[Evaluation of In Vitro Efficacy of Ceftazidime-Avibactam, Meropenem, and Colistin Single and Binary Combinations Against Carbapenem Resistant Klebsiella pneumoniae Strains Isolated from Various Clinical Specimens]. / Seftazidim-Avibaktam, Meropenem ve Kolistinin Tek Basina ve Ikili Kombinasyonlarinin Çesitli Klinik Örneklerden Izole Edilen Karbapenem Dirençli Klebsiella pneumoniae Suslarina Karsi In Vitro Etkinliginin Arastirilmasi.

It has been reported that many antibiotics used today, including the carbapenem group, fail to treat Klebsiella pneumoniae infections effectively. Despite many studies in recent years, the definitive treatment for carbapenem-resistant Klebsiella pneumoniae (CRKP) infections is still uncertain. In this study, it was aimed to investigate in vitro activities of colistin (COL) and meropenem (MEM), which are frequently used in the treatment of CRKP infections, and ceftazidime-avibactam (CZA), which is recently used in our country, alone or in combination against different CRKP isolates having different carbapenem resistance mechanisms andto analyze whether the presence of colistin resistance, which is an important problem in CRKP strains, influences the drug interaction results. This study was carried out in 42 K.pneumoniae isolates, which were isolated from various clinical samples as an infectious agent in Süleyman Demirel University Faculty of Medicine, Department of Medical Microbiology, Bacteriology Laboratory and whose carbapenem resistance was confirmed by carbapenemase inactivation test. The carbapenemase genes of the isolates were determined by the polymerase chain reaction (PCR) method. Antimicrobial susceptibilities of CRKP strains to CZA, MEM, and COL were determined by the broth microdilution method and in vitro synergy activities of dual combinations of these drugs were evaluated by checkerboard and time-kill methods. Statistical evaluation of categorical data was performed using Fisher's exact test, and p-value of less than 0.05 was considered statistically significant in terms of difference between the groups. Of the 42 CRKP isolates 34 (81%) were only OXA-48 positive, 5 (11.9%) were OXA-48+NDM and 3 (7.1%) were OXA-48+KPC positive. In the checkerboard test, synergy was detected against 97.6% of the isolates both with CZA+MEM and CZA+COL combinations, whereas this rate was 50% with MEM+COL. In the time-kill test, synergy was detected with CZA+MEM and CZA+COL combinations in the OXA-48 positive isolate and OXA-48+KPC positive isolate, while synergy was detected with CZA+COL and MEM+COL combinations in the OXA-48+NDM positive isolate. There was no significant relationship between whether the isolates were resistant to colistin or not and the checkerboard test results of antibiotic combinations (pCZA+MEM= 0.33, pCZA+COL= 0.11, pMEM+COL= 0.61). Results of our study revealed that the most common carbapenemase type in CRKP isolates was OXA-48 in our hospital, and the combinations of CZA with MEM and COL had high potential for synergism against these isolates.

Evaluation of biochemical, hematological, RIBA and PCR assays in predicting viremia in anti-HCV positive patients.

INTRODUCTION: The detection of HCV-RNA by PCR assays is considered to be the gold standard for confirming the presence of HCV viremia. However, high costs, long and laborious procedures limit their widespread usage. This retrospective study was conducted to assess the predictive performances of biochemical and hematological parameters, anti-HCV signal-to-cutoff (S/CO) ratios and RIBA assay for HCV viremia. METHODOLOGY: Medical records of 210 patients with positive anti-HCV results were analyzed. Samples were tested for anti-HCV by the Roche Elecsys assay. RIBA and PCR assays were performed with Inno-Lia HCV Score test, and Roche Cobas TaqMan HCV test, respectively. RESULTS: Anti-HCV positive patients were categorized into two groups: positive HCV-RNA(viremic) group (n = 94) and negative HCV-RNA(non-viremic) group (n = 116). All viremic patients had positive RIBA results, while in the non-viremic group, 80 (69%) patients had negative/indeterminate RIBA results and 36 (31%) patients had positive RIBA results. Compared with the non-viremic group, the viremic group had significantly higher alanine aminotransaminase (ALT), aspartate aminotransferase, gamma-glutamyl transferase, mean platelet volume, platelet distribution width and anti-HCV levels, and significantly lower platelet count and plateletcrit levels (p < 0.05). With multivariate logistic regression analysis, serum ALT and anti-HCV levels were found to be strong predictive factors for HCV viremia. A S/CO ratio of ≥ 12.34 was identified as the optimal anti-HCV level to predict viremia. CONCLUSIONS: An anti-HCV S/CO ratio of 12.34 can determine the necessity for PCR assay, when carefully evaluated together with the biochemical and hematological evidence. This approach may reduce the cost of diagnosis particularly in low-resource settings.

