Apoptosis-Related Gene Expression in an Adult Cohort with Crimean-Congo Hemorrhagic Fever.

Crimean-Congo Hemorrhagic Fever (CCHF) is a life threatening acute viral infection characterized by fever, bleeding, leukopenia and thrombocytopenia. It is a major emerging infectious diseases threat, but its pathogenesis remains poorly understood and few data exist for the role of apoptosis in acute infection. We aimed to assess apoptotic gene expression in leukocytes in a cross-sectional cohort study of adults with CCHF. Twenty participants with CCHF and 10 healthy controls were recruited at a tertiary CCHF unit in Turkey; at admission baseline blood tests were collected and total RNA was isolated. The RealTime ready Human Apoptosis Panel was used for real-time PCR, detecting differences in gene expression. Participants had CCHF severity grading scores (SGS) with low risk score (10 out of 20) and intermediate or high risk scores (10 out of 20) for mortality. Five of 20 participants had a fatal outcome. Gene expression analysis showed modulation of pro-apoptotic and anti-apoptotic genes that facilitate apoptosis in the CCHF patient group. Dominant extrinsic pathway activation, mostly related with TNF family members was observed. Severe and fatal cases suggest additional intrinsic pathway activation. The clinical significance of relative gene expression is not clear, and larger longitudinal studies with simultaneous measurement of host and viral factors are recommended.

Ceftaroline activity on certain respiratory tract and wound infection agents at the minimum inhibitory concentration level.

INTRODUCTION: The aim of this study was to investigate the effectiveness of ceftaroline against agents frequently isolated from respiratory tract and wound infections. METHODOLOGY: The study included a total of 250 strains isolated from various clinical specimens, among which were Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus dysagalactiae, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catharralis. The bacteria were identified using the matrix-assisted laser desorption/ionization time-of-flight method and conventional methods. The bacteria's antibiotic susceptibility was tested using appropriate broth microdilution. Mueller-Hinton broth with 4% lysed horse blood, Haemophilus test medium broth, and Mueller-Hinton broth were used. Ceftaroline fosamil results at the minimum inhibitory concentration (MIC) were evaluated using Clinical and Laboratory Standards Institute (CLSI) criteria. For quality assurance, E. coli ATCC 35218, S. aureus ATCC 29213, S. aureus ATCC 43300, S. pneumoniae ATCC 49619, H. influenzae ATCC 49766, H. influenzae ATCC 10211, and H. influenzae ATCC 49247 standard strains were used. RESULTS: According to CLSI criteria, resistance was not detected in any strains. Due to the absence of CLSI criteria for M. catharralis, the susceptibility state for this bacterium was not evaluated. The various strains' MIC50-MIC90 values were as follows: for S. pyogenes, 0.015-0.06; for S. agalactiae, 0.03-0.125; for S. dysagalactiae, 0.03-0.06; for S. pneumoniae, 0.06-0.125; for H. influenzae, 0.015-0.125; and for M. catharralis, 0.5-1. CONCLUSIONS: The results indicate that ceftaroline is quite effective against bacteria that are frequently isolated from respiratory tract and wound infections.

[Investigation of the presence of class 1, 2, 3 integrons and their relationships with antibiotic resistance in clinical Stenotrophomonas maltophilia isolates]. / Klinik Stenotrophomonas maltophilia izolatlarinda sinif 1, 2, 3 integron varliginin ve antibiyotik direnci ile iliskilerinin arastirilmasi

