Evaluation of Samples with DFS Staining Pattern Detected by Indirect Immunofluorescence Assay.

BACKGROUND: In this study, we aimed to determine the presence of anti DFS70 antibody by a specific IB method in samples showing the DFS pattern and to determine the distribution of DFS pattern in different patient groups. METHODS: 2,401 serum samples, which were received for ANA screening, were tested by IIF method at Akdeniz University Hospital Diagnostic Laboratory. Out of 139 samples with DFS pattern, 75 samples were tested for the presence of anti DFS70 antibody by IB and were included in the study. Patients' clinical diagnoses were obtained retrospectively from medical records. RESULTS: 63 (84%) of 75 samples, which showed DFS patern by IIF, were found to have anti DFS70 antibody by IB. Five of these patients were diagnosed with SARD while the rest (58) had diseases other than SARD. CONCLUSIONS: DFS pattern detected by IIF and isolated anti DFS70 antibody positivity detected by IB show high concordance. However IIF results should be confirmed because of the patterns that can be misidentified as DFS pattern. The presence of anti-DFS70 antibodies, which help to exclude SARD, prevent further unnecessary referral demands.

The role of IL-25 and IL-33 in chronic rhinosinusitis with or without nasal polyps.

The role of IL-25 and IL-33 in the aetiology and pathogenesis of nasal polyps has been controversial in the literature. The objective of the study is to detect serum and tissue levels of IL-25 and IL-33 in patients with (CRSwNP) or without (CRSsNP) nasal polyps using enzyme-linked immunosorbent assay (ELISA). Study group consisted of 20 CRSwNP and 20 CRSsNP patients. Control group comprised of 20 volunteers who had been operated with septum deviation without any additional sinonasal pathology, allergy, systemic disease, or medication use. All groups preoperatively underwent paranasal CT examinations and sinonasal pathologies were recorded based on Lund-Mackay radiological staging system. IL-25 and IL-33 levels in serum and tissue samples were analyzed using the ELISA method. Serum IL-25 and IL-33 levels in CRSsNP, CRSwNP, and control groups did not differ statistically significantly (p = 0.345 and p = 0.338). Any statistically significant difference was not detected in mean tissue IL-25 levels among CRSsNP, CRSwNP, and control groups (p = 0.698). Mean tissue IL-33 level in the CRSwNP group was statistically significantly lower when compared with those of CRSsNP and control groups (p < 0.001 and p < 0.001, respectively). A statistically significant negative correlation was detected between tissue IL-33 levels and Lund-Mackay CT scores (r = -0.436 and p = 0.005). In the present study, we conceivably contributed to scarce number of studies conducted on this issue and we think that further studies will better clarify the role of IL-25 and IL-33 in the development of nasal polyps.

Association of interleukin 18-607A/C and -137C/G polymorphisms with oxidative stress in renal transplant recipients.

Objectives IL-18 mediates various inflammatory and oxidative responses including renal injury, fibrosis, and graft rejection. It has been reported that the promoter -607 and -137 polymorphisms of IL-18 influence the level of IL-18. This prospective observational study investigated the association between oxidative stress with IL-18-607 and -137 polymorphisms in renal transplant recipients. Patients and methods This study included 75 renal transplant recipients (28 female, 47 male) from living-related donors. Blood samples were collected immediately before and after transplantation at day 7 and month 1. Serum IL-18, creatinine, cystatin C, CRP, and oxidative stress markers (TOS, TAC) were measured. The Oxidative Stress Index (OSI) was calculated. Polymorphisms of the promoter region of the IL-18 gene, IL18-607A/C, and -137C/G were determined by analysis of a "real-time PCR/Melting curve". Results Serum creatinine, cystatin C, CRP, IL-18, TOS, and OSI levels significantly decreased after transplantation. Post-transplant levels of serum TAC and estimated GFR demonstrated consistent significant increases. Serum IL-18 levels were significantly higher in patients with IL-18-137 GG and IL-18-607 CC genotypes before transplantation. Conclusion Our results indicate that the IL-18-137 GG and -607 CC genotypes contribute to higher IL-18 levels; however, the influence of these polymorphisms on oxidative stress has not been observed.

[Determination of cytomegalovirus glycoprotein B genotypes in different geographical regions and different patient groups in Turkey]. / Türkiye'de farkli cografi bölgelerde ve farkli hasta gruplarindaki sitomegalovirus izolatlarinin glikoprotein B genotiplerinin belirlenmesi.

