The relevance between anti-rods/rings antibody and different treatment regimens in chronic hepatitis C virus infection.

The antibodies and other issues associated with immunity in chronic hepatitis C virus (HCV) have been widely investigated, especially non-organ-specific antinuclear antibodies. Rods-rings (RR) antibody patterns are frequently observed due to pegylated IFN-α (PEG-IFN)/ribavirin (RBV) treatment by indirect immunofluorescence (IIF). We evaluated the relevance between anti-RR and PEG-IFN/RBV and/or direct-acting antiviral (DAA) regimens in chronic HCV. Sampling was done after achieving a sustained virological response (SVR) for 178 patients (aged >18 years). Patients were grouped according to treatment protocols (Group 1 [G1]: PEG-IFN/RBV [n = 53], Group 2 [G2]: PEG-IFN/RBV and Telaprevir or Boceprevir [n = 31], Group 3 [G3]: second- and third-wave DAA and previously received PEG-IFN/RBV (n = 38), and Group 4 [G4]: second- and third-wave DAA [n = 56]). Anti-RR was investigated by IIF (Euroimmun AG) test. Overall, 27 (15.16%) patients were anti-RR positive and received PEG-IFN/RBV. The numbers of anti-RR positivity for G1/2/3/4 (%) were 16/3/8/0 (30.2/9.6/21/0), respectively (p < .001). The anti-RR positivity rate for G1/2/3 was 22.13% (27/122, p = .088). Anti-RR was positive in 17.5% (11/63) of G1/2/3 patients who did not achieve SVR after the first treatment. This rate was 27.1% (16/59) in patients with SVR after the first treatment in G1/2 and there was no difference between these two classified groups in terms of antibody titers (p = .915). Anti-RR was detected up to 172 months after SVR. In summary, anti-RR was positive in high rates in patients receiving PEG-IFN/RBV therapy. Frequent monitoring is needed during patient follow-up to get more data on the relationship between anti-RR titer, treatment regimens, and SVR.

[Investigation of Regulatory T Cells and Secreted Immunomodulatory Cytokine IL-10 Levels in Patients with Hepatitis B]. / Hepatit B Hastalarinda Regülatör T Hücre ve Salinan Immünmodülatör Sitokin IL-10 Düzeylerinin Degerlendirilmesi.

Hepatitis B infection is still among the most important public health problems worldwide, even great improvements have been made in the treatment strategies. Hepatitis B virus (HBV) replicates itself by entering the liver cells and simultaneously with the antigen release, many antagonistic immune responses are induced by the regulatory cells including T cell (Treg), T helper 17 (Th17), T helper 1 (Th1) and T helper 2 (Th2) cells. The main function of Treg cells is to develop an appropriate immune response against infection and to suppress the immune response if it is not required. Tregs suppress the effector T cells via secreting immune system supressor cytokines such as Transforming Growth Factor-Beta and interleukin (IL)-10 or contact dependent way. Tregs protect cells from immunopathologic damage of HBV specific T cell immune response and also cause viral persistence, cirrhosis, hepatocellular carsinoma (HCC) and autoimmunity but the mechanisms are not clear, yet. In this study, we aimed to determine whether evaluation of Treg cells and cytokine IL-10 levels together in hepatitis B patients is useful that may indicate the disease survey and response to the treatment. The peripheral blood samples of ninety-one volunteers, including 61 HBV infected patients and 30 healthy controls selected from applicants of Infectious Diseases Outpatient/Clinic Service, were taken. Their CD4+CD25highFOXP3+CD152+CD127lowTreg cell distribution were measured by flow cytometry method, using the recently defined markers. The level of IL-10 cytokine released by immunomodulatory cells was determined by quantitative ELISA method. Treg cell percentages of the patients with acute hepatitis B were below the normal range (2-4%) (median= 1.50%, 0.6-3.5) and the difference was statistically significant (p= 0.005). Treg cell percentages of the patients with chronic hepatitis B were higher than the control group (p< 0.05), and it was found to be related to the parameters used in the diagnosis, staging and follow-up of the disease. IL-10 levels were significantly higher in all hepatitis B clinical stages compared to the healthy controls (median= 11.7, 17.3-44.9) (p< 0.05). Also, in parallel with Treg cells, IL-10 levels were correlated with HBV DNA load and HBsAg levels (r= 0.48, p< 0.02). Treg cells and the related cytokine IL-10 are thought to play an important role in the immunology of HBV infection and therefore, promising to follow up the disease and to develop new therapeutic strategies targeting the Treg cell.

