Humic Acid, a Polyphenolic Substance, Decreases Alveolar Bone Loss in Experimental Periodontitis in Rats.

The purpose of this study was to evaluate the biochemical, morphometric, and histopathological changes associated with experimental periodontitis in rats in response to local administration of humic acid. Thirty-eight Wistar rats were divided into 5 experimental groups: nonligated (NL) group, ligature-only (LO) group, and ligature + local administration of humic acid (20, 80, and 150 mg/kg body weight per day for 15 days, respectively; L-20, L-80, and L-150 groups). Changes in alveolar bone levels were clinically measured as the distance from the cementoenamel junction to the alveolar bone crest with a stereomicroscope. Tissues were histopathologically examined to assess the osteoclast numbers, osteoblastic activity, and inflammatory cell infiltration among the study groups. Enzyme-linked immunosorbent assay interleukin1ß (IL-1ß) and IL-10 levels in serum and gingival homogenates were evaluated. At the end of 15 days, the alveolar bone loss was significantly higher in the LO group compared to the NL, L-20, and L-150 groups (P < .05). The osteoclast number in the LO group was significantly higher than the NL, L-20, and L-150 groups (P < .05). Inflammatory cell infiltration was significantly higher in the LO and L-80 groups than the other groups (P < .05). The highest serum and gingival homogenate IL-10 levels were determined in the NL group (P < .05). The serum and gingival homogenate IL-1ß levels in LO group were significantly higher than the NL, L-20, and L-150 groups (P < .05). Within the limits of this study, it can be suggested that humic acid, when administered locally at 20 and 80 mg/kg doses, may prevent alveolar bone loss in the rat model.

The effects of IL-10 gene polymorphism on serum, and gingival crevicular fluid levels of IL-6 and IL-10 in chronic periodontitis.

OBJECTIVE: Anti-inflammatory cytokines play a crucial role in periodontitis by inhibiting synthesis of pro-inflammatory cytokines. The purpose of this study was to evaluate the effect of interleukin-10 (-597) gene polymorphism and genotype distributions on chronic periodontitis (CP) development and IL-6 and IL-10 levels in gingival crevicular fluid (GCF) and serum before and after non-surgical periodontal treatment. MATERIAL AND METHODS: The study population consisted of 55 severe generalized CP patients as CP group and 50 healthy individuals as control group. Plaque index, gingival index, probing depth and clinical attachment level were recorded and GCF and blood samples were taken at both the baseline and the sixth week after non-surgical periodontal treatment. PCR-RFLP procedure was used for gene analyses and cytokine levels were measured via ELISA. RESULTS: IL-10 genotype distribution was significantly different between CP and control groups (p=0.000, OR:7, 95%CI, 2.83-60.25). Clinical measurements significantly improved in the CP group after periodontal treatment (p<0.05). Periodontal treatment significantly decreased GCF IL-6 and IL-10 levels. No significant difference was found in clinical parameters between IL-10 AA and AC+CC genotypes at both the baseline and the sixth week (p>0.05). Sixth week GCF IL-10 levels were significantly lower in patients carrying IL-10 AC+CC genotype compared to the patients carrying IL-10 AA genotype (p<0.05). Serum IL-6 and IL-10 levels were lower in patients carrying the IL-10 AA genotype compared to patients with IL-10 AC+CC genotype, but the difference was not significant (p>0.05). CONCLUSION: IL-10 AA genotype carriers had lower IL-6 and IL-6/10 levels in serum; however, GCF IL-6/10 levels were similar in both genotypes. Within the limitations of our study, a possible association between IL-10(-597) gene polymorphism and CP might be considered.

The effects of IL-10 gene polymorphism on serum, and gingival crevicular fluid levels of IL-6 and IL-10 in chronic periodontitis

