Isolation, serological and molecular methods in screening of Burkholderia mallei in East Azerbaijan province, Iran.

Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and fourth quarters of 2020, throughout 2021, and in the first and second quarters of 2022, the complement fixation test (CFT) was performed on 350 horses. The malleination was used to confirm the positive CFT cases. Blood samples were taken for culture and for preparing serums to perform the enzyme-linked immunosorbent assay (ELISA). Deep eye discharge, nostril, cutaneous ulcers and lymph fluid swabs were cultured, and polymerase chain reaction (PCR) was carried out. Eleven horses were CFT-positive. Based on the malleination on the 11 horses, six were affected by glanders, five were not affected (false positive), and one horse was CFT-negative despite exhibiting clinical signs. It was positive by malleination, ELISA and PCR. A total number of seven positive cases of glanders were diagnosed. The B. mallei could not be isolated, but the Burkholderia cepacia complex was isolated in one case. Except for three cases, the results of the CFT, mallein and ELISA tests were consistent. The amount of confidence interval was 95.00%. It is suggested that ELISA could be used as a complement to CFT in screening and, if positive results are observed in one of the tests, the entire herd must be examined more accurately using the mallein and western blot confirmatory tests.

Methotrexate loaded in alginate beads for controlled drug release against breast cancer.

Methotrexate (MTX), as a folate antagonist is used for breast cancer chemotherapy, but its application due to the adverse side effects was limited. In this study, MTX were encapsulated in magnetic alginate beads coated with glutaraldehyde to control its release in order to reduce the side effects and improve its stability. The complex was characterized by physicochemical studies. The encapsulation efficiency was 75 % and the complex showed acceptable controlled release behavior. The cell cytotoxicity assessed using methylthiazol tetrazolium (MTT) method showed that magnetic alginate beads-MTX, in lower dosage has higher anticancer effect compared to the free MTX. The real-time polymerase chain reaction (PCR) was used to evaluate apoptotic factors Bcl2 associated X gene (Bax), B-cell lymphoma 2 (Bcl-2), and neuroinflammatory marker tumor necrosis factor-alpha (TNF-α) genes expression level on the treated cells. The findings demonstrated the significant increase of expression of Bax and a significant decrease in the expressions of Bcl-2 and TNF-α in Michigan cancer foundation-7 (MCF-7) cells. These results indicated that the developed drug can overcome the side effects of MTX and offer a controlled drug release for a sustained period with the long-term treatment of breast cancer.

Viola tricolor Hydroalcoholic Extract Improves Behavioral Deficiencies in Rats Exposed to Chronic Immobilization Stress

Abstract This study aims to investigate the effect of Viola tricolor extract on hippocampal neuronal death, interleukin (IL) -6 and IL-10 expression, spatial memory, anxiety, and depression in rats exposed to chronic immobilization stress. Rats were divided into groups Control, Viola300, Viola600, Stress, Stress-Viola300, and Stress-Viola600. Animals were placed in a restrainer (6 h / 21 days) to stress exposure. V. tricolor hydro-alcoholic extract was also administrated at doses of 300 and 600 mg/kg by gavage. The extract caused immobilized animals to spend more time in the target quadrant in the Morris water maze test. It also increased the percentage of entries into the open arm and the percentage of time spent in the open arm of the elevated plus-maze in immobilized rats. Treatment with the V. tricolor extract significantly reduced the immobility time of stressed rats in the forced swimming test. Furthermore, it significantly reduced neuronal death and expression of IL-6 in the hippocampus of immobilized animals but could not prevent the decrease of IL-10 expression. We concluded that V. tricolor protects rats from stress-induced behavioral damages, at least in part, by suppressing neuronal death and decreasing IL-6 expression.

Possible ameliorating effects of Glycyrrhiza Glabra (Licorice) on the sperm parameters in rats under high fat diet.

OBJECTIVES: Adverse effects of obesity, which is caused by an imbalance between the energy intake and expenditure, on the male reproductive system have been reported. Considering the anti-obesity effect of Glycyrrhiza Glabra (GC), we conducted this study to elucidate whether it can ameliorate the sperm parameters. METHODS: In this experimental study, male Wistar rats of 6-8 weeks old were divided into four groups: control, high fat diet (HFD), GC50 (HFD plus 50 mg/kg GC extract), and GC100 (HFD plus 100 mg/kg GC extract). During the 16 weeks of the study course, the rats consumed the extract through gavage, daily. Body mass index (BMI), body weight gain, serum lipid profile, leptin concentration, and sperm parameters were investigated. Data were analyzed by one-way analysis of variance (ANOVA) (post hoc Tukey) to express the significance of mean differences of variables between groups, and linear regression test was used to express the correlation model of variables. Both tests were performed by SPSS software; p≤0.05 was considered significant. RESULTS: BMI was significantly decreased by the GC50 and GC100 groups compared to HFD group. GC50 group considerably decreased leptin level compared to HFD group. A significant positive correlation between leptin and triglyceride levels was evident. GC50 and GC100 extensively increased the total sperm motility and ameliorated the sperm abnormal morphology and count compared to HFD group. CONCLUSION: Glycyrrhiza Glabra extract may exert its ameliorating effects on the sperm parameters through its anti-obesity impact. Both doses of the extract were effective, however, the GC100 was more effective in improving the sperm parameters.

