Further expanding the phenotype of anti-Ku antibody associated disease in children and adolescents.

Anti-Ku autoantibodies are associated with several autoimmune inflammatory diseases. We aimed to review our anti-Ku positive pediatric patients in this study. Four pediatric patients (all female) who had anti-Ku positivity were included (Patients 1-2-3 with idiopathic inflammatory myopathy (IIM); Patient 4 with chronic urticaria). Patient 1 (onset:10.5 years) had proximal muscle weakness, Raynaud phenomenon, sclerodactyly, hyperpigmentation, joint contracture, and tenosynovitis. The disease course was progressive despite treatment with corticosteroids, intravenous immunoglobulin (IVIG), plasma exchange, and 11 different immunosuppressive drugs. Patient 2 (onset:15 years) presented with proximal muscle weakness, fatigue, weight loss. She recovered normal muscle strength after treatment with corticosteroids, IVIG, methotrexate, cyclosporine A, mycophenolate mofetil. Patient 3 (onset:10 years) had juvenile dermatomyositis with proximal muscle weakness, Gottron's papules, and calcinosis. She also had anti-NXP2 positivity. Remission was achieved with corticosteroids, methotrexate, azathioprine, and infliximab. Muscle biopsy findings revealed a variable spectrum of necrosis, regeneration, perifascicular pattern, and inflammation. Patient 4 had only chronic urticaria (onset: 6.5 years). The striking features of this series were heterogeneity in clinical presentations including solely chronic urticaria and IIM; variable response to immunosuppressive treatments; and histopathology revealing a spectrum of necrosis, regeneration and inflammatory infiltration. Expanding the spectrum of anti-Ku positivity will allow better understanding of anti-Ku-associated phenotype clusters.

[The Importance of Gram-stained Smears in the Diagnosis of Nocardia Infections]. / Nocardia Enfeksiyonlarinin Tanisinda Gram Boyanmis Yaymalarin Önemi.

Nocardia species are low virulence bacteria found in nature. They can be an infectious agent, especially in patients with risk factors such as underlying immunosuppression, chronic lung disease, and malignancy. They can be easily overlooked because they are not seen frequently and has no pathognomonic symptoms. With this study, it was aimed to draw attention to the importance of microscopic examination of Gram-stained smears in the diagnosis of Nocardia infections in routine microbiology laboratories. Cases in which Nocardia spp. were detected in their clinical samples between November 2014-December 2015 in Hacettepe University Medical Faculty Hospital were included in the study. In the direct microscopic examination of Gram-stained smears of the samples arriving to the laboratory, the incubation periods of the cultures of the samples compatible with Nocardia spp. were extended. Then relevant colonies were identified by conventional microbiological methods and also by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS, bioMerieux, France) automated system. Species-level identification of Nocardia isolates was performed by 16S rRNA gene sequence analysis. To demonstrate the genetic relationship between Nocardia isolates, pulsed-field gel electrophoresis (PFGE) was performed. In vitro susceptibility of the isolates against amoxicillin-clavulanate (AMC), linezolid, moxifloxacin, trimethoprim-sulfamethoxazole (TMP-SXT), amikacin, imipenem, clarithromycin, cefepime, cefotaxime, ceftriaxone, and ciprofloxacin was determined using the gradient strip method (E-test). A total of 19 Nocardia spp. strains were isolated from eight patients. Four cases exhibited repeated growth of Nocardia spp. up to a period of nine months. The most frequently isolated species was N.cyriacigeorgica, which was identified in four cases. Other species isolated from patients were N.asteroides, N.transvalensis, N.farcinicia, and N.asiatica/arthritidis. When the results obtained with DNA sequence analysis and MALDI-TOF MS were compared, 16 (84.2%) of 19 isolates were correctly identified to the genus level and 9 (47.4%) to the species level with MALDI-TOF MS, while three (15.8%) isolates could not be identified, and seven (36.8%) isolates were misidentified. According to the PFGE results, it was determined that the strains isolated from the same patient were genetically identical. All isolates were susceptible to amikacin, cefepime, cefotaxime, ceftriaxone, imipenem, linezolid, and except one isolate to TMP-SXT. Among the study isolates, the most common resistance was against ciprofloxacin (62.5%), followed by clarithromycin (37.5%). N.cyriacigeorgica was determined as the most frequently detected and the most resistant species to antibiotics in the study population. Direct microscopic examination of clinical specimens is one of the most valuable methods for the identification of Nocardia-type bacteria, which is difficult to isolate in microbiology laboratories. With this study, the importance of examining Gram-stained clinical samples was emphasized in the identification of Nocardia species, which can emerge with a wide variety of clinical forms and can be easily overlooked. In addition, antibiotic susceptibility profiles of the isolated bacteria were determinedto contribute to species-specific susceptibility profiles. Accurate identification of Nocardia species will contribute to clinical and epidemiological studies.