[Investigation of antinuclear antibodies in chronic hepatitis B patients]. / Kronik hepatit B hastalarinda antinükleer antikorlarin arastirilmasi.

Antinuclear antibodies (ANA) facilitate the diagnosis and evaluation of patients in many systemic autoimmune conditions. Besides, ANA may also be detected in chronic infectious diseases. Although a number of investigations associated with autoantibody positivity in patients with chronic hepatitis C were reported, autoantibody positivity in patients with chronic hepatitis B remain rarely addressed in the literature. The aim of this study was to evaluate the antinuclear antibody (ANA), antimitocondrial antibody (AMA), anti-smooth muscle antibodies (ASMA) and anti-liver-kidney microsomal antigen (LKM) antibodies in chronic hepatitis B patients. Serum samples were obtained from adult patients with chronic hepatitis B diagnosis according to "European Association for the Study of the Liver (EASL)" criteria. Samples were taken from 47 patients (22 female, 25 male) with treatment-naive, histologically-proven chronic hepatitis B. Cases co-infected with HCV and/or HIV or that also had systemic autoimmune diseases were excluded. As a control group, 30 healthy blood donors were included in the study. Autoantibodies, including ANA, AMA, ASMA and LKM were detected with indirect immunofluorescence (IIF) method (Euroimmune, Lubeck, Germany) and evaluated by fluorescence microscope (Eurostar III plus, Germany). Positive results were graded into 4 levels ( "+", "++","+++" and "++++") from weak to strong Positive samples were studied with a immunoblotting method (ANA Profile 3, Euroimmun, AG) for the detection of extractable nuclear antigen (ENA). The positive results were detected in 8 (17%) of the HBV patients while all the samples were negative in the control group. The difference between the groups was significant (p< 0.05). Among the 47 serum samples tested, none of the patients were positive for AMA, ASMA, LKM. ANA was present in eight of the serum samples in which six were female and two were male patients. Among the IIF patterns of ANA positivity, one mixed pattern (homogeneous and nucleolar) and one cytoplasmic anti-golgi antibody pattern were detected. Positivity grade was ''++''. Other positive patterns were nucleolar (two patients), granular (two patients), ribozomal (one patient) and homogeneous (one patient) and positivity grade was ''+''. ENA was detected in three samples. Two of them was granular pattern positive samples. SS-A was borderline (±) in one and SS-B was borderline (±) in one of the samples. In the mixed pattern positive sample, histon was detected as ''+''. Autoantibody positivity between the patient and control groups were statistically significant (p< 0.05). The difference between autoantibody positivity and gender/age was not statistically significant. In conclusion, autoimmune manifestations may be detected in patients with chronic hepatitis B. Low level titer of antibodies such as ANA, AMA, ASMA or LKM may be present in such patients. An increased frequency of these autoantibodies may be associated with non-autoimmune conditions such as chronic viral infection even in treatment-naive patients.

Antimicrobial susceptibility and resistance mechanisms of methicillin resistant Staphylococcus aureus isolated from 12 Hospitals in Turkey.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important nosocomial pathogens and is also emerging in Turkish hospitals. The aim of this study was to determine the antimicrobial susceptibility profiles of MRSA isolated from Turkish hospitals. MATERIALS AND METHODS: A total of 397 MRSA strains isolated from 12 hospitals in Turkey were included to present study. Antimicrobial susceptibilities were tested using agar dilution method. Presence of ermA, ermB, ermC, msrA, tetM, tetK, linA and aac-aph genes were studied by PCR. RESULTS: All strains were susceptible to vancomycin and linezolid. The susceptibility rates for fusidic acid, lincomycin, erythromycin, tetracyclin, gentamycin, kanamycin, and, ciprofloxacin were 91.9%, 41.1%, 27.2%, 11.8%, 8.5%, 8.3% and 6.8%, respectively. Lincomycin inactivation was positive for 3 isolates. Of 225 erythromycin resistant isolates 48 had ermA, 20 had ermC, and 128 had ermA-C. PCR was negative for 15 strains. Of 3 isolates with lincomycin inactivation one had linA and msrA. Of 358 gentamycin resistant isolates 334 had aac-aph and 24 were negatives. Among 350 tetracyclin resistant isolates 314 had tetM. Of 36 tetM negative isolates 10 had tetK. CONCLUSION: MRSA isolates from Turkish hospitals were multiresistant to antimicrobials. Quinolone and gentamycin resistance levels were high and macrolide and lincosamide resistance were relatively low. Susceptibility rates for fusidic asid were high. Linezolide and vancomycin resistance are not emerged. The most common resistance genes were ermA, tetM and aac-aph. Evolution of antimicrobial susceptibilities and resistance genes profiles of MRSA isolates should be surveyed at regional and national level for accurate treatment of patients and to control dissemination of resistance genes.