Abstract Objective Anti-inflammatory cytokines play a crucial role in periodontitis by inhibiting synthesis of pro-inflammatory cytokines. The purpose of this study was to evaluate the effect of interleukin-10 (-597) gene polymorphism and genotype distributions on chronic periodontitis (CP) development and IL-6 and IL-10 levels in gingival crevicular fluid (GCF) and serum before and after non-surgical periodontal treatment. Material and Methods The study population consisted of 55 severe generalized CP patients as CP group and 50 healthy individuals as control group. Plaque index, gingival index, probing depth and clinical attachment level were recorded and GCF and blood samples were taken at both the baseline and the sixth week after non-surgical periodontal treatment. PCR-RFLP procedure was used for gene analyses and cytokine levels were measured via ELISA. Results IL-10 genotype distribution was significantly different between CP and control groups (p=0.000, OR:7, 95%CI, 2.83-60.25). Clinical measurements significantly improved in the CP group after periodontal treatment (p<0.05). Periodontal treatment significantly decreased GCF IL-6 and IL-10 levels. No significant difference was found in clinical parameters between IL-10 AA and AC+CC genotypes at both the baseline and the sixth week (p>0.05). Sixth week GCF IL-10 levels were significantly lower in patients carrying IL-10 AC+CC genotype compared to the patients carrying IL-10 AA genotype (p<0.05). Serum IL-6 and IL-10 levels were lower in patients carrying the IL-10 AA genotype compared to patients with IL-10 AC+CC genotype, but the difference was not significant (p>0.05). Conclusion IL-10 AA genotype carriers had lower IL-6 and IL-6/10 levels in serum; however, GCF IL-6/10 levels were similar in both genotypes. Within the limitations of our study, a possible association between IL-10(-597) gene polymorphism and CP might be considered.

IL-6 and IL-10 gene polymorphisms in patients with aggressive periodontitis: effects on GCF, serum and clinic parameters.

Genetic variations observed in cytokines affect periodontitis susceptibility. The aim of this study was to investigate interleukin(IL)-6(-174) and IL-10(-597) gene polymorphisms in generalized aggressive periodontitis (GAgP) patients. Also, we aimed to evaluate the effects of IL-6 and IL-10 gene polymorphisms on the clinical outcomes of non-surgical periodontal therapy and cytokine levels in gingival crevicular fluid(GCF) and serum. Fifty-three patients with GAgP and 50 periodontally healthy individuals were included in this study. Clinical parameters, GCF and blood samples were collected at baseline and at 6-week. Non-surgical periodontal therapy was performed in patients with GAgP. Gene analysis were determined by PCR-RFLP(polymerase chain reaction-restriction fragment length polymorphism) and cytokine levels were determined by enzyme-linked immunosorbent assay(ELISA).GAgP patients showed significant improvement on clinical parameters after periodontal therapy(p<0.05). In the GAgP group, IL-6 GG genotype and G allele frequency were higher than in the control group. GCF IL-6 level was also significantly lower at 6-week in the GAgP group. Higher GCF IL-10 levelswere observed in patients carrying the IL-6 GG genotype than in those carrying the GC+CC genotype at baseline. In conclusion, IL-6(-174) and IL-10(-597) gene polymorphisms were found to be associated with GAgP and genotype distribution did not affect the outcome of non-surgical periodontal therapy, while patients with IL-6(-174) GG genotype had higher levels of GCF IL-10 levels.

Effects of Curcumin on Alveolar Bone Loss in Experimental Periodontitis in Rats: A Morphometric and Histopathologic Study.

Background: Curcumin is found in the rhizomes of the turmeric plant that has been showed antioxidant and anti-inflammatory effect. The aim of this study was to evaluate the effects of systemic curcumin therapy on alveolar bone loss in an experimental periodontitis model in rats. Material and Methods: Thirty-two male Wistar rats were randomly divided to 4 groups: 75 mg/kg/daily curcumin (C75; n =8), 150 mg/kg/daily curcumin (C150; n =8), Control (n =8), and Ligated (n =8). Curcumin was administrated using gastric gavage. After 12 days, the rats were sacrificed. Right mandibles samples were histopathologically examined. Alveolar bone loss was measured. Interleukin 1ß (IL-1ß) and interleukin 10 (IL-10) were evaluated in the serum samples and gingival homogenates. Results: The measurements of alveolar bone loss in the mandibular molars revealed significantly higher bone-loss values in the Ligated group than the Control, C75 and C150 groups. The IL-1ß levels in the gingival homogenates were significantly increased in the Ligated group compared to those of the Control, C75 and C150 groups. The serum IL-1ß levels in the Ligated group were significantly higher than the Control group. The mean osteoblast numbers in the Ligated group were lower than those of the Control, C75 and C150 groups. The C150 groups showed significantly more osteoblasts than the Control group. The osteoclast numbers in the Ligated group increased significantly compared to the C75, C150 and control groups. Conclusion: This study demonstrates that systemic administration of curcumin at the 75 and 150mg/kg doses reduced alveolar bone loss in the periodontal disease in rats. Keywords: Alveolar bone loss, Antioxidant, Curcumin, Ligature-induced, Histomorphometric, Micronutrition.