Cytomegalovirus (CMV), a common virus found all around the world, usually causes asymptomatic infections in immunocompetent hosts, however it may lead to serious complications in immunodeficient patients and in the fetus. CMV is divided into four genotypes according to the polymorphisms in UL55 gene that encodes for envelope glycoprotein B. Nucleotide polymorphisms of CMV gB gene can affect the cell tropism of the virus and host immune response and believed to have important changes in the pathogenesis of CMV. The aim of this study was to determine the gB genotypes of CMV isolates from different patient groups selected from different regions of Turkey. A total of 136 clinical specimens from patients (66 female, 70 male; age range: 0-65 years, mean age: 24.03 ± 17.17) who were diagnosed to have CMV infection by polymerase chain reaction (PCR) and/or antigenemia tests, between 2001-2014, in the medical school hospitals of Akdeniz, Ege, Istanbul Cerrahpasa and Erciyes Universities (located at Mediterranean, Aegean, northwest and central Anatolia regions, respectively), were included in the study. The patient group consisted of 80 renal transplant (RT) recipients, 35 stem cell transplant (SCT) recipients, 13 newborns, seven heart transplant (HT) recipients and one pregnant woman. CMV gB genotypes were determined by PCR-RFLP (restriction fragment length polymorphism) method, and DNA sequencing and phylogenetic analysis were performed for the randomly selected 15 isolates with different genotypes. Among 136 (135 plasma, 1 amnion fluid) samples, the most frequent genotype was gB1 (n= 44, 32.4%), followed by gB2 (n= 39, 28.6%), gB3 (n= 36, 26.5%) and gB4 (n= 8, 5.9%); however nine (6.6%) samples could not be genotyped. When analysis were interpreted according to the patient groups, it was determined that the genotypes in RT recipients were gB1 32.3%, gB2 28.7%, gB3 26.5% and gB4 5.9%; in SCT recipients gB1 34.3%, gB2 28.6%, gB3 22.9% and gB4 5.7%; in HT recipients gB3 57.1%, gB1 14.3% and gB2 14.3%; in newborns gB1 38.4%, gB3 30.8%, gB2 15.4% and gB4 7.7%, and gB2 genotype in the pregnant woman. As our study was a descriptive study to determine the genotypes of CMV gB, the relationship between the genotypes and the variants such as viral load, symptomatic disease and prognosis were not analyzed. As a result, the isolation of different gB genotypes in various case groups from four distinctive provinces, underlines the diversity of CMV gB genotypes in Turkey.

[Monitoring of cytomegalovirus-specific CD4+ and CD8+ T cell responses by cytokine flow cytometry in renal transplant recipients]. / Böbrek nakil hastalarinda sitomegalovirusa özgül CD4+ ve CD8+ T hücre yanitinin sitokin akim sitometri yöntemiyle izlemi.

In spite of the improvements in the clinical management of solid organ transplant (SOT) recipients provided by immunosuppresion and universal prophylaxis, human cytomegalovirus (CMV) infections continue to be one of the most leading causes of morbidity and mortality. Cell-mediated immunity specific to CMV (CMV-CMI) plays an important role in the control of CMV replication. Therefore, monitoring of CMV-specific T-cell response can be used to predict individuals at increased risk of CMV disease. The aim of this study was to investigate the levels of CMV-specific interferon (IFN)-γ producing CD4(+) and CD8(+) T cells in kidney transplant recipients before and after the transplantation, by cytokine flow cytometry. A total of 21 kidney transplant recipients (14 male, 7 female; age range: 18-66 years, mean age: 34.5 ± 9.9) who were all CMV seropositive have been evaluated in the study. Blood samples from the patients were obtained before and at the 1(st), 3(rd) and 6(th) months after transplantation. CMV seropositive healthy kidney donors (n= 20) constituted the control group. The main stages of our procedure were as follows; isolation of peripheral blood mononuclear cells from whole blood, freezing and storing of the samples, later on thawing the samples, ex vivo stimulation of lymphocytes with pooled CMV peptides and counting CMV-specific IFN- producing CD4(+) and CD8(+) T cells by flow cytometry following surface and intracellular cytokine staining. Monitoring of the viral load (CMV-DNA) was performed in 10 days intervals in the first 3 months followed by 3 week intervals until 6 months using COBAS AmpliPrep/COBAS TaqMan CMV test system (Roche Diagnostics, USA). The frequencies of pretransplant CMV-specific IFN-γ producing CD8(+) T cells in patient (3.53 ± 4.35/µl) and control (4.52 ± 5.17/µl) groups were not statistically different (p= 0.266). The difference between the number of virus-specific CD4(+) T cells in patients (8.84 ± 9.56/µl) and those in the control group (8.23 ± 11.98/µl) was at the borderline of significance (p= 0.057). The age and gender of the patients and type of antiviral prophylaxis protocols [valgancyclovir (n= 4); valacyclovir (n= 17)] did not have any significant effect on CMV-CMI (p> 0.05). Similarly, induction therapy administered to four patients did not show any effect on CMV-CMI (p> 0.05). CMV-specific immune responses of patients who received different immunosuppression protocols [tacrolimus + mycophenolate mofetil (MMF) + steroid (n= 17); cyclosporine + MMF + steroid (n= 2); mTOR inhibitor + MMF + steroid (n= 2)] were not different (p> 0.05). The number of CMV-specific CD4(+) T cells in all patients were significantly decreased in the 3rd month compared to the 1st month after the transplantation (p=0.003), indicating a relationship with the period of immunosuppressive therapy. In one of the patients who did not have CMV-specific CD4+ T-cell response but had cytotoxic T-cells (CD8(+) T= 0.6%) before transplantation, CD4(+) T-cell response have developed during monitorization (1.4%, 1.5% and 0.5% in 1st, 3rd and 6th months, respectively), and no viral reactivation was detected. Out of the two patients who had no CD4(+) and CD8(+) T cell response in the 3rd month, one of them developed low level viremia (150 copies/ml) in the 6th month. In this patient the level of CMV-CMI in the 6th month (CD4(+)T + CD8(+)T= 0.9%), have reached higher values than the values obtained before the transplantation (CD4(+) T + CD8(+) T= 0.5%). The viremia was cleared spontaneously in this patient and no antiviral therapy was required. In conclusion, our results suggested that pretransplant and posttransplant monitoring of CMV-specific T-cell responses might be helpful as well as viral load in the clinical management of CMV infection in SOT patients.