Evaluation of hepatitis delta virus (HDV) infection in blood donors in western Turkey.

BACKGROUND: Hepatitis B virus (HBV) and hepatitis delta virus (HDV) infections are potentially dangerous complications of transfusion therapy. OBJECTIVE: The aim of this study was to determine the prevalence of HDV markers examined by serological and molecular methods in hepatitis B surface antigen (HBsAg)-reactive sera among blood donors. MATERIALS AND METHODS: Samples from 88 HBsAg-reactive blood donors were investigated for total anti-delta antibody (anti-HDV) and HDV-RNA between April 2010 and February 2011. HBsAg screening tests were performed by "microparticle enzyme immunoassay" (MEIA) method using the AxSYM system (Abbott Laboratories, USA), and total anti-delta antibody tests were performed by MEIA method using the Alisei system (Radim, Italy). HDV-RNA was quantified using the polymerase chain reaction (PCR) method. Viral nucleic acid isolation system (Anatolia Geneworks) was used with Bosphore HDV quantification kit. RESULTS: HBsAg reactivity was determined as 1% (124/12.423) among blood donors as a whole. Eighty-eight of these 124 samples were investigated further for HDV. Three (3.4%) of the 88 HBsAg-reactive serum samples were total anti-delta antibody-reactive. Of the 3 anti-HDV-reactive sera, 2 were reactive for HDV-RNA. Therefore, HDV-RNA reactivity was determined as 2.3% (2/88) in HBsAg-reactive donors as a whole. The 2 HDV-RNA-reactive donors were brothers. CONCLUSIONS: Investigation of HDV is important because HBV infection is endemic in Turkey. Intrafamilial transmission is important in HDV transmission.

[Comparison of Cefoxitin Disk Diffusion Test, Automated System and Chromogenic Medium for Detection of Methicillin Resistance in Staphylococcus aureus Isolates]. / Staphylococcus aureus Suslarindaki Metisilin Direncinin Belirlenmesinde Sefoksitin Disk Difüzyon Testi, Otomatize Sistem ve Kromojenik Besiyerinin Karsilastirilmasi

The mecA gene is responsible for the development of methicillin resistance in staphylococci however accurate detection of methicillin resistance is not feasible evermore because of heterogenous expression of mecA gene. Although mecA gene determination by polymerase chain reaction (PCR) is considered as the gold standard method, molecular tests are not easily applied in all routine laboratories. Thus, for the rapid and accurate diagnosis of MRSA strains, easy and practical phenotypic tests are still required. This study was aimed to compare the performance of mecA gene analysis by gel bases multiplex PCR with dual primer (Seeplex, Seegene Inc, Korea), cefoxitin disc diffusion method (30 µg, Oxoid, UK), automated system (Phoenix 100, Becton Dickinson, USA) and chromogenic medium CHROMagar MRSA (CHROMagar Microbiology, Salubris, Turkey) for the detection of methicillin resistance in staphylococci. It was found that 60 of the 98 Staphylococcus aureus strains carried the mecA gene. Methicillin resistance was observed by cefoxitin disc diffusion test in 59 isolates, by automated system in 61 isolates, and by CHROMagar MRSA in 65 isolates. When mecA gene analysis was considered as the reference method, the sensitivity, specificity, positive and negative predictive values of the tests that were used for the detection of methicillin resistance were found as 98.3%, 100%, 100% and 97.4% for cefoxitin disc diffusion (CDD) method; 100%, 97.4%, 98.4% and 100% for automated system; 96.7%, 81.6%, 89.2% and 93.9% for chromogenic medium CHROMagar MRSA, respectively. The highest sensitivity and negative predictive values were obtained by the automated system, and the highest specificity and positive predictive values were obtained by the CDD test. Although the sensitivity of chromogenic medium was found to be similar with the CDD test at the end of 48 hours, the specificity of chromogenic medium was lower than the other tests at the end of each incubation period. Likewise, positive and negative predictive values of the chromogenic medium were determined low compared to other tests. In laboratories that cannot perform molecular analysis, the determination of methicillin resistance should be done by the CDD test which is known to be a better inducer of the mecA gene expression of staphylococci. Determination of minimum inhibitory concentration (MIC) with automated systems can be the second choice especially in laboratories with intensive work loads. As a result chromogenic media can be particularly used for screening in laboratories that have a heavy workload and insufficient personnel number. However, due to its low specificity and the possibility of false positive results, it was recommended that positive strains should be confirmed by other methods such as disc diffusion or microdilution.