Investigation of Carbapenem resistance mechanisms in Klebsiella pneumoniae by using phenotypic tests and a molecular assay.

INTRODUCTION: The aim of this study was to investigate the presence of carbapenemase production and carbapenem resistance mechanisms in 47 carbapenem resistant Klebsiella pneumoniae isolates by phenotypic confirmatory tests and molecular assay. METHODOLOGY: Carbapenem resistance genes KPC, OXA-48 and NDM were investigated with the BD MAX CRE assay kit in the BD MAX real time PCR instrument. Modified Hodge test, MBL gradient strip test, D70C Carbapenemase Detection Set, Temocillin gradient strip test methods were used as phenotypic confirmatory tests. Clonal relationship between study isolates was investigated with pulsed-field gel electrophoresis. RESULTS: Analysis with BD MAX CRE assay revealed OXA-48 positivity in 17 (36%) strains, NDM positivity in 6 (13%) strains and coexistence of OXA-48 + NDM positivity in 8 (17%) strains. In 16 (34%) strains, none of the KPC, OXA-48 and NDM genes were detected. While MHT was the most sensitive phenotypic confirmatory test, D70C disc set had not been considered as a useful tool to assist the search for carbapenemase production. Temocillin gradient test alone could not be considered as sufficient to detect the presence of OXA-48. PFGE analyses revealed that 23 of 31 carbapenemase producing strains were in three major PFGE genotypes (A, B and C). CONCLUSIONS: This study revealed that carbapenem resistance observed in K. pneumoniae isolates was mainly due to OXA-48 and NDM genes and the increase of carbapenem resistance among K. pneumoniae strains in our hospital was due to the interhospital spread of especially 3 epidemic clones.

[Evaluation of eosinophil cationic protein levels with clinical symptoms and laboratory findings of patients with cystic echinococcosis]. / Kistik ekinokokkozisli hastalarda eozinofilik katyonik protein düzeylerinin klinik semptomlar ve laboratuvar bulgulariyla birlikte degerlendirilmesi.

Among proteins secreted from activated eosinophil granulocytes, eosinophil cationic protein (ECP) is the most useful tool for the follow-up of inflammatory diseases. Since ECP level reflects the eosinophil activation, it gives valuable information about disease activity. In this study, we aimed to investigate the possible relation between ECP levels and symptoms and laboratory findings of cystic echinococcosis (CE) and to evaluate the role of this protein in the diagnosis of CE. The study which was conducted at Clinical Microbiology Laboratory of Suleyman Demirel University Medical Faculty, Isparta, Turkey, included 58 patients with a pre-diagnosis of CE and 32 healthy individuals as control group. The diagnosis of CE was established serologically by modified enzyme-linked immunosorbent assay (ELISA) and indirect hemagglutination (IHA) test. The quantitative determination of ECP levels was done by fluoro-enzyme immunoassay (FEIA; Uni-CAP ECP, Pharmacia-Upjohn). The mean ECP level was 31.6 +/- 37 microg/ml in the patient group and 9.1 +/- 2.1 microg/ml in the control group, the difference being statistically significant (p = 0.001). Significant differences were also detected for erythrocyte sedimentation rate (ESR) (p = 0.001), total IgE level (p = 0.001), eosinophile count (p = 0.05) and CRP (p = 0.001) between the patient and the control groups. ECP was detected to be high in 35 (60%), IgE in 37 (63%), CRP in 29 (50%) and eosinophile count in 9 (15.5%) patients. While age, gender, ESR, IgE and CRP levels of patients with high ECP levels were not significantly different from levels of patients with normal ECP levels, significantly different eosinophil counts were detected among patients with high ECP values when compared to patients with normal ECP values. Furthermore, a correlation was detected between ECP levels and eosinophil rate, IgE and CRP levels of patients with CE (p = 0.01), while there was no correlation between ECP and ESR levels. Although high ECP level patients exhibited higher ALT and AST levels, no correlation was determined between liver enzyme levels and ECP levels (p > 0.05). The most common symtoms among CE patients were abdominal pain (41%), other gastrointestinal complaints (38%), shortness of breath (12%) and fever (10%). No statistically significant difference in terms of symptoms was detected between patients with high ECP levels and normal ECP levels. However, statistically significant difference was detected between ECP levels of patients with symptoms (except shortness of breath) and patients without symptoms (p < 0.05). In conclusion, ECP seems to be associated with the symptoms and signs of CE and it can be used as a valuable marker besides the other laboratory tests for the evaluation of patients with CE.