Stenotrophomonas maltophilia is an opportunistic emergent pathogen causing hospital-acquired infections. It is resistant to majority of the broad spectrum antibiotics due to several mechanisms which significantly limit the treatment options. Although the relationship between integrons, mobile genetic elements which play role in transferring resistance genes, and the antibiotic resistance in different gram-negative bacteria have been investigated, the data are limited in Turkey especially for S.maltophilia. The aims of this study were to detect the presence of different classes of integrons and plasmids in clinical isolates of S.maltophilia and to investigate the antibiotic resistance profiles of those isolates. One hundred S.maltophilia strains isolated from various clinical samples (32 sputum, 25 tracheal aspirates, 9 urine and blood, 7 exudates and catheters, 4 sterile body fluids and wounds, 2 CSF, 1 conjunctiva) in our microbiology laboratory during January 2011-September 2012, were included in the study. The isolates were identified by VITEK2 Compact (BioMerieux, France) or Phoenix 100 (BD, USA) automatized systems, and the susceptibilities of the strains to levofloxacin, chloramphenicol, ceftazidime and trimethoprim/sulfamethoxazol (SXT) were evaluated via broth microdilution method according to the CLSI recommendations. Class 1 (intI-1), class 2 (intI-2), class 3 (intI-3) integron gene cassettes and integron 5'-3' conserved gene regions (intI-5'-3'CS) were investigated by polymerase chain reaction (PCR) using specific primers in all of the strains. Nucleotide sequence analysis of PCR products was performed in case of positive result, and the presence and size of plasmids were further investigated. The susceptibility rates of S.maltophilia strains to ceftazidime, chloramphenicol, SXT and levofloxacin were found as 24%, 66%, 93% and 95%, respectively, while MIC(50) and MIC(90) values were 64-128 µg/ml, 8-16 µg/ml, 1/19-2/38 µg/ml and 1-2 µg/ml, respectively. In PCR amplification with intI-1, intI-2 and intI-3 primers, 12%, 2% and 10% of the isolates yielded expectative bands, respectively. DNA sequence analysis of the amplified products revealed five isolates to harbour intI-1 gene, while intI class 2 and class 3 genes were not detected in any of the strains. Furthermore in PCR amplification with intI-5'CS and 3'CS primers, 20% of the strains yielded expected bands. Sequence analysis of these amplicons revealed the presence of quaternary ammonium compound resistance protein genes (qacL) in two, aminoglycoside adenyltransferase gene (aadA) in one and integron-associated recombination site (attI1) genes in five strains. Additionally, the presence of plasmids have been detected in 9 (9%) of the strains, however all of them was integron-negative. The sizes of plasmids were 2340, 1350, 2760, 18600, 20000, 3570-2540, 2510 and 5000-2540 base pairs, respectively. When the antibiotic susceptibility patterns of strains were compared with the presence of intI gene regions, no statistically significant relationship was observed (p> 0.05). In conclusion, the demonstration of integron class 1 genes and plasmids among clinical S.maltophilia strains is regarded as a warning data to indicate the potential for spread of those resistant strains in our hospital.

[NDM-1 cases and outbreaks in Turkey]. / Ülkemizdeki NDM-1 olgu ve salginlari

We are grateful to Hatipoglu and Turhan [Mikrobiyol Bul 2014; 48(1): 188-9] for their interest in our study published in Mikrobiyol Bul 2013; 47(2): 382-4. As Hatipoglu and Turhan mentioned in their comment, ertapenem is more sensitive than other carbapenem antibiotics for the identification of New Delhi Metallo-beta-lactamase (NDM-1) producers among carbapenem-resistant strains being studied. However, its low specificity [Dortet et al. Biomed Res Int 2014; 2014: 249856] makes it equal with other carbepenems. Since all the isolates in our study were not tested for ertapenem susceptibility, we used the susceptibility data for three carbapenems to increase the sensitivity of our study regarding isolate selection. We agree Hatipoglu and Turhan about the Modified Hodge Test (MHT) and we did not use MHT at all in our study. However we couldn't understand how they came to a conclusion that we used MHT and didn't mention in Material and Methods section. ZnSO4 supplemented MHT which was recommended by the authors [Dortet et al. Biomed Res Int 2014; 2014: 249856] has a sensitivity rate of about 85%. Thus we used molecular methods instead of MHT not to miss any single isolate. Hatipoglu and Turhan mentioned about previously reported four NDM-1 positive isolates without any international relation in Turkey. However, since this mentioned study [Alp et al. J Hosp Infect 2013; 84(2): 178-80] was published after the appeal, acceptance and publication of our study, eventually we didn't have the opportunity to discuss the data of Alp's report. In the same study authors stated that NDM-1 producing isolates were isolated from pediatric patients and had no connection with patients from Indian peninsula. At the same time Poirel et al. [Antimicrob Agents Chemother 2014; 58(5): 2929-33] reported in their study that NDM-1 producing isolates from pediatric patients had clonal relation with Enterobacter cloacae strains and subject to an outbreak. The evaluation of the previous reports about NDM-1 indicated that NDM-1 was initially originated from foreign sources before exhibiting endemicity in a country. Thus the situation in our region was not an exception. In conclusion, medical facilities taking care of foreign patients should pay particular attention to identification of NDM-1 isolates and establishment of appropriate control measures.

