Diversity of Lyme borreliosis spirochetes isolated from ticks in Serbia.

Spirochetes from the Borrelia burgdorferi sensu lato (s.l.). (Spirochaetales: Spirochaetaceae) species complex, including the causative agents of Lyme borreliosis, have been isolated from ticks, vertebrate reservoirs and humans. Previous analyses based on direct molecular detection in ticks indicated a considerable diversity of B. burgdorferi s.l. complex in Serbia. The present study aimed (a) to isolate borrelia strains from Serbia; (b) to determine their genotypic characteristics; and (c) to establish a collection of viable B. burgdorferi s.l. strains for further biological, ecological and genetic studies. For the present study, 231 adult Ixodes ricinus (Ixodida: Ixodidae) ticks from 16 ecologically different localities in Serbia were individually processed to cultivate B. burgdorferi s.l. This led to the isolation of 36 strains. A hbb gene quantitative real-time polymerase chain reaction (PCR) based on melting temperature determination and ospA gene sequencing were used to genotype the isolated spirochetes. The species identified based on the hbb gene real-time PCR were: Borrelia lusitaniae (44.4%), Borrelia afzelii (36.1%), Borrelia garinii (13.9%) and Borrelia valaisiana (5.6%), whereas the ospA sequence analysis revealed the occurrence of Borrelia bavariensis. This is the first report of the isolation of B. lusitaniae, B. garinii, B. bavariensis and B. valaisiana strains in Serbia.

Ticks (Acari: Argasidae, Ixodidae) parasitizing bats in the central Balkans.

Ticks parasitizing bats have been largely understudied, especially in the central part of the Balkan Peninsula, where the last data from the field research date from almost 25 years ago. Bats are hosts to a large number of ectoparasites, including ticks, which can act as vectors of zoonotic agents. For this reason, it is important to identify the distribution of ticks and their relationship to different hosts, including wild animals, bats in particular. The present research was conducted at 16 localities throughout Serbia, Montenegro, Bosnia and Herzegovina, and Former Yugoslav Republic of Macedonia (FYROM). We examined 475 individuals of bats belonging to 13 species. A total of three tick species were identified, I. simplex being the most numerous and widespread, followed by I. vespertilionis and A. vespertilionis. To the best of our knowledge, the presented data include the first records of I. simplex in Serbia and Montenegro, I. vespertilionis for Montenegro and A. vespertilionis in FYROM. Also, we identify a new possible host/parasite association between I. simplex and Rhinolophus euryale.

Clinical application of autologous fibroblast cell culture in gingival recession treatment.

BACKGROUND AND OBJECTIVE: Gingival recession is defined as soft and hard tissue displacement resulting in root surface exposure. The optimal outcome of gingival recession treatment is complete, predictable and long-lasting root coverage with a significant level of tissue regeneration. Tissue engineering, which applies active regeneration principles, presents the contemporary treatment approach in the restitution and regeneration of lost tissues. The objective of the present study was to evaluate and compare the clinical results of application of an autologous fibroblast cell culture (AFCC) on a collagen matrix and a connective tissue graft (CTG) placed under a coronally advanced flap (CAF), in the treatment of single and multiple gingival recessions. MATERIAL AND METHODS: Eighteen patients from the Department of Periodontology, School of Dentistry, University of Belgrade, were randomly enrolled in this study. Inclusion criteria were the bilateral presence of Miller Class I or II single or multiple maxillary gingival recessions. A split-mouth design was used in the study. The experimental group was treated with AFCC on a collagen scaffold, which was placed under a CAF. The control group received a combination of CTG and CAF. Clinical parameters such as gingival recession coverage, keratinized tissue width, clinical attachment level and gingival index were recorded at baseline and at 12 mo postoperatively. The oral hygiene level was assessed by plaque index evaluation. Postoperative healing was evaluated through the healing index, recorded 1, 2 and 3 wk postoperatively. The final esthetic outcome was assessed using the mean root coverage esthetic score (RES). RESULTS: Statistically significant improvement of all parameters assessed was found compared with baseline. A statistically significant difference between groups was observed only in keratinized tissue width. Greater keratinized tissue width is still obtained with the use of CTG. Regarding the tissue-healing results, no statistically significant difference was achieved. The RES results were similar for both groups. CONCLUSIONS: Within the limitations of the present study, both procedures proved to be efficient in gingival recession treatment. AFCC, as a novel tissue-engineering concept and living cell-based therapy, proved to be a reliable and successful treatment concept.

Salivary antioxidants as periodontal biomarkers in evaluation of tissue status and treatment outcome.

