Antimicrobial activity of Stachys rupestris Montbret et Aucher ex Benth. and inactivation of the pathogens inoculated on lab-made skin by the essential oil.

The aim of this study was to investigate the antimicrobial activity of Stachys rupestris essential oil and inactivation of the pathogens on lab-made skin the oil in the fight against Escherichia coli, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus faecalis, Candida albicans and Candida tropicalis. S. rupestris EO was extracted by hydrodistillation, and its contents were analyzed by GCMS. Logarithmic reduction of the pathogens inoculated on the artificial skin surface by S. rupestris EO was studied for the first time. The highest inhibition zone was 22.1 mm on C. tropicalis, while the lowest IZ was 0.1 mm on E. coli. The other zones were 20.01 mm for Acinetobacter baumannii, 20.02 mm for Enterococcus faecalis, 20.01 mm for Staphylococcus aureus, 22.03 for Candida albicans (p < 0.05). As a result, S. rupestris essential oil was effective on most of the microorganisms and might be increased to use in the treatment of skin infections in the future.

Possible relationship between IL28B rs12979860 and TLR2 -196 to -174 del/ins polymorphisms and the liver fibrosis stage in hepatitis C patients.

It has been shown that host factors play an important role in the progression of hepatitis C virus (HCV) infection. Toll-like receptor 2 (TLR2) del and interleukin 28B (IL28B) T alleles can mediate liver inflammation and pathogenesis of hepatocellular carcinoma. In the present study, the possible relationship between the IL28B rs12979860 C/T and TLR2 -196 to -174 del/ins gene variants and different fibrosis stages and host factors in hepatitis C patients was investigated. IL28B and TLR2 polymorphisms in the blood of 50 hepatitis C patients at different stages of fibrosis (24 mild/moderate, 26 advanced) and 24 healthy controls were examined by RT-qPCR. The highest frequency of the TLR2 del (26.9%) and IL28B T (46.2%) alleles was found in hepatitis C patients with the most advanced fibrosis, and the lowest frequency was found in healthy controls. There was a statistically significant difference between hepatitis C patients with advanced fibrosis and healthy controls in terms of the TLR2 del (p = 0.0062) and IL28B T (p = 0.0017) allele frequencies. However, no statistically significant difference was found between the mild/moderate fibrosis and severe fibrosis patient groups in terms of genotype or IL28B and TLR2 polymorphisms (p > 0.05). In addition, there was a significant difference between patients with mild/moderate or advanced fibrosis who carried the TLR2 del allele together with the IL28B CT genotype and healthy controls. The present study emphasizes that the TLR2 and IL28B gene variants cannot be single biomarkers for the determination of fibrosis stage in hepatitis C infection but together can play an important role in predicting severe disease.

hsa-miR-17-5p: A Possible Predictor of Ombitasvir/Paritaprevir/Ritonavir + Dasabuvir ± Ribavirin Therapy Efficacy in Hepatitis C Infection.

Although persistent sustained viral response rates are increased in hepatitis C infection following administration of direct-acting antiviral (DAA) agents, the pre-use predictive parameters of these antivirals and the clinical progression in patients post-treatment remain unknown. To obtain data pertaining to the predictive parameters prior to the use of ombitavir/paritaprevir/ritonavir + dasabuvir and the clinical progression in patients following antiviral treatment. The expression profiles of miR-223-3p, miR-17-5p, miR-24-3p, and TLR2 - 196 to - 174 del/ins polymorphisms from the blood/serum of 34 hepatitis C virus (HCV)-infected patients pre- and post-ombitavir/paritaprevir/ritonavir + dasabuvir treatment were determined by RT-qPCR. The expression levels of miR-17-5p (P < 0.001) and miR-24-3p (P = 0.011) were significantly downregulated post-treatment as compared with those pre-treatment; however, there was no significant difference between these two groups in terms of miR-223-3p expression. In addition, there was no significant difference in TLR2 genotype or allele distribution between pre-and post-treatment (P > 0.05); nevertheless, the TLR2 del allele was decreased post-treatment (16.2%) as compared with that pre-treatment (19.1%), although the difference was not statistically significant. Moreover, a significant difference was found between the mRNA levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and HCV RNA pre-and post-treatment (P < 0.05). Further, miR-17-5p expression correlated with both ALT and AST mRNA levels post-treatment (P.