Association between early chronic methicillin-susceptible Staphylococcus aureus colonization and lung function in children with cystic fibrosis.

BACKGROUND: Our aim was to determine whether early chronic methicillin-susceptible Staphylococcus aureus (MSSA) colonization in children with cystic fibrosis (CF) is associated at 8 years of age with poorer lung function, poorer nutritional status, and increased exacerbation frequency. METHODS: In this retrospective cohort study, a total of 52 children with chronic MSSA colonization were included. Of them, 26 were chronically colonized with MSSA before the age of 4 years (early onset), and 26 were chronically colonized from 4 years to 6 years of age (late-onset). At the age of 8 years, lung function, body mass index (BMI) as an indicator of nutritional status and frequency of pulmonary exacerbations were compared between two groups. RESULTS: At 8 years of age, BMI was similar between the early-onset and late-onset groups (15.0 [min-max: 12.9-26.8] vs. 15.7 (min-max: 13.0-24.9), p = 0.327]. Percentage of forced expiratory volume in 1 s (FEV1) and forced expiratory flow between 25% and 75% of vital capacity (FEF25-75) in 8-year-old children were significantly lower in the early onset group compared to the late-onset group (98 [min-max: 44-139] vs. 106.5 [min-max: 82-131], p = 0.047; 84.9 ± 25.5 vs. -102.3 ± 26.3, respectively; p = 0.019], although the percentage of forced vital capacity did not differ significantly between two groups (93.5 [min-max: 45-131] vs. 103 [min-max: 84-119], respectively; p = 0.092). Exacerbation frequency between the ages of 6 and 8 years in the early onset group was higher compared to the late-onset group according to the Poisson regression model [1 (min-max:0-10) vs 0 (min-max:0-4), respectively; p = 0.044]. CONCLUSIONS: Early chronic MSSA colonization is associated with poorer lung function and frequent exacerbations in children with CF. However, further studies are needed to reveal the cause-and-effect relationship between early chronic MSSA colonization and pulmonary outcome.

Vancomycin Containing PDLLA and PLGA/ß-TCP Inhibit Biofilm Formation but Do Not Stimulate Osteogenic Transformation of Human Mesenchymal Stem Cells.

Aims: Chronic osteomyelitis, including implant-related prosthetic joint infection, is extremely difficult to cure. We develop vancomycin containing release systems from poly(d,l-lactide) (PDLLA) and poly(d,l-lactide-co-glycolide) (PLGA) composites with beta-tricalcium phosphate (ß-TCP) to treat methicillin-resistant Staphylococcus aureus osteomyelitis. We ask whether vancomycin containing PDLLA/ß-TCP and PLGA/ß-TCP composites will prevent early biofilm formation, allow cell proliferation and osteogenic differentiation, and stimulate osteogenic signaling molecules in the absence of an osteogenic medium. Methods: Composites were produced and characterized with scanning electron microscopy. In vitro vancomycin release was assessed for 6 weeks. Biofilm prevention was calculated by crystal violet staining. Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) and osteosarcoma cell (SaOS-2) proliferation and differentiation were assessed with water soluble tetrazolium salt and alkaline phosphatase (ALP) staining. Real-time quantitative polymerase chain reaction defined osteogenic signaling molecules for hBM-MSCs. Results: Totally, 3.1 ± 0.2 mg and 3.4 ± 0.4 mg vancomycin released from PDLLA/ß-TCP and the PLGA/ß-TCP composites, respectively, and inhibited early biofilm formation. hBM-MSCs and SaOS-2 cells proliferated on the composites and stimulated ALP activity of cells. Runt-related transcription factor 2 (RUNX2) and SRY-Box transcription Factor 9 (SOX9) expressions were, however, lower with composites when compared with control. Conclusion: Vancomycin containing PDLLA/ß-TCP and PLGA/ß-TCP composites inhibited early biofilm formation and proliferated and differentiated hBM-MSCs and SaOS-2 cells, but osteogenesis-related RUNX2 and SOX9 transcription factors were not strongly expressed in the absence of an osteogenic medium for 14 days.

Correlation of Treponemal Chemiluminescent Microparticle Immunoassay Screening Test Signal Strength Values With Reactivity of Confirmatory Testing.