Serosurvey of Coxiella burnetii in high risk population in Turkey, endemic to Crimean-Congo haemorrhagic fever virus.

BACKGROUND & OBJECTIVES: Q fever caused by Coxiella burnetii is a zoonotic infection that spreads to human beings from animals. This study was aimed to demographically examine the C. burnetii seroprevalence in the people living in villages where Crimean-Congo haemorrhagic fever virus (CCHFV) is endemic, in terms of various risk factors such as tick bites, tick contact, and occupational groups. METHODS: A total of 440 serum samples from those living in rural areas of Sivas and Tokat regions in Turkey were included in the study as a risk group; 387of them were serologically CCHFV positive (as confirmed in our previous research). Serums of the control group composed of 110 people living in urban areas. In all serum samples, IgG antibodies of C. burnetii against phase-I and phase-II antigens were diagnosed using the ELISA method. RESULTS: Coxiella burnetii seropositivity was detected in 19.09% of those living in rural areas and 4.55% of those living in urban areas (p < 0.001, OR = 4.96). In terms of their approach to the ticks, no statistical difference was observed between the risk groups in the chi-square test (p = 0.787). However, according to univariate analysis, the absorbance means of antibodies reactive to C. burnetii was statistically higher for the rural people who have made contact with ticks than those who have not (p = 0.017). No seroepidemiological relation was found between CCHFV and C. burnetii serology (p = 0.787), and the rate of co-seropositivity between them was 5.43% (21/387). INTERPRETATION & CONCLUSION: The findings of the study showed that C. burnetii infection is epidemic especially in the people living in rural areas. Contact with ticks in various ways might have resulted in the increased risk of C. burnetii infection in the study. Personal protective measures against tick bites may be important for reducing Q fever risk as in other tick-borne infectious disease.

Investigating the Effects of Systemically Administered Strontium Ranelate on Alveolar Bone Loss Histomorphometrically and Histopathologically on Experimental Periodontitis in Rats.

BACKGROUND: The aim of this study is to investigate effects of strontium ranelate (SR) on alveolar bone loss (ABL) in rats with experimental periodontitis. METHODS: Forty Wistar rats were randomly divided into five groups: 1) control (n = 8); 2) ligated (n = 8); 3) 300 mg/kg SR (SR300, n = 8); 4) 625 mg/kg SR (SR625, n = 8); and 5) 900 mg/kg SR (SR900, n = 8). To create experimental periodontitis, 4/0 silk ligatures were inserted submarginally around first molars at the right mandible. After 11 days, rats were sacrificed. ABL was calculated by measuring cemento-enamel junction and alveolar crest distance. Interleukin (IL)-1ß, osteoprotegerin (OPG), and bone-specific alkaline phosphatase (BALP) serum levels were determined by enzyme-linked immunosorbent assay. Histopathologic analysis was used to evaluate inflammatory cell infiltration, numbers of osteoblasts and osteoclasts, and receptor activator of nuclear factor-kappa B ligand (RANKL) activity. RESULTS: ABL was significantly lower in SR900 group than in the ligated group (P <0.05). Osteoclast numbers in ligated group were significantly higher than in the control, SR300, and SR900 groups (P <0.05). In ligated, SR625, and SR900 groups, significantly higher osteoblast numbers were detected than in control group (P <0.05). Osteoblast numbers in SR625 group were significantly higher than in the SR300 group (P <0.05). RANKL activities in SR900 and control groups were close to each other (P >0.05). Serum IL-1ß, OPG, and BALP levels revealed no significant difference (P >0.05). CONCLUSION: It can be concluded that SR can reduce RANKL activity and osteoclast numbers, as well as ABL.

Evaluation of IL-1ß, IL-1ra, and IL-10 levels and outcome of periodontal therapy in chronic periodontitis with familial Mediterranean fever.

