Belma Durupinar

Antimicrobial activity of plant extract Ankaferd Blood Stopper.

The in vitro antimicrobial activity of Ankaferd Blood Stopper (ABS) was assessed on 102 clinical isolates from both Gram negative and Gram positive bacteria and four standard strains, including MRSA ATCC 43300, MSSA ATCC 25923, P. aeruginosa ATCC 27853 and E. coli ATCC 35218. ABS was significantly active against all bacteria investigated.

T helper 1 type cytokines polymorphisms: association with susceptibility to Behçet’s disease

The pathogenesis of Behçet’s disease (BD) is not fully understood and immunological abnormalities and genetic factors have been investigated. Because serum concentrations of mainly T helper 1 (Th1) type cells have been reported to be increased in BD, we aimed to investigate whether certain cytokine polymorphisms might represent a risk factor for developing BD. We genotyped 80 patients with BD and 105 healthy controls for interleukin (IL)-1 α (T/C −889), IL-1 β (C/T −511, T/C +3962), IL-1R (C/T pst11970), IL-1RA (T/C mspa111100), IL-2 (T/G −330), IL-12 (C/A −1188), interferon (IFN)-γ (A/T UTR 5644), and TNF-α (G/A −238) polymorphisms. Analyses of cytokine polymorphisms were performed with PCR-SSP. The genotype and allele frequencies of the patients and controls were compared and the association between the polymorphisms of the cytokines with the clinical findings was investigated. Genotype distribution showed significant differences between the patients and the controls for the IL-1 α −889, IL-1 β −511, IL-1 β +3962, IL-1R, IL-12, IFN- γ, and TNF-α cytokines. We didn’t observe significant difference in genotypic frequencies of IL-1RA and IL-2 in our study. Comparison of the IL-1 α −889, IL 1 β −511, and IL 1 β +3962 genotype frequencies showed significant increase in CC genotype between the patients and the controls. The individuals with IL-1R TT polymorphism had a higher risk for BD compared to patients with CT/CC polymorphism. Comparison of IL-12, IFN- γ, and TNF-α, genotype frequencies showed significant increase in CA, AA, and AA genotypes between the patients and controls, respectively. The frequencies of genotypes according to the clinical features of the patients with BD did not show a significant difference (p > 0.05). Our study suggests that development of BD might be determined by various cytokine gene polymorphisms. However, further studies on larger numbers of cases are needed before definite conclusions can be drawn.

A Multidrug Efflux Pump Inhibitor Reduces Fluoroquinolone Resistance in Pseudomonas aeruginosa Isolates

In general, resistance to fluoroquinolones (FQs) in gram-negative bacteria is acquired either by mutations in DNA gyrase and topoisomerase IV or by active export of the agents via antibiotic efflux pumps. Reduced porin expression is also proposed to be another mechanism leading to resistance. In this study, interaction between levofloxacin, ofloxacin, and ciprofloxacin with MC-207,110 (multidrug efflux pump inhibitor) was investigated by a checkerboard assay using Pseudomonas aeruginosa. Levofloxacin, ofloxacin, and ciprofloxacin were tested at different concentrations (0.06–64 µg/ml) and MC-207,110 was tested at a concentration range of 4–128 µg/ml. In the presence of MC-207,110 (at 128, 64, 32, 16 µg/ml) resistance to FQs was inhibited significantly and MIC values were decreased, except at 8 and 4 µg/ml of MC-207,110. When MC-207,110 was used, resistance of P. aeruginosa to FQs in vitro was inhibited significantly, suggesting that MC-207,110 may be useful for use in clinical treatment protocols to overcome FQs resistance.