Confirmation of anti-DFS70 antibodies is needed in routine clinical samples with DFS staining pattern.

BACKGROUND: Recognition of nuclear dense fine speckled (DFS) pattern by indirect immunofluorescence (IIF) is not easy. Thus, confirming the presence of these antibodies might be needed. In this study, we aimed to determine the frequency of DFS pattern in our diagnostic laboratory and to investigate the presence of anti-DFS70 antibodies in samples showing DFS pattern by two commercially available research kits retrospectively. MATERIAL AND METHODS: Seventy-four sequential serum samples with DFS pattern on HEp2010 cell substrates by IIF were included in this study. The semiquantitative DFS70 ELISA Kit (MBL International Corporation, Woburn, UK) was used for detection of anti-DFS70 antibodies in these samples. Twenty selected samples were tested for the presence of anti-DFS70 antibodies using ANA Line Immunoassay (LIA) (Immco Diagnostics, New York, USA). RESULTS: Sixty-two (83.8%) of 74 serum samples were found positive with ELISA, when 15 U/ml was taken as a reference value. Among 18 samples that were found positive by ELISA, five were negative for anti-DFS70 antibodies by LIA, while 13 were found positive. The lowest ELISA result of the sample that was positive by LIA was found to be 45.3 U/ml. When 45.3 U/ml was considered as a reference value, 45 (60.8%) of 74 serum samples were positive by ELISA. Nineteen of 20 patients had no SARD, while one had systemic lupus erythematosus (SLE). CONCLUSIONS: DFS pattern should be confirmed with an objective method such as ELISA, LIA, or IB. We think that confirmation tests for detection of anti-DFS70 antibodies should be included in diagnostic algorithms.

[Distribution of hepatitis C virus genotypes among patients with chronic hepatitis C infection in Akdeniz University Hospital, Antalya, Turkey: a five-year evaluation]. / Akdeniz Üniversitesi Hastanesinde kronik hepatit C enfeksiyonu olan hastalarda hepatit C virus genotipleri: Bes yillik sonuçlarin degerlendirilmesi.