BACKGROUND: Automated chemiluminescent microparticle immunoassays (CMIAs) are the most common first step at high-volume laboratories for syphilis screening. If the initial screening test is reactive, 1 more treponemal test is required, resulting in increased cost. In this multicenter study, we aimed to determine the correlation between the CMIA signal-to-cutoff ratio (S/Co) and the confirmatory tests to reduce unnecessary confirmatory testing. METHODS: Eight hospitals from 5 provinces participated in this study. All laboratories used Architect Syphilis TP CMIA (Abbott Diagnostics, Abbott Park, IL) for initial screening. Treponema pallidum hemagglutination (TPHA), rapid plasma reagin (RPR), and fluorescent treponemal antibody absorption (FTA-ABS) were used as confirmatory tests according to the reverse or European Centre for Disease Prevention and Control algorithms. A receiver operating characteristic analysis was used to determine the optimal S/Co ratio to predict the confirmation results. RESULTS: We evaluated 129,346 serum samples screened by CMIA between January 2018 and December 2020. A total of 2468 samples were reactive; 2247 (91%) of them were confirmed to be positive and 221 (9%) were negative. Of the 2468 reactive specimens, 1747 (70.8%) had an S/Co ratio ≥10.4. When the S/Co ratios were ≥7.2 and ≥10.4, the specificity values were determined to be 95% and 100%, respectively. In a subgroup of 75 CMIA-positive patients, FTA-ABS was performed and 62 were positive. Among these FTA-ABS-positive patients, 24 had an S/Co ratio <10.4, and negative TPHA and RPR. CONCLUSIONS: We propose a potentially cost-effective reverse screening algorithm with a treponemal CMIA S/Co ratio ≥10.4, obviating the need for secondary treponemal testing in about 71% of the screening-reactive samples. This would substantially reduce the confirmatory testing volume and laboratory expenses. However, in high-risk group patients with CMIA positive results, S/Co ratio <10.4, and negative TPHA and RPR, FTA-ABS may be used for confirmation.

Effectiveness of different eradication treatment protocols for new-onset Pseudomonas aeruginosa in children with cystic fibrosis.

OBJECTIVES: While eradicating new-onset Pseudomonas aeruginosa in children with cystic fibrosis is an important issue, there is no clear evidence about the best treatment approach. This retrospective observational cohort study aims to compare the effectiveness of intravenous therapy versus inhalation with/without oral therapy in the eradication of new-onset P. aeruginosa, determine the factors affecting the treatment success and assess lung function at baseline and posttreatment. METHODS: Of 399 children, 110 (140 episodes) with either the first P. aeruginosa isolation or a new isolation after at least 1 year free of infection were included. Different eradication regimens (intravenous therapy or inhaled tobramycin or inhaled tobramycin plus oral ciprofloxacin) were compared. Eradication success was accepted as remaining free of infection with a negative culture for 12 months. Demographic, clinical, and microbiological characteristics of children, effectiveness of different eradication strategies, time to a new P. aeruginosa isolation, and the relationship between lung function and the type of eradication regimen were determined. RESULTS: Of 140 episodes, intravenous therapy was administered in 53 and inhalation therapy (in combination with or without oral ciprofloxacin) in 87. Total success rate of eradication was 60.7%. Eradication was achieved in 56.6% of children with intravenous therapy, 59.7% with inhaled tobramycin therapy, and 72% with inhaled tobramycin plus oral ciprofloxacin therapy. Success rates of different eradication regimens did not differ significantly (p = 0.419). Lung function by the end of the first year was worse in the intravenous group compared to the inhalation group (p = 0.017 for forced expiratory volume in 1 s, p = 0.015 for forced vital capacity). CONCLUSION: No advantage of intravenous therapy was demonstrated compared to inhalation therapy in terms of eradication success.

Comparison of serum anti-neuronal antibody levels in patients having autism spectrum disorder with and without regression.

BACKGROUND: Autism Spectrum Disorders (ASD) may present with a delay in social and communication development, or less frequently, with regression in social and language skills. The reasons for this difference in clinical presentation are unknown, and the regressive symptoms in the second group suggest an acquired process. METHODS: We investigated serum autoantibodies in these two types of ASD in a cross-sectional design in a total of 50 children, 24 with autistic regression and 26 with classical ASD according to the DSM-5 criteria. Clinical assessment by the Childhood Autism Rating Scale (CARS) and Ankara Developmental Screening Test (ADST), parental questionnaires consisting of the Aberrant Behavior Checklist (ABC) and Autism Behavior Checklist (AuBC) were completed. Serum samples were tested for anti-neuronal antibodies including anti-N-methyl-Daspartate receptor (NMDAR), anti-contactin-associated protein (CASPR2), anti-leucine rich glioma inactivated 1 (LG1), anti-glutamate type 2-amino-3-propionic Acid (AMPA) 1-2, anti-gamma amino butyric acid (GABA) B, anti-dipeptidyl aminopeptidase-like protein 6 (DPPX) and anti-glutamic acid decarboxylase 65(GAD). RESULTS: Serum anti-GAD antibodies were at detectable levels in five (20.8%) patients with autistic regression, of whom three had 2 to 4-fold increased titers, and in none of the patients with classical ASD. The age of the father at the patient`s birth and the duration of autistic regression correlated with anti-GAD IgG levels (P: 0,045, P: 0.855 respectively) in the ASD-regression group. No other antibodies were detected in either group. CONCLUSIONS: Our results do not suggest a causative role of anti-neuronal antibodies, but the possibility of an autoimmune process accompanying regressive symptoms in a small subgroup of ASD.

