Two-site jumps in dimethyl sulfone studied by one- and two-dimensional 17O NMR spectroscopy.

Polycrystalline dimethyl sulfone is studied using central-transition oxygen-17 exchange NMR. The quadrupolar and chemical shift tensors are determined by combining quantum chemical calculations with line shape analyses of rigid-lattice spectra measured for stationary and rotating samples at several external magnetic fields. Quantum chemical computations predict that the largest principal axes of the chemical shift anisotropy and electrical field gradient tensors enclose an angle of about 73°. This prediction is successfully tested by comparison with absorption spectra recorded at three different external magnetic fields. The experimental one-dimensional motionally narrowed spectra and the two-dimensional exchange spectrum are compatible with model calculations involving jumps of the molecules about their two-fold symmetry axis. This motion is additionally investigated by means of two-time stimulated-echo spectroscopy which allows for a determination of motional correlation functions over a wider temperature range than previously reported using carbon and deuteron NMR. On the basis of suitable second-order quadrupolar frequency distributions, sin-sin stimulated-echo amplitudes are calculated for a two-site model in the limit of vanishing evolution time and compared with experimental findings. The present study thus establishes oxygen-17 NMR as a powerful method that will be particularly useful for the study of solids and liquids devoid of nuclei governed by first-order anisotropies.

Ankaferd blood stopper enhances healing after osseous grafting in patients with intrabony periodontal defects.

BACKGROUND AND OBJECTIVE: The aim of this clinical study were to compare the clinical efficacy of ankaferd blood stopper (ABS) when used in combination with autogenous cortical bone graft (ACB) in the treatment of intrabony periodontal defects. MATERIAL AND METHODS: The study was planned as a split-mouth design. Fifteen patients with chronic periodontitis at 30 sites (six men, nine women; 42 ± 7 years) were included. Treatment sites had probing pocket depths (PPD) of ≥ 6 mm and osseous defect depths of ≥ 4 mm as radiographically assessed. Following the initial periodontal therapy, patients were randomly assigned to two treatments in contralateral areas of the dentition: ACB + ABS or ACB alone. At baseline and 6 mo after surgery, clinical parameters of plaque index, gingival index, PPD, clinical attachment level and gingival recession (GR) were recorded. The primary outcome variable was the change in clinical attachment level between baseline and 24 wk after surgery. Gingival crevicular fluid samples were collected immediately before surgery and at 2, 4, 6, 12 and 24 wk after the surgery. Gingival crevicular fluid volume was calculated and vascular endothelial growth factor levels in gingival crevicular fluid were measured. RESULTS: PPD decreased, clinical attachment level improved and gingival index decreased significantly in response to both modes of treatment (p < 0.05). Both treatment modalities resulted in a significant gain in radiographic bone levels compared to baseline (p < 0.05). Intergroup comparisons showed that there was a significantly higher gain in clinical attachment level in the ABS/ACB group compared to ACB group (p < 0.05) with significantly less GR (p < 0.05). Similarly, vascular endothelial growth factor concentration in gingival crevicular fluid was significantly higher in the ABS/ACB group at postoperative weeks 2 and 4 compared to the ACB group (p < 0.01). CONCLUSIONS: The findings suggest that ABS enhances the soft tissue healing during the periodontal defect fill by the ACB by stimulating angiogenesis and vascular endothelial cell function, prevents GR and thereby increases the clinical attachment gain.

Evaluation of gingival alterations in rats medicated with cyclosporine A, tacrolimus and sirolimus: a stereological study.