[Determination of reverse transcriptase inhibitor nucleoside analogue resistance profile in pretreatment phase of patients with viral hepatitis B]. / Viral hepatit B hastalarinda ilk tedavi öncesi revers transkriptaz inhibitörü nükleozid analogu ilaç direnç profilinin belirlenmesi.

HBV is a DNA virus and the causative agent of hepatitis B infection. Hepatitis B is a contagious disease and is still a major health problem all over the world. When the infection become chronic, it may cause serious diseases such as fibrosis, cirrhosis and/or hepatocellular carcinoma. Interferon/pegylated interferon by intravenous route and nucleoside/nucleotide (NA) analogues such as lamivudine, adefovir, entecavir, telbivudine and tenofovir given by oral route are used in the treatment. Antivirals given by oral route are mostly preferred in the treatment. However, because of the replication strategy and biological properties of HBV, mutations that cause antiviral resistance against these drugs can occur at different rates, although they can vary from drug to drug over time. It is possible that drug resistant virus may transmit from patient to healthy individuals. Therefore, there is a possibility of infection with drug-resistant HBV before treatment. Antiviral resistance mutations are divided into four categories; i) Nucleos(t)ide analog resistance (NAr)-related mutations, ii) primary drug resistance mutations, iii) secondary/compensatory mutations, iv) putative antiviral resistance mutations and pre-treatment variations. Recent studies have focused particularly on putative mutations and pre-treatment variations. The aim of this study was to better understanding of the antiviral resistance profiles of chronic hepatitis B (CHB) patients treated and untreated with NA, and help to prevent unnecessary drug use, minimize the side effects and economic damages. A total of 124 patients who have received nucleoside analog (NA) drug treatments (n= 72) and patients without NA treatment (n= 52) were included in the study. Viral DNA was isolated from the plasma samples of the patients. A DNA fragment, which is 551 bp, was amplified and sequenced including the binding side of all nucleoside analogs containing the B, C and D domains located in the reverse transcriptase region in the HBV genome. Different types of mutations were detected in 13 (18.05%) of 72 treated patients and in 18 (34.61%) of 52 untreated patients (p< 0.05). Primary drug resistance mutations such as rtI169T, rtA181T/V, rtT184A/C/F/G/I/L/M/S, rtA194T, rtS202C/G/I, rtM204I/V/S, rtN236T, rt M250I/L/V and rtV173L were not detected in any of the patient samples. However, potential drug resistance mutations such as rtR164R, rtG165D/A, rtG172Q, rtS176N, rtF178V, rtA181G, rtS185N/G/C, rtV207M, rtQ215H/S, rtL231V, rtI233K, rtN238S, rtV253T, rtC256G/S and rtI266R/V were detected in untreated patient samples in B, C, D and D domains of reverse transcriptase region. Our results have suggested that the detection of pretreatment variations could be helpful for choosing the correct antiviral drug for the better treatment management.

Serum microRNAs; miR-30c-5p, miR-223-3p, miR-302c-3p and miR-17-5p could be used as novel non-invasive biomarkers for HCV-positive cirrhosis and hepatocellular carcinoma.

Recently, serum miRNAs have been evolved as possible biomarkers for different diseases including hepatocellular carcinoma and other types of cancers. Investigating certain serum miRNAs as novel non-invasive markers for early detection of HCV-positive cirrhosis and hepatocellular carcinoma (HCC). The expression profiles of 58 miRNA were analyzed in patient's plasma of chronic hepatitis C (CHC), HCV-positive cirrhosis and HCV-positive HCC and compared with control group samples. Totally 94 plasma samples; 64 patient plasma (26 CHC, 30 HCV-positive cirrhosis, 8 HCV-positive HCC) and 28 control group plasma, were included. The expression profiles of 58 miRNAs were detected for all patient and control group plasma samples by qRT-PCR using BioMarkTM 96.96 Dynamic Array (Fluidigm Corporation) system. In CHC group, expression profiles of miR-30a-5p, miR-30c-5p, miR-206 and miR-302c-3p were found significantly deregulated (p < 0.05) when compared versus control group. In HCV-positive cirrhosis group, expression profiles of miR-30c-5p, miR-223-3p, miR-302c-3p, miR-17-5p, miR-130a-3p, miR-93-5p, miR-302c-5p and miR-223-3p were found significantly deregulated (p < 0.05). In HCV-positive HCC group, expression profiles of miR-17-5p, miR-223-3p and miR-24-3p were found significant (p < 0.05). When all groups were compared versus control, miR-30c-5p, miR-223-3p, miR-302c-3p and miR-17-5p were found significantly deregulated for cirrhosis and HCC. These results imply that miR-30c-5p, miR-223-3p, miR-302c-3p and miR-17-5p could be used as novel non-invasive biomarkers of HCV-positive HCC in very early, even at cirrhosis stage of liver disease.