Hepatitis C virus (HCV) is one of the major causes of chronic hepatitis. It is important to know the genotypes of HCV in the decision of the HCV related chronic hepatitis therapy. The aim of this study was to evaluate the HCV genotypes determined at the Microbiology Laboratory of Akdeniz University Hospital, and to evaluate the changes in the distribution of the genotypes within the last five years. A total of 422 blood samples from HCV-RNA positive chronic hepatitis C patients (219 male, 203 female; age range: 8-79 yrs, mean age 46.3 ± 15.5 yrs) which were sent to our laboratory for genotyping between 2009-2013 period, were analyzed retrospectively. HCV-RNA extractions were performed in an automated system (EZ1 Virus Mini Kit v2.0, Qiagen, Germany), and a commercial reverse hybridization line probe-based assay (LIPA; GEN-C RT-PCR, Italy) was carried out for genotyping, For viral load determinations, a real-time PCR method (Cobas TaqMan HCV, Roche Diagnostics, Germany) was used. Demographic data of the patients were obtained from the hospital information systems and electronic patients' files. Out of the 422 patients, genotype 1b was detected in 63.3% (n= 267), genotype 1a in 14.7% (n= 62), genotype 3a in 11.1% (n= 47), genotype 2b in 0.9% (n= 4), genotype 4e in 0.2% (n= 1). The subtypes couldn't be determined for 5.4% (n= 23), 2.6% (n= 11) and 1.4% (n= 6) of the patients infected with genotype 1, 2 and 4, respectively. One (0.2%) patient, was coinfected with genotype 1 and 4. Of the patients, 40 were foreign-born (16 cases from Russia; 4 of each from Ukraine and Georgia; 3 of each from Turkmenistan, Kyrgyzstan, and Germany; one of each from Tajikistan, Azerbaijan, Uzbekistan, Chechnya, Moldova, Switzerland and Romania) and among these patients genotype 3a (19/40; 47.5%) was the most common genotype followed by genotype 1b (17/40; 42.5%). Median values of HCV viral load were 668.500 IU/ml (range: 2.000-9.630.000) in the whole group; while it was 732.000 IU/ml (range: 2.000-9.630.000) in patients infected with genotype 1 and 444.000 IU/ml (range: 2.650- 8.330.000) in patients infected with the other genotypes (p> 0.05). Patients infected with genotype 1 were found to be older than those infected with other genotypes (47 ± 15.7 and 39.5 ± 12.2, respectively; p< 0.001). Among patients infected with different genotypes, there was no statistically significant difference in terms of genders (p> 0.05). In conclusion, the determination of HCV genotypes is of crucial importance for treatment decision-making of chronic HCV infection. Besides, it also allows monitoring the changes in the epidemiology of HCV. In this study, although genotype 1b was determined as the most common HCV genotype, the detection of other genotypes was remarkable. This finding was attributed to the presence of many foreign national people in Antalya region which was a high capacity tourism area in Turkey.

Detection of human bocavirus in children with upper respiratory tract infection by polymerase chain reaction.

BACKGROUND: The aim of this study was to investigate whether in children with middle ear effusions (MEE), adenoid and tonsil tissues are associated with human bocavirus (HBoV). MATERIALS AND METHODS: A total of 124 patients (56 females (45.2%) and 68 males (54.8%)) with chronic adenotonsillitis and serous otitis media under the age of 15 were recruited. Two hundered four samples (113 adenoid (55.4%), 68 tonsil (33.3%), and 23 middle ear effusion (11.3%)) were analyzed for the presence of HBoV using polymerase chain reaction (PCR). RESULTS: HBoV was detected in only 6 (4.8%) adenoid tissue samples each belonging to a different patient. CONCLUSIONS: Our findings are consistent with the results of other studies, reporting approximately 5 - 10% of the samples being positive for HBoV. To understand the detailed role of HBoV in the etiology of RTI in children, further studies would be needed.

Serotype distribution of Streptococcus pneumoniae and pneumococcal vaccine coverage in adults in Turkey between 2015 and 2018.

OBJECTIVE: To evaluate the serotype distribution and antibiotic resistance in pneumococcal infections in adults and to provide a perspective regarding serotype coverage of both current and future pneumococcal vaccines. PATIENTS AND METHODS: This passive surveillance study was conducted with the Streptococcus pneumoniae strains isolated from the specimens of patients with pneumonia (materials isolated from bronchoalveolar lavage), bacteraemia, meningitis, pleuritis and peritonitis between 2015 and 2018. Serogrouping and serotyping were performed by latex particle agglutination and by conventional Quellung reaction using commercial type-specific antisera, respectively. The strains were analysed for penicillin, cefotaxime, erythromycin and moxifloxacin susceptibilities by E-test. RESULTS: In the whole study group (410 samples from adults aged ≥18 years), the most frequent serotypes were 3 (14.1%), 19 F (12%) and 1 (9.3%). The vaccine coverage for PCV13, PCV15, PCV20 and PPV23 was 63.9%, 66.6%, 74.1% and 75.9%, respectively, in all isolates. Penicillin non-susceptibility in invasive pneumococcal disease (IPD) was 70.8% and 57.1% in the patients aged <65 and ≥65 years, respectively. About 21.1% and 4.3% of the patients with and without IPD had cefotaxime resistance. Non-susceptibility to erythromycin and moxifloxacin was 38.2% and 1.2%, respectively. CONCLUSIONS: The results revealed that novel PCV vaccines may provide improved coverage as compared with the currently available vaccine, PCV13. The significant antibiotic resistance rates imply the need to extend the serotype coverage of the vaccines. Continuing the surveillance in pneumococcal diseases is critical to explore the serotype distribution and incidence changes of IPD cases in the population and to inform policy makers to make necessary improvements in the national immunization programmes.Key messagesThis multicentre study demonstrated the most recent serotype distribution and antibiotic resistance in adult population in Turkey.Shifting from PCV13 to novel conjugated vaccines will significantly increase the coverage.Continuing the surveillance in pneumococcal diseases is critical to explore the serotype distribution changes and the incidence of cases with invasive pneumococcal disease in the population.