[t030 is the most common spa type among methicillin-resistant Staphylococcus aureus strains isolated from Turkish hospitals]. / t030, Türkiye'deki Hastanelerden Izole Edilen Metisiline Dirençli Staphylococcus aureus Izolatlari Arasinda En Yaygin spa Tipidir.

Staphylococcus aureus is one of the most frequent agents causing hospital infections. S.aureus has a great ability to adapt itself to variety of conditions and successful clones can be epidemic and even pandemic by its ability spread from one continent to another. The aims of this study were to detect spa types of 397 methicillin-resistant S.aureus (MRSA) strains isolated from 12 centers in different geographical regions of Turkey from 2006 to 2008, and to investigate their clonality by PFGE and MLST typing. Additionally, 91 MRSA from four of those 12 centers isolated during 2011 were also studied for their spa types. PFGE profiles indicated the presence of a major pulsotype, namely pulsotype A with a rate of 91.4% (363/397), followed by pulsotype B (n= 18, 4.5%) and pulsotype C (n= 11, 2.8%). Among isolates tested 363 (91.4%) were SCCmec type III, 30 (7.6%) were SCCmec type IV. Sequence analysis of representative isolates revealed that ST239 (85.1%) was the most common MLST type followed by two MLST types ST737 (4%), and ST97 (2.8%), both SCCmec type IV. Two isolates were ST80 with SCCmec type IV. Of 397 isolates, 338 (85.1%) were t030, followed by t005 (2.5%) and t632 (2%). Among MRSA isolated during 2011, 64 (70.3%) of 91 were t030, 4 (4.4%) were t005, 2 (2.2%) were t015, and 2 (2.2%) were t1094. Among centers the t030 prevalence of 2006-2008 isolates ranged from 59-100%. The highest t030 prevalence was found in Ankara (100%) and lowest in Trabzon (59%) provinces which are located at central and northestern Anatolia, respectively. In Istanbul province, the prevalence of t030 was 94.5% among 2006-2008 isolates which decreased to 55.5% among 2011 isolates. Also a decrease in t030 rates was observed among samples from Konya and Trabzon but not from Aydin. Our results showed that the most common MRSA clone in Turkey is ST 239-SCCmec type III, t030 which persisted during the six years of the study period. Presence of PVL toxin gene was tested by PCR and 5 (3%) isolates found to be positive, of them two were SCCmec Type IV-ST80 and three were SCCmec Type III-ST239. This study is the largest epidemiological survey ever done in Turkey which showed presence of a hospital Turkish clone TR09 (ST239-SCCmecIII-t030) and a community clone TR10 (ST737-SCCmecIV-t005) largely disseminated in Turkey.

Infection and autoimmune disease.

Most infectious agents, such as viruses, bacteria and parasites, can trigger autoimmunity via different mechanisms. The development of an autoimmune disorder after infection tends to occur in genetically susceptible individuals. Some parameters, such as genetic predisposition, feature of the infectious agent and sometimes protective effect of the infections, have a significant role in this process. These parameters and various pathogens that could lead to enhancement or exacerbation of autoimmune disease were examined in this review. Recent studies were reviewed from a microbiological perspective.

[Linezolid-resistant Enterococcus faecium: the first G2576T mutation in Turkey]. / Linezolide Dirençli Enterococcus faecium: Türkiye'deki Ilk G2576T Mutasyonu.