[Characterization of basal core promoter/precore gene mutations in chronically infected patients with hepatitis B virus genotype D in Mersin Province, Turkey]. / Mersin ilinde hepatit B virus genotip D ile kronik enfekte hastalarda bazal kor promotor/prekor gen bölgesi mutasyonlarinin karakterizasyonu.

The basal core promoter (BCP) and precore (PC) gene regions of hepatitis B virus (HBV) genome are important for the viral replication and synthesis of "e" antigen. Genetic variability has been described in PCP and PC gene regions, commonly in HBeAg negative patients. The aim of this study was to determine the frequency of the predominant mutation patterns of BCP/PC gene regions and their correlations with HBeAg status, HBV-DNA levels, and liver biochemical profiles in chronic hepatitis B (CHB) patients infected with genotype D, in Mersin province which is located at Mediteranean part of Turkey. A total of 54 CHB patients (33 male, 21 female; mean age: 40.05±12.91 years) infected with HBV genotype D were enrolled in the study. Serum HBV-DNA levels, serological markers (HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc) and biochemical profiles (ALT and AST) were analyzed in all patients. BCP and PC gene regions were determined by polymerase chain reaction (PCR) and mutations of these regions were determined by direct sequencing of PCR products then aligned with known wild-type HBV sequences. BCP [nucleotide (nt.) 1753-1762/1764] and/or PC (nt. 1896) mutations were detected in 87.75% (43/49) of the patients. Mutation rates were detected as 97.1% (33/34) and 66.7% (10/15) in the HBeAg negative and in HBeAg positive patient groups, respectively (p=0.008). PC nt. G1896A mutation was more common in HBeAg negative samples than in HBeAg positive samples (73.5% vs. 20%, p=0.001), however there was no significant differences in the occurrence of BCP mutations between the two groups (p=0.331). No correlation was found between the presence of BCP and/or PC mutations and serum HBV-DNA or ALT-AST levels. Our study reveals that significant number of chronically infected patients with genotype D HBV have BCP and PC variants. G1896A stop codon mutation in precore region seems to have a significant role in the loss of HBeAg in our patients. The results of our study provided important data about the frequency and the genetic heterogeneity of different kinds of mutations occurring at BCP and PC gene regions.

Effects of 5,14-HEDGE, a 20-HETE mimetic, on lipopolysaccharide-induced changes in MyD88/TAK1/IKKß/IκB-α/NF-κB pathway and circulating miR-150, miR-223, and miR-297 levels in a rat model of septic shock.