Role of migratory birds in spreading Crimean-Congo hemorrhagic fever, Turkey.

We investigated migratory birds' role in spreading Crimean-Congo hemorrhagic fever virus (CCHFV) through attached ticks. We detected CCHFV RNA in ticks on migratory birds in Turkey. Two isolates showed similarity with CCHFV genotype 4, suggesting a role for ticks in CCHFV epidemics in Turkey and spread of CCHFV by birds.

Tuberculosis or tularemia? A molecular study in cervical lymphadenitis.

BACKGROUND: Over the last two to three decades there has been a marked decrease in certain bacterial infections in Turkey. One of them is tuberculosis. Of note, statistics published by the Turkish Ministry of Health (MoH) show decreasing pulmonary tuberculosis (PTB), but on the other hand, increasing extrapulmonary tuberculosis (EPTB). The most common form of EPTB is tuberculous cervical lymphadenitis (TCL). The increase in the number of TCL cases despite the decline in cases of PTB is seen as a paradoxical issue. In contrast there has been an increase in the number of oropharyngeal tularemia cases in the last decade in Turkey. The aim of this study was to draw attention to the importance of differentiating between TCL and tularemia lymphadenitis, because these diseases may have a similar histopathological appearance. METHODS: Thirty-two cases diagnosed as TCL were identified from the archives of a pathology laboratory (Patomer Pathology Laboratory, Bursa, Turkey). PCR tests for Francisella tularensis and Mycobacterium tuberculosis were carried out on the paraffin tissue blocks of these cases. At the same time, statistical data on PTB and EPTB for the period 1996-2010 were retrieved from the MoH and reviewed. Statistics related to tularemia, which has been diagnosed since 1988 and has been increasing in the last 10 years, were obtained from the Department of Zoonoses of the MoH. RESULTS: Six out of 32 (19%) cases who had previously been diagnosed with 'casseifying granulomatous lymphadenitis consistent with tuberculosis' were found to be positive for tularemia by PCR test of the cervical lymph nodes. CONCLUSIONS: Oropharyngeal tularemia should be kept in mind in the differential diagnosis of cervical lymphadenitis in widespread geographic regions of the Northern Hemisphere, including the Asian continent. In particular, if granulomatous inflammation with necrosis is seen histopathologically, tularemia should be excluded before making the diagnosis of TCL.

Distribution of nontuberculous Mycobacteria strains.

AIM: Mycobacteria other than tuberculosis (MOTT) cause increasingly serious infections especially in immunosuppressive patients by direct transmission from the environment or after colonization. However, identification of these species is difficult because of the cost and difficulties in defining to species level. Identification and distribution of these species can help clinician in the choice of treatment. MATERIALS AND METHODS: A total of 90 MOTT strains obtained from four different centers were included in the study. These strains were identified by sequence analysis of 16S rRNA and Hsp65 genetic regions. RESULTS: Accordingly, within the 90 MOTT strains, 17 different species were identified. In order of frequency, these species were M. gordonea (n = 21), M. abscessus (n = 13), M. lentiflavum (n = 9), M. fortuitum (n = 8), M. intracellulare (n = 6), M. kumamotonense (n = 6), M. neoaurum (n = 5), M. chimaera (n = 5), M. alvei (n = 5), M. peregrinum (n = 3), M. canariasense (n = 3), M. flavescens (n = 1), M. mucogenicum (n = 1), M. chelona (n = 1), M. elephantis (n = 1), M. terrae (n = 1) and M. xenopi (n = 1). Most frequently identified MOTT species according to the geographical origin were as follows: M. abscessus was the most common species either in Istanbul or Malatya regions (n = 6, n = 6, consequently). While M. kumamotonense was the most frequent species isolated from Ankara region (n = 6), M. gordonea was the most common for Samsun region (n = 14). CONCLUSION: Our study revealed that frequency of MOTT varies depending on the number of clinical samples and that frequency of these species were affected by the newly identified species as a result of the use of novel molecular methods. In conclusion, when establishing diagnosis and treatment methods, it is important to know that infections caused by unidentified MOTT species may vary according to the regions in Turkey. The results of the study showed that there were differences in the frequency of MOTT species in the different geographical regions of Turkey.