Linezolid which is the first member of oxazolidinone class of synthetic antimicrobial agents, was licensed for the treatment of gram-positive coccal infections in Turkey in 2006. In recent years, multidrug-resistant pathogens, especially vancomycin-resistant enterococci (VRE), have emerged rapidly worldwide and linezolid exhibited good clinical efficacy against VRE. However, linezolid-resistant bacteria have been reported from Turkey. In April 2011, a 66-year-old paraplegic woman was admitted to our hospital because of an infected decubitis ulcer and empirical antibiotic therapy was started. Since the patient's condition worsened during treatment, she was moved to the intensive care unit. Klebsiella pneumoniae, methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium were recovered from tracheal aspirates and blood, respectively. Following three weeks of linezolid therapy blood culture yielded linezolid and vancomycin resistant E.faecium. Linezolid resistance in the VRE strain was confirmed by polymerase chain reaction and sequence analysis, subsequently. Linezolid-resistant two isolates were identified as E.faecium by 16S rRNA sequencing and both isolates had G2576U mutation in 23S rRNA gene. Linezolid resistance which was identified in a vancomycin-resistant E.faecium isolate is a new problem for Turkey. Last year another mutation related to linezolid resistance was reported from Istanbul, Turkey. The isolate had G2576T mutated 23S rRNA genes. Resistance should be considered and closely followed-up during linezolid treatment.

[In vitro activity of daptomycin against methicillin-resistant Staphylococcus aureus strains isolated from blood cultures]. / Daptomisinin Kan Kültürlerinden Izole Edilen Metisiline Dirençli Staphylococcus aureus Suslarina In Vitro Etkinligi.

The aim of this study was to detect the in vitro activity of daptomycin against methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from blood cultures at Ataturk Training and Research Hospital, Izmir, Turkey between 2006-2010. A total of 64 MRSA clinical isolates were included in the study, and daptomycin susceptibility were investigated by E-test (AB bioMerieux, Sweden). The identification of the MRSA isolates was based on conventional microbiological methods and an additional automated identification system (Phoenix 100, BD Diagnostic Systems, USA). Etest strips were applied to the surface of Mueller-Hinton agar plates and incubated at 35°C in ambient air for 18 to 24 hours. Strains with a MIC value of ≤ 1 µg/ml were accepted as susceptible to daptomycin. In our study all of the 64 MRSA isolates were found susceptible to daptomycin (MIC ≤ 1 µg/ml). The MIC50, MIC90 and MIC ranges were detected as 0.125, 0.5 and 0.125-0.5 µg/ml, respectively. Only a single isolate yielded MIC value of 1 µg/ml. As a result daptomycin was found to be very active against MRSA strains in vitro. Our findings suggested that daptomycin might be a suitable alternative agent for treating bacteremia caused by MRSA. However, further large-scaled studies and clinical trials are necessary to support these in vitro data.

Antinuclear antibodies (ANAs) detected by indirect immunofluorescence (IIF) method in acute COVID-19 infection; future roadmap for laboratory diagnosis.

INTRODUCTION: As in other viral infections, anti-nuclear antibodies (ANAs) are observed in SARS-CoV-2 infection. We investigated the presence of autoantibodies in acute COVID-19 and the association with early laboratory findings. MATERIALS AND METHODS: We examined 50 sera (>18 years, 25 Female) from patients with acute COVID-19. ANAs (HEp-20-10 liver biochip), anti-neutrophil cytoplasmic antibody (ANCA, Europlus Granulocyte Mosaic 32) and anti-double stranded DNA were investigated with product of Euroimmune AG (Luebeck, Germany) by indirect immunofluorescence (IIF) method. Also, antibody against cyclic citrullinated peptide (anti-CCP) was examined by a chemiluminisens assay (Euroimmun AG, Luebeck, Germany). Samples from 50 blood bank donors collected before the COVID-19 pandemic were used as controls. RESULTS: The IIF-ANA test was positive in 18% (N = 9/50) of the patients. The median time of sample collection was 7 days (range: 1-28 days) after diagnosis. ANA was positive in only one (2%) control sample. Five (55.5%) patients were ANA positive with a strong titer (3+). There was no relationship between antibody titration and time of sample collection (p = 0,55). Anti-CCP was detected in a nucleolar (3+) positive patient (2%). ANA was detected in 14.28% (N = 1/7, rods-rings (±), p = 0,78) of patients in the intensive care unit(ICU). Patients treated in the clinic have more and higher titers of ANA, mostly in nucleolar patterns, than ICU patients. CONCLUSIONS: The variety of antibodies detected in acute COVID-19 and the uncertainty of how long they persist can lead to confusion, especially in the diagnosis of systemic autoimmune rheumatic diseases for IIF-ANA testing in immunology laboratories. Improvements in cell lines and methods will facilitate the diagnostic process.