OBJECTIVES: We have previously demonstrated that a stable synthetic analog of 20-hydroxyeicosatetraenoic acid (20-HETE), N-(20-hydroxyeicosa-5[Z],14[Z]-dienoyl)glycine (5,14-HEDGE), which mimics the effects of endogenously produced 20-HETE, prevents vascular hyporeactivity, hypotension, tachycardia, inflammation, and mortality in a rodent model of septic shock. The present study was performed to determine whether decreased renal and cardiovascular expression and activity of myeloid differentiation factor 88 (MyD88)/transforming growth factor-activated kinase 1 (TAK1)/inhibitor of κB (IκB) kinase ß (IKKß)/IκB-α/nuclear factor-κB (NF-κB) pathway and reduced circulating microRNA (miR)-150, miR-223, and miR-297 expression levels participate in the protective effect of 5,14-HEDGE against hypotension, tachycardia, and inflammation in response to systemic administration of lipopolysaccharide (LPS). METHODS: Conscious male Wistar rats received saline (4 ml/kg) or LPS (10 mg/kg) at time 0. Blood pressure and heart rate were measured using a tail-cuff device. Separate groups of LPS-treated rats were given 5,14-HEDGE (30 mg/kg) 1 h after injection of saline or LPS. The rats were killed 4 h after LPS challenge and blood, kidney, heart, thoracic aorta, and superior mesenteric artery were collected for measurement of the protein expression. RESULTS: LPS-induced fall in blood pressure and rise in heart rate were associated with increased MyD88 expression and phosphorylation of TAK1 and IκB-α in cytosolic fractions of the tissues. LPS also caused an increase in both unphosphorylated and phosphorylated NF-κB p65 proteins in the cytosolic and nuclear fractions as well as nuclear translocation of NF-κB p65. In addition, serum miR-150, miR-223, and miR-297 expression levels were increased in LPS-treated rats. These effects of LPS were prevented by 5,14-HEDGE. CONCLUSIONS: These results suggest that downregulation of MyD88/TAK1/IKKß/IκB-α/NF-κB pathway as well as decreased circulating miR-150, miR-223, and miR-297 expression levels participate in the protective effect of 5,14-HEDGE against hypotension, tachycardia, and inflammation in the rat model of septic shock.

Profiles of serum microRNAs; miR-125b-5p and miR223-3p serve as novel biomarkers for HBV-positive hepatocellular carcinoma.

Recently, circulating miRNAs have been reported as promising biomarkers for various pathologic conditions including cancer. Certain microRNAs (miRNAs) have been shown early diagnostic potential for many types of cancer. The objective of this study was to investigate the potential of certain serum/plasma miRNAs as novel non-invasive biomarkers for early diagnosis of hepatitis B virus (HBV) related hepatocellular carcinoma (HCC). For this reason, the expression levels of 24 miRNA (let-7c, miR-92a-3p, 423-5p, 150-5p, 223-3p, 125b-5p, 342-3p, miR-206, 122-5p, 375, 223-5p, 10a-5p, 23b-5p, 99a-5p, 23a-5p, 10a-3p, 122-3p, 125b-1-3p, 23b-3p, 125b-2-3p, 23a-3p, 92a-1-5p, 92a-2-5p, 99a-3p) were analyzed in plasma of patients with chronic hepatitis B, HBV-positive cirrhosis and HBV-positive HCC and compared with control group samples. Totally 94 plasma samples; 28 control and 66 patient plasma (24 CHB, 22 HBV-positive cirrhosis, 20 HBV-positive HCC) and were included in this study. The expression levels of 24 miRNAs were detected for all control and patient group plasma samples by qRT-PCR using BioMark™ 96.96 Dynamic Array (Fluidigm Corporation) system. The expression levels of miR-125b-5p were detected 2.85 fold, 2.46 fold and 1.89 fold (p = 0.01513, p = 0.0009440, p = 0.0001446) up regulated in CHB, HBV-positive cirrhosis and HBV-positive HCC, respectively when compared versus control group individually by Mann-Whitney U test. The expression levels of miR-223-3p were detected 5.55 fold, 13.88 fold and 12.65 fold (p = 0.01513, p = 0.0009440, p = 0.0001446) down regulated in same comparisons. When all groups were compared versus control group by one-way ANOVA test, the expression levels of miR-223-3p were also found statistically significant (p < 0.05). Although not statistically significant, miR-125b-5p tended to be upregulated. (p = 0.07192). These results significantly imply that miR-125b-5p and miR223-3p could be used as novel non-invasive biomarkers of HBV-positive HCC in very early, even at CHB stage of liver disease.

Contribution of iNOS/sGC/PKG pathway, COX-2, CYP4A1, and gp91(phox) to the protective effect of 5,14-HEDGE, a 20-HETE mimetic, against vasodilation, hypotension, tachycardia, and inflammation in a rat model of septic shock.

