[In Vitro Activity of Ceftazidime-avibactam and Colistin Against Carbapenem-Resistant Klebsiella pneumoniae Clinical Isolates]. / Karbapenem Dirençli Klebsiella pneumoniae Klinik Izolatlarina Karsi Seftazidim-avibaktam ve Kolistinin In Vitro Etkinligi.

Infections caused by multi drug-resistant gram-negative bacilli are increasingly reported worldwide. Colistin, tigecycline and aminoglycosides are almost the only and last choice antibiotics in the treatment of infections caused by carbapenem-resistant Enterobacterales members. Ceftazidime-avibactam is a novel antibiotic combination consisting of a broad-spectrum cephalosporin and avibactam with good antimicrobial activity against carbapenem-resistant Enterobacterales members. The aim of this study was to assess the in vitro activity of ceftazidime-avibactam and colistin against carbapenem-resistant Klebsiella pneumoniae isolates and to obtain local antimicrobial surveillance data. A total of 150 carbapenem-resistant K.pneumoniae isolates obtained from various clinical samples of the patients hospitalized in our hospital between 2018-2021 were included in the study. Duplicate isolates were excluded from the study. The isolates were recovered from blood (n= 72), tracheal aspirate (n= 40), wound (n= 20), biopsy and abscess (n= 10), steril body fluid (n= 5), and peripheral venous catheter (n= 3) samples. Isolates were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS, Bruker Daltonics, Germany). The minimum inhibitory concentration (MIC) values of the isolates for meropenem, colistin, ceftazidime, and ceftazidime-avibactam were determined by broth microdilution method. Susceptibility of the isolates to the tested antibiotics was evaluated by the European Committee of Antimicrobial Susceptibility Testing (EUCAST) criteria. The presence of carbapenemases (VIM, IMP, NDM, KPC, and OXA-48) was investigated by polymerase chain reaction (PCR) using specific primers. The mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 genes were evaluated by PCR for plasmid-mediated colistin resistance. All K.pneumoniae isolates were found to be positive for at least one of the carbapenemase genes evaluated in the study. The blaOXA-48 gene was detected in 107 (71.3%), blaKPC gene in 25 (16.7%); blaNDM gene in 7 (4.7%), co-production of blaOXA-48 and blaKPC genes in 10 (6.7%), co-production of blaOXA-48 and blaNDM genes in 1 (0.6%) isolate. None of the isolates harbored the blaVIM and blaIMP genes. None of the mcr genes screened in the study were detected among the isolates. The susceptibility of the isolates to ceftazidime-avibactam and colistin was 92.7% (139/150) and 48% (72/150), respectively. The MIC50 and MIC90 values for meropenem, ceftazidime, ceftazidime-avibactam, and colistin of the isolates were determined as 32/256, > 128/> 128, 1/8, and 4/16 µg/ml, respectively. Of the ceftazidimeavibactam resistant isolates, seven were positive for blaNDM, three for blaKPC, and one for both blaOXA-48 and blaNDM genes. High ceftazidime-avibactam MIC levels (> 128 µg/ml) were detected in metallo-betalactamase producing isolates. Consequently, our data suggested that ceftazidime-avibactam exhibited as a good alternative therapeutic choice for carbapenem-resistant K.pneumoniae isolates. It is noteworthy that high rate of colistin resistance was detected in K.pneumoniae isolates. Another notable finding of this study is the increase in K.pneumoniae isolates producing blaKPC for our country. To prevent the development of resistance which is observed even in last-choice therapeutic antibiotics, the principles of rational antibiotic use should be followed. The appropriate antimicrobial susceptibility testing should be routinely performed for surveillance of ceftazidime-avibactam and colistin.

Impact of methods and duration of surgical hand scrub on bacterial count: A randomized controlled trial.

