Short-Term Humoral Immune Response of the pcDNA4-G Plasmid Expressing the Bovine Ephemeral Fever Virus G Gene in BALB/c Mice.

Bovine ephemeral fever (BEF) is an arthropod­borne viral disease characterised by a short­term clinical expression that can lead to significant losses in high­yielding cattle and water buffaloes. In this study, we aimed to generate a recombinant plasmid expressing the glycoprotein (G) of the BEF virus (BEFV) and to stimulate a humoral immune response to this protein in BALB / c mice immunised with the recombinant plasmid. Expression of the encoded protein was demonstrated by western blotting and immunoperoxidase tests. The suitable plasmids were intramuscularly administered to BALB/c mice on days 0, 14 and 21. The antibody response in the immunised mice was measured by a plaque reduction neutralization test (PRNT) and enzyme­linked immunosorbent assay (ELISA). According to BEFV ELISA, only two of the seven animals in these groups exceeded the cut­off value. A significant difference was observed in the mean OD values at 450 nm absorbance in the pcDNA4­G­immunised group when compared with those in the plasmid control group at 30 days (p < 0.05). According to PRNT50 results, a 1:20 (p < 0.05) antibody response was obtained at 30 days in pcDNA4­G (100 µg)­immunised mice, whereas this ratio was 1:80 (p < 0.001) in BEFV­immunised mice (1,000 PFU/0.5 ml). We conclude that the humoral immune response was stimulated in experimental mice immunised with the recombinant plasmid. However, disappointingly, the antibody response was markedly low in pcDNA4­G­immunised mice.

Detection and molecular characterisation of feline viruses from swab samples.

Feline calicivirus (FCV), feline alphaherpesvirus 1 (FHV-1) and feline panleukopenia virus (FPLV) as well as retroviral agents such as feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) are important viral pathogens of cats. The aim of this study was to detect and characterise FHV-1, FPLV, FeLV, FIV and feline foamy virus (FFV) in oropharyngeal, nasal and conjunctival swabs from 93 cats that had been screened for FCV previously. We wanted to determine the possible risk factors for infection with these viruses. The prevalence was found to be 12.9% for FHV-1 and 9.7% for FPLV. FIV was detected only in two samples and FeLV in one sample, whereas the presence of FFV was not demonstrated in any of the clinical samples. The statistical analysis of the results showed that breed, age, health status, and lifestyle are important predisposing factors to FHV-1 (P < 0.05). For FPLV, only clinically unhealthy animals were found to be at risk (P < 0.001). Sequence analysis revealed that the two FIV-positive samples in this study contained different (A and B) subtypes of the virus. This is the first report on the occurrence of subtype A FIV in Turkey.

Increased Circulatory Extrarenal 1α,25-Dihydroxyvitamin D3 in Bilaterally Nephrectomized Rats.

BACKGROUND: Extrarenal 1α,25-dihydroxyvitamin D3 (1,25-D) locally produced by immune cells plays crucial roles in the regulation of the immune system. However, in vivo status of extrarenal 1,25-D and 25-hydroxyvitamin D (25-D) in acute inflammatory conditions are unknown. OBJECTIVE: The aim of this study was to determine the extrarenal 1,25-D level in circulation in bilaterally nephrectomized rats, induced by low-dose lipopolysaccharide (LPS). METHODS: Renal 1,25-D synthesis was terminated through bilateral nephrectomy in rats. The rats received intraperitoneal LPS (50 µg/kg BW) three times and the experiment was ended 24 hours after nephrectomy. Serum 1,25-D, 25-D, calcium, phosphorus, intact parathyroid hormone, and calcitonin levels were measured and immunohistochemistry was applied to detect the sources of extrarenal 1,25- D synthesis. RESULTS: Circulatory 1,25-D concentration remarkably increased in both LPS-treated and non-treated bilaterally nephrectomized rats. Elevated circulatory 1,25-D did not have hypercalcemic endocrinal effects. The increased 1,25-D level also resulted in a concurrent rapid and dramatic depletion of circulatory 25-D. CONCLUSION: Extrarenal 1,25-D could enter into the systemic circulation and, therefore, might have systemic effects besides its autocrine and paracrine functions.

