[Investigation of carbapenemases in carbapenem-resistant Escherichia coli and Klebsiella pneumoniae strains isolated in 2014 in Turkey]. / Türkiye'de 2014 yili içinde izole edilen karbapeneme dirençli Escherichia coli ve Klebsiella pneumoniae izolatlarinda karbapenemaz varliginin arastirilmasi.

Carbapenems are the choice of treatment in infections caused by multidrug resistant Enterobacteriaceae. In recent years carbapenem-resistant Enterobacteriaceae isolates due to carbapenemases have been increasingly reported worldwide. Multicenter studies on carbapenemases are scarce in Turkey. The aim of this study was to determine the distribution of carbapenemases from different parts of Turkey as a part of the European Survey of Carbapenemase Producing Enterobacteriaceae (EuSCAPE) project. Beginning in November 2013, carbapenem-resistant isolates resistant to at least one of the agents, namely imipenem, meropenem, and ertapenem were sent to the coordinating center. Minimum inhibitory concentrations for these carbapenems were determined by microdilution tests following EUCAST guidelines. Production of carbapenemase was confirmed by combination disk synergy tests. Types of carbapenemases were investigated using specific primers for VIM, IMP; NDM, KPC and OXA-48 genes by multiplex polymerase chain reaction. In a six month period, 155 suspected carbapenemase-positive isolates were sent to the coordinating center of which 21 (13.5%) were E.coli and 134 (86.5%) were K.pneumoniae. Nineteen (90.5%) strains among E.coli and 124 (92.5%) strains among K.pneumoniae were shown to harbour at least one carbapenemase gene by molecular tests, with a total of 92.3% (143/155). Carbapenemases were determined as a single enzyme in 136 strains (OXA-48: 84.6%; NDM: 6.3%; VIM: 2.8%; IMP: 1.4%) and as a combination in seven isolates (OXA-48 + NDM: 2.1%; OXA-48 + VIM: 2.1%; VIM + NDM: 0.7%). KPC was not detected in any of the isolates. According to the microdilution test results, resistance to imipenem, meropenem and ertapenem in OXA-48 isolates were 59.5%, 52.9% and 100%, respectively. The combination disk synergy test was 100% compatible with the molecular test results. As most of the OXA-48 producing isolates were susceptible to meropenem but all were resistant to ertapenem, ertapenem seems to be the most sensitive agent in screening carbapenemases in areas where OXA-48 is prevalent and phenotypic combination tests can be useful in centers where molecular tests are not available.

[Investigation of hydrophobic characteristics of biofilm producer and non-producer Staphylococcus aureus clinical isolates]. / Biyofilm olusturan ve olusturmayan Staphylococcus aureus klinik izolatlarinin hidrofobik ozelliklerinin arastirilmasi.

The ability of staphylococcus to adhere certain structures and to form biofilm (slime) layer plays an important role in the pathogenesis of staphylococcal infections. Hydrophobic interactions and hydrogen bonds are important factors that play role in adherence. This study was designed to compare the hydrophobic properties of slime positive and negative Staphylococcus aureus strains isolated from blood cultures. Ten methicillin-resistant S. aureus isolates (five of them being slime positive) obtained from blood cultures of patients at intensive care unit of a university hospital, between May 2006 and June 2007, were included in the study. Slime production of the isolates was determined by Christensen's method. Methicillin resistance was determined by cefoxitin disc test and oxacillin salt agar test. It was determined that the test strains did not exhibit any autoaggregation. The adherence of strains to the three different hydrocarbons as solid phases (butyl-sepharose, octyl-sepharose and phenyl-sepharose; Amersham Bioscience, Sweden) were studied by using hydrophobic interaction chromatography (HIC) method. After butyl- and octyl-sepharose chromatography, it was determined that slime negative S. aureus strains were separated into three fractions eluted with phosphate buffered saline (PBS), 40% and 96% ethanol, while slime positive strains were separated into two fractions eluted with 40% and 96% ethanol, respectively. By phenyl-sepharose chromatography analysis; both slime negative and positive strains were separated into two fractions eluted in 40% and 96% ethanol. Hydrophobicity tests were repeated at 4 degrees C and pH 6-9 to evaluate the effect of changing conditions on hydrophobicity. However, no changes we re observed at these temperature and pH values. According to these analysis it was concluded that; (a) S. aureus strains consist heterogeneous fractions with distinct hydrophobic binding strengths; (b) hydrophobic surface protein secretion may be different in heterogeneous groups, and (c) slime positive S. aureus strains were more hydrophobic than non-slime producing strains. Further research is required in order to characterise the eluted fractions and to evaluate their pathogenic capacities.