We have previously demonstrated that a stable synthetic analog of 20-hydroxyeicosatetraenoic acid (20-HETE), N-[20-hydroxyeicosa-5(Z),14(Z)-dienoyl]glycine (5,14-HEDGE), prevents vascular hyporeactivity, hypotension, tachycardia, and inflammation in rats treated with lipopolysaccharide (LPS) and mortality in endotoxemic mice. These changes were attributed to decreased production of inducible nitric oxide (NO) synthase (iNOS)-derived NO, cyclooxygenase (COX)-2-derived vasodilator prostanoids, and proinflammatory mediators associated with increased cyctochrome P450 (CYP) 4A1-derived 20-HETE and CYP2C23-dependent antiinflammatory mediator formation. The aim of this study was to determine whether decreased expression and activity of iNOS, soluble guanylyl cyclase (sGC), protein kinase G (PKG), COX-2, gp91(phox) (NOX2; a superoxide generating NOX enzyme), and peroxynitrite production associated with increased expression of COX-1 and CYP4A1 and 20-HETE formation in renal and cardiovascular tissues of rats contributes to the effect of 5,14-HEDGE to prevent vasodilation, hypotension, tachycardia, and inflammation in response to systemic administration of LPS. Mean arterial pressure fell by 28mmHg and heart rate rose by 47beats/min in LPS (10mg/kg, i.p.)-treated rats. Administration of LPS also increased mRNA and protein expression of iNOS and COX-2 associated with a decrease in COX-1 and CYP4A1 mRNA and protein expression. Increased NOS activity, iNOS-heat shock protein 90 complex formation (an index for iNOS activity), protein expression of phosphorylated vasodilator stimulated phosphoprotein (an index for PKG activity), gp91(phox), p47(phox) (NOXO2; organizer subunit of gp91(phox)), and nitrotyrosine (an index for peroxynitrite production) as well as cGMP (an index for sGC activity), 6-keto-PGF1α (a stable metabolite PGI2) and PGE2 levels (indexes for COX activity), and nitrotyrosine levels by LPS were also associated with decreased CYP hydroxylase activity as measured by 20-HETE formation from arachidonic acid in renal microsomes of LPS-treated rats. These effects of LPS, except iNOS mRNA and COX-1 protein expression, were prevented by 5,14-HEDGE (30mg/kg, s.c.; 1h after LPS). A competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid (30mg/kg, s.c.; 1h after LPS) reversed the effects of 5,14-HEDGE, except iNOS and COX-1 mRNA and protein expression as well as expression of CYP4A1 mRNA. These results suggest that increased CYP4A1 expression and 20-HETE formation associated with suppression of iNOS/sGC/PKG pathway, COX-2, and gp91(phox) participate in the protective effect of 5,14-HEDGE against vasodilation, hypotension, tachycardia, and inflammation in the rat model of septic shock.

Dishwashers are a major source of human opportunistic yeast-like fungi in indoor environments in Mersin, Turkey.

The natural habitat of opportunistic fungal pathogens is outside of the host; therefore, it is crucial to understand their ecology and routes of transmission. In this study, we investigated the presence of black and filamentous fungi in moist indoor environments in the city of Mersin in subtropical Turkey. In total, 177 private dwellings were screened and 893 samples obtained using cotton swabs and moistened with physiological saline from dishwashers, washing machines, refrigerators, bath-tubs, bathroom walls, and shower heads. These were then inoculated onto malt extract agar supplemented with chloramphenicol, followed by incubation at 37°C. Thirty samples (3.4%) were positive for fungi, which were then identified by sequencing the rDNA internal transcribed spacer region. Exophiala dermatitidis was the most common species (23), followed by E. phaeomuriformis (three), Magnusiomyces capitatus (two), and Candida parapsilosis (two). Genotype A of E. dermatitidis (14) was more prevalent than genotypes B (eight) and C (one) and E. phaeomuriformis was also represented by two genotypes. Our findings suggest that dishwashers are a major indoor niche for thermophilic black yeasts. The occurrence of the opportunistic filamentous fungus M. capitatus in dishwashers is consistent with a recent report.

5,14-HEDGE, a 20-HETE mimetic, reverses hypotension and improves survival in a rodent model of septic shock: contribution of soluble epoxide hydrolase, CYP2C23, MEK1/ERK1/2/IKKß/IκB-α/NF-κB pathway, and proinflammatory cytokine formation.