BACKGROUND: There is no standard protocol for surgical scrubbing. This study aims to compare the effectiveness of surgical hand scrub duration and method by analyzing their effects on bacterial count. METHODS: The study was conducted on 180 surgical nurses and surgeons. While the duration of surgical hand scrub in Groups I and II was one minute, participants in Group I used a nail brush, whereas Group II did not. Similarly, the duration of surgical hand scrub in Groups III and IV was two minutes, but Group III used a nail brush, whereas Group IV did not. Bacterial count on the hands of all participants was measured before and after the surgical hand scrub and after the surgery by using the glove juice method. RESULTS: Bacterial count on the hands of the participants in Group III after surgical hand scrub was significantly higher than Group IV (P < .001). We did not find any statistically significant difference between Group II and Group IV in terms of bacterial count on the hands immediately after surgical hand scrub and after the surgery (P = .401, P =.658, respectively). CONCLUSIONS: This study found that brushing during surgical hand scrub increased the number of bacteria on the hand. Besides, one-minute surgical hand scrub was equally effective as two-minute scrub to reduce the number of bacteria on the hand.

An Investigation into Bacterial Bloodstream Infections and Antibiotic Resistance Profiles in a Tertiary Hospital for a Ten-Year Period.

BACKGROUND: Bloodstream infections are one of the major causes of healthcare-associated morbidity and mortality. The present study aims to investigate the prevalence of the microorganisms isolated from blood cultures and to evaluate susceptibilities to antimicrobial agents in a tertiary center, Gulhane Training and Research Hospital, Ankara, Turkey. METHODS: Blood cultures (BCs) were incubated in BACTEC/9050 (Becton Dickinson, USA) (2007 - 2015) and BacT/ALERT (bio-Merieux, France) (2014 - 2016) automated systems. PhoenixTM 100 system (Becton Dickinson, USA) (2007 - 2014), MALDI-TOF MS (Bruker, USA) (2015 - 2016) and conventional techniques were used for the identification of isolated microorganisms. According to CLSI (2007 - 2014) and EUCAST (2015 - 2016) criteria, Kirby-Bauer disc diffusion method, PhoenixTM system, and broth microdilution were applied for antimicrobial susceptibility testing. Two five-year periods were statistically compared regarding antibiotic resistance. RESULTS: From the overall evaluated 31,380 BCs, 7,367 cultures (23.5%) were positive, excluding 503 BCs (6.4%), which were interpreted as contamination. Of 7,367 isolated microorganisms, 3,680 (50.0%) were gram-negative, 3,303 (44.8%) were gram-positive bacteria, and 384 (5.2%) were fungi. Coagulase-negative staphylococci (CoNS) were predominantly isolated (n = 2,075; 28.2%) among gram-positives. E.coli (n = 978; 13.3%) was the most frequently isolated gram-negative species. Between the first and the last five-year period, three genera (Enterococcus spp., Acinetobacter spp., Streptococcus spp.) showed significant differences when isolated, and only Enterococcus spp. showed increased isolation rates. In total, 90.3% of CoNS and 32% of S. aureus were methicillin-resistant. Only 75 strains of Enterococcus spp. (12.1%) were vancomycin-resistant. ESBL was detected in 40.6% of E. coli and 30.7% of Klebsiella spp. isolates. Carbapenem resistance showed a significant increase, particularly in K. pneumoniae (> 20%). CONCLUSIONS: The findings suggest that there was a threatening condition in antimicrobial resistance rates, especially for some antimicrobials between two periods. Although antimicrobial resistance is usually associated with MRSA, carbapenem resistance, ESBL, and VRE, the problem is far beyond these definitions, consisting of not just medicine, but also commercial companies, food industry, veterinarians, and other areas.

False resistance after artemether-lumefantrine treatment in a falciparum malaria patient in Turkey: A case report.

INTRODUCTION: Artemisinin-based combination therapy (ACT) is recommended by the World Health Organization as first-line treatment of uncomplicated Plasmodium falciparum malaria. ACT treatments failures among travellers returning from Africa to non-endemic countries are considered to be caused by resistance. CASE PRESENTATION: We report on a case of artemether-lumefantrine treatment failure in a Turkish traveller with uncomplicated P. falciparum malaria returning from Bamako, Mali. CONCLUSIONS: Information on returning travellers, includes ensuring that the patients receive supervised treatment with the recommended dose of a quality controlled medicine, routine follow-up of all cases, assessment of adequate absorption of the drug, and/or testing the prevalence of molecular markers of drug resistance if validated, can be an important source of an early warning system for emerging resistance.