Investigation of dense fine speckled pattern and anti-dense fine speckled 70 antibody by a single step assay.

The indirect immunofluorescence (IIF) method is the gold standard for identifying anti-nuclear antibodies (ANAs). It is recommended that ANA, including the dense fine speckled (DFS) pattern, should be verified with a highly specific confirmatory test after a sensitive screening test. Although methods such as ELISA and LIA are often used to confirm the presence of anti-DFS70 antibodies, new IIF methods have been developed in recent years to prevent the difficulties in the recognition of the DFS pattern and to carry out the confirmatory test in a single step. In this study, we evaluated CytoBead (Generic Assays, Germany) test, which contained both HEp-2 cell substrate and beads coated with DFS70 antigen in one well, in comparison to the routine two-step test strategy. Five hundred forty-one samples were studied by conventional IIF assay, LIA, and CytoBead assay; 264 samples were studied by ELISA. The Bead component of the CytoBead test was found to be reliable as a confirmational test when compared with ELISA and LIA (total agreement values were 85.6% and 87.6%, respectively). The CytoBead ANA DFS70 might be a promising test in the future, allowing both screening and confirmation in a single step, saving time and being easier than two-step testing.

DNA methylation or histone modification status in metastasis and angiogenesis-related genes: a new hypothesis on usage of DNMT inhibitors and S-adenosylmethionine for genome stability.

Metastasis is a leading cause of mortality and morbidity in cancer. This process needs angiogenesis. The biology underlying cancer, metastasis, and angiogenesis has been investigated so as to determine the therapeutic targets. Invasive and metastatic cancer cells have undergone numerous genetic and epigenetic changes, manifested by cytoskeletal changes, loss of adhesion, and expression of proteolytic enzymes that degrade the basement membrane. Additionally, in endothelial cells, some epigenetic modifications occur during the formation of angiogenesis. Researchers have used some methylation inhibitors, histone deacetylase inhibitors, or methylating agents (such as S-adenosylmethionine, SAM) against cancer and angiogenesis. Although they are effective to beat these diseases, each one results in differentiation or changes in genome structure. We review epigenetically modified genes related with angiogenesis and metastasis in cancer and endothelial cells, and suggest a new proposal. This hypothesis has discussed the importance of the usage of DNA methylation inhibitors together with SAM to prevent tumor progression and genome instability or changes resulting in additional diseases.

Inhibition of angiogenesis by S-adenosylmethionine.

Metastasis is a leading cause of mortality and morbidity in cancer. One of the steps in metastasis process is the formation of new blood vessels. Aberrant DNA methylation patterns are common in cancer cells. In recent studies, S-adenosylmethionine (SAM), which is a DNA methylating agent, has been found to have inhibitory effects on some carcinoma cells in vivo and in vitro. In the present study, we have used SAM to investigate whether it is effective against angiogenesis in vitro. Our results have shown that SAM can reduce the formation and organization of capillary-like structures of endothelial cells in tumoral environment. Besides, we have found SAM can block endothelial cell proliferation and the migration of cells towards growth factors-rich media. In conclusion, our study suggests that SAM may be used against angiogenesis as a natural bio-product.

Evaluation of in vitro activities of tigecycline and various antibiotics against Brucella spp.

Brucellosis is a zoonosis with a worldwide distribution and remains a significant public health problem mainly in the developing world. In this study we evaluated the in vitro activities and synergistic effects of antibiotic combinations against blood culture isolates of Brucella spp. In vitro susceptibilities of 76 blood culture isolates of Brucella melitensis and one blood culture isolate of Brucella abortus to doxycycline, streptomycin, gentamicin, trimethoprim-sulfamethoxazole, moxifloxacin, rifampin, ciprofloxacin, and tigecycline were examined by Etest method. For 37 patients with Brucella spp. isolates (36 B. melitensis, 1 B. abortus), antibiotic combinations used for treatment were identified with those tested in vitro for synergy using Etest method. Trimethoprim-sulfamethoxazole and tigecycline were the most active of the compounds tested with MIC90 value of 0.094 mg/l. Among antibiotic combinations only streptomycin-rifampin combination was synergistic for one Brucella spp. isolate. The other antibiotic combinations revealed antagonistic or indifferent activity. Complete clinical response was achieved in all patients. Further studies are required to determine the correlation between the antimicrobial susceptibility and synergy test results with the clinical course of patients. Brucellosis can be adequately treated with existing regimens in our region.