OBJECTIVES: This study aimed to examine the IL-1ß, IL-1ra, and IL-10 cytokine levels in gingival crevicular fluid (GCF) and serum of familial Mediterranean fever (FMF) and chronic periodontitis (CP) patients, and their response to nonsurgical periodontal therapy. MATERIALS AND METHODS: A total of 50 patients, 15 FMF patients with generalized chronic periodontitis (FMF-CP), 15 systemically healthy patients with generalized chronic periodontitis (CP), ten systemically and periodontal healthy controls (HC), and ten periodontally healthy FMF patients (FMF-HC) were enrolled in the study. The cytokine levels in GCF and serum were determined by ELISA. Probing depth, clinical attachment level, and gingival and plaque indices in each participant were also measured. The GCF and clinical parameters at baseline and 6 weeks were recorded. RESULTS: The study indicated statistically significant healing of the clinical parameters in both FMF-CP and CP groups after periodontal treatment. GCF IL-1ß levels at 6 weeks in FMF-CP group were significantly lower than the CP group (p < 0.05), and GCF IL-1ra levels were significantly decreased at 6 week in the FMF-CP group (p < 0.05). GCF IL-10 levels were significantly higher in the FMF-CP group than in the other groups at baseline and 6 weeks (p < 0.05). There were no significant differences in serum-IL-1ß, IL-1ra, and IL-10 levels either FMF-CP or CP groups at baseline or 6 weeks (p > 0.05). CONCLUSIONS: The results of our study suggested that there was a positive correlation between gingival inflammation and serum cytokine levels in FMF patients and also colchicine treatment showed protective effects on GCF cytokine levels in FMF-CP group. CLINICAL RELEVANCE: Following treatment, GCF IL-1ß and GCF IL-1ra levels were decreased in FMF-CP group. GCF IL-10 levels were increased in FMF-CP group compared to other groups. Also, the serum cytokine levels associated with periodontal inflammation in FMF patients.

IL-6 and IL-10 gene polymorphisms in patients with aggressive periodontitis: effects on GCF, serum and clinic parameters

Abstract Genetic variations observed in cytokines affect periodontitis susceptibility. The aim of this study was to investigate interleukin(IL)-6(-174) and IL-10(-597) gene polymorphisms in generalized aggressive periodontitis (GAgP) patients. Also, we aimed to evaluate the effects of IL-6 and IL-10 gene polymorphisms on the clinical outcomes of non-surgical periodontal therapy and cytokine levels in gingival crevicular fluid(GCF) and serum. Fifty-three patients with GAgP and 50 periodontally healthy individuals were included in this study. Clinical parameters, GCF and blood samples were collected at baseline and at 6-week. Non-surgical periodontal therapy was performed in patients with GAgP. Gene analysis were determined by PCR-RFLP(polymerase chain reaction-restriction fragment length polymorphism) and cytokine levels were determined by enzyme-linked immunosorbent assay(ELISA).GAgP patients showed significant improvement on clinical parameters after periodontal therapy(p<0.05). In the GAgP group, IL-6 GG genotype and G allele frequency were higher than in the control group. GCF IL-6 level was also significantly lower at 6-week in the GAgP group. Higher GCF IL-10 levelswere observed in patients carrying the IL-6 GG genotype than in those carrying the GC+CC genotype at baseline. In conclusion, IL-6(-174) and IL-10(-597) gene polymorphisms were found to be associated with GAgP and genotype distribution did not affect the outcome of non-surgical periodontal therapy, while patients with IL-6(-174) GG genotype had higher levels of GCF IL-10 levels.

Inhibitors of the Cysteine Synthase CysM with Antibacterial Potency against Dormant Mycobacterium tuberculosis.

Cysteine is an important amino acid in the redox defense of Mycobacterium tuberculosis, primarily as a building block of mycothiol. Genetic studies have implicated de novo cysteine biosynthesis in pathogen survival in infected macrophages, in particular for persistent M. tuberculosis. Here, we report on the identification and characterization of potent inhibitors of CysM, a critical enzyme in cysteine biosynthesis during dormancy. A screening campaign of 17 312 compounds identified ligands that bind to the active site with micromolar affinity. These were characterized in terms of their inhibitory potencies and structure-activity relationships through hit expansion guided by three-dimensional structures of enzyme-inhibitor complexes. The top compound binds to CysM with 300 nM affinity and displays selectivity over the mycobacterial homologues CysK1 and CysK2. Notably, two inhibitors show significant potency in a nutrient-starvation model of dormancy of Mycobacterium tuberculosis, with little or no cytotoxicity toward mammalian cells.