Comparison of various surgical methods in the treatment of implant-related infection.

The treatment of implant-related infections is troublesome. This study was conducted to compare the effectiveness of three different surgical modalities in the treatment of implant-related infection. A total of 32 Wistar albino rats were randomised into four groups after the establishment of implant-related infection: no treatment, surgical débridement, antibiotic-loaded bone cement and antibiotic-loaded autogenous bone. Microbiological colony counts were made at the sixth week in order to evaluate the effectiveness of of the treatments. The antibiotic-loaded bone cement group revealed superior results compared with the other groups in terms of reduction of microbiological colonies. Three animals in the bone cement group revealed extensive infection. Although antibiotic-loaded bone cement showed superiority over other treatment modalities, it should be employed after an unsuccessful trial of débridement because of the risk of extensive infection.

Adiponectin as a biomarker of systemic inflammatory response in smoker patients with stable and exacerbation phases of chronic obstructive pulmonary disease.

BACKGROUND AND OBJECTIVE: Adiponectin is an adipose tissue-derived specific protein that has a role in energy homeostasis, that has a protective role against the development of insulin resistance and atherosclerosis and that exhibits anti-inflammatory properties. We investigated serum adiponectin as a biomarker of systemic inflammatory response and its relation with leptin, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and nitric oxide (NO) in chronic obstructive pulmonary disease (COPD) patients. MATERIAL AND METHODS: We studied 36 male patients with COPD (15 stable and 21 exacerbated) and 17 age and sex-matched healthy subjects. The adiponectin and leptin levels were measured by enzyme-linked immunosorbent assay. Serum CRP levels were measured using the nephelometric method. ESR was determined using the Westergren method and NO by the cadmium reduction method. RESULTS: Adiponectin levels in COPD patients were significantly higher than those in control subjects (p<0.001), whereas there were no differences in leptin or NO levels. Serum levels of CRP, ESR and adiponectin were significantly higher in the exacerbated COPD patients compared to the stable group (p<0.001, p = 0.033 and p = 0.024, respectively), whereas the differences in leptin and NO levels were not significant. Serum levels of adiponectin were not correlated with FEV(1), FEV(1)/FVC, dyspnoea score, BMI or other inflammatory parameters in the stable COPD group. CRP and ESR correlated negatively with FEV(1) in the stable COPD group. CONCLUSIONS: Adiponectin may be a marker of low-grade systemic inflammatory response in COPD. A further rise in serum adiponectin in the exacerbation period denotes that this may also be a biomarker of the exacerbation phase as well as CRP and ESR.

[Evaluation of a rapid trehalase test for the identification of Candida glabrata]. / Candida glabrata'nin tanisinda hizli trehalaz testinin degerlendirilmesi.

Candida species which cause local infections, may also lead to fatal systemic infections. The increasing incidence of non-albicans Candida, especially fluconazole susceptible or resistant dose-dependent C. glabrata, increased the importance of rapid and accurate species level identification for Candida. Rapid and correct identification of C. glabrata is essential for the initiation of the appropriate antifungal therapy. This study was conducted to evaluate the performance of the rapid trehalase test in the diagnosis of C. glabrata isolates. A total of 173 Candida strains isolated from various clinical specimens and identified according to germ tube test, growth on cornmeal Tween 80 agar and the colony morphologies on Mast-CHROMagar Candida medium (Mast Diagnostics, UK), were included to the study. The identification of non-albicans Candida species were also confirmed by API 20CAUX (BioMerieux, France) system. Accordingly 86 (50%) of the isolates were identified as C. glabrata, 48 (28%) C. albicans, 17 (10%) C. krusei, 13 (8%) C. tropicalis, 5 (3%) C. parapsilosis, 3 (2%) C. kefyr and 1 (1%) Cutilis. In order to detect the presence of trehalase enzyme in Condida strains, all isolates were grown on Sabouraud dextrose agar containing 4% glucose and then one yeast colony was emulsified in 50 microl of citrate buffer containing 4% (wt/vol) trehalose for 3 h at 37 degrees C. Presence of glucose which emerged after the action of trehalase on trehalose, was detected by a commercial "urinary glucose detection dipstick" (Spinreacta, Spain). All C. glabrata strains yielded positive result by trehalase test. None C. glabrata isolates were found negative by trehalase test except for one strain of C. tropicalis. In this study, the trehalase test allowed identification of C. globrata with 100% sensitivity and 98.9% specificity. It was concluded that trehalase test is a rapid, cost-effective and simple test that can be used for the accurate identification of C. glabrata.