HLA-B27 polymorphism in Turkish patients with ankylosing spondylitis

The aim of this study was to determine the distribution of human leukocyte antigen (HLA)-B27 subtypes in serologically HLA-B27-positive ankylosing spondylitis (AS) patients and healthy controls from the Turkish population and to compare this with other reports from other populations. We subtyped HLA-B27 in 38 HLA-B27-positive Turkish patients with AS and 47 HLA-B27-positive healthy controls without AS by polymerase chain reaction with specific sequence primers (PCR-SSP). The results demonstrated that: B*2705 was the predominant subtype among both of the patients (71.1%) and controls (68.0%). B*2702 was observed in 26.3% and 32.0% of the patients and controls, respectively. B*2708 subtype was found in 2.6% of the patients but not among the control group. When the distribution of B27 subtypes in Turkish populations was compared with that in other populations, similar frequencies with the Caucasian–Europe groups were noted. However, this should be interpreted carefully because of the small number of individuals in our study.

Drug susceptibility testing of Mycobacterium tuberculosis by the broth microdilution method with 7H9 broth

In this study, we have evaluated the broth microdilution method (BMM) for susceptibility testing of Mycobacterium tuberculosis. A total of 43 clinical isolates of M. tuberculosis and H37Rv as a control strain were studied. All isolates were tested by the proportion method and the BMM for isoniazid (INH), rifampicin (RIF), streptomycin (STR), and ethambutol (ETM). The proportion method was carried out according to the National Committee for Clinical Laboratory Standards (NCCLS) on Löwenstein-Jensen (LJ) medium. The BMM was carried out using 7H9 broth with 96 well-plates. All strains were tested at 3.2-0.05 micro g/ml, 16-0.25 micro g/ml, 32-0.5 micro g/ml, and 32-0.5 micro g/ml concentrations for INH, RIF, STR, and ETM, respectively. When the BMM was compared with the proportion method, sensitivity was 100, 100, 96.9, and 90.2%, while specificity was 100, 85.7, 90.9, and 100% for INH, RIF, STR, and ETM, respectively. The plates were examined 7, 10, 14, and 21 days after incubation. The majority of the result were obtained at 14th days after incubation, while the proportion method result were ended in 21-28 days. According to our results, it may be suggested that the BMM is suitable for early determining of multidrug-resistance-M. tuberculosis strains in developed or developing countries.

Two New Colorimetric Methods for Early Detection of Vancomycin and Oxacillin Resistance in Staphylococcus aureus

ABSTRACT We developed two colorimetric methods for the detection of vancomycin- and oxacillin-resistant Staphylococcus aureus in ≤6 h: (i) a nitrate reductase assay and (ii) a resazurin microplate method. MICs agreed with results obtained by CLSI methods for oxacillin. However, detection of vancomycin resistance required a larger inoculum. These methods may be recommended for the detection of vancomycin- and oxacillin-resistant S. aureus.

Human leucocyte antigens and cytokine gene polymorphisms and tuberculosis

PURPOSE Several genes encoding different cytokines and human leucocyte antigens (HLA) may play crucial roles in host susceptibility to tuberculosis (TB). Our objective was to investigate whether these genes might be associated with protection from or susceptibility to TB. MATERIALS AND METHODS Genomic DNA from patients with TB (n = 30) and ethnically matched controls (n = 30) was genotyped by using sequence-specific primers-polymerase chain reaction and sequence-specific oligonucletid methods. RESULTS Our results demonstrated that HLA-CwFNx0101 [P = 0.05, odds ration (OR) (95% confidence interval) = 2.269 (1.702-3.027)] allele frequency was significantly more common in TB patients than in healthy controls, and HLA-CwFNx0101 may be associated with susceptibility to TB. Analysis of cytokine allele frequencies showed that interleukin (IL)-10, -819 C and -592 C alleles was significantly more common in TB patients than in controls (pc: 0.038 and 0.017, respectively). From the IL-10 cluster, a positive significant difference was found at positions -1082 and -592 C/C (pc: 0.027 and 0.054, respectively) genotypes. Although these differences could be explained by the highest frequency of C/C and G/G homozygous patients with TB, in contrast to the control group, statistically significant differences for the C/C genotype however were lost after Bonferroni correction of the P-values. CONCLUSION Altogether, our results suggest that the polymorphisms in HLA (class I) and cytokine (IL-10) genes may affect the susceptibility to TB and increase the risk of developing the disease.

