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    Differential expression of inflammasome regulatory transcripts in periodontal disease
    (Wiley, 2020) Aral, Kubra; Berdeli, Eynar; Cooper, Paul Roy; Milward, Michael Robert; Kapila, Yvonne; Unal, Beyza Karadede; Aral, Cuneyt Asim
    Background The inflammasome modulates the release of key proinflammatory cytokines associated with periodontal disease pathogenesis. The aim of this study was to evaluate the expression of proteins that regulate the inflammasome, namely pyrin domain-only proteins (POPs), caspase activation recruitment domain (CARD)-only proteins, and tripartite motif-containing (TRIM) proteins, in periodontal diseases. Methods A total of 68 participants (34 males and 34 females) were divided into four groups, including periodontal health (H), gingivitis (G), chronic periodontitis (CP), and aggressive periodontitis (AgP) based on clinical parameters. Gingival tissue samples were obtained from all participants for reverse transcription polymerase chain reaction (RT-PCR)-based gene expression analyses of molecules that regulate the inflammasome, including apoptosis-associated speck-like protein (ASC) containing CARD, caspase-1, interleukin-1 beta (IL-1 beta), interleukin-18 (IL-18), nucleotide-binding domain, leucine rich family (NLR) pyrin domain containing 3 (NLRP3), NLR family pyrin domain containing 2 (NLRP2), AIM2 (absent in melanoma 2), POP1, POP2, CARD16, CARD18, TRIM16, and TRIM20 by RT-PCR. Results NLRP3 and IL-1 beta were upregulated in the G, CP, and AgP groups compared with group H (P < 0.05). AIM2 was downregulated in the CP group compared with the H, G, and AgP groups (P < 0.05). TRIM20, TRIM16, and CARD18 were downregulated in the G, CP, and AgP groups compared with the H group (P < 0.05). POP1 and POP2 were downregulated in the CP and AgP, and AgP and G groups, respectively (P < 0.05). Conclusion Active periodontal disease may result in downregulation of inflammasome regulators that may increase the activity of NLRP3 and IL-1 beta in periodontal disease.
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    Evaluation of psychological stress and cortisol levels in males with and without gingivitis
    (2020) Aral, Kubra; Milward, Michael R; Cooper, Paul R; Celik Guler, Ozge
    Aim: Gingivitis is a prevalent health problem seen most commonly in males. Psychological stress is also associated with periodontal disease. Thus, to evaluate the possible role of stress in gingivitis, this study investigated the psychological stress and cortisol levels in males with and without gingivitis. Material and Methods: Sixty systemically healthy males between 18 and 28 years were divided into two groups, including those with generalized gingivitis (G) (n=30) and periodontally healthy controls (H) (n=30). Gingival crevicular fluid (GCF) and saliva samples were obtained for determination of cortisol levels using enzyme-linked immunosorbent assay. Plaque (PI) and gingival index (GI), bleeding on probing (BOP), probing depth (PD) and clinical attachment level (CAL) were noted. In addition, BECK depression inventory (BDI), Perceived Stress Scale (PSS), and Oral Health Impact Profile-14 (OHIP-14) were recorded from all participants. Results: PI, GI, BOP (p0.01), and PD (p0.05). Cortisol levels in saliva and GCF were similar between groups (p>0.05). Conclusion: Psychological stress and cortisol levels in GCF and saliva were found to be similar in males with and without gingivitis.
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    Evaluation of the role of mitofusin-1 and mitofusin-2 in periodontal disease
    (Wiley, 2024) Kirmiziguel, Omer Alperen; Sabanci, Arife; Disli, Faruk; Yildiz, Sedat; Milward, Michael R.; Aral, Kubra
    BackgroundMitochondria and endoplasmic reticulum are key cellular organelles and create contact sites (mitochondria-endoplasmic reticulum contact [MERC]), which plays a major role in calcium metabolism, apoptotic processes, and inflammation. Previously, proteins that have been associated with these MERC contact sites mitofusin-1 (MFN1) and mitofusin-2 (MFN2) have been found to be downregulated in periodontal disease in vitro. Therefore, the aim of the current study was to evaluate MFN1 and MFN2 in gingival crevicular fluid (GCF) of patients with periodontal disease compared with healthy controls clinically. MethodsA total of 48 participants were divided into three groups including periodontally healthy (n = 16), patients with gingivitis (n = 16), and patients with stage 3 grade B periodontitis (n = 16). GCF levels of MFN1, MFN2, calcium (Ca), caspase-1, and tumor necrosis factor-alpha (TNF-& alpha;) were determined via enzyme-linked immunosorbent assay (ELISA). Results were calculated as total amount and concentration. ResultsMFN1 levels (total amount) were significantly higher in patients with periodontitis and gingivitis when compared with healthy controls (p < 0.05). However, concentration levels of MFN1, MFN2, Ca, caspase-1, TNF-& alpha; significantly decreased in periodontal disease groups compared with healthy controls (p < 0.05). A positive correlation was detected among all evaluated markers (p < 0.05). ConclusionThe MERC protein MFN1 may have a role in the pathogenesis of periodontal disease due to its increase in GCF of patients with periodontitis and gingivitis.
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    Gene expression profiles of mitochondria-endoplasmic reticulum tethering in human gingival fibroblasts in response to periodontal pathogens
    (Pergamon-Elsevier Science Ltd, 2021) Aral, Kubra; Milward, Michael R.; Cooper, Paul R.