BACKGROUND AND OBJECTIVE: It has previously been shown that both cyclosporine A and tacrolimus cause gingival overgrowth in the rat. We proposed that sirolimus may play an important role in decreasing the severity of gingival overgrowth. Therefore, the aim of this study was to evaluate the gingival changes induced by immunosuppressants, in the presence and absence of sirolimus, using histopathology and stereological methods. MATERIAL AND METHODS: Thirty-six male Sprague-Dawley rats were distributed into six treatment groups, each containing six rats, as follows: (i) cyclosporine A for 8 wk; (ii) tacrolimus for 8 wk; (iii) sirolimus for 8 wk; (iv) cyclosporine A + sirolimus for 8 wk; (v) tacrolimus + sirolimus for 8 wk; and (vi) distilled water for 8 wk. Histomorphometric analyses included measurements of epithelial thickness and connective tissue width and height. Stereological analyses included measurements of volumetric densities of fibroblasts (Vf ), collagen fibers (Vcf ) and blood vessels (Vbv ). RESULTS: Connective tissue width and height were significantly increased in cyclosporine A, tacrolimus and cyclosporine A + sirolimus groups compared with the control group (p < 0.05), and epithelial thickness was significantly increased in the cyclosporine A group and tacrolimus group compared with the control group (p < 0.05). Vf was significantly increased in the cyclosporine A group and the tacrolimus group compared with the control group (p < 0.05), whereas Vcf and Vbv were significantly increased in the cyclosporine A, tacrolimus and cyclosporine A + sirolimus groups compared with the control group (p < 0.05). CONCLUSION: The results of the study suggest that sirolimus seems not to be associated with gingival overgrowth, and combined usage of sirolimus and immunosuppressants decreases the severity of gingival overgrowth.

Assessment of periostin levels in serum and gingival crevicular fluid of patients with periodontal disease.

BACKGROUND AND OBJECTIVE: Periostin, a secreted adhesion molecule essential for periodontal tissue integrity, is highly expressed in the periodontal ligament and plays a critical role in tooth and bone development. The purpose of this study was to investigate periostin levels in the gingival crevicular fluid and serum of patients with periodontal disease and compare them with those of healthy individuals. MATERIAL AND METHODS: Eighty individuals (41 males and 39 females; age range: 25-48 years) were enrolled in the study. Individuals were divided into three groups following clinical and radiographic examinations: the periodontal-healthy group (n = 20), gingivitis group (n = 30) and chronic periodontitis group (n = 30). Gingival crevicular fluid and serum samples were collected and periostin levels were determined using the enzyme-linked immunosorbent assay. RESULTS: The total amount and concentration of periostin decreased in gingival crevicular fluid with the progression and severity of the disease from healthy controls to gingivitis and to chronic periodontitis groups and differed significantly (p < 0.05). However, there was no significant difference in serum periostin concentration within all groups (p > 0.05). Periostin in gingival crevicular fluid negatively correlated with the gingival index in the periodontal disease groups, whereas it is inversely correlated with the clinical attachment level only in the periodontitis group (p < 0.05). When all the clinical groups were examined together, the periostin concentration negatively correlated with clinical attachment level and gingival index; moreover, total periostin positively correlated with periostin concentration and clinical attachment level (p < 0.05). CONCLUSIONS: The periostin levels in gingival crevicular fluid decreased proportionally with the progression and severity of periodontal disease, and negatively correlated with the clinical parameters. Within the limits of the study, the periostin level in gingival crevicular fluid can be considered a reliable marker in the evaluation of periodontal disease susceptibility and activity.

The role of phosphatase and tensin homolog in drug-induced gingival overgrowth.

BACKGROUND AND OBJECTIVE: We proposed that phosphatase and tensin homolog (PTEN) might be one of the signaling proteins that alter the balance between cell growth and cell death in drug-induced gingival overgrowth. The aim of this study was to investigate the expression of PTEN in subjects using cyclosporine A and to analyze the relationship between PTEN and cell proliferation marker, proliferating cell nuclear antigen (PCNA), in cyclosporine A-induced gingival overgrowth. MATERIAL AND METHODS: In total, samples from 36 subjects, i.e. 24 cyclosporine A-mediated renal transplant patients with gingival overgrowth (n = 12) or without gingival overgrowth (n = 12) and 12 matched periodontally healthy subjects, were included in the study. PTEN and PCNA expressions in gingival tissues were analyzed using immunohistochemistry, PTEN expression was also analyzed by western blot. PTEN immunoreactivity was calculated with a histologic score (HSCORE) value and PCNA immunoreactivity was calculated with the PCNA-proliferative index. RESULTS: Phosphatase and tensin homolog HSCORE for the group with gingival overgrowth was found to be significantly lowest compared to the group without gingival overgrowth and the control group (p < 0.001) while the highest PTEN HSCORE was found in the control group. In addition, the PTEN HSCORE for the group without gingival overgrowth was significantly lower compared to controls (p < 0.001). The highest PCNA-proliferative index score was observed in the group with gingival overgrowth while the lowest score was observed in the control group (p < 0.001). The immunoblot signal for PTEN was significantly decreased in the group with gingival overgrowth compared to the group without gingival overgrowth and the control group (p < 0.001). Western blot results were different from immunohistochemistry and revealed there was no significant difference between the without gingival overgrowth and the control group (p > 0.05). CONCLUSION: Our results showing decreased PTEN levels in patients with gingival overgrowth supported with increased PCNA expression suggested that PTEN might take part in the imbalance between cell proliferation and death in drug-induced gingival overgrowth.