[Investigation of the presence of New Delhi metallo-beta-lactamase-1 (NDM-1) by PCR in carbapenem-resistant gram-negative isolates]. / Karbapeneme dirençli gram-negatif izolatlarda New Delhi metallo-beta-laktamaz-1 (NDM-1) varliginin PCR ile arastirilmasi

Bacteria producing New Delhi metallo-beta-lactamase-1 (NDM-1) exhibit high level resistance to beta-lactams including carbapenems. This broad-spectrum resistance limits treatment options for infections caused by NDM-1 producers. NDM-1 was first isolated from an Indian patient in Sweden; since then, NDM-1 producing isolates have been identified in many countries including Turkey. In this study, we investigated the presence of NDM-1 by PCR method in various gram-negative isolates recovered from clinical specimens in tertiary care hospitals in Samsun, Turkey. A total of 210 carbapenem-resistant gram-negative isolates (132 Acinetobacter baumannii, 54 Pseudomonas aeruginosa, 5 Pseudomonas putida, 8 Enterobacter cloacae, 3 Enterobacter aerogenes, 3 Klebsiella pneumoniae, 2 Providencia rettgeri, 2 Escherichia coli and 1 Citrobacter freundii) were included in the study. Identification and antibiotic susceptibility testing of the isolates were performed by using Vitek-2 Compact (bioMerieux, France) and BD Phoenix (BD Diagnostic Systems, MD) automated systems. The results of antibiotic susceptibility testing were interpreted according to the CLSI recommendations. In our study, NDM-1 gene was not detected in any of the clinical isolates by PCR. There was only one case study that reported the presence of NDM-1 in clinical isolates from Turkey [Poirel L et al. Antimicrob Agents Chemother 2012;56:2784]. Our data, together with the others, indicated that the existence of NDM-1 in clinical isolates is not common in Turkey. However, since NDM-1 is a plasmid-encoded enzyme, there is always a risk of spread of this resistance through the bacterial strains in our country. Therefore, continuous surveillance and investigation of carbapenem-resistant isolates with resistance patterns suggestive of NDM-1 may enable to identify NDM-1 producing isolates. Meanwhile special care should be given on rational antibiotic use and establishment of appropriate infection control policies to prevent the spread of NDM-1 producers and other potential resistant strains.

Comparison of Culture, Real-Time DNA Amplification Assay and Ehrlich-Ziehl-Neelsen for Detection of Mycobacterium tuberculosis.

OBJECTIVE: Mycobacterium tuberculosis is still a substantial health problem universally. Although culture is the gold standard method, reliable, rapid and new methods are required for effective struggle with disease. We retrospectively compared the results of Ehrlich-Ziehl-Neelsen (EZN) stain and real-time DNA amplification assay (BD ProbeTec ET system) with culture. STUDY DESIGN: Retrospective study. MATERIAL AND METHODS: A total of 703 samples, 182 pulmonary and 521 extra pulmonary, collected from 630 patients between May 2008 and February 2011 were evaluated. Culture was considered the gold standard. RESULTS: For pulmonary specimens, sensitivity, specificity, positive predictive and negative predictive values of BD ProbeTec ET and EZN were calculated to be 100%, 98.8%, 87.5%, 100% and 71.4%, 98.8%, 83.3%, 97.6%, respectively. For extra pulmonary specimens, sensitivity, specificity, positive predictive and negative predictive values of BD ProbeTec ET and EZN were calculated to be 80%, 98.7%, 76.9%, 98.9% and 24%, 98.3%, 42.8%, 96.2%, respectively. CONCLUSION: According to these results, we suggest that the BD ProbeTec ET system is more reliable than EZN. In addition, the BD ProbeTec ET system produces faster results. Based upon these results, we consider that the BD ProbeTec ET system may be employed in the diagnosis of M. tuberculosis.

Mediterranean spotted fever in the Trakya region of Turkey.