[A pneumonia case caused by Cedecea lapagei]. / Cedecea lapagei'nin neden oldugu bir pnömoni olgusu.

Cedecea spp. which are the members of Enterobacteriaceae family, are mostly isolated from sputum and their clinical importance is not yet demonstrated. This report presents a pneumonia case caused by Cedecea lapagei. A 38-years-old male patient admitted to Inonu University Faculty of Medicine Emergency department with prediagnosis of subarachnoid haemorrhage was operated and transferred to Intensive Care Unit of Reanimation where he underwent artificial ventilation. On the third day of hospitalization his temperature was 39 degrees C, white blood cell count was 27.000/ml and he was still unconscious. He had a history of chronic obstructive pulmonary disease. Chest X-ray revealed opacities in the right lower lobe and mucoid tracheal secretion ensued following tracheal entubation performed after operation. Direct microscopic examination of bronchoalveolar lavage (BAL) fluid yielded abundant number of leukocytes and gram-negative bacilli. Bacteria isolated from BAL specimen were identified as C. lapagei by Phoenix 100 (Becton Dickinson, USA) automated system and also by API 20E kit (Biomerieux, France). Upon the initiation of intravenous amikacin (1 x 1 g) and meropenem (3 x 1 g), the signs of infection decreased in intensity, however, the patient was lost due to subarachnoid hemorrhage on the 12th day of hospitalization. In this case it was estimated that C. lapagei pneumonia originated from the aspiration of upper airway secretion owing to unconsciousness of the patient. Although there were reports of Cedecea infections in the literature, this was the first documented case of C. lapagei pneumonia when the accessible related literature was concerned.

Molecular epidemiology of methicillin-resistant Staphylococcus aureus isolates from clinical specimens of patients with nosocomial infection: are there unnoticed silent outbreaks?

Bacteriological and epidemiological studies were carried out on 90 isolates of methicillin-resistant Staphylococcus aureus (MRSA) at Turgut Ozal Medical Center of Inönü University, (Malatya/Turkey). MRSA isolates were obtained from patients with nosocomial infections. Staphylococcus aureus clinical isolates were collected between May 2004-May 2005. Isolates were tested for resistance to methicillin. Antimicrobial susceptibility testing and slime production evaluation was performed. Genotype studies were carried out by arbitrarily primed polymerase chain reaction (AP-PCR) and consequent cluster analysis. All of the isolates were mecA-positive in a PCR-based assay; all exhibited resistance to oxacillin, by agar dilution (MICs > or = 4 mg/L) and disc diffusion methods, and multiple antibiotics. Most MRSA isolates were collected in intensive care units. Of 90 samples, 53 were found to be unrelated to the others while the remaining 37 strains were either identical or closely related. Dendrogram analysis identified nine major clusters. These data support the opinion that MRSA are significant nosocomial pathogens in intensive care units and that resistant clones may be transmitted between patients. Molecular epidemiological tools are helpful for understanding transmission patterns and sources of infection, and are useful for measuring outcomes of intervention strategies implemented to reduce nosocomial MRSA.

Evaluation of 23 cutaneous anthrax patients in eastern Anatolia, Turkey: diagnosis and risk factors.

BACKGROUND: Anthrax is a potentially fatal zoonotic disease. The diagnosis of cutaneous anthrax (CA) may be very difficult, particularly in atypical presentations and nonendemic regions. AIM: To evaluate the clinical features and diagnostic difficulties of 23 anthrax cases seen between May 2004 and September 2006. METHODS: Twenty-three patients with CA were included in this study. The diagnosis of CA was based on clinical findings and/or microbiologic procedures. RESULTS: All patients with a diagnosis of CA were followed up. One patient experienced toxemic shock. Twenty-two patients had a history of animal contact. Only one patient did not recall any history of suspicious contact. The clinical presentation of CA was typical in 20 patients (87%). Two patients were initially misdiagnosed with insect bites and one patient with angioedema. Cultures from the lesions were positive for Bacillus anthracis in seven cases (30.4%). Gram stain from the lesions revealed Gram-positive rods in eight cases (34.8%). Fifteen patients (65.2%) were diagnosed by clinical presentation and a history of contact with sick animals and/or contaminated animal products. CONCLUSION: CA is a very contagious and important infectious disease worldwide. Early and accurate diagnosis dramatically affects the prognosis of the disease. The diagnosis of CA may be difficult, especially in atypical presentations and nonendemic areas. Thus, CA should be kept in mind, especially in these situations.

Fluvastatin reduced liver injury in rat model of extrahepatic cholestasis.