Activity of platelet activating factor acetylhydrolase following phase I periodontal therapy.

OBJECTIVE: Elevated levels of platelet activating factor (PAF), a potent inflammatory mediator in periodontal disease and decreased PAF levels following periodontal surgical therapy have been previously detected in gingival tissues and gingival crevicular fluid (GCF). Platelet activating factor acetylhydrolase (PAF-AH) is a calcium-independent phospholipase A2 that catalyses the hydrolysis of PAF, thereby inactivating this mediator The hypothesis, a relationship between activity of PAF-AH and healing following periodontal therapy, was tested by detecting activity of PAF-AH in GCF samples collected from sites that had undergone phase I periodontal therapy with generalized chronic periodontitis. METHODS: Twenty patients with generalized chronic periodontitis were divided into two groups (n = 10): group 1 with probing pocket depth (PPD) 4-5 mm and group 2 with PPD > or = 6-8 mm. Clinical parameters were recorded and GCF was sampled before phase I periodontal therapy and at the 2nd, 7th, 14th, 21st and 28th day follow-up evaluation visits. Activity of PAF-AH in GCF was analysed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Probing pocket depth at the 21st and 28th day in group 1, and PPD at the 14th, 21st and 28th day in group 2 were significantly decreased when compared to the baseline values (p < 0.001). Activity of PAF-AH (micromol/ml) was significantly decreased at the 7th, 14th, 21st and 28th day following phase I periodontal therapy in both groups 1 and 2 compared to the baseline values (p < 0.05). CONCLUSION: Platelet activating factor acetylhydrolase is detectable in GCF by ELISA and showed a continuous decrease following phase I periodontal therapy. Changes in the PAF-AH activity would be a progressive marker of periodontal healing to evaluate the success of periodontal therapies.

Detection of avian metapneumovirus subtypes in turkeys using RT-PCR.

This study investigated the prevalence of avian metapneumovirus (aMPV) and the detection of molecular subtypes of field strains of the virus using RT-PCR in clinically healthy turkeys and those showing signs of respiratory disease. In the RT-PCR examination of 624 tracheal tissue samples collected from a local turkey abattoir, 2.9 per cent (18/624) of samples tested positive. In the examination of tracheal swab samples collected from flocks with respiratory problems, 18 of 20 samples tested positive. When the results were assessed at flock level, aMPV infection was detected in only one of the 23 clinically healthy turkey flocks, whereas all four flocks with respiratory problems were infected. Molecular typing using primers specific to the attachment glycoprotein (G) gene showed that all 36 positive samples belonged to subtype B. Partial sequence analysis of DNA samples showed 95 per cent homology between the field types and the reference strain aMPV subtype B. Whereas clinically healthy turkeys had been vaccinated with a subtype A virus vaccine, the flocks with respiratory problems had been vaccinated with a subtype B virus vaccine. Despite four blind passages of RT-PCR-positive samples on Vero and chicken embryo fibroblast cells, no cytopathic effect was detected by microscopic examination.

Development of secondary ovarian lesions after hysterectomy without oophorectomy versus unilateral oophorectomy for benign conditions: a retrospective analysis of patients during a nine-year period of observation.