Comparative effects of riboflavin, nicotinamide and folic acid on alveolar bone loss: A morphometric and histopathologic study in rats.

Introduction: Periodontitis is a chronic inflammatory and osteolytic disease. Vitamin B complex is a class of water-soluble vitamins that play important roles in cell metabolism. Objective: The aim of this study was to evaluate the effects of riboflavin (RBF), nicotinamide (NA), and folic acid (FA) on alveolar bone loss in experimental periodontitis rat model. Methods: Sixty-four male Wistar rats were randomly divided into the following eight groups: Control, Ligated, RBF50 (RBF, 50 mg/kg daily), NA50 (NA, 50 mg/kg daily), FA50 (FA, 50 mg/kg daily), RBF100 (RBF, 100 mg/kg daily), NA100 (NA, 100 mg/kg daily), and FA100 (FA, 100 mg/kg daily). Periodontitis was induced using silk ligature around the right first mandibular molar. After 11 days the rats were sacrificed. Mandible and serum samples were collected. Changes in alveolar bone levels were measured clinically, and periodontal tissues were examined histopathologically. Serum IL-1ß (pg/ml) levels were analyzed by using ELISA. Results: Mean alveolar bone loss in the mandibular first molar tooth revealed to be significantly lower in RBF100 group than in the Control group. In the Ligated group, alveolar bone loss was significantly higher than in all other groups. The ratio of presence of inflammatory cell infiltration in the Ligated group was significantly higher than in the Control group. The differences in the serum IL-1ß levels between the groups were not statistically significant. Osteoclasts that were observed in the Ligated group were significantly higher than those of the Control and FA100 groups. The osteoblastic activity in the Ligated group, RBF100, and NA100 groups were shown to be significantly higher than those in the Control group. Conclusion: This study has demonstrated that systemic administration of RBF, NA, and FA in different dosages (50­100 mg/kg) reduced alveolar bone loss in periodontal disease in rats.

Crystal structures of the kinase domain of the sulfate-activating complex in Mycobacterium tuberculosis.

In Mycobacterium tuberculosis the sulfate activating complex provides a key branching point in sulfate assimilation. The complex consists of two polypeptide chains, CysD and CysN. CysD is an ATP sulfurylase that, with the energy provided by the GTPase activity of CysN, forms adenosine-5'-phosphosulfate (APS) which can then enter the reductive branch of sulfate assimilation leading to the biosynthesis of cysteine. The CysN polypeptide chain also contains an APS kinase domain (CysC) that phosphorylates APS leading to 3'-phosphoadenosine-5'-phosphosulfate, the sulfate donor in the synthesis of sulfolipids. We have determined the crystal structures of CysC from M. tuberculosis as a binary complex with ADP, and as ternary complexes with ADP and APS and the ATP mimic AMP-PNP and APS, respectively, to resolutions of 1.5 Å, 2.1 Å and 1.7 Å, respectively. CysC shows the typical APS kinase fold, and the structures provide comprehensive views of the catalytic machinery, conserved in this enzyme family. Comparison to the structure of the human homolog show highly conserved APS and ATP binding sites, questioning the feasibility of the design of specific inhibitors of mycobacterial CysC. Residue Cys556 is part of the flexible lid region that closes off the active site upon substrate binding. Mutational analysis revealed this residue as one of the determinants controlling lid closure and hence binding of the nucleotide substrate.

Structure-guided design of novel thiazolidine inhibitors of O-acetyl serine sulfhydrylase from Mycobacterium tuberculosis.

