Semen analysis was done for all participants. Hormonal profile including FSH, LH, serum prolactin, total testosterone and estradiol was performed as well as assessment of serum and seminal levels of Gal-1 and Gal-3 by ELISA commercial kits. Finally, scrotal Duplex was done in standing and supine position. Results: Serum and seminal levels of Gal-1 and Gal-3 were statistically significant higher in azoospermic patients compared with normal individuals (p < 0.001 for all). In addition, in healthy individuals there were statistically significant positive correlations between serum levels of Gal-1 and age, FSH, LH levels (r = 0.296, p = 0.005; r = 0.333, p = < 0.001; r = 0.312, p = 0.003, respectively) and serum levels of Gal-2 and FSH and LH (r = 0.436, p < 0.001; r = 0.350, p < 0.001, respectively), whereas serum Gal-3 showed a borderline positive correlation with age (r = 0.2, p = 0.059). Additionally, statistically significant positive correlations between seminal levels of Gal-1 and Gal-3 and free testosterone in healthy individuals were reported (r = 0.205, p = 0.053; r = 0.219, p = 0.038, respectively). On the other hand, there were negative correlations between serum and seminal levels of Gal-1 and Gal-3, total and progressive sperm motility, sperm count and abnormal sperm forms in healthy individuals (r = -0.382, p < 0.001; r = -0.405, p < 0.001; r = -0.376, p < 0.001; r = -0.364, p < 0.001) (r = -0.394, p < 0.001; r = -0.467, p < 0.001; r = -0.413, p < 0.001; r = -0.433, p < 0.001); (r = -0.372, p < 0.001; r = -0.377, p < 0.001; r = -0.317, p = 0.002; r = -0.311, p = 0.003)(r = -0.445, p < 0.001; r = -0.498, p < 0.001; r = -0.453, p < 0.001; r = -0.463, p < 0.001, respectively). Furthermore, statistically significant positive correlations between serum levels of Gal-1 and Gal-3 and age in azoospermic patients were reported (r = 0.511, p < 0.001; r = 0.390, p = 0.008, respectively). On the other hand, there were negative correlations between seminal Gal-1 and estradiol (E2) and seminal Gal-3 and FSH and LH in azoospermic patients (r= -0.318, p = 0.033; r = -0.322, p = 0.031; r = -0.477, p < 0.001, respectively). Also, negative correlations between serum Gal-3 and total and free testosterone in azoospermic patients were detected (r = -0.396, p = 0.007; r = -0.375, p = 0.011, respectively). Conclusions: Elevated serum and seminal levels of Gal-1 and Gal-3 have detrimental effects on spermatogenesis. Furthermore, the current study demonstrated potential regulatory effects of reproductive hormones on Gal-1 and Gal-3. Thus, future studies are needed to confirm such findings

Association of MiRNA-146a, MiRNA-499, IRAK1 and PADI4 Polymorphisms with Rheumatoid Arthritis in Egyptian Population

Background/Aims: Rheumatoid arthritis (RA) is a systemic autoimmune disease affecting up to 1% of the population worldwide. The aim of the present study was to investigate whether miRNA-146a rs2910164, miRNA-499 rs3746444, IRAK1 rs3027898 and PADI4 rs1748033 polymorphisms are associated with susceptibility to RA in Egyptians and whether they influence disease severity and activity. Methods: The study was performed on 104 unrelated RA patients and 112 healthy subjects. RA patients were further subdivided into active and inactive RA groups. Polymorphisms were genotyped by using real-time polymerase chain reaction with TaqMan allelic discrimination assay. Results: Significant differences in the frequency of miRNA-146a rs2910164, miRNA-499 rs3746444, IRAK1 rs3027898 and PADI4 rs1748033 alleles and genotypes were observed between RA patients and controls. Only CA and AA genotypes of IRAK1 rs3027898 shows a significant difference between active and inactive subgroups. MiRNA-146a rs2910164 and IRAK1 rs3027898 polymorphisms were a risk factor for predisposition to RA in codominant and dominant tested inheritance models, while, the miRNA-499 rs3746444 and PADI4 rs1748033 polymorphisms were a risk factor in codominant and recessive one. CG and GG genotypes of miRNA-146a rs2910164 were associated with positive erosions. CA genotype of IRAK1 rs3027898 was associated with low disease activity and negative erosions, while, the AA genotype was associated with high disease activity. CC genotype of PADI4 rs1748033 was associated with negative rheumatoid factor. Conclusion: The 4 studied SNPs were likely to play an important role in the susceptibility to RA and can influence disease severity and activity in Egyptian population

Evaluation of serum and salivary transforming growth factor beta, vascular endothelial growth factor and tumor necrosis factor alpha in oral lichen planus.