The relationship between obesity and dental caries according to life style factors in schoolchildren: a case-control study.

Objectives: To evaluate the association between obesity and dental caries in terms of life-style habits and socio-economic status in Turkish school children.Materials and methods: This case-control study was undertaken with 178 children aged 6-11 years in a Turkish government children's hospital. The case group consisted of 86 obese children who were categorized in ≥95 percentile according to the BMI. The control group consisted of 90 non-obese children that would not raise any doubts about the teeth disease. Data of demographic features and life-style habits were obtained by a questionnaire at the dental examination. Caries experience was measured with DMFT and dmft indices.Results: No difference was found between obesity and caries prevalence in primary dentition (p = .957); however, there was a statistically significant association in the permanent dentition (p = .002). Also, no differences were found in children with healthy natural teeth between the study groups according to education level, family income and food consumption during TV viewing in primary dentition (p = .297; p = .652; p = .023).Conclusions: It can be concluded that obesity appears to be not a possible risk factor for dental caries in primary dentition, but would be a probable endangerment in the permanent dentition.

GENETIC DIVERSITY of OVINE HERPESVIRUS 2 STRAINS OBTAINED FROM MALIGNANT CATARRHAL FEVER CASES in EASTERN TURKEY.

Malignant Catarrhal Fever (MCF) is a generalized, definitive lethal disease affecting the epithelial and lymphoid tissues of the respiratory and digestive tract, mainly cattle and some wild ruminants such as deer, buffalo or antelope. The sheep-related form of MCF is known to be present in Turkey and is caused by ovine herpesvirus 2 (OvHV-2). The aim of this study was to reveal the genetic diversity of OvHV-2 strains obtained from MCF cases in Eastern Turkey where the livestock industry has an important impact on economic activities. For this purpose, RTA (Replication and transcription activator), FGARAT (formylglycineamide ribotide amidotransferase) and some of glycoprotein genes (Ov7, Ov8 ex2, ORF27 and Ov9.5) were investigated in blood samples from 24 cattles, clinically diagnosed with MCF. Genomic data of chosen samples were furthermore used to characterize and undergo combined phylogenetic analysis to determine possible alleles and subvariants. The results showed that high level of OvHV-2 diversity existed in selected genes and strains carrying allelic variants might circulate both in two geographically distinct regions and in a region itself. Moreover, three different OvHV-2 types and various subtypes were identified based on multi locus approach. This study provides important data to epidemiological research and thereby helps to determine the source of the virus and understand the spread of the disease.

Multidisciplinary management of a fused maxillary central incisor moved through the midpalatal suture: A case report.

Fusion of teeth is a developmental anomaly. It occurs at the stage of tooth formation, which determines the shape and size of the tooth crown, when one or more teeth fuse at the dentin level during the morphodifferentiation of the dental germs. Such teeth show macrodontia and may cause crowding, as well as esthetic and endodontic problems. In this article, we report a rare case of a maxillary central incisor fused to a supernumerary tooth showing labial and palatal talon cusps, which was orthodontically moved across the midpalatal suture. A 13-year-old Caucasian boy sought treatment for the unesthetic appearance of his maxillary central incisor and anterior crowding. He was rehabilitated successfully via a multidisciplinary approach involving orthodontic, nonsurgical endodontic, periodontal, and prosthodontic treatments. After a 26-month treatment period, the patient's macroesthetics and microesthetics were improved. The overall improvement of this macrodontic tooth and its surrounding tissues through multidisciplinary treatment was documented using cone-beam computed tomography.

Complete genome analysis of highly pathogenic bovine ephemeral fever virus isolated in Turkey in 2012.