The cysteine biosynthetic pathway is absent in humans but essential in microbial pathogens, suggesting that it provides potential targets for the development of novel antibacterial compounds. CysK1 is a pyridoxalphosphate-dependent O-acetyl sulfhydrylase, which catalyzes the formation of l-cysteine from O-acetyl serine and hydrogen sulfide. Here we report nanomolar thiazolidine inhibitors of Mycobacterium tuberculosis CysK1 developed by rational inhibitor design. The thiazolidine compounds were discovered using the crystal structure of a CysK1-peptide inhibitor complex as template. Pharmacophore modeling and subsequent in vitro screening resulted in an initial hit compound 2 (IC50 of 103.8 nM), which was subsequently optimized by a combination of protein crystallography, modeling, and synthetic chemistry. Hit expansion of 2 by chemical synthesis led to improved thiazolidine inhibitors with an IC50 value of 19 nM for the best compound, a 150-fold higher potency than the natural peptide inhibitor (IC50 2.9 µM).

Some acute phase reactants and cholesterol levels in serum of patient with Crimean-Congo haemorrhagic fever.

The purpose of this study is to determine erythrocyte sedimentation rate (ESR), C - reactive protein (CRP), serum amyloid-A (SAA) and cholesterol levels in patients with Crimean-Congo Hemorrhagic Fever (CCHF) and determine the relationship of these parameters with the severity of disease. By polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) method 40 patients were diagnosed as CCHF and 39 volunteer without any systemic disease whose blood were taken and their serum separated. SAA, CRP and ESR were measured with ELISA, nephelometry and Mix-Rate x100 vital diagnostic device, respectively, in serum samples. High density lipoprotein (HDL), low density lipoprotein (LDL) and total cholesterol levels were determined by using autoanalyzer HDL, LDL and total cholesterol kit (Syncron LX20). Statistically significant difference was determined between patients and controls in terms of the levels of SAA, CRP, HDL, LDL and total cholesterol (p<0.05). However, there was no significant difference between the groups in terms of the levels of ESR. In addition, neither SAA, CRP, ESR nor HDL, LDL and total cholesterol levels varied with the severity of disease in the cases assessed (p>0.05). Using of CRP and SAA together might increase the sensitivity of diagnosis of CCHF infection. However, none of the parameters investigated in this study were found to be a proper marker of the prognosis in CCHF. Cholesterol levels were significantly decreased in patients with CCHF, which was suggested to be associated with the increased serum levels of SAA in the patient group.

Discovery of novel inhibitors targeting the Mycobacterium tuberculosis O-acetylserine sulfhydrylase (CysK1) using virtual high-throughput screening.

Cysteine biosynthesis in Mycobacterium tuberculosis (MTB) is crucial for this pathogen to combat oxidative stress and for long term survival in the host. Hence inhibition of this pathway is attractive for developing novel drugs against tuberculosis. In the present study, the crystal structure of the mycobacterial enzyme O-acetylserine sulfhydrylase CysK1 bound to an oligopeptide inhibitor was used as a framework for virtual screening of the BITS-Pilani in-house database to identify new scaffolds as CysK1 inhibitors. Thirty compounds were synthesized and evaluated in vitro for their ability to inhibit CysK1, activity against M. tuberculosis and cytotoxicity as steps towards the derivation of structure-activity relationships (SAR) and lead optimization. Compound 8-nitro-4-(2-(trifluoromethyl)phenyl)-4,4a-dihydro-2H-pyrimido[5,4-e]thiazolo[3,2-a]pyrimidine-2,5(3H)-dione (4n) emerged as the most promising lead with an IC(50) of 17.7 µM for purified CysK1 and MIC of 7.6 µM for M. tuberculosis, with little or no cytotoxicity (>50 µM).

[Investigation of Chlamydophila pneumoniae seropositivity and risk factors in patients with atherosclerotic vascular disease in Sivas, Turkey]. / Sivas Yöresinde Aterosklerotik Damar Hastaligi Olan Kisilerde Chlamydophila pneumoniae Seropozitifligi ve Risk Faktörleri ile Iliskisi.