Mediterranean spotted fever (MSF) is caused by a tick-borne pathogen, Rickettsia conorii subsp. conorii, belonging to the spotted fever group (SFG) rickettsiae. The aim of the present study was to evaluate the cases with confirmed diagnosis of MSF from 2003 to 2009 in the Trakya region of Turkey. Patients with high fever, maculopapular rash (involving the palms or soles) and/or a black inoculation eschar at the site of the tick bite (tache noire) were included in the study. Before doxycycline treatment, skin biopsy specimens, preferably from the eschar or from the maculopapular rash, were obtained for DNA extraction. Immunofluorescence assay (IFA) was performed to detect IgM and IgG antibodies against R. conorii in acute and convalescent sera. Afterwards, a standard PCR reaction using primers suitable for hybridisation within the conserved region of genes coding for outer membrane protein A (ompA) and citrate synthase (gltA) and DNA sequencing were performed. There were 128 patients with confirmed MSF diagnosis. Using IFA, seroconversion or a fourfold or greater rise in titre was observed in 97 (77%) patients, whereas a single high titre was demonstrated in 16 (12.7%) patients. According to PCR analysis, 77 (72.6%) of 106 biopsy samples showed positive results. Of these, 58 (73%) of 79 biopsy specimens were from the eschar and 19 (70%) of 27 specimens were from the maculopapular rash. No significant difference was found between the rate of positive skin biopsies taken from the eschar and the maculopapular rash. DNA sequence analysis was performed to all PCR-positive cases, and R. conorii conorii (type strain: Malish, ATCC VR-613) was detected in each of them. MSF is prevalent, but has been underdiagnosed and underreported so far in Turkey. It is a potentially severe and even fatal disease resembling viral haemorrhagic fevers that has to be included in the differential diagnosis of febrile illness associated with thrombocytopenia, even in the absence of an eschar or a tick bite. While IFA allows for retrospective diagnosis in MSF, advanced molecular techniques provide the rapid detection of rickettsia in all skin samples, including eschar and maculopapular rash.

[Investigation of the frequency of PER-1 type beta-lactamase and antimicrobial resistance rates in nosocomial isolates of Pseudomonas aeruginosa]. / Hastane Kaynakli Pseudomonas aeruginosa Izolatlarinda PER-1 Tipi Beta-Laktamaz Sikliginin ve Antibiyotiklere Direnç Oranlarinin Arastirilmasi

Pseudomonas aeruginosa which is a common cause of nosocomial infections, usually leads to treatment difficulties due to multi-drug resistance. PER-1 type extended-spectrum beta-lactamase (ESBL) producing bacteria are shown to be common in Turkey. Since limited number of antibiotics such as antipseudomonal penicillins, cephalosporins, aminoglycosides, fluoroquinolones and carbapenems are available for the treatment of P.aeruginosa infections, it is essential to monitor and eventually control the spread of antibiotic resistance genes. The aims of this study were to investigate the presence of PER-1 type ESBLs in nosocomial P.aeruginosa isolates and to evaluate their resistance to some commonly used antibiotics. A total of 110 P.aeruginosa strains isolated from clinical samples [40 urine, 26 exudate, 20 blood, 24 others (sputum, tracheal aspirate, tissue biopsy, cerebrospinal fluid, pleural fluid, conjunctiva)] of the inpatients who were proven to have nosocomial infections in Ondokuz Mayis University Faculty of Medicine Hospital between May 2002-June 2003 were included in the study. Identification of the isolates was performed by ATB system ID 32 GN (bio-Merieux, France). Antibiotic susceptibilities were detected by standard disk diffusion method and PER-1 type ESBL was searched by polymerase chain reaction using PER-1 and PER- 2 primers. PER-1 positivity was detected in 62 of 110 (56.4%) P.aeruginosa isolates and 51 of 65 (78.5%) ceftazidime-resistant strains. The highest susceptibility rate was detected for ciprofloxacin (76.4%), while the lowest susceptibility rate was for ticarcillin-clavulanic acid (22.7%). Rates of resistance to beta-lactam agents (excluding piperacillin/tazobactam), amikacin and gentamicin were statistically significantly higher for PER-1 positive strains than PER-1 negative ones. Resistance rates to ceftazidime, cefepime, aztreonam, piperacillin and ticarcillin-clavulanic acid in PER-1 positive isolates versus negative ones were as 82.3% vs. 29.2% (p< 0.01), 75.8% vs. 25% (p< 0.01), 83.9% vs. 30.4% (p< 0.01), 73.8% vs. 52.2% (p< 0.05), 85.5% vs. 66.7% (p< 0.05), respectively. Considering resistance rates to piperacillin-tazobactam and ciprofloxacin, PER-1 positive isolates versus negatives were 35.5% vs. 31.3%, and 19.4% vs. 29.2%, respectively, revealing no statistical significance (p> 0.05). As a result, PER-1 type ESBL frequency and beta- lactam and aminoglycoside resistance rates were found remarkably high in nosocomial P.aeruginosa strains isolated in our hospital. It was concluded that antibiotic resistance should be continously monitorized and necessary measures to prevent further increase in resistance should be promptly established.