We have previously demonstrated that a stable synthetic analog of 20-HETE, N-[20-hydroxyeicosa-5(Z),14(Z)-dienoyl]glycine (5,14-HEDGE), restores vascular reactivity, blood pressure, and heart rate in endotoxemic rats. The aim of this study was to determine whether decreased renal expression and activity of soluble epoxide hydrolase (sEH), MEK1, ERK1/2, IKKß, IκB-α, and NF-κB as well as systemic and renal proinflammatory cytokine production associated with increased expression and activity of CYP2C23 contributes to the effect of 5,14-HEDGE to prevent hypotension, tachycardia, inflammation, and mortality in response to systemic administration of lipopolysaccharide (LPS). Blood pressure fell by 33 mmHg and heart rate rose by 57 beats/min in LPS (10 mg/kg, i.p.)-treated rats. Administration of LPS also increased mRNA and protein expression of sEH associated with a decrease in CYP2C23 mRNA and protein expression. Increased activity of sEH and p-MEK1, p-ERK1/2, p-IκB-α, NF-κB, and p-NF-κB protein levels as well as TNF-α and IL-8 production by LPS were also associated with a decreased activity of AA epoxygenases. These effects of LPS were prevented by 5,14-HEDGE (30 mg/kg, s.c.; 1 h after LPS). Treatment of endotoxemic mice with 5,14-HEDGE also raised the survival rate of animals from 84% to 98%. A competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa-6(Z),15(Z)-dienoic acid, 20-HEDE (30 mg/kg, s.c.; 1 h after LPS) prevented the effects of 5,14-HEDGE on blood pressure, heart rate, expression and/or activity of sEH, CYP2C23, and ERK1/2 as well as TNF-α and IL-8 levels in rats treated with LPS. These results suggest that decreased expression and/or activity of sEH and MEK1/ERK1/2/IKKß/IκB-α/NF-κB pathway as well as proinflammatory cytokine production associated with increased CYP2C23 expression and antiinflammatory mediator formation participate in the protective effect of 5,14-HEDGE against hypotension, tachycardia, inflammation, and mortality in the rodent model of septic shock.

Determination of plasma microRNA for early detection of gastric cancer.

Gastric cancer is the fourth most prevalent malignancy worldwide and remains the second most common cause of cancer-related death globally. Understanding the molecular structure of gastric carcinogenesis might identify new diagnostic and therapeutic strategies for this disease. Thus, early detection of gastric cancer is a key measure to reduce the mortality and improve the prognosis of gastric cancer. There have recently been several reports that microRNAs (miRNAs) circulate in highly stable, cell-free forms in blood. Because serum and plasma miRNAs are relatively easy to access, circulating miRNAs also have great potential to serve as non-invasive biomarkers. Although a number of miRNAs associated with gastric cancer have been identified, the underlying mechanism of these miRNAs in tumorigenesis and tumor progression remains to be investigated. The purpose of this study is to identify the potential of serum miRNAs as biomarkers for early detection of gastric cancer patients. RNA was isolated using the High Pure miRNA Isolation Kit (Roche) following the manufacturer's protocol. cDNA and preamplification protocols were obtained from the isolated plasma miRNAs. The BioMark™ 96.96 Dynamic Array (Fluidigm Corporation) for real-time qPCR was used to simultaneously quantite the expression of 740 miRNAs. All statistical analyses were performed using the Biogazelle's qbase PLUS 2.0 software. In this study, among 740 miRNAs that we analyzed only miR-195-5p was significantly (p < 0.05, fold changes = 13, 3) down-regulated in gastric cancer patients compared with control. We demonstrated that miR-195-5p is a novel tumor suppressor miRNA and may contribute to gastric carcinogenesis. The miRNA expression profile described in this study should contribute to future studies on the role of miRNAs in gastric cancer.

Epidemiology of dermatophytosis in junior combat and non-combat sports participants.

Participation in competitive sports is popular and widely encouraged worldwide. Herein, we investigated 252 male and 67 female sports players, aged 16.4 ± 1.3 years, active in 15 different types of combat (n = 143) and non-combat (n = 176) sports. Of the 319 participants in this study, 11 (3.5%) players, including six wrestlers, four football players and one handball player, all of whom were men, harboured dermatophytic fungi. Briefly, Trichophyton tonsurans was present in three athletes, who were scalp carriers of the fungus. Furthermore, T. rubrum (4), T. interdigitale (3) and Arthroderma simii (1) were recovered from eight participants with tinea inguinalis (4), tinea pedis (2) or both (1). One patient was a trunk carrier of concomitant tinea pedis. All dermatophytic fungi were identified using both direction sequence of the rDNA regions spanning the internal transcribed spacers (ITS1 and ITS2) and 5.8 rRNA gene. Although sports-active individuals are active and sweat more, we observed a low prevalence of dermatophytosis, both in combat (5.2%) and non-combat sports participants (3.4%) (P > 0.05). However, dermatophyte infections require more attention and appropriate management to eradicate the infection and to prevent possible outbreaks. This study also documents the first case of zoophilic A. simii in Turkey.

