Aynur Karadenizli

Outbreak of tularaemia in Golcuk, Turkey in 2005: Report of 5 cases and an overview of the literature from Turkey

Tularaemia was diagnosed by TaqMan RT-PCR and microagglutination tests in 5 patients, all from a new settlement constructed after the earthquake of 1999. During the follow-up, 129 more cases were found in this settlement (data from the local Health Care Authority). In this study, clinical features of 5 cases are presented briefly, and the Turkish literature on past outbreaks of tularaemia is reviewed.

Genetic and Enzymatic Properties of Metallo-β-Lactamase VIM-5 from a Clinical Isolate of Enterobacter cloacae

ABSTRACT A VIM-5-producing Enterobacter cloacae isolate (EDV/1) was identified in a collection of clinical strains stored before 2002. The gene, blaVIM-5, was located on a 2,712-bp BamHI-HindIII fragment of a 23-kbp (approximately) nonconjugative plasmid (pEDV5) in a class 1 integron as a single gene cassette.

A novel application of Fourier-transformed infrared spectroscopy: classification of slime from staphylococci.

Abstract It has been proposed that the virulence of nosocomial Staphylococcus infections associated with indwelling medical devices is related to the ability of the bacterium to colonise these materials by forming a biofilm composed of multilayered cell clusters embedded in a slime matrix. However, the pathogenic role of exopolysaccharide biofilms is not fully understood. A new method was sought for differentiating the structure of slime from two closely related bacterial strains, Staphylococcus aureus and Staphylococcus epidermidis. Using PCR it was confirmed that these strains were positive for the icaA and icaD genes and the complete ica operon (2.7 kb). Monosaccharide analysis by thin-layer chromatography revealed an identical profile for both strains, with xylose and glucose present among the four visible bands. Using Fourier-transformed infrared spectroscopy and hierarchical cluster analysis, three of four S. aureus samples (75%), and four of five S. epidermidis samples were grouped according to species. A novel FTIR approach in classifying slime produced by S. aureus and S. epidermidis is reported.

Piperacillin with and without Tazobactam against Extended-Spectrum Beta-Lactamase-Producing Pseudomonas aeruginosa in a Rat Thigh Abscess Model

Background: We compared the antibacterial effect of piperacillin, piperacillin-tazobactam and imipenem in a paired rat thigh abscess model. Methods: Two abscesses were provoked in the thighs of rats, one on the right with an extended-spectrum beta-lactamase (OXA-14)-producing Pseudomonas aeruginosa (Ps-162) and the other on the left thigh with a control strain. Results: The colony counts from the abscesses in log 10 colony-forming units per gram (mean ± SD) in the imipenem group were 2.78 ± 1.71 and 3.19 ± 1.66, in the piperacillin-tazobactam group 4.36 ± 0.23 and 2.44 ± 1.97, and in the piperacillin group 4.44 ± 0.21 and 3.71 ± 0.99 for Ps-162 and the control strain, respectively. The mean colony counts were significantly different (p < 0.05) between Ps-162 and the control strain in the piperacillin and piperacillin-tazobactam groups. Conclusion: These data showed that piperacillin and piperacillin-tazobactam were significantly less effective against extended-spectrum beta-lactamase-producing P. aeruginosa, while imipenem was equally effective against both Ps-162 and the control strain in this abscess model.

A case of pneumonia caused by Bacillus anthracis secondary to gastrointestinal anthrax.

We present herein an unusual case of anthrax pneumonia secondary to gastrointestinal infection. In this case, severe abdominal pain occurred during the course of a stent placement procedure. The patient had undergone surgery with the prediagnosis of intestinal ischemia. On the second postoperative day, pneumonia developed and B. anthracis grew as the etiologic agent. Pathological examination of small-bowel sections revealed findings in accordance with anthrax.

N-ACETYLCYSTEINE IMPROVES INTESTINAL BARRIER IN PARTIALLY HEPATECTOMIZED RATS

Background:  Translocating enteric bacteria play an important role in the development of infections following partial hepatectomy. The intestine itself is the first line of defence against bacterial translocation (BT). We investigated the effect of N‐acetylcysteine (NAC) on BT and the intestinal wall.

Overcoming difficulties on synthesis of cardiac troponin-I

ABSTRACT Human cardiac troponin-I (cTnI) is one of the most sensitive and specific indicators, used in the diagnosis of myocardial infarction. To produce the protein efficiently, Escherichia coli and Pichia pastoris systems were used. Initial trials for the expression in E. coli were not successful, although different expression vectors with different promoters were tested. This led us to use P. pastoris for the expression. After several trials with two different expression strains of P. pastoris, it was concluded that P. pastoris was also not an optimal expression host for cTnI. Comprehensive analysis of the expression systems indicated that an efficient expression is only possible when the gene is optimized for expression in E. coli. For this purpose, the gene was optimized in-silico, but edited manually afterwards. It was synthesized and cloned into pQE-2 vector. Expression was performed using routine experimental conditions. Thus, cTnI could be efficiently expressed from the optimized gene in E. coli. The expression and purification were practical and may be used for commercial purposes since a total yield of 25µg highly pure protein per milliliter of culture could be obtained. The protein was in its ready-to-use form for many biological applications, including as a standard in diagnostic tests and an antigen for antibody production.