CTX-M type extended spectrum beta-lactamases in Escherichia coli isolates from community acquired upper urinary tract infections at a university in the European part of Turkey.

Extended spectrum beta-lactamase (ESBL) producing Escherichia coli has been an emerging etiologic agent in the community acquired infections. We investigated the occurrence of ESBL producing E. coli isolated from patients admitted with community acquired urinary tract infection (UTI) to the hospital of the Trakya University, Turkey during 2006. Eleven single patient isolates of E. coli harboring ESBL were identified among 30 E. coli isolated from patients admitted with symptoms corresponding to upper UTI. CTX-M type ESBLs were detected in all 11 ESBL-producers by isoelectric focusing and polymerase chain reaction screening. Sequence analysis revealed CTX-M-1 in one isolate, CTX-M-3 in three isolates and CTX-M-15 in seven isolates. ESBL-producing E. coli isolated from community acquired UTIs are widespread in the European part of Turkey.

Vascular endothelial growth factor expression levels of gingiva in gingivitis and periodontitis patients with/without diabetes mellitus.

OBJECTIVE AND DESIGN: The aim of this study was to determine vascular endothelial growth factor (VEGF) mRNA expression levels in gingival tissues of gingivitis and periodontitis patients with diabetes mellitus and those without. The hypothesis tested is that expression of VEGF, considered the effective cytokine in the relationship between diabetes and periodontal disease, is differentially affected in gingivitis and periodontitis patients with or without diabetes mellitus compared to healthy controls. METHODS: Forty-five subjects were evaluated in five groups; individuals with gingivitis (group 1; n = 10), individuals with periodontitis (group 2; n = 10), individuals with gingivitis + type II diabetes (group 3; n = 10), individuals with periodontitis + type II diabetes (group 4; n = 10), and individuals without periodontal and systemic disease (group 5; n = 5). VEGF mRNA levels in gingival tissues were measured by quantitative real-time PCR using Lightcycler. RESULTS: Expression of VEGF mRNA was detected in all groups. There was no significant difference in expression levels of VEGF mRNA between groups (P > 0.05). CONCLUSIONS: VEGF expression is probably related to both maintenance of periodontal health and periodontal tissue destruction. It can be concluded that systemic condition in type II diabetes mellitus under good metabolic control does not seem to have additional effects on gingival tissue VEGF mRNA levels in gingivitis and periodontitis patients.

Detection of hepatitis B virus in used razor blades by PCR.

BACKGROUND AND AIMS: Hepatitis B virus (HBV) infection ranks among the most devastating health problems in the world.The most probable transmission routes of HBV are blood contact, sexual, and horizontal transfer. Other sources of HBV transmission are razor sharing, beauty treatments, tattooing, piercing, and manicures and other chiropody treatments.Many infections have been reported in South-East Asia, where barbers commonly share and reuse razors. Detection of HBV DNA in contaminated devices such as razor blades is important in the demonstration of transmission routes and indirect estimation of HBV prevalence in specific subpopulations such as barbershop clientele. Therefore, we aimed to detect the presence of HBV contamination on razor blades by nucleic acid testing. METHODS: Used razor blades (n = 151) were purchased from different barber's shops. Used razor blades purchased from chronic HBV patients (n = 8) were included as a positive control. The amplification and detection of HBV DNA was carried out by a semi-nested PCR method in a thermal cycler. RESULTS: The presence of HBV DNA was found in 10 (6.6%) used razor-blade samples by the detection of a specific positive band with agarose gel electrophoresis. CONCLUSIONS: In conclusion, used razor blades may be contaminated with HBV, and the practice of sharing used razor blades may pose a risk of transmission. Nucleic acid detection methods involving PCR can be used to detect HBV contamination of razor blades. HBV control and prevention programs should educate barbers about the importance of contagious diseases, proper sterilization techniques, and avoiding reuse and sharing of contaminated equipment and supplies such as razor blades. As an infection control measure, prohibition of razor reuse can reduce the spread of HBV infection in rural areas, where the practice is often common at barbershops.