[Shall we report the carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii strains detected by BD Phoenix system?]. / Pseudomonas aeruginosa ve Acinetobacter baumannii suslarinda BD Phoenix sistemi ile saptanan karbapenem direncini raporlayalim mi?

Imipenem and meropenem are broad spectrum antimicrobial agents that are especially useful in the treatment of nosocomially acquired Pseudomonas aeruginosa and Acinetobacter spp. infections. Previous reports have noted that susceptibility tests could show false resistance to imipenem. For this reason, Centers for Disease Control and Prevention has recommended that all carbapenem resistant or intermediate resistant isolates should be tested with an additional method to verify the results. This study was aimed to evaluate the imipenem and meropenem susceptibilities by disk diffusion, E-test and broth microdilution in P. aeruginosa and Acinetobacter baumannii strains found to be resistant or intermediate to imipenem-meropenem by BD Phoenix automated susceptibility testing system. Between January 2006-January 2007, 85 non-duplicate isolates of A. baumannii and 51 non-duplicate isolates of P. aeruginosa which were determined as resistant or intermediate resistant to imipenem and/or meropenem by BD Phoenix automated identification and susceptibility system (Becton Dickinson, Sparks, MD, USA) were collected in Akdeniz University Hospital Central Laboratory. All strains were tested by E-test (AB Biodisk, Sweden), disk diffusion and reference broth microdilution (BMD) method following CLSI recommendations. All 51 isolates of P. aeruginosa determined as imipenem and/or meropenem resistant or intermediate resistant by BD Phoenix, were found to be imipenem and/or meropenem resistant or intermediate resistant by the reference BMD method. Minor error rates were same for all testing systems (1.9%) except for the meropenem results of BD Phoenix system (5.9%). No major errors were produced by any system. For A. baumannii, only one very major error was detected for meropenem by BD Phoenix system. Number of minor errors determined for meropenem by all testing systems compared to the reference test, ranged from 2 (2.4%) to 3 (3.5%). It was concluded that carbapenem susceptibility test results obtained by BD Phoenix system for P. aeruginosa and A. baumannii isolates, could be reported without an additional susceptibility testing method unless indicated on case basis.

[Investigation of ESBL types in community acquired urinary Escherichia coli isolates by isoelectric focusing and polymerase chain reaction]. / Toplum kaynakli üriner sistem enfeksiyonu etkeni Escherichia coli suslarinda GSBL enzim tiplerinin izoelektrik odaklama ve polimeraz zincir reaksiyonu yöntemleri ile arastirilmasi

The aim of this study was to determine the extended-spectrum beta-lactamase (ESBL) types by isoelectric focusing (IEF) and polymerase chain reaction (PCR) methods in 56 Escherichia coli strains isolated from urine samples of patients with community-acquired urinary tract infection and determined as ESBL positive with the phenotypic screening tests (E test and combined disk method). IEF revealed that most of the strains produced 1 to 3 different bands, mostly at the isoelectric points 8.2 (n= 44, 79%) compatible with CTX-M. Twenty four (43%) isolates had CTX-M and TEM enzyme bands together, 16 (29%) isolates had only CTX-M enzyme bands, 3 (5%) isolates had CTX-M, TEM, SHV bands, one had CTX-M and SHV enzyme bands together, and one had only TEM band. Eleven E.coli strains did not yield any enzyme bands. PCR analysis revealed that 93% (n= 52) of the isolates had CTX-M, 64% (n= 36) had TEM and 11% (n= 6) had SHV, while 29 (52%) had CTX-M + TEM, three had CTX-M + SHV, and three had CTX-M + TEM + SHV genes together. PER-1 type beta-lactamases were not detected by PCR method. PCR analysis of the eleven strains that yielded no band in IEF showed that 5 strains had CTX-M + TEM, 3 had CTX-M and 3 had TEM enzyme genes. The consistency between IEF and PCR methods for the determination of CTX-M, TEM and SHV enzymes was 85%, 78% and 67%, respectively. Genes encoding ESBL's are usually located on transferrable plasmids that may also carry other resistance determinants. Thus detection of beta-lactamase enzyme types in ESBL positive bacteria is important for the choice of appropriate antimicrobial agents for treatment.

Changes in substance P levels of inferior turbinate in patients with mucosal contact headache.