[Investigation of the relationship between serum nitric oxide levels, HBV-DNA and ALT levels in chronic hepatitis B patients]. / Kronik hepatit B hastalarinda serum nitrik oksit düzeyi ile HBV-DNA ve ALT düzeyleri arasindaki iliskinin arastirilmasi.

It has been reported that increased nitric oxide (NO) production by the hepatocytes during chronic inflammatory processes, plays an important role in the pathogenesis of chronic hepatitis B. The aim of this study was to investigate the relationship between serum levels of NOx (nitrite + nitrate) with the viral load and alanine aminotransferase (ALT) levels in chronic hepatitis B (CHB) patients. A total of 93 CHB patients (67 male, 26 female; mean age: 47.3 +/- 10.9 years) and 53 healthy control subjects (17 male, 36 female; mean age: 58.6 +/- 2.1 years) followed-up during 2006-2007 period were included to the study. Hepatitis B virus (HBV) serologic markers, viral load and ALT levels were studied by chemiluminescence method (Ortho-Clinical Diagnostics, USA), by real-time polimerase chain reaction (PCR) (ABI PRISM 7700, Applied Biosystem, CA), and by Aeroset System (Abbott Laboratories, USA), respectively. NOx levels were determined by a method which was based on the reduction of nitrate to nitrite by cadmium. Mean levels of ALT and HBV-DNA of the patients were found as 98.7 +/- 138.4 IU/I and 1.6 x 10(9) +/- 4.0 x 10(9) copies/ml, respectively. In the evaluation of mean levels of NOx in patient and control groups, the difference was found statistically significant (30.6 +/- 21.7 micromol/l and 23.7 +/- 5.2 micromol/l, respectively; p< 0.05). In view of the relationship between the parameters, a positive correlation was detected between viral load and ALT levels (r= 0.768; p< 0.001), besides the significant correlations between NOx and viral load, and NOx and ALT (r= 0.346, p= 0.001 and r= 0.314, p= 0.002, respectively). As a result, although the NOx levels in chronic hepatitis patients were found higher than those in the control group, and significant correlations were detected between NO, viral load and ALT, the exact role of NO in the disease pathogenesis and outcome needs to be studied further at cellular level.

Investigation of anti-DFS70 antibody in patients with systemic autoimmune rheumatic diseases.

INTRODUCTION: Dense fine-speckled 70 (DFS70) antibody is defined as an antinuclear antibody (ANA) pattern in indirect immunofluorescence (IIF). The presence of anti-DFS70 antibody has been shown as a potential marker for the exclusion of systemic autoimmune rheumatic diseases (SARD) (without any other SARD-associated autoantibodies). We aimed to investigate the frequency of anti-DFS70 antibodies in patients with SARD and in the blood bank donors (BD). MATERIALS AND METHODS: The study group consists of 418 rheumatoid arthritis (RA), 101 systemic lupus erythematosus (SLE), 71 Sjogren's syndrome (SS), 43 ankylosing spondylitis (AS), 36 systemic sclerosis-scleroderma (SSc), 2555 undifferentiated connective tissue disease (UCTD), and 507 BD. All samples were tested on the HEp-2 IIF-ANA assay. Samples that showed DFS70 pattern in IIF were confirmed by a specific DFS70 antibody enzyme-linked immunosorbent assay (ELISA). RESULTS: The DFS70 pattern was detected in 43 (1.33%) in SARD and four (0.78%) in BD. The anti-DFS70 antibody was detected in three (0.59%) in BD, six (1.43%) in RA, three (2.97%) in SLE, one (1.40%) in SS, and 25 (0.97%) in UCTD, however, it was not detected in AS and SSc by ELISA. There was no significant difference between BD and SARD (p = 0.28). Distinctly, the frequency of anti-DFS70 was significantly different for SLE in SARD (p = 0.02). CONCLUSION: Anti-DFS70 antibody was more prevalent in the subsets of SARD than BD. This result may be related to the demographic formation of study groups and individual immunological status. More comprehensive studies are needed to investigate the importance of the anti-DFS70 antibody for SARD.Key Points• This study draws attention to the importance of anti-DFS70 antibodies in the diagnostic algorithm in systemic autoimmune rheumatic diseases.• This study emphasizes the further investigation of anti-DFS70 antibodies in undifferentiated connective tissue diseases.• This study emphasizes the need to verify the DFS70 pattern detected in IIF-ANA test for definitive diagnosis with additional confirmation methods.