PURPOSE: The effect of retained one or both ovaries on the de novo ovarian pathologies required re-operation after hysterectomy due to benign gynecologic conditions were investigated retrospectively. This study was done to determine the occurrence of disease in retained ovaries after hysterectomy. METHODS: A retrospective analysis of patient charts was performed, comparing the patient reports of women who had secondary ovarian lesions those whose previously undergone total abdominal hysterectomy with unilateral oophorectomy or without oophorectomy in our Department during the nine year period of observation (2000-2009). The study included 1242 women with at least one ovary saved after hysterectomy for benign indications. RESULTS: De novo ovarian disease was established in 5.1% of patients of hysterectomy without oophorectomy and in 17.6% of patients of at least one ovary saved after hysterectomy for benign indications (p = 0.005). Ovarian pathology requiring re-operation developed in 3.8% of patients who underwent hysterectomy without oophorectomy and in 5.9% of patients who underwent hysterectomy with unilateral oophorectomy (p = 0.536). CONCLUSION: Women with unilateral oophorectomy at the time of hysterectomy had more than twice the risk of secondary ovarian lesions, compared with those without oophorectomy at hysterectomy. Determinants, such as age, parity and gravidity must be considered when deciding whether or not to perform oophorectomy at hysterectomy.

Cardiac troponin levels in pregnant women with severe pre-eclampsia.

Pre-eclampsia is a multisystem disorder that involves vascular endothelial dysfunction and diffuse inflammatory response. The cardiac troponin (cTn-I) levels in pre-eclampsia are controversial. The objective of this study was to compare the cTn-I levels between normal pregnant women and those with severe pre-eclampsia. A total of 78 patients who underwent caesarean section were included in the study. The patients were assigned into two groups as the severe pre-eclamptic pregnant group (study group, n = 36) and normotensive pregnant group (control group, n = 42).The cTn-I levels of all patients were measured preoperatively and postoperatively. A statistically significant difference was not determined between the preoperative and postoperative cTn-I levels (p > 0.05) between the two groups. In the present study, a relation was not determined between pre-eclampsia and increased cTn-I levels. If high cTn-I levels are determined in pre-eclamptic patients, other pathologies that may cause myocardial damage should be investigated.

Detection of contagious caprine pleuropneumonia in East Turkey.

A study was implemented to investigate the presence of contagious caprine pleuropneumonia (CCPP) in East Turkey. This study was based on clinical surveillance in the field, surveillance at regional slaughterhouses and regular submission of suspected lesions to regional laboratories. The results showed that the agent of CCPP, Mycoplasma capricolum subspecies capripneumoniae (Mccp), could be detected by culture and specific polymerase chain reaction from 37.5% (12/32) of lung samples taken from goats of ten different herds. This agent was also isolated from two of 13 sheep samples (one from the lung and the other from a nasal swab). Mycoplasma capricolum subsp. capripneumoniae was isolated in pure culture and characterised at a finer molecular level. The East Turkish isolate was found to be closely related to another strain of Turkish origin, as well as to Mccp strains isolated in Tunisia. The isolation of Mccp from sheep lung lesions brings the strict host-specificity of this pathogen into question. It may also indicate that Mccp presents a risk for wildlife in the region. Such results, the authors believe, demonstrate that adequate risk assessments should be undertaken in Turkey and neighbouring countries.

Detection of mycoplasma species in turkeys by culture and polymerase chain reaction.

The purpose of the present study was to investigate the presence of pathogenic mycoplasma species in the turkey population of Turkey. Tracheal samples randomly collected from a total of 624 apparently healthy meat-type turkeys at a commercial abattoir located in the north of the country were examined by culture and genus- and species-specific polymerase chain reaction (PCR) assays for mycoplasma. In the direct plating onto solid specific media, mycoplasma growth was observed from 1.4% (9/624) of the samples, which were confirmed to belong to the Mycoplasma genus by genus-specific PCR. Mycoplasma iowae (MI) and M. meleagridis (MM) were identified by the species-specific PCR from eight and one of the samples, respectively. However, genus-specific PCR amplification was obtained from 2.6% (16/624) of the samples which produced turbidity in the liquid media. Interestingly, these positive samples were different from those obtained from solid agar and mycoplasma growth was not observed when the broth samples were inoculated onto solid media. In the species-specific PCR analysis of the broth samples, MI, MM and M. gallisepticum were identified from twelve, two and two samples, respectively. The inconsistency between the results obtained from liquid and solid media raises questions about the efficiency of isolation procedures for mycoplasma and this warrants further investigation.