Intracranial infections: lessons learned from 52 surgically treated cases.

OBJECTIVE: Intracranial infections are serious and life-threatening health problems. They may present as subdural empyemas or intracerebral abscesses. Surgical drainage and subsequent antibiotic treatment is the main technique for a satisfactory clinical outcome. The aims of this study were to present a 10-year intracranial infection series and discuss the surgical characteristics in the light of literature. METHODS: Fifty-two patients with intracranial infection underwent surgical treatment between 2008 and 2018. Eleven patients were female and 41 patients were male. The mean age was 40.46 years (range 10-75 years). Eighteen patients had intracerebral abscesses, and 34 had subdural empyemas. All patients underwent surgical treatment as well as an antibiotic regimen. RESULTS: No etiological agent was isolated in 29 (56%) cases. Bacterial agents were detected in 20 cases, while fungi were observed in 3 cases. Staphylococci species were the most common agents and were isolated in 8 (15%) cases. Endoscopic aspiration was performed in 3 cases, while surgical drainage and capsule resection via craniotomy was performed in 49 cases. An associated intracranial tumor was diagnosed in 2 patients with brain abscesses. Four (8%) patients died despite surgical and medical treatments. CONCLUSIONS: Surgical treatment via craniotomy is an older method, but it is still the best to treat the intracranial infections not only for decompression of the brain but also to attain an accurate diagnosis. The abscess wall should always be histologically examined after surgery to rule out any intracranial tumor.

Moxifloxacin-induced neutropenia in 26-year-old man.

Moxifloxacin is a fourth generation widely used fluoroquinolone antibiotic. There are three cases of moxifloxacin-induced neutropenia reported in the literature and we report the fourth case. A 26-year-old man with pneumonia was treated with moxifloxacin because of penicillin allergy. On the second day of therapy, leukopenia [White blood cell (WBC) count 2.7×10³/µL] and neutropenia (neutrophils 1.21×10³/µL) occurred. Rothia mucilaginosa was isolated in sputum culture. On the fourth day of hospitalization moxifloxacin treatment was stopped and clarithromycin 500 mg PO twice daily was started. Leukopenia and neutropenia resolved one day after discontinuation of moxifloxacin that WBC and neutrophil count rose 4.5×10³/µL and 1.97×10³/µL, respectively. On the sixth day of hospitalization, WBC and neutrophil count was 4.3×10³/µL and 2.29×10³/µL, respectively. The immunomodulatory effects of moxifloxacin may result in the changes of WBC count like leukopenia with neutropenia. Moxifloxacin induced neutropenia may be more common and is an important adverse effect. More observational studies about safety profiles of moxifloxacin are needed.

The effects of different types of hair shaving on the body image and surgical site infection in elective cranial surgery.

AIMS AND OBJECTIVES: To investigate the effects of different types of shaving on body image and surgical site infection in elective cranial surgery. BACKGROUND: Hair shaving before cranial surgery is commonly performed in many countries. However, the impact of shaving on the patients' body image and surgical site infection is not, as yet, well investigated. DESIGN: A randomised-controlled design was used in this study. METHODS: The sample comprised 200 patients who underwent elective cranial surgery between March 2013-August 2014. The Center for Disease Control and Prevention criteria were applied for the preoperative preparation of patients and for the follow-up of surgical site infection. Wound swab cultures were obtained four times from all patients. The Social Appearance Anxiety Scale was used to assess changes in the body image of patients. FINDINGS: The rate of surgical site infection was 1% for each group and for all patients. There was no difference between the groups of surgical site infection. Coagulase-negative staphylococci and Staphylococcus epidermidis were mostly isolated in the swab cultures. The Social Appearance Anxiety Scale score decreased in patients who underwent strip shaving and increased in patients with regional shaving. CONCLUSION: There is no difference between strip shaving and regional shaving in the development of surgical site infection after cranial surgery. In addition, regional hair shaving negatively affects the patients' body image. RELEVANCE TO CLINICAL PRACTICE: Findings of this study provide useful evidence-based information for healthcare professionals. The development and implementation of effective interventions result in the prevention of surgical site infection and improvement of the patients' body image in elective cranial surgery.