Inhibitors of 3-hydroxy-3methylglutarly coenzyme A, reductase, namely statins, exert pleiotropic actions beyond lipid-lowering effects. In ex vivo and in vitro studies, statins have antioxidative and antiinflammatory effects. Herein, we sought to determine whether treatment with fluvastatin (FV) would be beneficial in a rat model of common bile duct ligation (BDL)-induced liver injury. Female rats were subjected to a sham (n=10) or BDL (n=20). Obstructive jaundice was induced in rats by the ligation and division of the common bile duct. Three days after operation, rats subjected to CBDL were randomized to receive treatment with either FV (10 mg/kg) or saline every day over a 10 days experimental period. High levels of alanine aminotransferase, aspartate aminotransferase, and gamma glutamyltransferase decreased significantly (P<0.05) in animals treated with FV with compared to saline-administrated BDL animals. Compared with sham-operated rats, CBDL rats showed significantly higher levels of total nitrite and nitrate, malondihaldehyde, tumor necrosis factor alpha, myeloperoxidase, and lower concentrations of glutathione, superoxide dismutase, and catalase in the liver tissue (P<0.001). All of these changes were significantly attenuated (P<0.05) by treatment with FV after CBDL. CBDL was associated with increased apoptosis and nuclear factor kappa beta expression in saline-treated rats. Treatment with FV also decreased these parameters. These data support the view that FV ameliorates hepatic inflammation, lipid peroxidation, and tissue injury in rats subjected to CDBL. FV warrants further evaluation as an adjunctive treatment to ameliorate liver injury from extrahepatic biliary obstruction.

Attenuation of ureteral obstruction-induced renal injury by polyenylphosphatidylcholine.

BACKGROUND: The cytoprotective, antioxidant and antifibrotic effects of polyenylphosphatidylcholine (lecithin, PPC) have been demonstrated both experimentally and clinically. The present study investigated whether PPC treatment has any beneficial effect on renal injury in unilateral partial ureteral obstruction (UUO) in rats. METHODS: Forty Wistar-Albino rats were split into three groups (sham-operated controls, untreated and treated rats). Rats of the untreated and treated groups (n = 15) underwent UUO with two-thirds of the left ureter embedded in the psoas muscle. In group 3, PPC was given orally at a dose of 100 mg/day for 30 days. At the end of the 30th day of the experimental period, obstructed kidneys and blood samples were harvested. To investigate the therapeutic efficacy of PPC treatment in UUO kidneys, oxidant and antioxidant enzyme levels, lipid peroxidation, proinflammatory cytokines (interleukin-1, interleukin-6, tumor necrosis factor alpha), transforming growth factor beta-1 (TGFbeta-1), alpha smooth muscle actin (alpha-SMA) and nuclear factor kappa beta (NF-kappabeta) expression, leukocyte infiltration (ED1, ED2, CD4 and CD8 immunohistochemistry), and tubulointerstitial damage in the obstructed kidneys were studied. RESULTS: Oxidative stress, neutrophil infiltration, release of cytotoxic mediators, TGFbeta-1 levels, tubulointerstitial damage, alpha-SMA and NF-KB expressions in kidney tissue were significantly increased in the UUO rats. PPC treatment attenuated oxidative stress, leukocyte infiltration, cytotoxic mediator, and TGFbeta-1 levels and also decreased expressions of alpha-SMA and NF-kappabeta. It was associated with decreased tubulointerstitial damage, compared with UUO alone. CONCLUSIONS: These results indicate that PPC treatment protects against UUO-induced renal injury in rats possibly through its antioxidant, anti-inflammatory and antifibrotic actions.

Sulfasalazine reduces inflammatory renal injury in unilateral ureteral obstruction.

The purpose of this study was to test whether sulfasalazine has a protective action against interstitial inflammation and the development of renal fibrosis in obstructive nephropathy. Female rats were subjected to a sham (n = 10) or unilateral ureteral obstruction (UUO, n = 30). UUO was induced in rats by ligating the left ureter. Three days after operation, rats subjected to UUO were randomized to receive tretment with either sulfasalazine (100 mg/kg) or vehicle every day for the last 7 days of the experiment. At 10 days following UUO, the obstructed kidney exhibited tubulointerstitial injury and leukocyte infiltration (mainly monocytes) that were associated with high levels of reactive oxygen species, cytokines, transforming growth factor (TGF)-beta1, myeloperoxidase (MPO), and lipid peroxidation. Ten days after UUO, the obstructed kidney was also associated with increased nuclear factor kappa beta (NF-kappabeta) expression in saline-treated rats. Compared with sham-operated rats, UUO rat kidneys showed lower concentrations of antioxidant enzymes in the obstructed kidney tissue. All of these changes were significantly attenuated by treatment with sulfasalazine in the obstructed kidney. Sulfasalazine protected against the renal interstitial inflammation and tissue damage elicited by ureteral occlusion. Inhibition of the NF-kappabeta-dependent pathway and inflammatory response and oxidative stress inhibition is likely to be involved in the beneficial effects of sulfasalazine.