Levels of platelet activating factor in gingival crevice fluid following periodontal surgical therapy.

BACKGROUND AND OBJECTIVE: Elevated levels of platelet activating factor (PAF), a potent inflammatory phospholipid mediator, have been previously detected in gingival tissues and gingival crevice fluid (GCF) in periodontal disease. However, the role of this mediator during wound healing after periodontal surgery remains unclear. The hypothesis, a relationship between PAF levels and periodontal healing, was tested by measuring PAF levels in GCF samples collected from sites that had undergone guided tissue regeneration (GTR) or flap surgery. MATERIAL AND METHODS: Using a split-mouth design, 30 intrabony defects were randomly assigned to treatment with GTR (group 1) or to flap surgery (group 2). GCF was sampled pre-operatively and at 6-, 12- and 24-wk follow-up evaluation visits. PAF levels in GCF were analyzed by high-performance liquid chromatography (HPLC). RESULTS: Both treatment modalities significantly reduced the probing pocket depth and improved the clinical attachment level (p < 0.01). Compared with pre-operative values, the GCF volume and PAF levels were significantly decreased at postoperative weeks 6, 12 and 24 in both groups (p < 0.01). There were also significant differences in GCF volume and PAF levels at all time points up to 24 wks in both groups (p < 0.01). No statistically significant differences were observed in any of the parameters investigated between the two groups (p > 0.05). CONCLUSION: PAF is detectable in GCF by HPLC and showed a continuous decrease at all the time points monitored following periodontal surgical therapy. This suggests that changes in the levels of this mediator in GCF might be useful for monitoring the progress of periodontal repair and regeneration.

Evaluation of immunomagnetic separation-polymerase chain reaction in direct detection of Brucella abortus and Brucella melitensis from cheese samples.

The current study was carried out to assess the use of immunomagnetic separation-polymerase chain reaction (IMS-PCR) in direct detection of Brucella abortus and B. melitensis from soft cheese and to examine a relatively small number of field samples for the presence of these species. Two methodologies, one with IMS and the other without IMS, were employed for recovery of the Brucella species from cheese samples. IMS in conjunction with the PCR assay was determined to detect as low as 3x10(2) bacteria/mL, while the limit of detection with the other extraction procedure was 3x10(3) bacteria/mL. In the analysis of 40 cheese samples collected from various markets, only B. abortus was detected by PCR using both DNA extraction procedures in two (5%) samples. No positive results were obtained by culture and B. melitensis was not found in any cheese samples examined. The results suggest that this technique is promising owing to its pace and high sensitivity and should aid in direct detection of Brucella species from complex food samples.

Crohn's disease in people exposed to clinical cases of bovine paratuberculosis.

Mycobacterium avium subspecies paratuberculosis (Map), the cause of ruminant paratuberculosis, has been proposed as the causative agent of Crohn's disease. The objective of this study was to determine whether exposure to clinical cases of bovine paratuberculosis was a risk factor for Crohn's disease. A questionnaire was sent to dairy farmers living on premises where the occurrence or absence of clinical cases of bovine paratuberculosis had previously been determined. The prevalence of Crohn's disease was found to be similar to that reported in other studies in the United Kingdom and showed no association with bovine paratuberculosis. There was, however, a univariate association with geographical region. Ulcerative colitis showed univariate associations with age, frequency of contact with cattle and with smoking. The results do not support the hypothesis that Map plays a causative role in the aetiology of Crohn's disease.

The heterogeneity of Campylobacter jejuni and Campylobacter coli strains isolated from healthy cattle.