Relatively high prevalence and mortality rates of bovine ephemeral fever (BEF) have been reported in recent epidemics in some countries, including Turkey, when compared with previous outbreaks. A limited number of complete genome sequences of BEF virus (BEFV) are available in the GenBank Database. In this study, the complete genome of highly pathogenic BEFV isolated during an outbreak in Turkey in 2012 was analyzed for genetic characterization. The complete genome of the Turkish BEFV isolate was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. It was found that the complete genome of the Turkish BEFV isolate was 14,901 nt in length. The complete genome sequence obtained from the study showed 91-92% identity at nucleotide level to Australian (BB7721) and Chinese (Bovine/China/Henan1/2012) BEFV isolates. Phylogenetic analysis of the glycoprotein gene of the Turkish BEFV isolate also showed that Turkish isolates were closely related to Israeli isolates. Because of the limited number of complete BEFV genome sequences, the results from this study will be useful for understanding the global molecular epidemiology and geodynamics of BEF.

Effects of lapatinib and trastuzumab on vascular endothelial growth factor in experimental corneal neovascularization.

BACKGROUND: The aim of this study was to compare the effects of lapatinib and trastuzumab on vascular endothelial growth factor on experimental corneal neovascularization. METHODS: A total of 35 Wistar albino rats were randomly divided into five groups, each containing seven animals. Corneas of rats in the control group were not cauterized and did not receive any treatment. A silver nitrate pencil was applied on the right corneas of rats in the non-control groups to induce corneal neovascularization. Rats in the sham, lapatinib, trastuzumab and lapatinib + trastuzumab groups were administered systemic saline, 50 mg/kg lapatinib once a day orally by gavage, 4 mg/kg trastuzumab once a day intraperitoneally, or 50 mg/kg lapatinib once a day orally by gavage together with 4 mg/kg trastuzumab once a day intraperitoneally, respectively, for 7 days. Rats were sacrificed on the eighth day, and corneas were excised using a 4-mm punch trephine. Vascular endothelial growth factor immunostaining in the corneal epithelial and stromal layers was evaluated. Staining intensities were determined semi-quantitatively, and corneal vascular endothelial growth factor levels were measured by enzyme-linked immunosorbent assay. RESULTS: The mean immunostaining intensities of epithelial and stromal vascular endothelial growth factor in the control group were significantly lesser than those in the sham group (P < 0.05). The mean immunostaining intensities of epithelial and stromal vascular endothelial growth factor and vascular endothelial growth factor enzyme-linked immunosorbent assay levels of corneas in all treatment groups (lapatinib, trastuzumab and lapatinib + trastuzumab groups) were lesser than those in the sham group (P < 0.05); however, it was similar to those in the control group (P > 0.05). CONCLUSIONS: It is suggested that systemically administered lapatinib is more effective than systemically administered trastuzumab in preventing corneal neovascularization.

A large-scale outbreak of bovine ephemeral fever in Turkey, 2012.

Regional cases of bovine ephemeral fever (BEF) were documented previously in Turkey. Previous cases were confirmed in South-East Turkey with low cow mortality. Recent BEF-suspected outbreaks with high mortality were documented in many regions of Turkey in 2012. The aim of study was the epidemiological examination of the outbreak and molecular characterization of the viruses detected from the outbreak. For this reason, blood samples were collected from BEF-suspected outbreak regions. From the results of RT-PCR, high rate of BEF-suspected samples (48/60 or 80%) was found positive for BEF virus (BEFV) RNA. The nucleotide sequences of the G1 region of G gene of BEFV in the current study during the 2012 outbreak were grouped into cluster II of BEFV. It was suggested that BEFV may be spread out to other neighbor countries in the future years.

Antibodies to Schmallenberg virus in domestic livestock in Turkey.

This, partly retrospective study, was designed to determine the seroprevalence of Schmallenberg virus (SBV), a new Orthobunyavirus first reported in Germany in late 2011, in domestic ruminants from the Middle Black Sea, West, and Southeast regions of Turkey. An indirect enzyme-linked immunosorbent assay was used to screen serum samples collected from slaughterhouse animals between 2006 and 2013. The overall seroprevalence was 335/1,362 (24.5 %) with 325/816 (39.8 %), 5/307 (1.6 %), 3/109 (2.8 %), and 2/130 (1.5 %) recorded in cattle, sheep, goats, and Anatolian water buffalo, respectively. This is the first study to demonstrate the presence of antibodies to SBV in Turkish ruminants; it indicates that cattle are more susceptible to infection than sheep, goats, or buffalo and that exposure of domestic ruminants to SBV in Turkey may have occurred up to 5 years prior to the first recorded outbreak of the disease in 2011.