Evaluation of Effect of Topical Tacrolimus Treatment on Herpetic Stromal Keratitis in a Rat Model.

Objectives: To investigate the effectiveness of topical tacrolimus treatment on herpetic stromal keratitis (HSK) in a rat model. Methods: The development of HSK was monitored for 14 days after the inoculation of rats with herpes simplex type 1 virus. Rats that developed HSK were divided into four groups as follows: (1) topical antiviral treatment (control), (2) topical antiviral and 1% prednisolone acetate, (3) topical antiviral and 0.03% tacrolimus ointment, and (4) topical antiviral plus 0.1% tacrolimus ointment. After 14 days of treatment, the severity levels of HSK were scored and compared with the levels before the treatment. The expression of CD3, CD4, and CD8 was evaluated by flow cytometry. The development of the disease was evaluated clinically and histologically. Results: Significant improvement in vascularization was observed in the groups with the drug treatment in addition to the antiviral agent (P<0.05), but there was no obvious difference within groups 2, 3, and 4 in the vascularization severity. The regression of corneal edema was 8.05%±6% in group 1, 25.17%±14.55% in group 2 (P=0.01), 36.40%±21.69% in group 3 (P=0.03), and 46.39%±14.96% in group 4 (P=0.00). A significant decrease in the number of inflammatory cells in the groups with the drug treatment was evaluated by immunohistochemical staining and confirmed by flow cytometry analysis. Conclusions: Topical tacrolimus treatment caused a significant decrease in corneal vascularization accompanied by a lower number of inflammatory cells in the experimental HSK corneal edema model. Therefore, topical tacrolimus has the potential to be used in the treatment of HSK.

Tularemia in a Kidney Transplant Recipient

To the Editor: Tularemia, a zoonotic bacterial illness caused by Francisella tularensis, is extremely infectious and potentially fatal, especially in immunocompromised hosts such as bone marrow or solid organ transplant recipients. Tularemia in the posttransplant period has been reported rarely in the literature. Here, we present a case of glandular tularemia in a 24-year-old male kidney transplant recipient who presented 12 months after transplant with cervical lympadenopathy. Pathologic examination of the lymph node biopsy showed chronic necrotizing granulomatous inflammation, which suggested tuberculous lympadenitis. However, because the patient lived in a region in which tularemia is endemic, a real-time polymerase chain reaction–based test for tularemia was performed on the lymph node specimen and found to be positive. When subjected to an F tularensis microagglutination test, the patient’s serum also was positive at a titer of 1:320. Aminoglycosides were not administered due to possible nephrotoxicity; instead, he was started on treatment with doxycycline, 100 mg, twice daily for 4 weeks. At the latest follow-up, cervical lymphadenopathy had resolved and the patient was asymptomatic. Tularemia rarely has been reported in transplant recipients. We could identify only 4 published cases, including a case in a liver transplant recipient, 2 cases in bone marrow transplant recipients, and one prior case in a kidney transplant recipient (Table 1). Three of the 5 patients had systemic presentation of tularemia and one died. A high level of suspicion, prompt diagnosis, and institution of effective antibiotic therapy are required for the management of tularemia in the posttransplant period, especially in endemic areas.

Improved Production of Highly Active and Pure Human Creatine Kinase MB

Human creatine kinase MB (hCKMB) is one of the most preferred biomarkers used for the diagnosis of acute coronary syndrome due to its high sensitivity and specificity. The increasing need for highly purified and biologically active hCKMB in the field of diagnostics makes its production valuable. Currently, the production of hCKMB is mainly achieved in methylotrophic yeast, Pichia pastoris, because the production in Escherichia coli is challenging and generally yields an inactive enzyme with a low quantity. With the aim of finding the best way for the high-yield production of active hCKMB in E. coli, an efficient strategy was developed using a construct allowing tandem expression of each subunit with 2 different tags. The strategy allowed the efficient expression and separate characterization of each subunit and 1-step purification of the heterodimeric protein into homogeneity. The heterodimeric protein displayed more than 11-fold greater specific activity than the commercially available one. The production strategy described in this study shows a clear advantage over the currently used ones and can be made available not only for laboratory scale production but also for commercial production. Our study is also a well-suited example for the studies in which novel protein expression strategies are needed to achieve greater yields with higher purities.