AIMS: To identify campylobacters isolated from clinically healthy cattle at species level by a multiplex polymerase chain reaction (m-PCR). The heterogeneity among Campylobacter jejuni and Campylobacter coli isolates was also investigated by using a restriction fragment length polymorphism (RFLP) analysis of flagellin (flaA) gene. METHODS AND RESULTS: Samples of intestinal contents, gall bladders, liver and faeces were collected from a total number of 1154 healthy cattle. The samples were inoculated onto Preston enrichment broth and agar. Of 1154 samples, 301 (26.1%) were positive for Campylobacter spp. Using an m-PCR assay for species identification, 179 (59.5%) were positive with C. jejuni specific primers while 30 (10%) were positive with C. coli specific primers. None of the liver samples examined was positive for C. jejuni or C. coli by mPCR. All the isolates identified as C. jejuni and C. coli were successfully subtyped by flaA typing. Of the 209 isolates tested, 28 different flaA types were found. Twenty-three flaA types were identified among 179 C. jejuni isolates and the remaining five from C. coli isolates. CONCLUSIONS: Although the overall results suggest that the degree of heterogeneity among the flaA genes of thermophilic Campylobacter strains isolated from healthy cattle is relatively high, they should be treated cautiously as the number of band types for C. coli was low and band type 8 in C. jejuni was represented by a high percentage (%58). SIGNIFICANCE AND IMPACT OF THE STUDY: The findings of the present study suggest that healthy cattle can play role in the contamination of environment and human food chain by Campylobacter spp.

Prevalence and distribution of tropical theileriosis in eastern Turkey.

This study was carried out to determine the prevalence and distribution of tropical theileriosis in cattle in eastern Turkey by microscopical, serological and molecular methods. A total of 1561 whole blood, 1505 serum and 1483 blood smear samples were collected from cattle of various breeds and ages in 11 towns of Eastern Turkey. Theileria annulata piroplasm DNA extracted from cattle blood was amplified by polymerase chain reaction (PCR) using species-specific primers. Serum antibodies against T. annulata were investigated by indirect fluorescence antibody test (IFAT). Blood smears were examined for Theileria piroplasms by microscopical examination (ME). In the examination of DNA extracted from 1561 blood samples, an amplicon with the size of 721bp was obtained in 37.8% (590/1561) of these samples. Serum antibodies against T. annulata and piroplasm of Theileria spp. were detected in 34.9% (526/1505) and 19.7% (293/1483) of the samples, respectively. The differences between ME and PCR results and between ME and IFAT results were statistically significant (P < 0.05). In contrast, there was no significant difference between the PCR and IFAT results. A total of 179 ticks (136 female; 43 male) belonging to Hyalomma spp. were collected from cattle from three towns. Ticks were identified to be Hyalomma anatolicum anatolicum on the basis of morphological features.

Investigation of arcobacters in meat and faecal samples of clinically healthy cattle in Turkey.

AIMS: To investigate the presence of Arcobacter spp. in minced beef meat (n = 97) and rectal faecal samples (n = 200) collected from cattle immediately after slaughter at a local abattoir in Turkey. METHODS AND RESULTS: Meat samples were examined using three different isolation procedures (CAT-supplemented media, de Boer arcobacter isolation method and membrane filtration method), but only one method (CAT-supplemented media) was employed for faecal samples. The isolated Arcobacter strains were identified by genus- and species-(multiplex) specific PCR assays. Arcobacter spp. were isolated from 5 and 9.5% of meat and faecal samples respectively. Although the only Arcobacter sp. found in meat samples was Arcobacter butzleri, all three pathogenic species--A. butzleri, A. cryaerophilus and A. skirrowii--were detected in the rectal swabs. No Arcobacter was isolated when the de Boer method was used for minced meat samples but the same five meat samples were found positive for arcobacters when CAT-supplemented media and membrane filtration method were used. CONCLUSIONS: The membrane filtration method was found to be superior to the CAT-supplemented media, because it led to a reduction in competing microflora. However, the necessity for one filter and medium for each sample makes this method somewhat expensive. The multiplex-PCR (m-PCR) assay shortened significantly the time required for the identification of Arcobacter spp. and also removed the possibility of false positive results due to other campylobacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports the isolation of Arcobacter spp. in cattle for the first time in Turkey. The m-PCR assay enables the identification and differentiation of all arcobacters simultaneously in one-step PCR.