Evaluation of the sedative and physiological effects of xylazine, detomidine, medetomidine and dexmedetomidine in goats.

BACKGROUND: Many α2 -agonists are commonly used for sedation and analgesia in ruminants. INTRODUCTION: The present study aims to compare the sedative and physiological effects of intravenous (IV) administration of xylazine, detomidine, medetomidine and dexmedetomidine in goats. METHODS: Ten healthy goats aged 6 ± 1 months and weighing 15 ± 2 kg were used in experimental, crossover Latin square, randomised and blinded study. Animals were assigned to five IV treatments: control (normal saline); xylazine (100 µg kg-1 ); detomidine (50 µg kg-1 ); medetomidine (20 µg kg-1 ) and dexmedetomidine (5 µg kg-1 ). The degree of sedation was investigated using a numerical ranking scale of 0-10. Sedation scores were compared at each time using nonparametric (Kruskal-Wallis and Mann-Whitney U) tests. RESULTS: Heart rate (HR), respiratory rate (RR), rectal temperature (RT), ruminal motility and capillary refill time (CRT) were performed before (baseline) and after drug administration. Animals in α2 -adrenergic agonist treatments were sedated at 5-60 min. There were no significant differences among α2 -adrenergic agonist treatments at 5-60 min in sedation scores. HR significantly decreased from baseline 5-90 min after α2 -adrenergic agonists' administration. Ruminal motility was decreased in α2 -adrenergic agonist treatments at 5, 90 and 120 min and absent at 10-60 min. A significant decrease from baseline in RR was detected between 30 and 90 min after α2 -adrenergic agonists' administration. RT was unchanged in any treatment for 120 min. CRT was less than 2 s at all time points following each treatment. CONCLUSIONS: The duration of sedation was up to 60 min after IV administration of xylazine (100 µg kg-1 ), detomidine (50 µg kg-1 ), medetomidine (20 µg kg-1 ) and dexmedetomidine (5 µg kg-1 ) in goats in this study. No significant differences were detected between xylazine, detomidine, medetomidine and dexmedetomidine in goats.

Effect of Two Whitening Toothpastes on Composite Resin Restorations Discolored by 0.2% Chlorhexidine Mouthwash.

Objectives: This study compared the effect of two whitening toothpastes on composite specimens discolored with 0.2% chlorhexidine (CHX). Materials and Methods: Twenty-four composite specimens were fabricated from Charisma Diamond composite resin. The initial color of specimens was measured according to the CIE L*a*b* color system using a spectrophotometer. The specimens were immersed in 0.2% CHX twice a day for 1 minute each time, for 2 weeks. The color of specimens was measured again, and the specimens were assigned to three groups (n=8). The control group specimens were immersed in distilled water. The two test group specimens were brushed twice daily for 21 days with Oral B toothbrush and Signal White Now and Crest 3D White whitening toothpastes each time for 30 seconds. The color of specimens was measured again. Data were analyzed by one-way ANOVA and t-test. Results: CHX increased the a, b, and L color parameters in all groups. There were no significant differences in ΔL (P=0.10), Δa (P=0.24), and Δb (P=0.07) among the study groups. The a, b, and L parameters decreased after brushing the specimens discolored with 0.2% CHX with the whitening toothpastes. There were significant differences in ΔL (P=0.03), Δa (P=0.02), and Δb (P=0.01) among the three study groups after using the whitening toothpastes. The highest ΔL, Δa, Δb, and ΔE values were recorded in Crest 3D White group, followed by Signal White Now group. Conclusion: Crest 3D White whitening toothpaste had higher efficacy to resume the original color of composite specimens discolored with 0.2% CHX.

Induction of Immunogenic Response in BALB/c Mice by Live and Killed Form of Recombinant Lactococcus lactis Displaying EG95 of Echinococcus granulosus.

Background: Cystic echinococcosis is a zoonotic parasitic infection caused by Echinococcus granulosus worldwide and is associated with economic losses among livestock animals. EG95 is an immunogenic antigen from the E. granulosus. Lactococcus lactis has been prested as a safe vehicle for antigen delivery. The goal of this study was to design a novel L. lactis strain displaying EG95 as a vaccine delivery system. Methods: The eg95 encoding gene fragment fused to the M6 anchoring protein was cloned into the pNZ7021 vector, and L. lactis NZ9000 displaying recombinant EG95 was constructed. The expression of an approximately 32-kDa EG95 protein was confirmed by Western blotting and immunofluorescence analysis. The immune responses were evaluated in BALB/c mice immunized orally and subcutaneously with the live and killed recombinant L. lactis, respectively. Results: Total IgG level in mice immunized with heat-killed recombinant L. lactis (pNZ7021-eg95) significantly increased compared to the control group. Mucosal IgA was significantly higher in mice received live recombinant L. lactis (pNZ7021-eg95) compared to the control mice. Splenic lymphocytes from immunized mice represented the high levels of IFN-γ and the low-levels of IL-4 and IL-10. Conclusion: Our results indicate that immunization with EG95-expressing L. lactis can induce both specific humoral and cellular immune responses in mice.