A new utilization area for hyperbaric oxygen? Improving quality of stored blood.

BACKGROUND/AIM: Since blood bags have the ability for diffusion of gases, we investigated whether hyperbaric oxygen (HBO) exposure affects several vital parameters of stored blood. MATERIALS AND METHODS: Bloods obtained from the same persons were used as both control and HBO groups and stored in pediatric bags with citrate-phosphate-dextrose solution. HBO administration was performed at 2.5 atm for 90 min, started 1 day after blood collection and repeated every 2 days for a total of 10 times. The study was terminated on the 21st day. Complete blood count, glucose, pH, and osmotic fragility values were measured every week. RESULTS: Glucose and pH levels decreased in stored blood. In the HBO-exposed group, these decreases were less than in the control. In addition, mean corpuscular and platelet volumes tended to increase during storing process, but with HBO, these indexes remained lower, near physiologic levels. Another interesting finding of the study was the relative stable osmotic fragility ratio in the HBO group compared to the control blood. CONCLUSION: HBO exposure has positive effects on pH, stability of erythrocytes, and energy source (glucose) of the medium. Thus, we concluded that HBO may be a useful application for life and quality of stored blood.

Effect of 2 elastomeric ligatures on microbial flora and periodontal status in orthodontic patients.

INTRODUCTION: The aim of this study was to compare the effects of a nonconventional elastomeric ligature (Slide; Leone, Florence, Italy) with those of a conventional elastomeric ligature (Ormco, Orange, Calif) on microbial flora and periodontal status in orthodontic patients. METHODS: A total of 13 orthodontic patients scheduled for fixed orthodontic treatment were selected for this study. The use of Slide and conventional elastomeric ligatures in fixed orthodontic appliances was tested. Microbial and periodontal records were obtained before bonding and 1 and 5 weeks after bonding. For the statistical analysis and calculations, SPSS software (version 15.0; SPSS, Chicago, Ill) was used. In the statistical decisions, P <0.05 values were accepted as significantly different. RESULTS: No significant differences between Slide and conventional elastomeric ligatures were evident at 1 week or 5 weeks after bonding, with regard to gingival index, plaque index, gingival bleeding index, or pocket depth scores (P >0.05). Similarly, aerobic and anaerobic bacteria counts did not differ significantly on the surface or on the elastics (P >0.05). CONCLUSIONS: Although the Slide ligatures cover the total surface of the bracket, they do not cause significantly more plaque accumulation or periodontal problems than do the conventional elastomeric ligatures.

Effectiveness of N-butyl cyanoacrylate-based microbial skin sealant on the prevention of surgical site infections.

BACKGROUND: Surgical site infections (SSIs) are a serious concern in health care, and wound contamination by endogenous skin flora is a major factor in the development of SSIs. Despite preventive tactics in pre-operative skin care, antibiotic prophylaxis, surgical technique, and post-operative incision care, complete sterilization of the skin is not possible. Recently developed microbial skin sealant forms a continuous but breathable barrier that prevents migration of endogenous skin flora into the incision. The skin sealant closes dermal microabrasions, preventing re-colonization of potential pathogens at the incision. The purpose of this study was to determine the effect of an N-butyl cyanoacrylate-based microbial skin sealant in reducing the occurrence of SSIs in an experimental rodent model. METHODS: This was a randomized, controlled animal trial. Forty-eight Wistar albino rats were divided into six groups of eight rats each. Three groups received application of sealant against specific bacteria, and three matched control groups received only the bacteria without the sealant. Group one underwent pre-operative hair removal, followed by application of skin sealant, then abdominal incision and closure. Group two (control) simply underwent hair removal, followed by incision and closure, with no skin sealant applied. Group three received an application of cage swabs (containing a mixture of urine, stool and sawdust from the animals' cages) before application of skin sealant, and group four (control) received cage swabs without subsequent skin sealant. Group five received methicillin-resistant Staphylococcus aureus (MRSA) followed by skin sealant, and group six (control) received MRSA without skin sealant. Seven days after surgery, the animals were sacrificed. Samples were taken from the abdomen of each rat and placed in culture medium. Proliferation of the following bacteria were observed: Coagulase-negative staphylococci (CoNS), gram-positive bacilli (GPB), Pseudomonas aeruginosa, and MRSA. RESULTS: There was a statistically significant difference between the median number of GPB in the group that received cage swabs+sealant and the group that received cage swabs without sealant (median, GPB count 29,430 colony-forming units [CFU]/g vs 359,100 colony-forming units [CFU]/g; p<0.05). The study results showed that microbial skin sealant was not as effective in preventing CoNS or MRSA contamination as it was in preventing GPB contamination. CONCLUSIONS: Use of a microbial skin sealant before surgery can lower the rate of SSIs by reducing the migration of some specific bacterial agents. Additional data are needed to validate its use in clinical practice.