Diverse clinical characteristics of Aspergillus growth in patients with cystic fibrosis.

BACKGROUND AND OBJECTIVES: Patients with cystic fibrosis (CF) have a varying spectrum of clinically significant Aspergillus disease in addition to allergic bronchopulmonary aspergillosis (ABPA). Here we aimed to review the different clinical phenotypes related with Aspergillus growth on the airway culture of patients with CF, we also aimed to investigate the effect of Aspergillus growth on lung function tests. METHOD: The medical records of 100 patients with CF who had Aspergillus growth on airway culture within the period of April 2001 and June 2016 were retrospectively analyzed. Age, gender, symptoms, physical examination findings, pulmonary function tests, the diagnosis of ABPA, and airway culture results were recorded for every visit. Patients with Aspergillus growth on airway cultures were classified into different groups as ABPA, Aspergillus sensitization, Aspergillus colonization and Aspergillus bronchitis. RESULTS: Medical records of 83 patients and 147 sputum cultures were attained from 100 patients. The mean age of the patients was 17.6±7.6 years and the mean age of the first Aspergillus growth in sputum culture was 12.5±6.7 years. At first isolation, Aspergillus fumigatus SC was the most common Aspergillus SC in sputum (76.3%) and 14.5% of these patients required hospitalization. Aspergillus sensitization was diagnosed in 3.6% (n= 3) of the patients. Aspergillus colonization was diagnosed in 18.1% (n= 15) of all patients and led to a decline in FEV1%, FVC% and FEF25-75% which was not statistically significant, furthermore. ABPA was detected in 9.6% (n= 8) of all patients and led to a statistically significant decline in FEV1% (p= 0.02); nonsignificant decline in FVC% and FEF25-75%. Aspergillus bronchitis was detected in 43.4% (n= 36) of all patients and led to nonsignificant decline in FEV1%, FVC% and FEF25-75%. CONCLUSION: ABPA is recognized as the most common Aspergillus associated disorder in CF patients and is related to deteriorated pulmonary function tests; however Aspergillus colonization and bronchitis may also be associated with worsening lung function.

RELATIONSHIP BETWEEN STAGES OF DIABETIC RETINOPATHY AND LEVELS OF BRAIN-DERIVED NEUROTROPHIC FACTOR IN AQUEOUS HUMOR AND SERUM.

PURPOSE: The aim of the study was to determine aqueous humor and serum levels of brain-derived neurotrophic factor (BDNF) in diabetic patients with and without retinopathy. METHODS: The study included diabetic patients with or without retinopathy, who had an indication for cataract surgery. The study groups were diabetic patients without retinopathy (Group 2), with nonproliferative diabetic retinopathy (Group 3), and with proliferative retinopathy (Group 4). To quantitatively determine the amount of BDNF in samples, the RayBio Human BDNF ELISA kit (Norcross, GA), based on an enzyme-labeled immunosorbent assay was used. RESULTS: The median serum BDNF levels were significantly lower in all the study groups than in the control group (P values: 0.038 Group 2, 0.02 Group 3, and 0.002 Group 4). Serum BDNF was lower in Group 4 than in Group 3 (P = 0.030), and in Group 3 than in Group 2 (P = 0.04). The median aqueous humor BDNF levels were significantly decreased in all groups (P values: 0.047 Group 2, 0.021 Group 3, and 0.007 Group 4). There was no significant difference between Groups 2, 3, and 4 (P = 0.214). CONCLUSION: The serum and aqueous humor BDNF levels decreased in patients with diabetes mellitus (DM) before the emergence of clinical signs of retinopathy.

[Investigation of role of anaerobic bacteria in cystic fibrosis patients]. / Kistik fibrozisli hastalarda anaerop bakterilerin rolünün arastirilmasi.

INTRODUCTION: Repetitive pulmonary infections are the main cause of morbidity and mortality in cystic fibrosis (CF) patients. In recent years, non-culture dependent metagenomic studies showed complex dynamics of the pulmonary environment of CF patients and pointed out the importance of anaerobic bacteria. Molecular-based studies indicate that anaerobic bacteria can be found more than aerobic or facultative anaerobic bacteria in CF lung environment. However, limited number of studies are far away to clarify the importance of anaerobic bacteria in CF pulmonary disease. MATERIALS AND METHODS: The aim of this study was to evaluate the role of anaerobic bacteria in CF patients admitted to Hacettepe University, Pediatric Respiratory Diseases Department, by using quantitative culture method for both aerobic and anaerobic bacteria. Anaerobic bacteria were identified by conventional and semi-automated methods. Antibiotic susceptibilities were performed by agar dilution method. RESULT: Seventy-seven anaerobic bacteria were isolated from 35 (81.4%) of 43 patients. The total count of anaerobes and facultative bacteria (mean 16 x 106), was higher than aerobes and facultative bacteria (mean 14.1 x 106). If anaerobe culture were not performed merely 63.65% of all species could be obtained. In patients whose samples yielded intermediate or high numbers of PMNLs, significantly more obligate anaerobic bacteria were isolated (p= 0.046). Patients older than 18 years were colonized with higher number of anaerobic bacteria. Susceptibilities of 72 isolates out of 77, against ampicillin, sulbactam-ampicillin, piperacillin, piperacillin-tazobactam, moxifloxacin, metronidazole, imipenem, and clindamycin were also evaluated. Clindamycin was found to be the least effective antibiotic among all. None of the isolates was resistant to imipenem. CONCLUSIONS: This is the first study to show the role and importance of anaerobic bacteria in CF patients in our country. The resistance rates in anaerobic bacteria isolated from CF patients is concerning. Therefore, intermittent anaerobic culture and follow-up of resistance rates will be helpful in the follow-up of these patients.