[Investigation of the presence of autoantibodies in patients with toxocariasis]. / Toksokariazisli hastalarda otoantikor varliginin arastirilmasi.

Immunopathologic reactions may occur during toxocariasis due to tissue invasion and destruction by the secretions of larvae containing various enzymes with broad spectrum. The aim of this study was to search for autoantibodies such as anti-nuclear (ANA), anti-mitochondrial (AMA), anti-smooth muscle (ASMA), anti-neutrophil cytoplasmic (ANCA), anti-myeloperoxidase (MPO) and liver-kidney microsomal type 1 (LKM-1) antibodies in patients with toxocariasis, in order to investigate the role of toxocariasis as a trigger factor for autoimmune reactions. Forty patients (22 were male; mean age: 35.6 +/- 10.7 years) diagnosed as toxocariasis by clinical findings (abdominal pain, allergic symptoms and/or eosinophilia, without detection of any other causative agents, and without liver dysfunction, diabetes mellitus, cardiac or renal failure, and autoimmune disease) and in-house ELISA positivity and 32 healthy controls (16 were male; mean age: 40.7 +/- 11.2 years) were included to the study. ANA (screen), dsDNA, SS-A, SS-B, Scl-70, LKM-1, MPO and M2 autoantibodies have been investigated by ELISA (Euroimmun, Germany), while ANCA, AMA and ASMA antibodies by indirect immunofluorescence (IMMCO, NY) methods. Autoantibody positivity was detected in 18 (45%) patients of whom 11 yielded a single type, and 7 yielded > or = 2 types of autoantibodies. This rate was 12.5% for control group (two subjects were positive for ANA-Screen, one for anti-M2 and one for anti-LKM-1). The difference between the total positivity rates in patient and control groups was found statistically significant (chi2 = 5.72, p = 0.004). The most frequent autoantibody type among patients were ASMA (n = 6), followed by anti-dsDNA (n = 5), anti-M2 (n = 5), anti-SS-B (n = 4), anti-LKM-1 (n = 3), anti-SS-A (n = 2), ANCA (n = 2) and anti-MPO (n = 1). Positivity rate for ASMA was found statistically significant in patients' group compared to controls (chi2 = 12.24, p = 0.03), while there was no significant difference between the groups in terms of other autoantibody rates (p> 0.05). These data could be related to the possible release of autoantigens following muscle tissue injury during toxocariasis and/or antigenic mimicry of parasitic products during the infection in which muscle invasion is frequently seen. In conclusion, since autoantibodies are frequently detected in toxocariasis, this situation should be taken into consideration in the presence of autoantibodies.

[The relationship between viral load and malondialdehyde and antioxidant enzymes in patients with hepatitis C virus infection]. / Hepatit C virus enfeksiyonu olan hastalarda viral yük ile malondialdehid ve antioksidan enzimlerin iliskisi.