Recombinant Lactococcus Lactis Displaying Omp31 Antigen of Brucella melitensis Can Induce an Immunogenic Response in BALB/c Mice.

Since Brucella infection mostly occurs through the mucosal surfaces, immune response induced by vaccine that is delivered by a way of mucosal route can be drastically enhanced to control the brucellosis. Omp31is the major outer membrane protein of Brucella, and is considered as a protective antigen against Brucella infection. Accordingly, Lactococcus lactis has been used as an antigen-delivering vector to develop a vaccine-induced mucosal response for having a safer vaccination against brucellosis. A designed omp31 gene fused to the usp45 signal peptide and M6 cell wall anchor was sub cloned in the pNZ7021 expression vector, and a recombinant L. lactis displaying Omp31 was constructed. Omp31 protein expression was confirmed using Western blotting and immunofluorescence analysis. Animals were orally and intraperitoneally immunized with live or killed L. lactis expressing Omp31, respectively. The humoral and cellular immune responses were evaluated by measuring the specific cytokines and antibodies. sIgA, serum IgA, IgM, and total IgG antibodies significantly increased in the mice immunized with live recombinant L. lactis expressing Omp31 and also serum IgM, and total IgG antibodies significantly increased in mice immunized with killed recombinant L. lactis expressing Omp31. Among IgG subtypes, IgG2a response was significantly higher in both groups compared to IgG1. In mice groups immunized with recombinant L. lactis, the IFN-γ and IL-10 level elevated; however, there was no change in the level of IL-4. These results indicated that recombinants L. lactis induce both humoral and cellular immune responses in mice, and also vaccines based on L. lactis-derived live carriers are promising interventions against Brucella melitensis infections.

Helicobacter pylori Peptidyl Prolyl Isomerase Expression Is Associated with the Severity of Gastritis.

PURPOSE: Helicobacter pylori secretory peptidyl prolyl isomerase, HP0175, is progressively identified as a pro-inflammatory and pro-carcinogenic protein, which serves to link H. pylori infection to its more severe clinical outcomes. Here, we have analyzed host HP0175-specific antibody responses in relation to the severity of gastritis. METHODS: The HP0175 gene fragment was PCR-amplified, cloned, expressed and purified by Ni-NTA affinity chromatography. Serum antigen-specific antibody responses of non-ulcer dyspeptic patients (N = 176) against recombinant HP0175 were detected by western blotting. The infection status of these subjects was determined by rapid urease test, culture, histology, and serology. The grade of inflammation and stage of atrophy were scored blindly according to the OLGA staging system. RESULTS: The recombinant HP0175 (rHP0175) was expressed as a ~35 kDa protein and its identity was confirmed by western blotting using anti-6X His tag antibody and pooled H. pylori-positive sera. Serum IgG antibodies against rHP0175 segregated our patients into two similar-sized groups of sero-positives (90/176, 51.1 %) and sero-negatives (86/176, 48.9 %). The former presented with higher grades of gastric inflammation (OR = 4.4, 95 % CI = 1.9-9.9, P = 0.001) and stages of gastric atrophy (OR = 18.3, 95 %CI = 1.4-246.6, P = 0.028). CONCLUSION: Our findings lend further support to the pro-inflammatory nature of H. pylori peptidyl prolyl isomerase (HP0175) and recommends this antigen as a non-invasive serum biomarker of the severity of H. pylori-associated gastritis.

Distribution of cytokine gene single nucleotide polymorphisms among a multi-ethnic Iranian population.

BACKGROUND: Cytokine gene single nucleotide polymorphisms (SNPs) are widely used to study susceptibility to complex diseases and as a tool for anthropological studies. MATERIALS AND METHODS: To investigate cytokine SNPs in an Iranian multi-ethnic population, we have investigated 10 interleukin (IL) SNPs (IL-1ß (C-511T, T-31C), IL-2 (G-384T), IL-4 (C-590T), IL-6 (G-174C), IL-8 (T-251A), IL-10 (G-1082A, C-819T, C-592A) and tumor necrosis factor-alpha (TNF-α) (G-308A) in 415 Iranian subjects comprising of 6 different ethnicities. Allelic and genotypic frequencies as well as Hardy-Weinberg equilibrium (HWE) were calculated by PyPop software. Population genetic indices including observed heterozygosity (Ho), expected heterozygosity (He), fixation index (FIS), the effective number of alleles (N e) and polymorphism information content (PIC) were derived using Popgene 32 software. Multidimensional scaling (MDS) was constructed using Reynold's genetic distance obtained from the frequencies of cytokine gene polymorphism. RESULTS: Genotypic distributions were consistent with the HWE assumptions, except for 3 loci (IL-4-590, IL-8-251 and IL-10-819) in Fars and 4 loci (IL-4-590, IL-6-174, IL-10-1082 and TNF-α-308) in Turks. Pairwise assessment of allelic frequencies, detected differences at the IL-4-590 locus in Gilakis versus Kurds (P = 0.028) and Lurs (P = 0.022). Mazanis and Gilakis displayed the highest (Ho= 0.50 ± 0.24) and lowest (Ho= 0.34 ± 0.16) mean observed heterozygosity, respectively. CONCLUSIONS: MDS analysis of our study population, in comparison with others, revealed that Iranian ethnicities except Kurds and Mazanis were tightly located within a single cluster with closest genetic affinity to Europeans.