Spinacia oleracea L. extract attenuates hippocampal expression of TNF-α and IL-1ß in rats exposed to chronic restraint stress.

Background: Restraint stress causes inflammation in nervous system that leads to emersion of neurodegenerative diseases. Spinach (Spinacia oleracea L.) contains different agents with antioxidant, antiapoptosis, and hepatoprotective properties. This study examined the effect of spinach hydroalcoholic extract (SHE) on TNF-α and IL-1ß expression in hippocampus of male Wistar rats exposed to chronic restraint stress. Methods: Rats were divided into 6 groups of 5: (1) control (intact); (2) nS-S200; (3) nS-S400; (4) stress; (5) stress-S200; (6) stressS400. Groups 2 and 3 and groups 5 and 6 received S. oleracea leaf hydroalcoholic extract in 200 and 400 mg/kg doses for 21 consecutive days by gavage. Groups 4, 5 and 6 were put in a restrainer 6 hours per day for 21 consecutive days. Then, the expression of IL-1ß and TNF-α mRNAs and neuronal death in the hippocampus of rats were assessed by real time PCR and Nissl staining, respectively. Oneway analysis of variance was used for data analysis, and p<0.05 was considered statistically significant. Results: The results showed that the expression of IL-1ß and TNF-α was increased in hippocampus of rats exposed to stress compared to control groups (p<0.001). Furthermore, the expression of these proinflammatory cytokines was decreased in the stress-S200 and stress-S400 groups when compared to stress group (p<0.001). Immobility also caused neuronal death in CA1 region of hippocampus, and SHE reduced damage in CA1 pyramidal neurons layer in stressed rats. Conclusion: Spinach decreases neuroinflammation in hippocampus of stressed rats, which may be due to its abundant antiinflammatory and antioxidant phytochemicals. The results of this study suggest that spinach may be effective in the prevention and treatment of neurodegenerative diseases.

Study of association between interleukin-8 - 845 T/C and + 781 C/T polymorphisms with periodontitis disease among population from Western Iran.

Periodontitis is a chronic inflammatory disease that influences the protective tissues of teeth. IL-8, a member of the chemokine super-family, plays vital roles in the pathogenesis of periodontitis with activation and migration of neutrophils in inflammatory regions. The purpose of present study was to evaluate the association of interleukin-8 - 845 T/C and + 781 C/T polymorphisms with periodontitis in an Iranian population. A total of 65 patients with periodontitis including 18 patients with chronic periodontitis and 47 patients with aggressive periodontitis and 55 controls were enrolled into our study. Interleukin-8 - 845 T/C and + 781 C/T polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. For + 781C/T locus, in the dominant genetic model there was a significant difference between TT vs. CC + CT genotypes that significantly had a protective role against periodontitis disease with a value of 0.38 (95% CI 0.16-0.90, p = 0.02). Also, the analysis of results showed a significant positive association between the distribution of IL-8 - 845 T/C alleles and the risk of periodontitis disease (χ2 = 6.2, p = 0.01) that presence of C allele of IL-8 - 845 increased the risk of periodontitis disease by 9.08-fold [OR 9.08 (95% CI 1.14-72.12, p = 0.03)]. In conclusion, our results demonstrate a positive association between distribution of IL-8 - 845 T/C alleles and risk of periodontitis disease.

Evaluation of the Role of -137G/C Single Nucleotide Polymorphism (rs187238) and Gene Expression Levels of the IL-18 in Patients with Coronary Artery Disease.

OBJECTIVES: Interleukin-18 (IL-18) is a proinflammatory and proatherogenic cytokine, and its genetic variations may contribute to the development of coronary artery disease (CAD). We sought to investigate the role of -137G/C polymorphism and gene expression levels of IL-18 in patients with CAD. METHODS: The study population included 100 patients with angiographically proven CAD and 100 matched controls. Total RNA and DNA were extracted from leukocytes using appropriate kits. The genotype of -137G/C polymorphism and gene expression level of IL-18 was determined using allele-specific polymerase chain reaction (PCR) and real-time (RT)-PCR assay, respectively. RESULTS: The genotypic and allelic distribution of IL-18 -137G/C polymorphism was not significantly different between the two groups (p > 0.050). Moreover, the -137G/C polymorphism did not increase the risk of CAD in dominant and recessive genetic models (p > 0.050). However, subgroup analysis of CAD patients revealed that the IL-18 -137G/C polymorphism was significantly associated with increased risk of CAD in hypertensive patients (odds ratio (OR) = 7.51; 95% confidence interval (CI): 1.24-25.17; p = 0.019) and smokers (OR = 4.90; 95% CI: 1.21-19.70; p = 0.031) but not in the diabetic subpopulation (p = 0.261). The genotype distribution of IL-18 -137G/C genetic polymorphism was significantly different among patients with one, two, and three stenotic vessels (p < 0.050). The gene expression level of IL-18 was significantly higher in the CAD group than the control group (p < 0.001). Moreover, the carriers of CC genotype had significantly lower gene expression levels of IL-18 than carriers of GG genotype (p < 0.050). CONCLUSIONS: The -137G/C polymorphism of IL-18 may be associated with the CAD risk in hypertensive and smoker subgroup of CAD patients. The -137G/C polymorphism seems to play an important role in determining the severity of CAD. Increased IL-18 gene expression level is a significant risk factor for the development of CAD. The CC genotype of -137G/C polymorphism is associated with lower IL-18 gene expression levels.