[Multilocus sequence analysis, biofilm production, antibiotic susceptibility and synergy tests of Burkholderia species in patients with and without cystic fibrosis]. / Kistik fibrozisli ve kistik fibrozisi olmayan hasta grubunda Burkholderia türlerinin multilokus sekans analizi, biyofilm olusturma, antibiyotik duyarlilik ve sinerji testleri.

Burkholderia spp. emerged as important pathogens in the airways of immunocompromised humans, especially those with cystic fibrosis (CF). Failure of identification with conventional techniques, high intrinsic resistance to most antibiotics and biofilm formation can cause difficulties in the treatment of these infections. The aim of this study was to identify Burkholderia spp. strains isolated from CF and non-CF patients with with routine microbiological methods, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and multilocus sequence analysis (MLSA), to determine of the antibiotic susceptibility and synergies, and to evaluate biofilm formation of these isolates. A total of 38 Burkholderia spp. (25 CF, 13 non-CF) from 26 patients were identified by biochemical, phenotypical and matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and sequence types were revealed by multilocus sequence analysis (MLSA). Sequence types of isolates were identified using the PubMLST database. Characteristics of biofilm formation of clinical isolates were evaluated by microplate method. Antibiotic susceptibilities of ceftazidime, meropenem, trimethoprim-sulfamethoxazole (TMP-SXT) and levofloxacin were determined by broth microdilution method according to CLSI (2017) guidelines. Synergy tests were performed by checkerboard method. Clinical isolates were identified as Burkholderia cenocepacia (n= 16), Burkholderia contaminans (n= 11), Burkholderia gladioli (n= 4), Burkholderia dolosa (n= 4), Burkholderia multivorans (n= 2) and Burkholderia seminalis (n= 1). Sequence types of these isolates were determined as ST19, ST72, ST102, ST180, ST482, ST602, ST629, ST740, ST839 and ST1392. The correct identification at the species-level with MALDI-TOF MS was 94-100% for all isolates except B.contaminans. Biofilm formation among the identified species in the study was determined as 53% (n= 20). There was no statistical difference when the biofilm production was evaluated separately among Burkholderia species and biofilm production rates between CF (56%, 14/25) and non-CF (46%, 6/13) Burkholderia isolates (p> 0.05). Overall rates of resistance to ceftazidime, meropenem, TMP-SXT, and levofloxacin of the isolates were 35%, 66%, 50% and 40%, respectively. The antibiotic resistance against Burkholderia spp., isolates obtained from CF patients were more susceptible to ceftazidime, but no significant difference was found for other antibiotics. Synergy was determined between meropenem and TMP-SXT in two isolates. Antagonism was detected in 15 isolates, 12 of them were between meropenem and ceftazidime, three of them were between ceftazidime and TMP-SXT. Numerous resistance mechanisms may lead to higher resistance in this bacteria, whereas the antagonism between meropenem and ceftazidime in this study might be attributed to the expression of beta-lactamases. In this study, the distinctness of sequence types between Burkholderia spp. isolated from CF and non-CF patient, provided a better understanding about the importance of biofilm formation for the infections with these bacteria and emphasized that the management of therapy should be driven by the antibiotic test results.

The effect of trabeculectomy on serum brain-derived neurotrophic factor levels in primary open-angle glaucoma.

PURPOSE: We aimed to investigate the effect of trabeculectomy on serum brain-derived neurotrophic factor (BDNF) levels. Our secondary goal was to compare serum and aqueous humor (AH) BDNF levels in primary open-angle glaucoma (POAG) and control subjects. METHODS: This prospective, cross-sectional study consists of 20 eyes of with advanced-stage POAG who had trabeculectomy and 19 eyes of age- and sex-matched control healthy subjects who had cataract surgery. Serum and AH samples were obtained preoperatively in trabeculectomy group and control subjects. Serum samples were obtained at the third postoperative month in both groups. RESULTS: The aqueous humor and serum levels of BDNF at the surgery day were found to be strongly positive correlated (r = 0.868; p < 0.001). Serum and AH BDNF levels of POAG cases were significantly lower than control subjects at the surgery day (respectively p = 0.038, p = 0.011). In POAG cases, serum BDNF levels significantly increased at the third month after trabeculectomy while there was not a significant difference in control subjects with cataract surgery (p < 0.001; p = 0.717 respectively). CONCLUSION: Trabeculectomy was found to have a positive effect on serum BDNF levels in POAG cases.