Genotyping reveals no link between Candida albicans genotype and vaginitis severity in Turkish women.

Recent studies have clearly defined the vaginopathic Candida albicans strains that cause severe vulvovaginal candidiasis (VVC). Therefore, genotyping C. albicans isolates may predict the success of and assist in choosing the appropriate antifungal therapy. The purpose of this study was to compare the genotypes of C. albicans isolates causing VVC with those found in asymptomatic healthy pregnant and non-pregnant women in Adana, Turkey, as well as the antifungal susceptibility profiles of these isolates. A total of 216 independent C. albicans isolates were genotyped by allelic combination based on the microsatellite marker analysis of one such microsatellite, present in the promoter region of the elongation factor 3-encoding gene (CEF3) of C. albicans. The susceptibility testing profiles of all of the isolates against five antifungals and boric acid were obtained retrospectively from our laboratory records. We identified 20 genotypes on the basis of different allelic combinations at the CEF3 locus with a discriminatory power of 0.85. Genotypes 136-144 and 126-135 were present in 50 % of the isolates. No differences existed in the genotypic profiles of fungal isolates between pregnant and non-pregnant women. Remarkably, we did not find a single vaginopathic genotype. All of the isolates were susceptible to amphotericin B and 5-fluorocytosine, and the fluconazole and ketoconazole resistance rates were 0.9 and 3.7 %, respectively. Therefore, we did not find any correlation between genotype, severity of VVC, and antifungal resistance (P > 0.05). Even so, additional molecular data may provide new insights into the management of VVC.

[Investigation of the presence of human metapneumovirus in patients with chronic obstructive pulmonary disease and asthma and its relationship with the attacks]. / Kronik Obstrüktif Akciger Hastaligi ve Astimi Olan Hastalarda Insan Metapnömovirus Varliginin Arastirilmasi ve Hastalik Ataklari ile Iliskisi.

Human metapneumovirus (hMPV), an enveloped RNA virus classified in Paramyxoviridae family, was first characterized in 2001 from children with acute respiratory tract infection. Recent studies have suggested hMPV to play a role in chronic obstructive pulmonary disease (COPD) and asthma attacks. The aims of this study were to investigate the frequency of hMPV in patients with COPD and asthma, its effects on the severity of the attacks and the relationship between demographical and clinical factors. A total of 123 patients, including 66 with COPD (45 were in attack and 21 were stable) and 57 with asthma (33 were in attack and 24 were under control) diagnosed according to the criteria of Global Initiative for Chronic Obstructive Lung Disease and the Global Strategy for Asthma Management and Prevention, respectively, were included in the study. Nasopharyngeal lavage samples collected from all of the patients have been evaluated for the presence of hMPV-RNA by using a reverse transcriptase-polymerase chain reaction (RT-PCR) targeting F gene region of the virus. hMPV-RNA positivity rates in patients with COPD and asthma were observed as 30.3% (20/66) and 31.6% (18/57), respectively, and the difference between the groups were not statistically significant (p= 1.00). When patients were compared according to their disease status, hMPV was detected in 31.1% (14/45) of patients with COPD attack and 28.6% of stable patients (p> 0.05). These rates were found as 36.4% (12/33) and 25% (6/24) in patients with asthma attack and controlled asthma, respectively (p> 0.05). Although the virus detection rates in patients with COPD and asthma attacks (26/78; 33.3%) were higher than the patients with stable/controlled disease (12/45; 26.7%), the difference was not found as statistically significant (p= 0.57). The detection rate of hMPV-RNA was 26.1% in patients who can be treated at home and hospital without any need of intensive care and mechanical ventilation, while this rate was 36.4% in patients with COPD attack who require intensive care and mechanical ventilation (p= 0.67). Similarly, hMPV-RNA was detected more frequently in asthma patients with moderate and severe attacks (45%) than in patients with mild attacks (23.1%); however this difference was also not statistically significant (p= 0.28). No association of hMPV-RNA detection and demographical and clinical characteristics (age, gender, medical history, smoking status, allergy, COPD severity, asthma severity, the severity of attacks, using inhaled steroid, fever) of the patients could be demonstrated (p> 0.05), except the severity of the disease in patients with asthma (p= 0.02). In conclusion, further studies with large number of cases are needed to elucidate the role of hMPV in the occurrence and severity of COPD and asthma attacks.