[Investigation of various virulence factors among the hospital and community-acquired Staphylococcus aureus isolates by real-time PCR method]. / Hastane ve Toplum Kaynakli Staphylococcus aureus Izolatlarinda Çesitli Virülans Faktörlerinin Gerçek Zamanli PCR Yöntemiyle Arastirilmasi

Staphylococcus aureus is the most common cause of skin and soft tissue infections in the community and the most important cause of nosocomial infections. In this research, it was aimed to detect the presence of staphylococcal enterotoxin A to D (SEA, SEB, SEC and SED, respectively), toxic shock syndrome toxin-1 (TSST-1), Panton-Valentine leukocidin (PVL) and SCCmec phenotype in methicillin-resistant S.aureus (MRSA) isolates and to demonstrate the genotypic association between hospital acquired and community acquired isolates. In the study the virulence factors of 147 S.aureus strains isolated from various clinical samples at Gulhane Military Medical Academy Hospital Microbiology Laboratory between 2007 and 2010 were investigated by real-time polymerase chain reaction. MLVA (multiple locus variable number of tandem repeat analysis) method was used to demonstrate the genotypic association between hospital-and community-acquired isolates. Seventy-two (%48.9) of 147 S.aureus isolates were determined as community acquired and 75 (%51.1) as hospital acquired. Ninety-three (63.2%) isolates possesed at least one toxin (77 strains harboured one, and 16 strains harboured more than one). Of the isolates in which toxin was detected 59.1% (55/93) were hospital acquired, 40.9% (38/93) were community acquired. SEA was determined in 59 (40.1%), SEB in 8 (5.4%), SEC in 12 (8.1%), SED in 8 (5.4%), TSST- 1 in 17 (11.5%) and PVL in 6 (4.0%) of the isolates. Methicillin resistance was determined in 61.1% (44/72) of the hospital acquired isolates and 6.7% of the (5/75) community acquired isolates. In our study, SCCmec type III was detected in 90.9% (40/44) of hospital acquired MRSA isolates and SCCmec type IV in 40.0% (2/5) of community acquired MRSA isolates. Most of the strains (40/47; 85.1%) carrying SEA were hospital acquired, and they were determined as methicillin-resistant. According to MLVA, hospital and community acquired groups' clustering rates, number of clones, number of unique profile were determined as; 73.6% and 57.3%; 34% and 47%; 19% and 32%, respectively. It was concluded that high prevalence of SEA toxin in hospital acquired MRSA isolates indicated that there was a possible association between the presence of toxin and antimicrobial resistance.

[Comparison of endotracheal aspiration and mini-BAL culture results in the diagnosis of ventilator-associated pneumonia]. / Ventilatörle Iliskili Pnömoni Tanisinda Kullanilan Endotrakeal Aspirat Kültürü ile Mini-BAL Kültürünün Karsilastirilmasi