Oxidative stress is known to participate in the pathogenesis of HCV infection. The aim of this study was to investigate the relationship between antioxidant defence state, malondialdehyde (MDA) and viral load in patients with hepatitis C virus (HCV) infection. Fifty patients who were positive for serological and molecular markers of HCV infection, and 40 healthy volunteers as control group were included in the study. The patients were classified according to their viral loads, and the catalase, superoxide dismutase (SOD) and glutathione peroxidase (GP) activities of erythrocytes and MDA in sera of all groups were measured. These substances were detected by using the methods described by Aebi, Woolliams et al, Paglia and Valentine, Draper and Hadley, respectively. As a result, decrease in SOD and GP levels and increase in MDA and catalase levels have been detected in HCV infected patients when compared with healthy controls, and these differences were statistically significant (p<0.05, t=19.3),except for catalase. However, there were no statistically significant difference among groups classified according to viral load (p>0.05, t=1.6). Although our data in HCV infected patients demonstrate a significant decrease in antioxidant enzyme levels and a significant increase in MDA levels, a marker of oxidative stress, it could not possible to make a correlation between these differences and the viral loads of patients.

[The prevalence of hepatitis G virus in patients with hepatitis B and C virus infections]. / Hepatit B ve C virus enfeksiyonu olan hastalarda hepatit G virus prevalansi.

GB virus C/hepatitis G virus (GBV-C/HGV) is a recently identified flavivirus, which has been frequently detected in patients with chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. In this study, the prevalence of HGV-RNA and antibodies to E2 envelope antigen (anti-E2) which is a marker of past infection, have been investigated in the samples of patients with HCV and HBV infections, and the prevalence rates were compared with the control group. The study group consisted of 50 patients with HBV and 50 patients with HCV infections, who did not have any risk for parenteral transmission, and 60 healthy control subjects. Serum samples were tested for the presence of HGV-RNA by real time polymerase chain reaction (PCR) and for anti-E2 by a commercial enzyme-linked immunosorbent assay (ELISA). As a result, HGV-RNA and anti-HGV-E2 positivity rates in HBV and HCV infected patients were found as; 4% and 6%, 4% and 4%, respectively. Although the prevalence of HGV-RNA in patients with HBV and HCV infections were higher than the control group (1.66%), there was no statistically significant difference (p>0.05). The prevalence of anti-E2 antibodies in patients with HBV and HCV infections did not revealed any difference in comparison to the control group (6.66%) (p>0.05). In conclusion, GBV-C/HGV infection prevalence was found low in patients with HBV and HCV infections, supporting that although parenteral route is the most effective way, other routes such as sexual contact and intra-familial contact may also play role in HGV transmission.

Gastroduodenal Helicobacter pylori infection diagnosed by Helicobacter pylori stool antigen is related to atherosclerosis.

OBJECTIVE: The causative relation between Helicobacter pylori (H. pylori) and atherosclerosis has been determined as seropositivity or determination of H. pylori from atherome plaques by molecular methods. The site of entrance and the reservoir of the bacteria in the body is still a subject of discussion. In this study Helicobacter pylori stool antigen (HpSA) which shows gastrointestinal system colonization and infection with high specificity and sensitivity was determined in atherosclerotic, ectatic and angiographically normal groups. METHODS AND RESULTS: A total of 62 patients was categorized according to diagnostic coronary angiography as 12 had normal coronary arteries, eight had one, 18 had two, and 12 had three atherosclerotic coronary arteries. Twelve patients had ectatic vessels. There were 27 (44%) HpSA positive and 35 (56%) HpSA negative patients. There was a statistically significant relation between HpSA positivity and the degree of vessel involvement in coronary artery disease (CAD) patients, essentially between the group with three vessels (83%) obstructed and the normal group (25%). Ectatic vessel group had a higher incidence (50%) of HpSA positivity compared to the control group but not enough for statistical significance. CONCLUSIONS: The results indicate that gastrointestinal system H. pylori colonization increases the risk of atherosclerosis. We may speculate that the reservoir and spread of H. pylori is via gastrointestinal tract. Studies may be performed to detect whether gastrointestinal tract H. pylori infection treatment will decrease the risk of coronary artery damage caused by H. pylori.