    Objective: The current study aimed to elucidate the potential involvement of mitochondria-endoplasmic reticulum contact genes in the pathogenesis of periodontal disease by monitoring levels of contact associated genes including Mitofusion 1 (MFN1) and MFN2, inositol 1,4,5-trisphosphate receptor (IP3R), chaperone glucoseregulated protein 75 (GRP75), sigma non-opioid intracellular receptor 1 (SIGMAR1) and phosphate and tensin homolog induced putative kinase 1 (PINK1) in human gingival fibroblasts in response to periodontal pathogens Fusobacterium nucleatum (F. nucleatum) and Porphyromonas gingivalis (P. gingivalis) in vitro. Design: Primary human gingival fibroblasts were exposed to live cultures of P. gingivalis (W83; ATCC BAA-308) and F. nucleatum (subsp. Polymorphum; ATCC 10953) alone or in combination for 4 h at a 50 or 200 multiplicity of infection. Escherichia coli lipopolysaccharide (10 mu g/mL) exposure was used as a positive control. Gene expression levels of contact genes (MFN1, MFN2, IP3R, GRP75, SIGMAR1 and PINK1) as well as a proinflammatory cytokine, Tumor necrosis factor-alpha (TNF-alpha), and the apoptosis associated gene, Immediate early response 3 (IER3), were evaluated by reverse transcription polymerase chain reaction analysis. Results: MFN1, GRP75, IP3R and PINK1 were significantly upregulated by P. gingivalis with or without F. nucleatum. Only P. gingivalis with F. nucleatum caused a significant upregulation of SIGMAR1. TNF-alpha and IER3 gene expression positively correlated with the contact-associated gene expression changes. Conclusion: F. nucleatum and P. gingivalis alone or in combination may differentially dysregulate the gene expression levels of contact-associated genes in human gingival fibroblasts. These host-microbiome interactions may mechanistically be important in the pathogenesis of periodontal disease.
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    Oxidative stress, neutrophil elastase and IGFBP7 levels in patients with oropharyngeal cancer and chronic periodontitis
    (Wiley, 2020) Aral, Cuneyt Asim; Olcer, Sude Nur; Aral, Kubra; Kapila, Yvonne
    Objective The present study focused on investigating levels of oxidative stress, neutrophil elastase (NE), and insulin-like growth factor-binding protein 7 (IGFBP7) in oropharyngeal cancers (OC) with the presence and absence of periodontitis. Materials and Methods A healthy non-periodontitis group (H-NP; n = 20), a systemically healthy chronic periodontitis group (H-P; n = 20), a non-periodontitis group with OC (OC-NP; n = 12), and a chronic periodontitis group with OC (OC-P; n = 16) formed the study groups. The levels of NE and IGFBP7 were measured in gingival crevicular fluid (GCF) and saliva. In addition, oxidative status was determined by evaluating total oxidant status (TOS), total antioxidant status (TAS), and OSI (TOS/TAS). Results Gingival crevicular fluid NE was higher in all the groups compared with the H-NP group (p < .01). Salivary NE was higher in the OC-P and H-P groups compared with the H-NP and OC-NP groups (p < .05). Salivary IGFBP7 was significantly higher in the OC-NP and OC-P groups compared with the H-NP and H-P groups (p < .001). GCF TOS and OSI levels were significantly higher in all groups compared with the H-NP group (p < .05). Conclusions Gingival crevicular fluid NE levels were lower in healthy conditions compared with periodontal disease and OC. Salivary NE levels were higher in periodontal disease compared to states with no periodontal disease. Salivary IGFBP7 levels were higher in OC. Further analyses may help determine whether high salivary IGFBP7 levels distinguish OC from healthy conditions.
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    Six-month clinical outcomes of non-surgical periodontal treatment with antibiotics on apoptosis markers in aggressive periodontitis
    (Wiley, 2019) Aral, Kubra; Aral, Cuneyt A.; Kapila, Yvonne
    Objective Extrinsic and intrinsic pathways of apoptosis are involved in generalized aggressive periodontitis (GAgP). The aim of this study was to evaluate 6-month clinical outcomes relative to apoptosis of one-stage full-mouth disinfection (OSFMD) and systemic antibiotics (SA) in the treatment of GAgP. Methods Twenty-six patients with GAgP were included in this prospective follow-up intervention study. Gingival crevicular fluid (GCF) was collected from patients at baseline and 3 and 6 months after periodontal therapy, which consisted of OSFMD and SA (amoxicillin and metronidazole, 500 mg each for 7 days). The levels of p53, caspase-3, TNF-alpha, TRAIL, IL-1 beta, and IL-10 in GCF were measured via ELISA. Results Periodontal parameters were improved at 3 and 6 months compared to baseline (p < 0.05). p53 was decreased up to 6 months (p < 0.05). TRAIL, TNF-alpha, and IL-10 were similar at baseline and 3 and 6 months. Caspase-3 and IL-1 beta were decreased at 3 months (p < 0.05), but similar at 6 months compared to baseline (p > 0.05). Conclusion Although OSFMD plus SA improves clinical periodontal parameters up to 6 months, this treatment protocol differentially regulates the apoptosis markers caspase-3 at 3 months and p53 at 6 months without influencing TRAIL in GCF.