The objective of this study was to compare the results of cultures obtained by mini-bronchoalveolar lavage (BAL) and endotracheal aspiration (ETA) techniques, used for rapid and accurate determination of pathogens causing ventilator-associated pneumonia (VAP) in intensive care units. Of the 92 patients on mechanical ventilation followed at the emergency intensive care unit of our hospital between June 2010 and June 2011, 30 (32.2%) patients were diagnosed as VAP and they were included in this study. VAP diagnosis were based on the clinical and radiological findings. Clinical pulmonary infection score (CPIS) of > 6 was accepted as the clinical criteria of VAP. Initially ETA samples were collected from the patients followed by mini-BAL sampling 15 minutes later, together with urine and two blood cultures. Microbiological evaluation and identification were performed by conventional methods and Phoenix 100 (BD Diagnostic Systems, ABD) automated system. In quantitative culture analysis, > 10.000 cfu/ml for BAL and > 100.000 cfu/ml for ETA were accepted as the positive result. The mean ages of VAP-developed (n= 30; 18 were male) and nondeveloped (n= 62; 39 were male) patients were 68.23 ± 16.19 and 52.16 ± 10.41 years, respectively, and the mean durations of mechanical ventilation were 29.57 ± 15.78 and 12.11 ± 6.01 days, respectively. Multivariate logistic regression analysis showed that older age (p< 0.001) and duration of mechanical ventilation (p< 0.001) were independent risk factors for VAP development. There was also a statistically significant difference in CPIS values between patients who developed VAP and not (6.8 ± 1.15 and 2.71 ± 1.06, respectively; p< 0.001). The use of CPIS for VAP diagnosis was found to be useful in patients on mechanical ventilation. In our study, a total of 16 strains (six A.baumannii, three P.aeruginosa, one K.pneumoniae, six S.aureus) were isolated from ETA cultures, while 34 strains (16 A.baumannii, six P.aeruginosa, four K.pneumoniae, two E.coli, six S.aureus) were isolated from mini-BAL cultures of 30 VAP patients. The contamination rate for ETA cultures was found as 27% (8/30), however there was no contamination in mini-BAL samples. The rates of negative cultures for ETA and mini-BAL were 20% (6/30) and 7% (2/30), respectively. Seven (87.5%) of the eight contaminated ETA samples, yielded pathogenic bacterial growth (six A.baumannii, one K.pneumoniae) in mini-BAL samples. Similarly, of the six negative ETA samples, 5 (83%) yielded bacterial growth (two E.coli, two K.pneumoniae, one P.aeruginosa) in mini-BAL samples. Statistical analysis with Spearman test indicated no positive correlation between the culture results of mini-BAL and ETA (p= 0.464), and the concordance between the culture results of those methods was found as 50%. It was concluded that the use of mini-BAL instead of ETA samples for the isolation of causative microorganisms of VAP seemed to be more useful due to the high contamination risk in ETA culturing techniques and higher bacterial isolation rates in mini-BAL sampling.

[Investigation of West Nile virus RNA in blood donors by real-time RT-PCR]. / Kan Donörlerinde Gerçek Zamanli RT-PCR ile Bati Nil Virusu RNA'sinin Arastirilmasi

West Nile virus (WNV), a member of Flaviviridae family, is an enveloped, icosahedral symmetric RNA virus. Primary reservoir hosts of WNV are birds, but the virus can cause various infections in humans and other mammals. The most common and natural transmission way of WNV infections is mosquito bites, however, humans can be infected by different routes. The most important non-mosquito transmission route is contaminated blood and blood products. In this study, we aimed to investigate the risk of WNV transmission through blood and blood products in Ankara, Turkey. The presence of WNV RNA was investigated by in house real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in serum samples obtained from 729 healthy blood donors (mean age: 27.7 years; 711 were male), regardless of the donor's seropositivity status since the virus can be transmitted at the early stages of infection when seroconversion has not yet developed. Serum samples were collected in August-September 2009, the period when these infections are more frequent due to mosquito activity. The vast majority of donors (n= 702, 96.3%) have been inhabiting in Ankara and 569 (78%) of donors have had risk factors for arboviral infections (e.g. outdoor activity, mosquito and tick bites). WNV RNA was not detected by real-time RT-PCR analysis in any serum sample included in this study. According to the results of our study, it can be said that the risk of WNV transmission through blood and blood products is low in Ankara. However, WNV seropositivity was detected within the range of 0.56 to 2.4% among blood donors in previous studies and probable and confirmed WNV infections have been reported in our region. In addition, WNV outbreaks have emerged in some countries neighbouring Turkey recently. Thus, the risk of WNV transmission through blood and blood products should not be ignored and blood donor questionnaires should be evaluated in detail.