INTRODUCTION: Mucosal contact headache is a referred pain that arises from contact between the nasal septum and the lateral nasal wall. Evidence supports the role of substance P in a contact headache such that release of substance P from sensory nerve endings causes inflammation and allergy. OBJECTIVES: This study aimed to determine possible differences in substance P levels in inferior turbinate hypertrophy creating a contact headache. METHODS: 28 patients who had contact headaches (study group) and 16 volunteers with no complaints were included in the study. Substance P levels in the inferior turbinate tissue samples were quantified using a commercially available substance P EIA kit. RESULTS: In the study group average substance P levels were 2.65±0.27pg/mg tissue (range: 0.61-5.44) and in the control group it was 1.77±0.27pg/mg tissue (range: 0.11-4.35). The difference was statistically significant between the two groups (p=0.0215). Average preoperative headache group visual analog scale scores was 5.93±0.38 (2-9) and the turbinate volume was 6.56±0.35cm3 (3.50-10.30). The control group turbinate volume was 4.71±0.39cm3 (2.50-7.70). We found a correlation between the visual analog scale scores and substance P levels such that substance P levels were higher in visual analog scale scores above 5 (p=0.001). CONCLUSION: This study demonstrates the relationship between intranasal contact headaches and increased mucosal substance P levels. We also found that there is no correlation with substance P levels and volume of the inferior turbinate.

Serotype distribution of Streptococcus pneumonia in children with invasive disease in Turkey: 2015-2018.

Objectives: To determine the serotype distribution of pneumococcus causing invasive pneumococcal disease (meningitidis, bacteremia and empyema) in children in Turkey, and to observe potential changes in this distribution in time to guide effective vaccine strategies. Methods: We surveyed S. pneumoniae with conventional bacteriological techniques and with real-time polymerase chain reaction (RT-PCR) in samples of cerebrospinal fluid (CSF), blood and pleural fluid. S. pneumoniae strains were isolated from 33 different hospitals in Turkey, which are giving health services to approximately 60% of the Turkish population. Results: A total of 167 cases were diagnosed with invasive pneumococcal disease between 2015 and 2018. We diagnosed 52 (31.1%) patients with meningitis, 104 (62.2%) patients with bacteremia, and 11 (6.6%) patients with empyema. Thirty-three percent of them were less than 2 years old and 56% less than 5 years old. Overall PCV13 serotypes accounted for 56.2% (94/167). The most common serotypes were 19 F (11.9%), 1 (10.7%) and 3 (10.1%). Conclusions: Besides the increasing frequency of non-vaccine serotypes, vaccine serotypes continue to be a problem for Turkey despite routine and high-rate vaccination with PCV13 and significant reduction reported for the incidence of IPD in young children. Since new candidate pneumococcal conjugate vaccines with more serotype antigens are being developed, continuing IPD surveillance is a significant source of information for decision-making processes on pneumococcal vaccination.

[Resistance to newer beta-lactams and related ESBL types in gram-negative nosocomial isolates in Turkish hospitals: results of the multicentre HITIT study]. / Türkiye'de hastane izolati gram-negatif bakterilerde yeni beta-laktam antibiyotiklere direnç ve GSBL tipleri: çok merkezli HITIT sürveyansinin sonuçlari.

Increasing resistance due to extended-spectrum beta-lactamases (ESBLs) and multiple resistance mechanisms in gram-negative hospital isolates restrict the role of beta-lactam antibiotics in empirical treatment of serious infections. As the prevalence of ESBL producing strains and resistance rates to antimicrobial agents can vary in each center, local surveillance studies are required to guide therapy. In this study, in vitro rates of resistance to ceftriaxone, ceftazidime, cefepime, imipenem, cefoperazone/sulbactam and piperacillin/tazobactam were evaluated in 1196 gram-negative hospital isolates in a multicenter in vitro study with the participation of six different centers in Turkey between the period of June 2004-January 2005. The isolates included Escherichia coli (n= 457), Klebsiella pneumoniae (n= 390), Pseudomonas aeruginosa (n= 194) and Acinetobacter boumannii (n= 155). In addition, frequency of ESBL production and types of enzymes were determined in blood isolates of E. coli and K. pneumoniae. MICs and ESBL production were investigated by E-test (AB Biodisk, Solna) and the results were evaluated by using CLSI breakpoints. PCR analysis was used for typing of the ESBLs. In E. coli, 26% and in K. pneumoniae 32% of the isolates were ESBL producers. Among the blood isolates of E. coli and K. pneumoniae, 31.7% and 33.3% produced ESBLs, respectively. CTX-M (71.4%) was the most prevalent enzyme, followed by TEM (49.4%) and SHV (46.7%) derived enzymes. CTX-M-15 (69.4%) was the most frequent CTX-M type in blood isolates followed by CTX-M-3 (28.6%) and CTX-M-1 (2%). Resistance to imipenem was not observed in E. coli isolates, however it was 1.3% in K. pneumoniae, 28.9% in P. aeruginosa and 52.2% in A. baumannii strains. Resistance to cefoperazone/sulbactam was found as 6%, 17.7%, 27.9% and 41.3% in E. coli, K. pneumoniae, P. aeruginosa and A. baumannii isolates, respectively, whereas resistance rates to piperacillin/tazobactam were 10.2%, 22.3%, 22.7% and 78.7%, respectively. These results indicate that ESBL production and rates of resistance to beta-lactam antibiotics are high in hospital isolates of gram-negative bacteria in Turkey, however, they show variations in different hospitals and CTX-M enzymes are prevalent in these isolates.