Core mutations in patients with acute episodes of chronic HBV infection are associated with the emergence of new immune recognition sites and the development of high IgM anti-HBc index values.

Acute exacerbations in HBeAg negative patients with chronic hepatitis B virus (HBV) infection are invariably associated with concurrent increases in the index of IgM class antibodies against the core protein (anti-HBc) of the virus. This study aimed to investigate whether this was related to the clearance of variants from the quasispecies pool and the appearance of new ones, with aminoacid substitutions in well recognized B-cell epitopes. In this study, 5 HBeAg negative patients (A to E) with 13 sequential serum samples (A1-A2, B1-B2-B3, C1-C2, D1-D2-D3, E1-E2-E3) were investigated after amplification of the entire core encoding region followed by cloning/sequencing studies. The sequences at different time points were compared with those from a single HBeAg positive patient with no apparent acute exacerbations. The results from sequence comparison showed that virus variants emerged in all (A2, B3, C2, D3, E2, and E3) but two (B2 and D2) subsequent sera with amino-acid substitutions affecting B-cell epitopes. It is concluded that the rise in the values of IgM anti-HBc may be attributed to the alteration of the antigenic epitopes leading to new antibody production in the majority of the cases. However, it appears that increases in IgM anti-HBc indexes in a few cases may relate to other possible mechanisms which are discussed.

Emergence and spread of carbapenem-resistant Acinetobacter baumannii in a tertiary care hospital in Turkey.

An intensive care unit (ICU)-based OXA-23-producing multiple-drug resistant Acinetobacter baumannii (MDRAB) outbreak was detected between October 2005 and October 2006. A total of 47 patients were infected/colonized with the outbreak strain. Clinical data were available from 37 patients. The all-cause mortality rate among the patients exposed to the epidemic strain was 35% (13/37). The outbreak strain and the resistance determinants were characterized both by microbiological methods and by molecular techniques. Cloning and sequencing experiments identified ISAbaI-associated bla(oxa-23) on the chromosome. Screening of imipenem-resistant Acinetobacter isolated from the ICU during the outbreak period with PCR identified 97 isolates as positive for the ISAbaI-bla(oxa-23) structure. Pulsed-field gel electrophoresis and plasmid analyses with selected nonrepetitive isolates revealed the clonality. Disk diffusion on cloxacillin-supplemented agar media and the real-time PCR experiments showed that outbreak isolates are overexpressing the ampC enzyme. This study highlights the occurrence of OXA-23-producing and ampC-overexpressing MDRAB in ICUs.

[A case of cat scratch disease]. / Kedi tirmigi hastaligi olgusu.

Cat scratch disease (CSD) which is usually seen in children and young adults and characterized by self limited lymphadenopathy, is caused by Bartonella henselae. In this report, a 30-year-old male patient who was admitted to the outpatient clinic with the complaint of lymphadenopathy, was presented. Erythrocyte sedimentation rate and C reactive protein values of the patient were normal, and anti-HIV, Brucella agglutination and anti-toxoplasma IgM and IgG tests were negative. No bacterial growth was detected in the cultures of repetetive blood samples and biopsy specimens taken from the right axillar lymph node. This might probably be due to the prior antibiotic therapy (ciprofloxacin and cefazolin) given by another health center. Polymerase chain reaction performed with biopsy specimen by using 27f and 1525r primers, also yielded negative result. CSD was diagnosed depending on the history of direct contact with a cat (being scratched and bitten), together with the clinical features and histopathologic findings (necrotizing granulomatous inflammation concordant with CSD). Lymphadenopathies of patient had completely resolved after five-days oral azithromycin therapy. Since CSD is a rare infection of adults, it should be taken into consideration in the patients who suffer from silent lymphadenopathy and present with the history of direct contact with cats.