Introduction: Lichen planus is one of the most common oral mucosal lesions. Transforming growth factor-β (TGF- β) has a marked effect on epithelial–mesenchymal transition and immune cells function. Vascular Endothelial Growth Factor (VEGF) is a key regulator of vasculogenesis and angiogenesis. Tumor necrosis factor-α (TNF-α) mediates T-lymphocyte homing and apoptosis of epithelial cells. Objetive: The present study was conducted in order to compare the expression of serum and salivary TGF- β, VEGF, TNF-α between OLP patients and control individuals to investigate if saliva can be used as an alternative to serum for diagnostic purposes and for monitoring disease. Materials and Methods: 23 OLP patients and 23 control individuals were included to evaluate serum and salivary TGF-β, VEGF, TNF-α using ELISA kits. Five milliliters of venous blood was collected and unstimulated saliva was collected by the spitting method. Results: Serum and salivary levels of TGF- β, VEGF, TNF-α are higher in OLP patients compared to normal controls. Mean difference is higher in saliva than serum. Moreover, there was a significant difference in serum and salivary VEGF and TNF-α between symptomatic and asymptomatic groups. Conclusions: Saliva can be a used as a substitute for serum to evaluate levels of the assessed biomarkers.Introducción: El liquen plano oral es una de las lesiones de la mucosa oral más comunes. El factor de crecimiento transformante β (TGF-β) tiene un efecto marcado sobre la transición epitelial-mesenquimal y la función de las células inmunes. El factor de crecimiento endotelial vascular (VEGF) es un regulador clave de la vasculogénesis y la angiogénesis. El factor de necrosis tumoral α (TNF-α) media la localización de los linfocitos T y la apoptosis de las células epiteliales. Objetivo: El presente estudio se realizó con el fin de comparar la expresión en suero y saliva de TGF-β, VEGF, TNF-α entre pacientes con OLP y personas de control para investigar si la saliva se puede utilizar como alternativa al suero para fines de diagnóstico y monitoreo de la enfermedad. Material y Métodos: Se incluyeron 23 pacientes con OLP y 23 individuos control para evaluar los niéveles en suero y en saliva de TGF-β, VEGF, TNF-α utilizando kits ELISA. Se recogieron cinco mililitros de sangre venosa y se recogió saliva no estimulada por el método de escupir. Resultado: Los niveles séricos y salivales de TGF-β, VEGF, TNF-α son más altos en pacientes con OLP en comparación con los controles normales. La diferencia media es mayor en saliva que en suero. Además, hubo una diferencia significativa de VEGF y TNF-α en suero y saliva entre los grupos sintomáticos y asintomáticos. Conclusion: La saliva puede usarse como un sustituto del suero para evaluar los niveles de los biomarcadores estudiados

Serum interferon-related microRNAs as biomarkers to predict the response to interferon therapy in chronic hepatitis C genotype 4.

MicroRNAs are messengers during interferon-virus interplay and are involved in antiviral immunity, however, little is known about interferon-related microRNAs regarding their detection in serum and their potential use as non-invasive diagnostic and prognostic biomarkers in chronic hepatitis C (CHC). To elucidate some of the molecular aspects underlying failure of pegylated interferon-α/ribavirin therapy, we investigated pretreatment expression profiles of seven selected interferon-related microRNAs (miR-146a, miR-34a, miR-130a, miR-19a, miR-192, miR-195, and miR-296) by quantitative RT-PCR custom array technology in serum of Egyptian CHC genotype 4 patients and whether their pretreatment levels would predict patient response to the combination therapy. One hundred and six CHC patients and forty matched healthy controls were included. Patients were divided into sustained virological response (SVR) and non-responder (NR) groups. Serum miR-34a, miR-130a, miR-19a, miR-192, miR-195, and miR-296 were upregulated, whereas serum miR-146a was downregulated in CHC compared to controls. Significant correlations were found between expression levels of studied microRNAs and also with clinical data. Pretreatment levels of miR-34a, miR-130a, and miR-195 were significantly higher, whereas miR-192 and miR-296 levels were significantly lower in SVR than NR patients. miR-19a and miR-146a levels were not significantly different between the two groups. miR-34a was superior to differentiate CHC from controls, whereas miR-296 was superior to discriminate SVR from NR patients by receiver operating characteristic analysis. Multivariate logistic analysis revealed miR-34a and miR-195 as independent predictors for SVR and miR-192 as an independent variable for non-response. In conclusion, pretreatment expression profiles of five interferon-related microRNAs are associated with treatment outcome in CHC. Of these, miR-34a, miR-195, and miR-192 could predict treatment response. The profiling results could be used as novel non-invasive diagnostic and prognostic pharmacogenetic biomarkers for treatment personalization in CHC and could help to identify new microRNA-based antivirals

Single Nucleotide Polymorphism in SMAD7 and CHI3L1 and Colorectal Cancer Risk

Colorectal cancer (CRC) is one of the leading cancers throughout the world. It represents the third most common cancer and the fourth in mortality. Most of CRC are sporadic, arise with no known high-penetrant genetic variation and with no previous family history. The etiology of sporadic CRC is considered to be multifactorial and arises from the interaction of genetic variants of low-penetrant genes and environmental risk factors. The most common well-studied genetic variation is single nucleotide polymorphisms (SNPs). SNP arises as a point mutation. If the frequency of the sequence variation reaches 1% or more in the population, it is referred to as polymorphism, but if it is lower than 1%, the allele is typically considered as a mutation. Lots of SNPs have been associated with CRC development and progression, for example, genes of TGF-β1 and CHI3L1 pathways. TGF-β1 is a pleiotropic cytokine with a dual role in cancer development and progression. TGF-β1 mediates its actions through canonical and noncanonical pathways. The most important negative regulatory protein for TGF-β1 activity is termed SMAD7. The production of TGF-β can be controlled by another protein called YKL-40. YKL-40 is a glycoprotein with an important role in cancer initiation and metastasis. YKL-40 is encoded by the CHI3L1 gene. The aim of the present review is to give a brief introduction of CRC, SNP, and examples of some SNPs that have been documented to be associated with CRC. We also discuss two important signaling pathways TGF-β1 and CHI3L1 that influence the incidence and progression of CRC