Bone metabolism and mineral density in patients with beta-thalassemia major.

OBJECTIVE: To evaluate bone metabolism in patients with beta-thalassemia major and to determine the factors associated with the development of osteoporosis. METHODS: We studied 25 patients with thalassemia major with a mean age of 18.4 years (range 5-31) and aged and gender matched 24 healthy controls who were attending the outpatient physical medicine and rehabilitation clinic of Akdeniz University Hospital between January 2004 and March 2004 in Turkey. Bone mineral density (BMD) of lumbar spine (L1-L4) and proximal femur were determined using dual x-ray absorptiometry (DXA). Venous blood samples were obtained for determination of blood cell count and markers of bone formation and resorption. RESULTS: The BMD values, both at lumbar and femoral neck levels were significantly lower in patients compared to controls. Serum N-telopeptide level was slightly higher, whereas osteocalcin was slightly lower in patients; however, these values were not statistically significant. Plasma levels of insulin like growth factor-1 (IGF-I) and insulin like growth factor binding protein-3 (IGFBP-3) were significantly lower in patients. Also, serum levels of estradiol and progesterone in females, luteinizing hormone and follicle-stimulating hormone in both gender were significantly lower in patients. Serum levels of free testosterone and total testosterone were lower in patients, but not statistically significant. Patients also had significantly higher serum phosphorus levels, and lower serum calcitonin levels compared to controls. CONCLUSION: The BMD is decreased in thalassemic patients. Growth retardation, growth hormone / IGF-I / IGFBP-3 axis dysfunction, gonadal dysfunction and hypothalomo-pituitary-gonadal axis dysfunction may be responsible for the development of osteoporosis in the patients with beta-thalassemia major.

[Investigation of cytomegalovirus seroepidemiology in Antalya with a population-based cross-sectional study and review of related data in Turkey]. / Antalya'da sitomegalovirus seroepidemiyolojisinin toplum kaynakli kesitsel bir çalisma ile arastirilmasi ve Türkiye verilerinin derlenmesi.

Since there are numerous studies on CMV seroprevalence in various groups in Turkey, the number of population based, age-stratified cross-sectional studies which include epidemiological characteristics of the virus are limited. The aim of the study was to investigate the age-stratified seroprevalence and epidemiological characteristics of CMV infection in Antalya (a province located in Mediterranean region of Turkey). Study group was selected by cluster sampling method. The sample size was calculated as 360 subjects (151 male, 209 female; age range: 1-49 years, mean age: 22.5 +/- 14.4 years), with an expected prevalence rate of 80%, at a confidence level of 95% and a sample error less than 5%. With the thought of the presence of maternal antibodies, 0-1 year age group was not included to the study. Serum samples have been screened for CMV-IgG, and those given negative results were also searched for CMV-IgM by a commercial microELISA (Radim, Italy) test. The overall seroprevalence of CMV-IgG was found as 93.6% (337/360) in Antalya municipality and IgM positivity was not detected in CMV-IgG negative sera. An increase in the seroprevalence rates was observed with age (p < 0.001), and the rate was found quite high (93.3%) for the first year of life. The seropositivities in the age groups of 1-6, 7-14 and 14-49 years were detected as 82.1%, 92% and 97.8%, respectively. The seroprevalence rate of 82.1% before the age of seven has rised to 96.8% after that age, and being > or =7 years old was found statistically significant in terms of CMV infection (p < 0.001, OR: 6.635). Ages one and seven were found to be the critical ages for CMV infection in our region. CMV seropositivity was 97.4% in woman at childbearing age (15-49 years). Gender, marital status, education, living area, residence, income, history of sexually transmitted diseases, surgery, blood transfusion and day care attendance did not contribute independently to the seroepidemiology of CMV (p > 0.01). In addition, the data of this study were evaluated and discussed together with the results obtained from the other Turkish studies, as far as accessible. In conclusion, since CMV seroepidemiology in Turkey differs as the socioeconomic changes occur, the changes in CMV serostatus and dire consequences of high seroprevalence rates on public health should be evaluated with prospective, population based studies in further years.