The effect of caffeic acid phenethyl ester on bacterial translocation and intestinal damage in cholestatic rats.

We investigated the effect of caffeic acid phenethyl ester in rat ileum injury induced by chronic biliary obstruction. Swiss albino rats were divided into three groups: Group 1, sham (n = 7); Group 2, common bile duct ligation (n = 7); and Group 3, common bile duct ligation plus caffeic acid phenethyl ester (n = 7). In the caffeic acid phenethyl ester-treated rats, ileum tissue levels of malondialdehyde and myeloperoxidase were significantly lower than those of the bile duct-ligated rats (P < 0.001). The levels of tumor necrosis factor-alpha, interleukin-6, and interleukin-1alpha in the caffeic acid phenethyl ester group were significantly lower than those in the bile duct ligation group (P < 0.03, P < 0.01, and P < 0.02 respectively). The present study demonstrates that intraperitoneal administration of caffeic acid phenethyl ester in bile duct-ligated rats reduces intestinal oxidative stress. This effect may be useful in the preservation of intestinal damage in cholestasis.

The NF-kappaB inhibitors attenuate hepatic injury in bile duct ligated rats.

Cholestasis-induced liver injury during bile duct obstruction causes an inflammatory response and this inflammatory process may be an important source of tissue injury. We hypothesized that NF-kappaB inhibition would decrease liver injury in a rat model of extrahepatic biliary obstruction. A total of 40 female rats of Sprague-Dawley strain were allocated to four groups. First group was sham operated control. The second group underwent common bile duct ligation (BDL) and was monitored for 10 days. Third group of rats underwent BDL and received pyrrolidine dithiocarbomate (PDTC) at a dose of 100 mg/kg/day intraperitoneally. Fourth group underwent BDL and received sulfasalazine at a dose of 100 mg/kg b.w. Both inhibitors were administered once a day throughout last 7 days of the experimental period. Rats were terminated 10 days after sham operation or BDL. Aspartate aminotransferase, alanine aminotransferase, gamma-glutamil transpeptidase, and tumor necrosis factor-alpha levels were elevated in the BDL group as compared to the control group, while this increase was significantly decreased by treatment with PDTC and sulfasalazine (P < 0.05). Hepatic GSH, SOD and catalase levels were significantly depressed by BDL, but were elevated back to control levels in NF-kappaB inhibitor-treated BDL groups. Increases in tissue free radical and MDA levels and MPO activity due to BDL were reduced back to control levels by NF-kappaB inhibitor treatment (P < 0.05). Similarly histological damage in the BDL rats was reduced by treatments. These results indicate that inhibitors of NF-kappaB activity such as PDTB and sulfasalazine exert a therapeutic effect on cholestatic liver injury in rats with BDL through anti-inflammatory and antioxidant actions.

Protective effect of resveratrol against oxidative stress in cholestasis.

BACKGROUND: We investigated the protective role of resveratrol in rat liver injuries induced by chronic biliary obstruction. MATERIALS AND METHODS: Secondary biliary cirrhosis was induced by bile duct ligation for 28 days. Swiss albino rats were divided into the three following groups: group 1: sham (n = 7); group 2: bile duct ligation (n = 7); group 3: bile duct ligation plus resveratrol (n = 7). Bile duct ligation plus resveratrol group received 10 mg/kg dose of resveratrol intraperitoneally daily for 28 days. Liver damage and cholestasis were determined by the biochemical and the pathologic examination. RESULTS: The present data showed a decrease in both plasma bilirubin levels and aspartate aminotransferase and alanine aminotransferase levels in the resveratrol-treated rats, when compared with bile duct ligation group (P < 0.05). In the resveratrol-treated rats, tissue levels of malondialdehyde and nitric oxide were significantly lower than that of the bile duct ligation (P < 0.002). The levels of glutathione in resveratrol-treated rats were significantly higher than that in bile duct ligation group (P < 0.004). The levels of interleukin-1alpha, interleukin-6, and tumor necrosis factor-alpha in resveratrol group were significantly lower than that in bile duct ligation group (P < 0.004, P < 0.001, P < 0.001, respectively). Administration of resveratrol in the rats with biliary obstruction resulted in inhibition of ductular proliferation and lymphocytic inflammation. CONCLUSION: The present study demonstrates that intraperitoneal administration of resveratrol in bile duct ligated rats maintained antioxidant defenses and reduces liver oxidative damage and ductular proliferation. This effect of resveratrol may be useful in the preservation of liver function in cholestasis.