Clinical Strains of Chryseobacterium and Elizabethkingia spp. Isolated from Pediatric Patients in a University Hospital: Performance of MALDI-TOF MS-Based Identification, Antimicrobial Susceptibilities, and Baseline Patient Characteristics.

Our objective was to evaluate the performance of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for identification of the Chryseobacterium and Elizabethkingia spp. isolated from pediatric patients at Hacettepe University Hospital using 16S rRNA gene sequencing as the gold standard and to determine the antimicrobial susceptibility patterns of the isolates and baseline characteristics of patients. All stored Chryseobacterium and Elizabethkingia spp. isolated from various clinical specimens (sputum, blood, and urine) of pediatric patients at Hacettepe University Hospital between 2012 and 2016 were included in this study. Minimum inhibitory concentrations of 10 antimicrobial agents were determined by Etest for all isolates. To determine the baseline characteristics of patients, medical records of all patients were retrospectively reviewed. In total, 18 isolates of Chryseobacterium spp. (16 C. indologenes, 2 C. gleum) and 5 isolates of Elizabethkingia spp. (3 E. meningoseptica, 2 E. anophelis) were identified by 16S rRNA sequencing. MALDI-TOF MS correctly identified 19 (82.6%) isolates to the species level. The quinolones (ciprofloxacin and levofloxacin), trimethoprim/sulfamethoxazole and piperacillin/tazobactam showed the highest spectrum of activity against the overall collection of isolates. Cystic fibrosis (CF) was the underlying disease in 81.8% of patients. To our knowledge, this study includes the largest number of Chryseobacterium spp. isolated from clinical specimens of pediatric patients in Turkey. In this study, we also report the first clinical isolate of E. anophelis in Turkey. Since, the majority of strains were isolated from patients with CF; larger, prospective clinical studies are needed to establish whether chryseobacteria could be considered as an emerging opportunistic pathogen in patients with CF.

[Accurate diagnosis of Pseudomonas luteola in routine microbiology laboratory: on the occasion of two isolates]. / Rutin mikrobiyoloji laboratuvarinda Pseudomonas luteola'nin dogru tanisi: Iki sus nedeniyle.

Pseudomonas luteola which was previously known as Chryseomonas luteola; is a gram-negative, non-fermentative, aerobic, motile, non-spore-forming bacillus. It is frequently found as a saprophyte in soil, water and other damp environments and is an opportunistic pathogen in patients with underlying medical disorders or with indwelling catheters. It has been reported as an uncommon cause of bacteremia, sepsis, septic arthritis, meningitis, endocarditis, and peritonitis. Thus, early and accurate identification of this rare species is important for the treatment and also to provide information about the epidemiology of P.luteola infections. This report was aimed to draw attention to the accurate identification of P.luteola in clinical samples, upon the isolation and identification in two cases in the medical microbiology laboratory of a university hospital. In February 2011, a 66-year-old man, with chronic obstructive pulmonary disease, coronary artery disease and aplastic anemia, was admitted to our hospital due to progressive dyspnea. A chest tube was inserted on the 20th day of admission by the reason of recurrent pleural effusion. Staphylococcus aureus and a non-fermentative gram-negative bacillus (NFGNB) with wrinkled, sticky yellow colonies were isolated from the pleural fluid sample obtained on the 9th day following the insertion of the chest tube. In February 2012, a 7-year-old male cystic fibrosis patient who had no signs and symptoms of acute pulmonary exacerbation was admitted to the hospital for a routine control. This patient had chronic colonization with Pseudomonas aeruginosa and S.aureus and his sputum sample obtained at this visit revealed isolation of P.aeruginosa, S.aureus, Aspergillus fumigatus and a wrinkled, sticky yellow NFGNB. Both of these NFGNB were identified as P.luteola by the Phoenix automated microbial identification system (BD Diagnostics, USA). To evaluate the microbiological characteristics of these two isolates, the strains were further analysed by VITEK MS (bioMerieux, France) and Microflex LT mass spectrometer (Bruker Daltonics, Germany). Both of the MALDI-TOF-MS systems identified the isolates as P.luteola and 16S rRNA gene sequencing (ABI PRISM 3100, Applied Biosystems, USA) also confirmed the identification. The strains had wrinkled, sticky yellow colonies which were oxidase-negative, catalase-positive and non-fermentative. The Gram stained smears of the colonies revealed clusters of gram-negative bacilli probably embedded into a biofilm matrix. Since there are no accepted standards for testing the antibiotic susceptibility of P.luteola strains, the standards determined by CLSI for "other non-Enterobacteriaceae" (non-fermentative bacteria excluding P.aeruginosa, Acinetobacter spp., Burkholderia cepacia, B.mallei, B.pseudomallei and Stenotrophomonas maltophilia) were used for the susceptibility testing. Gradient MIC method (E-Test, bioMerieux, France) revealed that the isolates were susceptible to gentamicin, piperacillin-tazobactam, ceftazidime, cefepime, meropenem, colistin and levofloxacin. Accurate and prompt identification of P.luteola which is identified as a rare pathogen in serious cases is of critical importance since it has been suggested that this organism is likely to become more frequent as a nosocomial pathogen since the interventional processes increase in current medical practice. This report supported that Phoenix automated phenotypic identification system (BD Diagnostics, USA) and the two MALDI-TOF-MS based systems (VITEK MS and Bruker Microflex LT mass spectrometer) were successfull in the accurate identification of P.luteola.