Vitamin D status in cattle with malignant catarrhal fever.

The aim of the present study was to determine the vitamin D status in cattle with malignant catarrhal fever (MCF). Twelve cattle diagnosed as MCF and 6 healthy cattle (controls) were used in the study. Serum 1,25-dihydroxyvitamin D(3) (1,25-D), 25-hydroxyvitamin D(3) (25-D), calcium, phosphorus and parathyroid hormone (PTH) levels were determined as 96.83 pg/ml, 30.0 ng/ml, 2.19 mmol/l, 1.57 mmol/l and 15.21 pg/ml in MCF group and 42.33 pg/ml, 37.0 ng/ml, 2.43 mmol/l, 1.96 mmol/l and 36.08 pg/ml in controls, respectively. Although serum 1,25-D level in the MCF group was increased (P<0.01), serum calcium (P<0.01) and PTH (P<0.05) levels were decreased compared to the controls. The results suggest that there might be an interaction between vitamin D status and MCF.

Effect of orthodontic appliances on oral microbiota--6 month follow-up.

OBJECTIVE: This study investigates the effect of fixed and removable orthodontic appliances among children on salivary Streptococcus mutans, Lactobacillus sp. and Candida albicans. STUDY DESIGN: The study was conducted on 69 patients aged between 6-17 years who used fixed or removable orthodontic appliances. Five ml samples of unstimulated saliva from each patient were collected at baseline and at the 1, 3 and 6 month periodic controls. Samples were diluted and plated on Mitis Salivarus Agar (MSA), Man Rogosa Sharp Agar (MRS) and Saboroud Dextrose Agar (SDA). The growths on the plates were examined under a stereomicroscope. Numbers of colony forming units (CFU) per plate were counted. For statistical analysis, the paired t test and Chi-Square were used. RESULTS: S mutans and Lactobacillus sp counts increased significantly 6 months after the insertion of fixed/removable orthodontic appliances in the oral cavity. A significant increase for C albicans presence was noted after 3 months compared with baseline for fixed appliances. CONCLUSION: Long-term utilization of orthodontic appliances may have a negative effect on microbial flora and increase the risk of new carious lesions and periodontal problems. Patients should be recalled within short time intervals to be motivated for oral hygiene during their orthodontic therapy.

Multidisciplinary approach to a nonsyndromic oligodontia patient using advanced surgical techniques.

Treatment of a woman suffering from oligodontia and multiple diastemata with insufficient alveolar ridges in both the maxilla and mandible is described in this clinical report. The total number of missing teeth was 11 (excluding the wisdom teeth). The treatment strategy consisted of an interdisciplinary team approach of orthodontic, surgical, and prosthodontic phases. Sinus floor augmentation, alveolar ridge augmentation via vertical alveolar distraction, and lateral augmentation with ramus graft procedures were performed after fixed orthodontic treatment and prior to dental implant placement. Oral rehabilitation of the patient was completed with the placement of fixed prostheses in the maxillary and mandibular posterior edentulous areas. Early dental intervention improved the patient's appearance and minimized the onset of emotional and psychosocial problems.

High cardiac troponin I plasma concentration in a calf with myocarditis.

A 15-day-old Brown Swiss calf, whose dam had suffered from foot-and-mouth disease, was presented with a history of depression and failure to suckle. The calf had an irregular cardiac rhythm and increased plasma cardiac troponin I (cTnI) detected with a commercial human immunoassay. The calf died the following day and myocarditis was detected. The cTnI assay may be useful in diagnosis of myocarditis in cattle.

Can intra-coronally bleached teeth be bonded safely after antioxidant treatment?