Age-Specific Gastric Cancer Risk Indicated by the Combination of Helicobacter pylori Sero-Status and Serum Pepsinogen Levels.

BACKGROUND: Serologic screening of gastric cancer (GC) by serum pepsinogens (sPG) levels and Helicobacter pylori (Hp) sero-status, though highly informative, has provided heterogeneous results. Here, we have evaluated the modifying effects of demographic factors on the risk impact of Hp sero-status/sPG levels in gastric cancer, with particular emphasis on age. METHODS: A cross-sectional study was carried out on 1341 individuals (GC = 578, healthy = 763), who were stratified into two age groups: 35-59 years (middle-aged, n = 830) and ≥ 60 years (60 years-plus, n = 511). Demographic factors and serological states (Hp sero-staus and sPG levels) were recorded by subject interview and serum ELISAs, respectively. Covariate-specific odds ratios were calculated by multivariable logistic regression. RESULTS: Hp infection was consistently associated with increased sPGI and sPGII levels in the 60 year-plus, but not the middle-aged group. The joint examination of the variable states of the three serum biomarkers (Hp serology, sPGI, and sPGI/II ratio), in the 60 year-plus age group, demonstrated a stepwise escalation of risk from the single (sPGI low; OR = 2.6), to double (sPGI low/sPGI/II low; OR = 3.55, and Hp positive/sPGI low; OR = 5.0) and ultimately triple (Hp positive/PGI low/PGI/II low; OR = 10.48) positive states, in reference to the triple negatives. However, this pattern was not exhibited in the middle-aged subjects. CONCLUSION: Age was clearly identified as a modifying factor on the risk projection of the combined states of Hp serology and sPG levels in gastric cancer screening, reflected by the augmented (~10.5 fold) risk of GC in the triple positive (Hp positive/sPGI low/sPGI/II low) 60 year-plus subjects, which was not evident in the middle-aged group.

Helicobacter pylori Omp18 and its application in serologic screening of infection.

Helicobacter pylori (Hp) is a major risk factor for gastrointestinal disorders including gastric cancer. We evaluated host serum antibody responses toward outer membrane protein18 in comparison with Urease A and B subunits. omp18 and ureA-ureB gene fragments were PCR amplified, cloned, and expressed in E. coli expression system. The expressed proteins were visualized on SDS-PAGE and confirmed by immuno-blotting. Purified proteins were applied in western blotting assays in comparison with local and foreign ELISA kits. ROC curve analysis identified the optimum cut-off points for each protein. rOmp18 represented the highest rates of sensitivity (94%), specificity (89%), PPV (97.4%), NPV (77.4%), and accuracy (93.2%) in comparison with urease A and B subunits. These immunologic indices were in "substantial" agreement (Κ = 0.7) with the gold standard tests for Hp detection. This study recommends Hp conserved Omp18 as a reliable serologic marker for accurate detection of Hp infection particularly for application in population screening approaches.

Multiple gene status in Helicobacter pylori strains and risk of gastric cancer development.

BACKGROUND/AIM: The identification of the vacA intermediate region has provided new insights into the role of vacA heterogeneity in relation to gastro-duodenal pathogenesis. The aim of this study was to assess vacA polymorphism in Iranian Helicobacter pylori strains and its association with cagA as a major virulence determinant, gastric histopathology and disease. METHODS: vacA polymorphism and serum antibody responses were studied in 207 H. pylori-infected (139 NUD, 34 PUD, and 34 GC) patients and correlated with gastric histopathology. RESULTS: Multivariate logistic regression analysis found intermediate region typing superior to signal or mid region typing for screening high risk patients. vacA i1 allele was identified as an independent predictor of dysplasia (OR = 9.044; 95% CI: 1.11-73.33). Possession of s1/i1/cagA(+) strains was also identified as a predictor of intestinal metaplasia (OR = 3; 95% CI: 1.13-7.95), dysplasia (OR = 9.9; 95% CI: 1.23-80.86) and risk of GC (OR = 6.9; 95% CI: 2.5-18.66) as well as induction of anti-VacA sero-positivity (OR = 5.04; 95% CI: 1.8-13.6). Anti-VacA serology correctly detected 83.8% of s1/i1/cagA(+) strains carried by high-risk patients. CONCLUSIONS: The current study emphasizes the implication of vacA polymorphic structure, especially the s1/i1/cagA(+) genotype, in increasing the risk of GC by revealing their association with gastric pre-neoplastic changes and their reflection in VacA sero-positivity which encourages the application of noninvasive procedures in population screening.