Effect of 1-(1-Naphtylmethyl)-piperazine, an Efflux Pump Inhibitor, on Antimicrobial Drug Susceptibilities of Clinical Acinetobacter baumannii Isolates

In this study, the effects of 1-(1-naphtylmethyl)-piperazine (NMP), an efflux pump inhibitor, on antimicrobial drug susceptibilities of 42 clinical Acinetobacter baumannii isolates were investigated by the disc diffusion method. The inhibition zone diameters of antibiotic discs were tested in the presence and absence of NMP and then these zone diameters were compared. Presence of NMP restored ciprofloxacin susceptibility in 15 intermediate and 2 resistant isolates. One ciprofloxacin resistant isolate became intermediate in the presence of NMP. One isolate resistant to gentamicin became intermediate with NMP. Interestingly, one isolate susceptible to meropenem became resistant in the presence of NMP. Although NMP increased the inhibition zone diameters of some of the tested antibiotics against the resistant isolates, the increase was not enough to restore susceptibility. In conclusion, the presence of NMP increases the zone diameters of ciprofloxacin and levofloxacin. Intermediate strains become susceptible but the resistant isolates do not.

Rapid Susceptibility Test for Mycobacterium tuberculosis to Isoniazid and Rifampin with Resazurin Method in Screw-Cap Tubes

Abstract Multi-drug resistant (MDR) Mycobacterium tuberculosis is still a serious public health problem all over the world. MDR tuberculosis (MDR-TB) caused by these strains has emerged within the last decade and rapid detection is critical for the effective treatment of patients. Recently, a resazurin microtiter assay plate for detecting MDR strains was developed. In this study, it was adapted to screw-cap tubes and the activity of isoniazid (INH) and rifampin (RIF) to 50 M. tuberculosis clinical isolates was tested by this method for the first time. Results were compared with the radiometric reference method for the susceptibility testing of M. tuberculosis complex. The results of both methods were in 100% and 96% agreement for RIF and INH, respectively. Specificity, sensitivity, positive predictive value and negative predictive value were 91.7%, 100%, 92.8% and 100% for INH, respectively. All of these values were 100% for RIF. Susceptibility testing results were obtained on the 8th day of incubation for 42 isolates and on the 9th day for the other eight strains. Our results indicate that this method is suitable for the early determination of INH and RIF resistance in developing countries because it is inexpensive, rapid and easy to perform.

Prevalence and Characterization of Salmonella Isolated from Chicken Meat in Turkey

This study was conducted in a Turkish province to investigate the presence of Salmonella spp. in 150 chicken meat samples using 2 phenotyping techniques: classic culture technique (CCT) and immunomagnetic separation (IMS). For the confirmation of the isolates at molecular levels, invA gene was detected in these isolates. The presence of invA, class 1 (Cls1) integrons, and integrase (Int1) genes was demonstrated by PCR assay; and the resistance of the isolated Salmonella spp. strains to antibiotics was determined by disk diffusion test. All the cultural and PCR results were evaluated together; Salmonella spp. were detected in a total of 64 (42.66%) chicken meat samples. Contamination rate was higher in carcasses (53.33%, n = 75) than in meat pieces (32%, n = 75). When results of standard culture were compared with IMS technique, IMS (n = 54) showed a clear superiority over the CCT (n = 38). A very high resistance rate (≥ 89.28%) to vancomycin, tetracycline, streptomycin, or nalidixic acid was found. Trimethoprim-sulfamethoxazole resistance was present in 32.14%. Relatively lower incidence of resistance (≤ 8.33%) to gentamicin, chloramphenicol, ampicillin, and ceftriaxone was observed. Concurrent resistance to at least 4 antibiotics was detected in 92.85% of the isolates. Cls1 integrons and Int1 were positive in 80.95% and 95.23% of the isolates, respectively. However, Int1 alone was detected in 15.47% (n = 13). In conclusion, the high prevalence of Salmonella spp. in chicken meat may pose a potential public health risk, and the presence of antibiotic-resistant Salmonella spp. isolate together with Cls1 integron and/or integrase might play an important role in horizontal antibiotic gene transfer.