The aim of this study was to compare the effects of antioxidant treatment and delayed bonding, following intra-coronal bleaching, on the shear bond strength (SBS) and bond failure site of brackets bonded to enamel. Eighty mandibular incisors were divided into four equal groups. After root canal filling, specimens in Group 1 were used as control and not bleached. For experimental groups 2 to 4, bleaching agent was placed into the rest of the cavity for a four-day period and this bleaching treatment was performed two times. In Group 2, specimens were bonded immediately after bleaching; in Group 3, specimens were bleached and then immersed in artificial saliva for 30 days before bonding; in Group 4, specimens were bleached, treated with an antioxidant agent, and then bonded. The SBS values of bonded brackets were measured in megapascal (MPa), while adhesive remnant index (ARI) scores were determined after the brackets failed. The SBS values of Group 1 (mean: 20.3+/-7.1 MPa) and Group 4 (mean: 18.2+/-6.1 MPa) were significantly higher (p>0.001) than those of Group 2 (mean: 4.9+/-3.2 MPa) and Group 3 (mean: 8.7+/-4.9 MPa). No significant differences in SBS were found between Groups 1 and 4, and between Groups 2 and 3.

The effects of Kisspeptin antibodies on delayed estrus in rats.

In recent years, numerous studies have been conducted on the effects of Kisspeptin/GPR54 system on sexual cycle, which proved that this system regulated gonadotropin release through GnRH. This study aims to determine the effects of hyperimmune serum containing antibodies produced against Kisspeptin on the sexual cycle and GnRH receptors in rat pituitaries. To this end, five Wistar female rats were passively immunised using a hyperimmune serum obtained from two Wistar female rats against Kisspeptin 10. Another five rats were selected as the control group. Anti-Kisspeptin antibodies of the hyperimmunised rats in the serum were identified by ELISA. The sexual cycles of the rats were followed by the measurements of vaginal irrigations and the estradiol concentrations in their blood samples. The number of follicles and corpora lutea in their ovaries was determined through histopathological tests, and the GnRH receptors in their pituitaries were identified by immunohistochemistry. Consequently, strong seropositivity was detected in all passively immunised rats and the hyperimmune serum. However, no difference was found between the groups with regard to the number of estrous cycles observed, the interval between estrous periods, estradiol concentrations, the number of follicles and corpora lutea and immunohistochemical results.

Evaluation of the shear bond strength of 3 curing bracket bonding systems combined with an antibacterial adhesive.

INTRODUCTION: The purpose of this in-vitro study was to investigate the shear/peel bond strength of metal brackets bonded to human enamel with differently cured bonded systems combined with an antibacterial adhesive component. METHODS: One hundred twenty extracted molars were divided into 3 groups. Stainless steel brackets were bonded with 1 of 3 systems: group 1, no-mix bonding adhesive (Unite, 3M Unitek, Monrovia, Calif) (n = 40); group 2, 2-paste chemically cured bonding resin (Concise, 3M Unitek) (n = 40); and group 3, light-cured adhesive (Transbond XT, 3M Unitek) (n = 40). Each bonding group was separated into experimental (n = 20) and control groups (n = 20). In the 3 experimental groups, an antibacterial self-etch adhesive (Clearfil Protect Bond, Kurary, Osaka, Japan) was also applied to the enamel. Specimens in the control groups were bonded only with their relevant bonding systems according to the manufacturers' instructions. All specimens were stored in distilled water for 24 hours and thermocycled before testing. Debonded specimens were scored with the adhesive remnant index. RESULTS: The mean bond strengths (in megapascals) were 9.8 (control, 15.7) in group 1, 12.0 (control, 18.5) in group 2, and 11.6 (control, 12.4) in group 3. Statistical analysis with t tests showed no difference between group 3 and its control (P = .178), whereas groups 1 and 2 were statistically different from their controls (P = .000). CONCLUSIONS: The results indicated that the newly developed antibacterial self-etch adhesive can be combined with various bonding systems; achieved bond strengths were clinically more than satisfactory.

Hemorrhagic syndrome-like disease in calves with bovine viral diarrhea and mucosal disease complex.