Effect of a self-etching adhesive containing an antibacterial monomer on clinical periodontal parameters and subgingival microbiologic composition in orthodontic patients.

INTRODUCTION: The aims of this study were to evaluate the effect of a self-etching adhesive system containing an antibacterial monomer on periodontal health and subgingival microbiologic composition in orthodontic patients and to compare it with a conventional adhesive system. METHODS: A split-mouth design was chosen, and 15 patients were included in the study. Brackets in contralateral quadrants were bonded with either a conventional adhesive system (control) or a self-etching adhesive system that contained an antibacterial monomer. Clinical periodontal parameters including plaque index, gingival index, probing depths, and bleeding on probing were determined. Subgingival plaque samples were collected before bracket placement (T0) and at the 6-month follow-up (T1). The real-time TaqMan polymerase chain reaction assay was used to determine the subgingival counts of Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Campylobacter rectus. For clinical periodontal parameters, analysis of covariance (ANCOVA) and, for bacterial counts, Wilcoxon tests were used for statistical comparisons at the P <0.05 level. RESULTS: Clinical periodontal parameters were not changed, and they were not different between the groups from T0 to T1. T forsythensis and F nucleatum increased during the treatment period in both groups (P <0.05). The majority of the bacteria were T nucleatum at T0 and T1 in both groups. Changes in bacterial load from T0 to T1 were not different between groups except for T forsythensis and F nucleatum (P <0.05). CONCLUSIONS: The use of an antibacterial monomer did not have an additional positive effect on clinical periodontal parameters. When used in bonding orthodontic brackets, the antibacterial monomer failed to reduce periodontopathogenic bacteria when compared with the conventional adhesive system during a 6-month treatment period.

Ozone therapy prevents renal inflammation and fibrosis in a rat model of acute pyelonephritis.

INTRODUCTION: Not only bacterial characteristics but also oxidative/nitrosative stress could play a significant role in renal parenchymal inflammatory processes in acute pyelonephritis (APN). This study was conducted to evaluate the effect of ozone therapy (OT), as an immunomodulator and antioxidant, on the renal function, morphology and biochemical parameters of oxidative stress in an experimental model of APN in rats. MATERIALS AND METHODS: Forty rats were divided equally into five groups as control, APN, APN + Antibiotic, APN + OT, and APN + Antibiotic + OT. APN was induced by 0.1 ml of freshly prepared Escherichia coli (ATCC 25922) solution containing 10(10) colony-forming unit/ml into the kidney. A control group was administered 0.1 ml of 0.9 % NaCl solution. Treatment was begun 72 h after bacterial inoculation. Control and APN groups were given 0.9% NaCl solution, APN + Antibiotic and APN + OT were given either antibiotic (ciprofloxacine 150 mg/kg intramuscular/twice daily) or OT. APN + Antibiotic + OT group was given both antibiotic and OT for five consecutive days. At the end of the seventh day, animals were killed via decapitation and trunk blood was collected. Both kidneys were harvested and one half of each kidney were immediately stored for antioxidant enzyme activity, tissue lipid peroxidation and protein carbonyl content. The remainder was fixed for histopathologic examination. RESULTS: E. coli-induced APN increased the renal glomerular and tubular dysfunction, oxidative stress parameters and antioxidant enzyme activities. Either antibiotherapy or OT markedly ameliorated renal dysfunction, the antioxidant status of the kidneys and histopathological injuries subjected to E. coli-induced APN. Interestingly, the combination of antibiotherapy and OT was much more effective than either of the treatment modalities alone. CONCLUSION: The combination of antibiotherapy and OT markedly ameliorated renal dysfunction and improved antioxidant status and histopathologic modalities in rats subjected to E. coli-induced APN than either antibiotherapy or OT treatment alone. Therefore, OT may be considered as an adjuvant therapy to classical antibiotherapy to prevent renal inflammation and fibrosis in APN.