[A herpes simplex virus encephalitis case with no clinical response to acyclovir treatment]. / Asiklovir tedavisine klinik yanit alinamayan bir herpes simpleks virus ensefaliti olgusu.

In spite of high rates of morbidity and mortality in herpes simplex virus (HSV) encephalitis, however, it is one of the exceptional viral infections with specific and effective therapy. In this report a HSV encephalitis case who was clinically unresponsive to acyclovir treatment, has been presented. An 11 months old girl patient has been brought to our clinic with the complaints of high fever and focal convulsions. Analysis of cerebrospinal fluid (CSF) revealed decreased glucose level and abundant red blood cells, despite it was not traumatic. The other CSF biochemical findings were found normal. Viral serology performed with CSF yielded negative result for HSV-1 IgG, positive result for HSV-2 IgG, and negative result for HSV-1/2 IgM, however, antibody index could not be estimated since it was not possible to obtain a simultaneous serum sample. Cranial magnetic resonance imaging (MRI) showed contrast material enhancement on bilateral temporal lobes. There was no growth in the CSF cultures. Acyclovir therapy (30mg/kg/day) was started with the prediagnosis of herpes encephalitis. In the third week of therapy CSF analysis was repeated because of the presence of partial paroxysmal attacts and absence of sufficient clinical improvement. In this CSF sample HSV-1 DNA was found positive by real-time polymerase chain reaction. Since CSF findings were still abnormal and the clinical picture worsened despite 21 days of therapy, the dose of acyclovir was increased to 60 mg/kg/day (3 weeks) with a possible drug resistance problem. Control brain MRI showed contrast enhancement on bilateral temporal lobes, with more intensivity in left, and encephalomalacia. Valproic acid and haloperidol were given to the patient for the treatment of permanent partial paroxysms and orofacial dyskinesis, developing in the follow-up period, respectively. After getting these complications under control, the patient was discharged and taken into follow-up. As a result, although it could not be possible to confirm the drug resistance by molecular methods, it was thought that this might be both a clinical and virological resistance phenomenon, because of the detection of HSV-DNA in the CSF sample during the period of severity of the illness.

High prevalence of OXA-51-type class D beta-lactamases among ceftazidime-resistant clinical isolates of Acinetobacter spp.: co-existence with OXA-58 in multiple centres.

OBJECTIVES: This study was designed to demonstrate the prevalence of the newly discovered carbapenem-hydrolysing class D enzymes, OXA-51-type and OXA-58, among clinical isolates of Acinetobacter spp. METHODS: A total of 72 isolates from six centres were studied. Isolates were screened by PCR with specific primers for bla(OXA-51-type) and bla(OXA-58). PCR products were sequence-analysed. Plasmids were digested with EcoRV and genomic DNAs were digested with PvuII. Hybridization experiments were done with digoxigenin-labelled specific probes. Macro-restriction analysis was done on SmaI-digested genomic DNAs. RESULTS: A total of 56 (77.8%) isolates were positive for bla(OXA-51-type) genes. Sequence analysis of the products from 23 selected isolates revealed the occurrence of multiple alleles in all contributing centres. The bla(OXA-58) gene was detected among 10 isolates from five centres. All were also positive for bla(OXA-51-type) genes. Among the bla(OXA-58)-positive isolates, two from the same centre were positive for a novel OXA-51 allele (OXA-86). Southern hybridization of plasmids and of genomic DNAs suggested that bla(OXA-51-type) genes are located on chromosomes whereas bla(OXA-58) genes are plasmid borne in these 10 isolates. Plasmid profiles and pulsed-field gel electrophoresis patterns indicated the spread of the bla(OXA-58) gene among multiple clones. The bla(OXA-51-type) and bla(OXA-58) co-carrier strains were mostly associated with a pandrug-resistant phenotype. CONCLUSIONS: This study indicated that bla(OXA-58)-bearing plasmids are readily spreading among multiple clones of the bla(OXA-51-type)-bearing clinical isolates of Acinetobacter spp. Since these isolates are highly resistant to antibiotics this finding indicates the existence of a significant problem in Turkish hospitals.