A Multicenter Analysis of Subjectivity of Indirect Immunofluorescence Test in Antinuclear Antibody Screening.

Objectives: This study aims to evaluate the interpretation of the antinuclear antibody (ANA)-indirect immunofluorescence (IIF) test results based on the interpreter-related subjectivity and to examine the inter-center agreement rates with the performance of each laboratory. Patients and methods: The ANA-IIF testing was carried out in a total of 600 sera and evaluated by four laboratories. The inter-center agreement rates were detected. The same results given by the four centers were accepted as gold standard and the predictive values of each center were calculated. Results: The inter-center agreement was reported for ANA-IIF test results from 392 of 600 (65.3%) sera, while 154 of 392 results were positive. Four study centers reported 213 (35.5%), 222 (37.0%), 266 (44.3%), and 361 (60.2%) positive test results, respectively. In terms of the patterns, the highest and lowest positive predictive values were 72.3% and 42.7%, respectively, while the highest and lowest negative predictive values were 99.6% and 61.5%, respectively. The agreement for semi-quantitative evaluation at three levels of fluorescence intensity stated by four centers was detected in 100 sera at 87% 3(+), while the other two levels were 6% and 7%. The highest predictive value for the highest fluorescence intensity of 3(+) was found to be 71.9%. Conclusion: Significant differences may be observed among laboratories in terms of qualitative results, patterns, and semi-quantitative determination of the fluorescence intensity in the ANA-IIF testing, particularly at low fluorescence intensity levels and in those with speckled patterns. In case of any discrepancy between ANA-IIF test and clinical prediagnosis, the test should be repeated in another laboratory, if necessary.

Evaluation of the results of Mycobacterium tuberculosis direct test (MTD) and Mycobacterial culture in urine samples.

Tuberculosis remains a public health problem in Turkey. Rapid detection of Mycobacterium tuberculosis plays a key role in control of infection. In this article, the Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD) was evaluated for detection of M. tuberculosis in urine samples. The performance of the MTD was very good and appropriate for routine laboratory diagnosis.

[In vitro evaluation of gentamicin and teicoplanin release from cancellous human bone]. / Insan süngerimsi kemiginden gentamisin ve teikoplanin saliniminin in vitro degerlendirilmesi.

OBJECTIVES: This study was designed to determine the in vitro antibacterial activity of gentamicin- or teicoplanin-impregnated human cancellous bone as a local antibiotic carrier. METHODS: The study samples were obtained from human cancellous bone within the femur head in seven patients who underwent partial or total hip arthroplasty. Bone specimens were processed and incubated with gentamicin or teicoplanin for an hour. Control bone specimens were soaked in sterile saline solution for the same duration. Antibiotic release of bone specimens was assessed by the disc diffusion technique after 1, 3, 7, 10, 14. 18, and 21 days of antibiotic impregnation, with seven samples in each group. The test strains were E. coli ATCC 25922 for gentamicin, and S. aureus ATCC 25923 for teicoplanin. In vitro antibiotic efficacy was defined as an inhibition zone diameter of = or >15 mm for gentamicin, and = or >14 mm for teicoplanin. RESULTS: Evaluation of inhibition zone diameters showed that bone-teicoplanin complexes had a longer duration of antibiotic release than that of bone-gentamicin complexes (12 to 18 days vs 7 to 10 days). There was no inhibition in the control group. There were no significant differences in inhibition zone diameters of teicoplanin- and gentamicin-treated specimens on the first and third days; however, teicoplanin exhibited significantly greater zone diameters on the seventh (p=0.008) and tenth (p=0.003) days. CONCLUSION: Our data show that, under appropriate conditions, human cancellous bone incorporates a considerable amount of teicoplanin and exhibits effective antibiotic release for approximately two weeks.