Identification and characterization of OXA-48 producing, carbapenem-resistant Enterobacteriaceae isolates in Turkey.

Carbapenem resistance in Enterobacteriaceae isolates has been reported from Turkey and is most often mediated by OXA-48 type carbapenemases. We report the identification and characterization of four carbapenem-resistant isolates (three Klebsiella pneumoniae and one Escherichia coli) among 515 clinical Enterobacteriaceae isolates collected during a 7-month study period in Ankara, Turkey. The four isolates were recovered from blood and urine specimens in patients with varied clinical manifestations. They had distinct pulsed-field gel electrophoresis patterns and harbored a variety of ß-lactamases including bla(TEM-1), bla(SHV-12) genes, bla(SHV-11), and/or bla(CTX-M-15). PCR and sequencing analysis revealed that the bla(OXA-48) gene was present in all four isolates. Our data indicated that the OXA-48-type carbapenemase was the only mechanism for carbapenem resistance in our hospital.

Simultaneous detection and differentiation of pathogenic and nonpathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay.

Leptospirosis, caused by pathogenic Leptospira, is one of the most important zoonoses in the world. Several molecular techniques have been developed for detection and differentiation between pathogenic and saprophytic Leptospira spp. The aim of this study was to develop a rapid and simple assay for specific detection and differentiation of pathogenic Leptospira spp. by multiplex real-time PCR (TaqMan) assay using primers and probes targeting Leptospira genus specific 16S ribosomal RNA gene, the pathogen specific lig A/B genes and nonpathogen Leptospira biflexa specific 23S ribosomal RNA gene. Sixteen reference strains of Leptospira spp. including pathogenic and nonpathogenic and ten other negative control bacterial strains were used in the study. While the 16S primers amplified target from both pathogenic and non-pathogenic leptospires, the ligA/B and the 23S primers amplified target DNA from pathogenic and non-pathogenic leptospires, respectively. The multiplex real-time PCR (TaqMan) assay detection limit, that is, the sensitivity was found approximately 1 x 10(2) cells/ml for ligA/B gene and 23S ribosomal RNA gene, and 10 cells/ml 16S ribosomal RNA. The reaction efficiencies were 83-105% with decision coefficients of more than 0.99 in all multiplex assays. The multiplex real-time PCR (TaqMan) assay yielded negative results with the ten other control bacteria. In conclusion, the developed multiplex real-time PCR (TaqMan) assay is highly useful for early diagnosis and differentiation between pathogenic and non-pathogenic leptospires in a reaction tube as having high sensitivity and specificity.

[Expression of hepatitis B virus core antigen gene region in yeast cell]. / Hepatit B virusu kor antijeni gen bölgesinin mayada ekspresyonu.

In this study, the core antigen (HBcAg) gene region of hepatitis B virus (HBV) was transformed and expressed into an eukaryotic expression vector by recombinant DNA technology in order to obtain the protein used in anti-HBc tests which is being one of the most important marker for the serodiagnosis of HBV infections. For this purpose, HBV-DNA positive patient sera were used as the source of viral nucleic acids, and the primers coding HBcAg gene region have been designed. After the amplification of HBcAg gene region by polymerase chain reaction (PCR), the amplicons purified by Invisorb Spin Rapid PCR Kit" (Invitek, Germany), were cloned to pYES2.1 plasmid via the TOPO TA expression kit (Invitrogen, USA) and this plasmid was transformed to competent bacteria (TOPO 10F' Escherichia coli) by CaCl2 method. After competent bacteria were grown on LB (Lysogeny Broth) agar media supplemented with ampicillin, the plasmid "pYES2.1 + HBcAg" were isolated and transformed to Saccaromyces cerevisiae via the "S.c. EasyComp Transformation Kit" (Invitrogen, USA). Finally, the expression of HBcAg by the yeast was confirmed with the use of in house ELISA method. Since the diagnostic kits used in our country for hepatitis B serology are usually imported products, this creates a great economical burden. Thus, the experience and knowledge that builds up following such studies will help to produce our own diagnostic products using our equity.