The role of various microorganisms including Chlamydophila (formerly Chlamydia) pneumoniae, have been frequently investigated in the pathogenesis of atherosclerosis. In our study, the relationship between C.pneumoniae seropositivity and risk factors for atherosclerosis have been evaluated. A total of 90 atherosclerotic patients (71 of them were male; age range: 45-87 years; mean age: 65.3 ± 8.7 years) and 90 control subjects without coronary diseases (41 of them were male; age range: 42-84 years; mean age: 61.6 ± 9.6 years) were included in the study. Both groups were also evaluated for the presence of risk factors such as age, gender, smoking, hypertension, diabetes, obesity, dyslipidemia, familial history and the high levels of ferritin, cholesterol (total, HDL and LDL) and HS (high sensitive)-CRP. The presence of C.pneumoniae IgG, IgM and IgA antibodies were investigated by micro-immunofluorescence (MIF) and ELISA methods using commercial kits (Euroimmun, Germany). The total antibody seropositivity rate was found 100% (90/90) in patient group by both MIF and ELISA methods, while this rate in control group was 94% (85/90) by MIF and 92% (83/90) by ELISA. When MIF test results were taken into consideration (since it is accepted as the reference method for C.pneumoniae serology), IgG, IgM and IgA positivity rates in patient and control groups were found as 100% (90/90) and 89% (80/90); 70% (63/90) and 59% (53/90); 3% (3/90) and 2% (2/90), respectively. Statistically significant difference between patient and control groups was detected only for IgG positivity (p< 0.05) and for total antibody positivities (100% and 94%, respectively) (p< 0.05). The evaluation of the risk factors revealed that age, hypertension, dyslipidemia and HS-CRP levels exhibited statistically significant differences between patient and control groups (p< 0.05 for each parameter tested). Statistically significant relation was detected only between high HS-CRP levels and C.pneumoniae seropositivity (p< 0.05). It was concluded that in areas with high C.pneumoniae infection prevalence, early diagnosis and specific treatment of C.pneumoniae infections, may prevent establishment of chronic infection and eliminate a risk factor for the development of atherosclerosis.

Design of a specific colonic mucus marker using a human commensal bacterium cell surface domain.

Imaging living cells and organs requires innovative, specific, efficient, and well tolerated fluorescent markers targeting cellular components. Such tools will allow proceeding to the dynamic analysis of cells and the adaptation of tissues to environmental cues. In this study, we have identified and synthesized a novel non-toxic fluorescent marker allowing a specific fluorescent staining of the human colonic mucus. Our strategy to identify a molecule able to specifically bind to the human colonic mucus was on the basis of the mucus adhesion properties of commensal bacteria. We identified and characterized the mucus-binding property of a 70-amino acid domain (MUB(70)) expressed on the surface of Lactobacillus strains. The chemical synthesis of MUB(70) was achieved using the human commensal bacterium Lactobacillus reuteri AF120104 protein as a template. The synthesized Cy5-conjugated MUB(70) marker specifically stained the colonic mucus on fixed human, rabbit, and guinea pig tissues. Interestingly, murine tissue was not stained, suggesting significant differences in the composition of the murine colonic mucus. In addition, this marker stained the mucus of living cultured human colonic cells (HT29-MTX) and human colonic tissue explants. Using a biotinylated derivative of MUB(70), we demonstrated that this peptide binds specifically to Muc2, the most abundant secreted mucin, through its glycosylated moieties. Hence, Cy5-MUB(70) is a novel and specific fluorescent marker for mammalian colonic mucus. It may be used for live imaging analysis but also, as demonstrated in this study, as a marker for the diagnosis and the prognosis of colonic mucinous carcinomas.

Influence of smoking on interleukin-1beta level, oxidant status and antioxidant status in gingival crevicular fluid from chronic periodontitis patients before and after periodontal treatment.

BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the impact of smoking on the relationship between interleukin-1 (IL-1ß) and oxidation in patients with periodontitis and response to nonsurgical periodontal therapy. MATERIAL AND METHODS: Data were obtained from 30 patients with generalized chronic periodontitis (15 smokers and 15 nonsmokers) and from 10 periodontally healthy controls. IL-1ß level, total oxidant status (TOS) and total antioxidant status (TAS) were recorded in gingival crevicular fluid. Probing depth, clinical attachment level, gingival and plaque indices and bleeding on probing were also measured. The gingival crevicular fluid and clinical parameters were recorded at baseline and 6 wk after periodontal treatment. RESULTS: The study showed statistically significant improvement of clinical parameters in both smokers and nonsmokers after periodontal treatment. Moreover, the baseline IL-1ß levels were significantly higher in smokers compared with nonsmokers (p < 0.05). After periodontal treatment, the IL-1ß levels were significantly reduced in both smokers and nonsmokers (p < 0.05). There were no significant differences in TOS and TAS between periodontitis patients and healthy controls at baseline and 6 wk after periodontal treatment. The level of IL-1ß in gingival crevicular fluid was positively correlated with TOS in both smokers and nonsmokers. CONCLUSIONS: Periodontal treatment improved the clinical parameters in both smokers and nonsmokers. The results confirm that periodontal therapy has an effect on IL-1ß levels in gingival crevicular fluid, but not on TOS and TAS.