Alzheimer disease, inflammation, and novel inflammatory marker: resistin.

BACKGROUND/AIM: Inflammation may play an important role in Alzheimer disease (AD) pathogenesis. A growing amount of evidence indicates that resistin has hallmark regulatory functions such as inflammatory states. The aim of this study was to determine whether plasma resistin levels would be useful in the diagnosis of patients with AD and to investigate the relationships between resistin and other inflammatory markers such as hs-CRP and TNF-α. Materials and methods: In this cross-sectional study, 38 AD patients and 32 control subjects with normal cognitive function aged 65 years and over were included. The diagnosis of AD was made according to DSM-IV and NINCDS-ADRDA criteria. Serum levels of resistin were measured with an enzyme-linked immunosorbent assay method using the human resistin E50 kit. Results: The median resistin level of AD patients was significantly higher than in the control group (86.3 vs. 70.8 pg/mL, P = 0.002). Overall accuracy of resistin in determining AD was 70.66%, with sensitivity, specificity, PPV, and NPV of 75.0%, 65.5%, 73.0%, and 67.9%, respectively. There was no statistically significant difference between AD patients and control subjects with respect to hs-CRP and TNF-α levels. Conclusion: Resistin levels may be considered as a predictor of AD and it may predict activation of the immune system in AD pathophysiology.

Screening for latent tuberculosis infection in psoriasis and psoriatic arthritis patients in a tuberculosis-endemic country: a comparison of the QuantiFERON®-TB Gold In-Tube test and tuberculin skin test.

Since the introduction of biologic therapies for tuberculosis (TB), screening for latent TB infection has increased in importance, especially in countries in which TB is endemic. The aim of this study was to evaluate the effect of psoriasis on tuberculin skin test (TST) results and to compare two TB screening tests, the TST and QuantiFERON(®)-TB Gold In-Tube (QFT-GIT) test, in psoriasis and psoriatic arthritis (PA) patients living in a TB-endemic country (Turkey). This prospective study included 61 psoriasis and 40 PA patients, and 58 healthy controls. Demographic data, medical history, human immunodeficiency virus (HIV) status, level of education, smoking status, exposure to TB, personal and family histories of TB, and bacillus Calmette-Guérin (BCG) vaccination status were recorded for all participants. The TST and QFT-GIT were performed in all participants. The mean ± standard deviation TST indurations in the patient and control groups were 12.6 ± 6.4 mm and 10.2 ± 6.5 mm, respectively (P = 0.051). The TST positivity rate was higher in patients than in controls (86.1% vs. 37.9%; P < 0.001), whereas QFT-GIT positivity did not differ significantly (patients: 20.8%; controls: 17.2%; P = 0.737). False positive results can lead to unnecessary prophylactic TB treatment; therefore, the cut-off point for TST positivity in psoriasis and PA patients should be re-evaluated, or other tests, such as the QFT-GIT, should be used.

The role of serum Pseudomonas aeruginosa antibodies in the diagnosis and follow-up of cystic fibrosis.

In cystic fibrosis (CF), if Pseudomonas aeruginosa (Pa) infection is not diagnosed and treated early, chronic colonization occurs, which causes rapid decline in pulmonary functions. The aim of this study was to evaluate Pa antibodies, compare them with Pa cultures and determine their role in early diagnosis and follow-up. Ninety CF patients were included; they were divided into chronic, intermittent, negative, and mucoid groups. They were evaluated every 3-6 months. In each visit, pulmonary function tests and sputum cultures were obtained, and Pa antibodies exotoxin A (ExoA), elastase (ELA) and alkaline protease (AP) were determined in the serum by enzyme-linked immunosorbent assay (ELISA). The most specific test that discriminated chronic colonized patients from noncolonized patients was Pa culture, and the presence of at least one antibody had the highest sensitivity. AP had the highest specificity, and ELA had the highest sensitivity. All antibodies were highest in the mucoid group. ELA was highest in chronic and lowest in the negative group. The presence of antibodies was much higher than positive Pa cultures in patients younger than five years of age. A negative correlation between forced expiratory volume in 1 second (FEV1) and AP was determined only in the mucoid group. In the two-year follow-up, antibody presence did not show a regular pattern. In CF, Pa antibodies can be early markers for diagnosis, especially in young children who cannot expectorate, but they should only be used together with sputum cultures for long-term follow-up and treatment.