BACKGROUND: Bovine viral diarrhea virus (BVDV) infection is one of the causes of hemorrhagic diathesis in cattle but there have been limited field studies about that condition. HYPOTHESIS: To identify the cause of hemorrhagic diathesis in calves and describe its clinical findings. ANIMALS: Five calves from a farm with 150 dairy cows. METHODS: Clinical examination of the calves was performed. After blood samples were obtained from 2 calves, whole blood, sera, and leukocyte samples were used for hematologic and hemostatic examinations, neutralization tests, virus isolation, and viral genome sequencing. RESULTS: The calves had moderate pyrexia, dullness, serous or mucous nasal discharge, and petechial and ecchymotic hemorrhages on mucosal surfaces. Severe thrombocytopenia and anemia were identified on hematologic examinations. All calves died within 10 days of the onset of clinical signs. Virologic examinations identified BVDV as the causative agent of the disease. CONCLUSIONS AND CLINICAL IMPORTANCE: This paper identifies a hemorrhagic syndrome-like disease in calves with bovine viral diarrhea and mucosal disease complex in Turkey.

Effect of an antioxidizing agent on the shear bond strength of brackets bonded to bleached human enamel.

INTRODUCTION: The purpose of this study was to investigate the effect of antioxidant treatment and delayed bonding on the shear bond strength of metal brackets bonded with composite resin to human enamel after bleaching with carbamide peroxide (CP). METHODS: Eighty recently extracted premolars were divided into an experimental group (n = 60), which was bleached with 10% CP, and a control group (n = 20), which was not bleached. The experimental group was further divided into 3 groups. Specimens in group 1 (n = 20) were bonded immediately after bleaching; specimens in group 2 (n = 20) were bleached, then treated with 10% sodium ascorbate, an antioxidant agent, and then bonded; group 3 specimens (n = 20) were bleached, then immersed in artificial saliva and held for 1 week before bonding. The specimens were debonded, and the enamel surfaces and bracket bases were examined with a stereomicroscope. The adhesive remnant index was used to assess the amount of resin left on the enamel surfaces after debonding. The shear bond strength data were subjected to 1-way analysis of variance. Multiple comparisons were performed with the Bonferroni test. The level of significance was established at P < .05 for all statistical tests. RESULTS: Shear bond strength of brackets bonded immediately after bleaching with 10% CP was significantly lower than that of brackets bonded to unbleached enamel (P < .05). No statistically significant differences in shear bond strength were noted when the antioxidant-treated and delayed bonding groups were compared with the control group (P > .05). CONCLUSIONS: Bleaching with 10% CP immediately before bonding reduces the bond strength of composite resin to enamel. Treating the bleached enamel surface with 10% sodium ascorbate or waiting 1 week reverses the reduction.

[Investigating the anti-inflammatory effect of dexamethasone in an asthma mouse model]. / Astim fare modelinde deksametazonun antiinflamatuvar etkisinin arastirilmasi.

We performed an asthma mice model in this study and aimed to investigate the levels of mediators in bronchoalveolar lavage fluid (BALF), and lung tissue, and the pathological changes response to the steroid treatment. BALB/c mice divided into three groups. PBS was applied to group 1 (control group). Asthma model was performed by exposing to ovalbumin in group 2 and 3. DEX was injected to group 3. After the last DEX dose all of the mice were killed by cervical dislocation. The samples of BALF and lung tissue were obtained. IL-4 and IL-5 levels of all samples were measured and inflammatory cells were counted in BALF. Evident eosinophilia was determined in BALF of group 2. Eosinophil numbers were lower in group 3 when compared with group 2 and this was statistically significant (p< 0.001). Inflammatory cell infiltration, eodema and hyperemia observed around the walls of bronchus and bronchiols in group 2. The lungs of group 3 had normal histological appearance. Both two cytokin levels of lung tissue were higher in group 2 than group 1, and this was statistically significant (for IL-4 p< 0.003, and for IL-5 p< 0.002). In group 3, both two cytokin levels were statistically lower than group 2 (for IL-4 p< 0.001, and for IL-5 p< 0.026). In BALF samples both two cytokin levels were higher in group 2 than group 1, and this was statistically significant (for IL-4 p< 0.004, and for IL-5 p< 0.001). In group 3, both two cytokin levels were lower than group 2, but it was not statistically significant (p> 0.05). In conclusion, it is thought that antiinflammatory effect of glucocorticoids occur by inhibiting the formation of IL-4, IL-5 and eosinophils.