BACKGROUND AND OBJECTIVE: One of the major pathologic patterns in periodontitis represents an imbalance among the production of free radicals and local antioxidants resulting in periodontal tissue destruction. The objective of the study was to investigate the influence of non-surgical periodontal treatment on salivary antioxidants and to evaluate their capacity as biomarkers reflecting periodontal tissue condition and therapy outcome. MATERIAL AND METHODS: Sixty-three systemically healthy non-smokers, including 21 periodontally healthy subjects (HC) and 42 patients with current chronic periodontitis fulfilled the inclusion criteria. Half of the patients received scaling and root planing (SRP) and the other half received only oral hygiene instructions. Full mouth clinical measurements, including gingival index (GI), plaque index (PI), periodontal pocket depth, clinical attachment level and saliva sampling were performed at baseline visit and 2 mo after treatment/baseline visit. Total antioxidant capacity (TAOC), albumins (ALB), uric acid (UA), superoxide dismutase (SOD) and glutathione peroxidase (GPX) were evaluated in saliva samples using commercial kits. RESULTS: All measured antioxidants were affected by treatment resulting in significant increase in TAOC (p < 0.005), ALB (p < 0.001), UA (p < 0.001) and GPX (p < 0.001) and decrease of SOD (p < 0.005) in response to SRP, where no differences were observed for any of parameters in the oral hygiene instructions group. Comparison of antioxidant levels between the HC and SRP group showed that before treatment ALB were significantly higher in HC when compared to the SRP group (p = 0.039), and GXP (p = 0.000) and SOD (p = 0.021) levels were significantly higher in the SRP group. Comparison of values after treatment showed that TAOC was significantly higher in the HC than in the SRP group (p = 0.001), but UA was, inversely, significantly higher in the SRP group (p = 0.034). All clinical parameters except clinical attachment level were significantly decreased after SRP and significant correlations were observed between SOD and GI (p = 0.017), SOD and PI (p = 0.011), GPX and GI (p = 0.003) and GPX and PI (p = 0.008). CONCLUSION: Non-surgical periodontal treatment affected salivary TAOC, ALB, UA, SOD and GPX; moreover, these biochemical parameters convincingly reflected periodontal status and tissue response on treatment.

Salivary interleukin-8 levels in children suffering from type 1 diabetes mellitus.

OBJECTIVE: The aim of this study was to investigate the differences between the salivary levels of IL-8 in patients with Type 1 diabetes mellitus (DM) with (DM+P) or without (DM-P) concomitant periodontitis and healthy subjects. The correlations between the levels of these cytokines and clinical periodontal parameters were also established. METHODS: Twenty children and adolescents with Type 1 DM (10 diagnosed with periodontitis, 10 presenting no signs of periodontitis) and a control group consisting of 20 healthy children and adolescents aged 7-18 years were recruited for this study. RESULTS: The Salivary IL-8 level was statistically significantly (p < 0.005) elevated in subjects with Type 1 DM (474.47 +/- 716.76) compared to non-diabetic control group (101.99 +/- 68.32). There was no difference (p > 0.05) in the salivary IL-8 level when subjects with Type 1 DM with concomitant periodontitis were compared to diabetics without periodontitis. When the salivary IL-8 level in subjects with Type 1 DM was correlated with the clinical parameters, no statistical significance was found. CONCLUSION: An elevated salivary IL-8 level in subjects with Type 1 DM without concomitant periodontitis plays a major role in the development of diabetic micro and macroangiopathy and pathogenesis of atherosclerosis. Consequently, this may offer a basis for the assessment of risk, prophylaxis and treatment of diabetic complications.

Seasonal and spatial occurrence of glycerol-3-phosphate dehydrogenase variability in Ixodes ricinus (Acari: Ixodidae) populations.

Previous results indicate that glycerol-3-phosphate dehydrogenase variability represents the adaptation of Ixodes ricinus L. (Acari: Ixodidae) to fluctuations of environmental conditions, particularly to temperature. Analysis of crucial polymorphisms in I. ricinus Gpdh gene was done by the restriction method, and three different haplotypes were obtained (GPDH441 1, GPDH441 2, and GPDH441 3), corresponding to GPDH alleles detected by allozyme electrophoresis. Differences in GPDH441 haplotype and genotype frequencies were found between samples from open and forest habitats. Significant seasonal variations of GPDH441 haplotype and genotype frequencies were detected in samples from the open habitats. No seasonal variations were observed at forest localities, probably because of the less pronounced amplitude of environmental factors. The possible role of host availability was discussed as an important factor that affects seasonal dynamics and genetic composition of tick populations.

Ki-67 expression in oral lichen planus.

PURPOSE: The monoclonal antibody Ki-67 detects a nuclear antigen that is present only in proliferating cells. This is of particular interest for the analysis of the proliferation rates of malignant tumors. The aim of this study was to investigate the malignant potential of oral lichen planus (OLP) on the basis of expression of Ki-67 in healthy individuals (HI), patients with OLP and patients with squamous cell carcinoma (SCC), and to see for any potential interdependence between Ki-67 expression and different clinical and histopathological parameters in OLP. METHODS: Immunohistochemistry for Ki-67 was carried out using an avidin-biotin peroxidase complex method. RESULTS: Ki-67 was more expressed in keratinocytes and lymphocytes of OLP patients compared with HI, but less compared with patients with SCC. Keratinocytes and lymphocytes stained with Ki-67 in OLP patients were significantly higher in males, and in OLP specimens showed less developed civatte bodies (CB) and thickening of the basal membrane (TBM). CONCLUSION: Ki-67 may not serve as prognostic biomarker in oral cancer development from the initially diagnosed OLP, but it could help selecting patients with higher need of follow up for prevention of malignancy.