The value of fecal calprotectin as a marker of intestinal inflammation in patients with ulcerative colitis.

BACKGROUND/AIMS: To assess intestinal inflammation, simple, inexpensive and objective tools are desirable in inflammatory bowel disease. This study aimed to evaluate fecal calprotectin as a marker of active disease in ulcerative colitis. MATERIALS AND METHODS: Sixty patients with a diagnosis of ulcerative colitis and 20 controls were recruited into the study. The disease activity of ulcerative colitis was determined by modified Truelove-Witts criteria and Rachmilewitz endoscopic index. The enzyme-linked immunosorbent assay was used to measure the concentrations of fecal calprotectin. C-reactive protein, erythrocyte sedimentation rate and hemogram were also measured, and inflammatory markers were compared with fecal calprotectin in determining disease activity. RESULTS: Fecal calprotectin concentration in the patients with active ulcerative colitis (n=30) was significantly higher than that in the inactive ulcerative colitis group (n=30) and in the controls (n=20) (95% confidence interval: 232.5 (0.75-625) vs 11.7 (0.2-625), 7.5 (0.5-512) mg/L, p<0.001). There was no significant difference between the patients with inactive ulcerative colitis and controls (p>0.05). The calprotectin concentration was greater in the patients with a more severe clinical index, higher endoscopic activity (>4), elevated C-reactive protein, leukocytosis, and extensive colitis (p<0.05). The areas under the curve of the receiver operating characteristics were 0.817, 0.809, 0.532, and 0.507 for C-reactive protein, fecal calprotectin, leukocyte count, and erythrocyte sedimentation rate, respectively. There was a significant correlation between the fecal calprotectin concentration and the endoscopic activity in ulcerative colitis (r = 0.548, p<0.001). CONCLUSIONS: Fecal calprotectin is a useful marker in the diagnosis of active disease and evaluation of clinical and endoscopic activity in ulcerative colitis.

Predominance of hospital-associated MRSA among cystic fibrosis patients in a Turkish reference cystic fibrosis centre.

Methicillin-resistant Staphylococcus aureus (MRSA) strains are the major causative agents of numerous hospital- and community-acquired infections. Increasing prevalence of MRSA in cystic fibrosis (CF) populations is reported all over the world. Although there are papers reporting the prevalence and genetic backgrounds of MRSA isolates from different settings in Turkey, there is no information regarding the situation in the CF community. This study was conducted to characterize the MRSA strains recovered from CF patients followed-up at a Turkish reference CF centre. Microbiological testing of isolates was performed via conventional microbiological techniques. Molecular characterization of MRSA isolates was carried out by SCCmec typing by multiplex PCR and PVL gene determination. Among a total of 604 CF patients included in the study, 325 patients were found to harbour S. aureus (53.8%). Of those 325 patients, 24 were positive for MRSA during their follow-up (7.4%). Thirty-two MRSA isolates from these patients were chosen for further assessment of molecular characteristics. Twenty-six MRSA isolates exhibited a pattern like SCCmec type III (81.2%) and six consecutive MRSA isolates of a single patient revealed SCCmec type IV (18.7%). Our findings definitely support the need for further surveillance studies for CF-MRSA strains and highlight the need for infection control measures in the setting of CF centres.

Utility of a commercial quantitative hepatitis C virus core antigen assay in a diagnostic laboratory setting.

In this study, the utility and impact of hepatitis C virus (HCV) core antigen (Cag) detection via a commercial assay have been evaluated in diagnostic laboratory conditions. In a total of 272 samples from 226 individuals, HCV RNA was detected in 81.3% and anti-HCV antibody prevalence was 86.4%. HCV Cag reactivity was identified in 59.9% of the samples and in 75.8% with detectable RNA. The sensitivity and specificity of HCV Cag assay have been calculated as 75.8% and 95.1%, respectively, and agreement between HCV RNA and HCV Cag was moderate (κ = 0.554). HCV Cag and RNA levels were highly correlated (r = 0.915 and 0.937). A viral load threshold of 10(3) IU/mL has been recognized, above which the correlation with RNA became statistically significant and sensitivity increased to 90.9%. Detection and quantification of HCV core antigen have been observed as a strong alternative to nucleic acid testing for HCV monitorization.