Enhanced antibacterial effect of azlocillin in conjugation with silver nanoparticles against Pseudomonas aeruginosa.

In recent years, the problems associated with bacterial resistance to antibiotics caused nanodrugs to be considered as a new way for infectious diseases treatment. The main purpose of this study was to develop a new agent against Pseudomonas aeruginosa, a very difficult bacterium to treat, based on azlocillin antibiotic and silver nanoparticles (AgNPs). Azlocillin was conjugated with AgNPs by chemical methods and its antimicrobial activity was studied against P. aeruginosa using well diffusion agar method. Then, minimum inhibitory concentration and minimum bactericidal concentration of the new conjugate was specified with macro-dilution method. The animal study showed the considerable enhanced antibacterial effect of azlocillin in conjugation with AgNPs against P. aeruginosa in comparison with azlocillin alone, AgNPs alone and azlocillin in combination with AgNPs.

Role of IL28-B Polymorphism (rs12979860) on Sustained Virological Response to Pegylated Interferon/Ribavirin in Iranian Patients With Chronic Hepatitis C.

BACKGROUND: The current medical treatment for hepatitis C virus (HCV) infection is pegylated interferon plus ribavirin, but just 50% of genotype 1 HCV patients and about 80% of HCV genotype 3 patients are treated completely. Recently, the rs12979860 C/T polymorphism, which is located 3 kb upstream of the IL28b gene that codes IFNλ3, shows a powerful association in response to the treatment in HCV patients. OBJECTIVES: The aim of this study was to evaluate the relationship between IL28b single nucleotide polymorphism (SNP) and treatment outcomes among chronic HCV patients in Iran. PATIENTS AND METHODS: In this cross-sectional study, 108 blood samples were collected from chronic patients in Iran; 50 unrelated healthy subject samples were also collected. Genomic DNA was extracted, and rs12979860 SNP was done by PCR-RFLP. Finally, products were detected on 12% polyacrylamide gel electrophoresis. RESULTS: The analysis of data for C/T SNP showed that the CC genotype is more common in the control group than in the group of patients. In contrast, the frequency of TT as a mutant genotype is more frequent in patients than in uninfected people. In addition, results showed a statistically significant relationship between CC, CT, and TT genotypes in sensitive and resistant groups (P value: < 0.001, Or: 0.003, CI: 0-0.047). This relationship was also examined in terms of allele frequency, to determine whether the possibility of resistance to treatment in patients with T allele is more than in patients who carry C allele (P value: < 0.001). CONCLUSIONS: These results showed a significant effect between rs12979860 SNP and sustained virological response (SVR) rate in Iranian patients with chronic HCV. To decrease the cost of long treatments and to prevent severe side effects, determining this polymorphism at the beginning of treatment can be very helpful for patients and physicians.

The relationship between vitamin D receptor (VDR) polymorphism and the occurrence of osteoporosis in menopausal Iranian women.

BACKGROUND: Osteoporosis, a multifactorial disease with reduced bone mineral density which increases the probability of bone fractures, is caused by calcium deficiency, and its incidence increases with age. It has been determined that mutations in functional regions of vitamin D receptor gene will affect the metabolism of minerals especially calcium and, therefore, bone density. The present study evaluates the relation between vitamin D receptor polymorphisms, TaqI (rs731236) and ApaI (rs7975232), and osteoporosis in menopausal Azari women in Zanjan province. MATERIALS AND METHODS: This case-control study has been conducted on 50 menopausal women suffering from osteoporosis and 50 menopausal women who did not suffer from osteoporosis in Zanjan province. The diagnosis of osteoporosis was confirmed using DEXA instrument. Peripheral blood was collected from the subjects and controls to extract DNA and assess the ApaI and TaqI polymorphisms using PCR-RFLP method. The results were interpreted using independent T-test, chi-square, and Pearson correlation coefficient with a p-value less than 0.05. RESULTS: There was not a significant difference between the frequency of ApaI (AA/Aa/aa) and TaqI (TT/Tt/tt) genotypes in cases (mean age 68.72) and controls (mean age 64.7) (p=0.37 and p=0.64, respectively). In addition, ApaI/TaqI allele haplotype in osteoporotic population showed non-significant relation (p value=0.563) compared with the control group. DISCUSSION AND CONCLUSION: The relationship between the genotypes and osteoporosis, cancers, and mineral metabolism disorders has been studied for a long time. Although there has been a significant relation between the aforementioned genotypes and osteoporosis or reduced mineral density-related bone fractures in some studied, some other studies have opposing results. Therefore, it is only possible to reach an acceptable conclusion by studying the haplotype of the polymorphisms in subjects.