Oral status in patients receiving 5-fluorouracil for colorectal cancer.

PURPOSE: Oral complications are frequent and troublesome symptoms for those undergoing chemotherapy for cancer. Several antineoplastic agents are proved to have stomatotoxic potential, among them 5-fluorouracil (5-FU). The aim of the present study was to evaluate the oral status and patient experiences during chemotherapy with 5-FU for colorectal cancer. METHODS: Twenty-eight patients treated with 5-day 5-FU plus leucovorin entered this study. Positive data about oral symptoms were taken by anamnesis. Mucositis severity index, gingival index, plaque index, probing pocket depth and bleeding on probing have been used to assess oral mucosa and periodontal status of the patients. Patients were examined prior to chemotherapy and 14 days after the start of the chemotherapy cycle. RESULTS: Mild to moderate subjective complaints concerning oral cavity were reported by 17.9% of patients before and 39.2% of patients after chemotherapy. Clinical examination revealed oral mucosa damage in 10.7% and 35.7% of patients, with mean mucositis score of 0.14 and 0.54 before and after chemotherapy, respectively. Although mean values of all periodontal indices were elevated after chemotherapy, only increase in gingival index was statistically significant (p=0.035). Mucositis was significantly correlated with oral pain (p=0.00), xerostomia (p=0.00), and plaque index (p=0.077), while the correlation between mucositis and the rest of the examined parameters was not significant. CONCLUSION: Oral complications were not highly expressed in this study. Although 5-FU is considered to exert significant stomatotoxic effect, severe mucositis was far less common in this study compared to studies reported elsewhere.

Bcl-2 expression and its possible influence on malignant transformation of oral lichen planus.

PURPOSE: The aim of this study was to investigate the malignant potential of oral lichen planus (OLP) on the basis of the expression of the Bcl-2 marker in healthy individuals (H), patients with OLP and patients with squamous cell carcinoma (SCC), and to establish potential interdependence between expression of Bcl-2 and the different clinical and histopathological parameters in H, OLP, and SCC. METHODS: Immunohistochemistry for Bcl-2 was carried out using an avidin-biotin peroxidase complex method. Tissue sections were treated with mouse monoclonal antibody against Bcl-2 (124, DAKO A/S, Denmark; dilution 1/100). Immunohistochemical parameters measured included total tissue area, total stained area and intensity of stain. RESULTS: Keratinocytes were not Bcl-2 positive in H and were sparsely positive in OLP. The number of lymphocytes stained with Bcl-2 was significantly lower in H as compared to patients with SCC and OLP. Bcl-2 staining was weak to moderate in OLP, and moderate to intense in SCC. Bcl-2-positive lymphocytes were more expressed in older OLP patients (>55 years), and in OLP specimens with orthokeratinized epithelium, less developed acanthosis and highest grade of lymphocyte expression. CONCLUSION: Bcl-2 may not serve as a prognostic biomarker in oral cancer development from OLP, but it could help in selecting patients with higher need of follow up to prevent malignancy.

Periodontal therapy improves gastric Helicobacter pylori eradication.

The oral cavity has been proposed as a reservoir for H. pylori that could be responsible for the refractoriness of gastric infection to triple therapy (antibiotics, antimicrobials, and proton pump inhibitors). The aim of this study was to evaluate the efficiency of combined periodontal and triple therapy vs. triple therapy alone, in gastric H. pylori eradication in persons with H. pylori in the subgingival biofilm. Individuals positive for H. pylori in their gastric and oral samples, as determined by nested PCR, were treated either with periodontal and triple therapy or with triple therapy alone. Our results indicate that 77.3% of those treated with the combined therapy exhibited successful eradication of gastric H. pylori, compared with 47.6% who underwent only triple therapy. Analysis of these data suggests that periodontal treatment in combination with systemic therapy could be a promising approach to increasing the therapy's efficacy and decreasing the risk of infection recurrence.

[Evaluation of periodontal and oral mucosa status in HIV seropositive individuals]. / Stanje oralne sluznice i paradoncijuma u infekcijama humanim virusom imunodeficijencije.

We present the results of clinical and laboratory studies of HIV seropositive individuals. Twenty individuals in different stages of HIV infection (CDC clasification) and 20 healthy individuals were included in this study. The number of leucocytes, differential leucocyte count, number of CD4+ T lymphocytes and CD4/CD8 index were determined. Periodontal staus was determined by using plaque index (PI), gingival index (GI) and periodontal pocket depth (PPD). Fourty percent of HIV seroposiotive patients showed oral eflorescences and 35% confirmed the presence of subjective disturbances in oral cavity. Mean value of PPD (3.10+/-0.68) was significantly higher in these subjects than in those of the control group (2.36+/-0.92). At the same time, PI and GI showed no significant differences between the two examinde groups. No significant correlation between PI, GI and PPD on one side and the studied laboratory parameters on the other side, was found. No significant difference was found between HIV seropositive individuals with oral eflorscence and those without it.