Crimean-Congo hemorrhagic fever virus in ticks collected from humans, livestock, and picnic sites in the hyperendemic region of Turkey.

During June and July 2007, about 3125 adult ticks were collected from humans, animals, and vegetation in a hyperendemic region (Sivas and Tokat) of Turkey. A total of 2193 ticks were pooled in 225 pools and screened for the Crimean Congo hemorrhagic fever virus (CCHFV) presence by antigen-capture enzyme-linked immunosorbent assay. Infection rates were calculated as the maximum likelihood estimation with 95% confidence intervals (CI). The dominant tick species was found to be Hyalomma marginatum with the following infestation rates in human, cattle and sheep, respectively: 47.43%, 66.07%, and 30.12%. Maximum likelihood estimation values of CCHFV in H. marginatum ticks collected from human, cattle, and sheep were 0.91% (CI 0.05-4.42), 2.10% (CI 1.12-3.64), and 3.11% (CI 1.18-6.87), respectively. CCHFV antigens were also demonstrated in Hyalomma excavatum, Haemaphysalis parva, and Boophilus annulatus ticks collected from cattle and Rhipicephalus bursa ticks from sheep. Our results suggest that the studied area might maintain its endemic properties in the near future unless effective tick control measures are implemented.

[Seroprevalence of tick-borne encephalitis virus (TBEV) among the residents of rural areas in Sinop, central Black-Sea region, Turkey]. / Sinop'ta kirsal kesimde yasayan kisilerde kene kaynakli ensefalit virusu (TBEV) seroprevalansi

Tick-borne encephalitis (TBE), caused by tick-borne encephalitis virus (TBEV) is one of the serious neurological infections seen especially in the Asian and North European countries. The principal vectors of TBEV are hard ticks belonging to Ixodes genus. The major vector of European TBEV subtype is I.ricinus and the major vector of Far-Eastern and Siberian subtypes is I.persulcatus. I.ricinus exists in many climatic regions of Turkey, especially in the coastal areas. The aim of this study was to investigate the TBEV seroprevalence among the residents of rural areas in Sinop (a province located at the coast of Central Black-Sea region of Turkey). A total of 273 blood samples have been collected from the subjects (age range: 11-83 years) inhabiting in 12 villages of the central district of Sinop, during the months of May and June in 2006 and 2007. The presence of TBEV IgG antibodies in serum samples were searched by a commercial indirect fluorescent antibody kit (Euroimmun, Deutschland). TBEV IgG positivity was detected in 2.9% (8/273) of the subjects at a screening titer (1/10) and 7 (2.6%) of them also yielded positive results at further dilutions (1/100). The rates of TBEV seropositivity were not found statistically significant (p > 0.05), with respect to gender (141 of the subjects were male), age (142 of the subjects were between 21-50 years old), occupation (17 foresters, 57 were shepherds, 199 were farmers/stockbreeders) and history of tick bite (169 of the subjects had been bitten by ticks). Presence of IgG antibodies against Borrelia burgdorferi, another agent which is transmitted by the same vector, were also investigated in TBEV seropositive 8 subjects by a commercial ELISA kit (Zeus Scientific, The Netherlands). Four of these subjects yielded B.burgdorferi IgG positivity, so the TBEV - B.burgdorferi coinfection rate was estimated as 1.5% (4/273). However, since the results obtained by the tests used in this study (TBEV IgG IFA and B.burgdorferi IgG ELISA) have not been confirmed by additional confirmational tests, these subjects were referred as "probable cases". In recent years the detection rates of vector-borne viral infections is in an increment trend in Turkey due to the developments in diagnostic tests and awareness for emerging infections. In conclusion since Sinop, which is placed in the northernmost point of Turkey, is located close to TBEV endemic areas, the presence of TBEV in Sinop and the Black Sea region should always be considered.