[Genotyping of Vaginal Candida glabrata Isolates Using Microsatellite Marker Analysis and DNA Sequencing to Identify Mutations Associated with Antifungal Resistance]. / Vajinal Candida glabrata Izolatlarinin Mikrosatellit Gösterge Analizi ile Genotiplendirilmesi ve DNA Dizi Analizi ile Antifungal Direnç ile Iliskili Mutasyonlarin Belirlenmesi.

Vulvovaginal candidosis is the second most common cause of vaginitis (17-39%) after bacterial vaginosis (22-50%). Since the diagnosis of vulvovaginal candidosis mainly depends on clinical findings without mycologic confirmatory tests and treated empirically, the actual incidence rate of vulvovaginal candidosis is unknown. Approximately 70-90% of vulvovaginal candidosis cases are caused by Candida albicans, however the increasing incidence of C.glabrata infections and its reduced susceptibility to azole drug therapy have generated increasing attention. The epidemiology and population structure of vulvovaginal candidosis due to C.glabrata are poorly characterized. This study was aimed to genotype the C.glabrata strains isolated from vaginal samples in Cukurova region, Turkey by microsatellite markers, to investigate the antifungal susceptibility profiles of the strains and to determine the molecular mechanisms leading to phenotypical azole resistance. A total of 34 unrelated vaginal C.glabrata strains isolated from patients with acute (n= 11) and recurrent (n= 14) vulvovaginal candidosis, control group (n= 9) without vaginitis symptoms, and a reference strain of C.glabrata CBS 138 (ATCC 2001) were included in the study. These isolates were genotyped using multiple-locus variable number tandem repeat analysis of three microsatellite markers (RPM2, MTI, and Cg6). Analysis of microsatellite markers was performed by fragment size determination of RPM2, MTI, and Cg6 PCR products through capillary electrophoresis. For each of the evaluated strains, DNA sequence analysis was performed for one gene (CgERG11) and four loci (CgPDR1, NTM1, TRP1, and URA3) to detect mutations possibly associated with antifungal resistance in each strain. In vitro susceptibility profiles of the strains to 13 antifungals and boric acid were determined according to CLSI document M27-A3 to investigate possible relationships between detected mutations and phenotypic resistance. C.glabrata CBS 138 strain was found to be susceptible to all the antifungals tested, while one of (%2.9) 34 vaginal C.glabrata isolates was found to be dose-dependent susceptible to fluconazole, 13 (38.2%) to itraconazole and 3 (8.8%) to voriconazole. No resistant strain were detected in the study population. Only three isolates were found to be resistant to clotrimazole (8.8%), however no relationship was identified between the genotypes and phenotypic resistance (p> 0.05). Thirteen genotypes were detected by microsatellite marker analysis, with high discrimination power (DP= 0.877). As a result, microsatellite marker analysis was validated as a rapid, reliable method for genotyping C.glabrata strains with good, but not optimal discriminatory power. Further studies examining larger numbers of isolates are needed to verify possible relationships between mutations and phenotypic resistance.

Detection of hepatitis B virus X gene and PreC promoter mutations from chronic hepatitis B patients in the south of Turkey.

Hepatitis B virus (HBV) infection is a global health problem with more than 2 billion infected individuals. HBV infection leads to diverse outcomes ranging from acute to chronic hepatitis, which may result in severe complications as liver cirrhosis and hepatocellular carcinoma (HCC). HBV is one of the most important human DNA viruses having strong oncogenic potential. Recently, many studies have reported on HBV X gene and PreC promoter mutations associated with HCC. In order to detect the prevalence of HBx gene and PreC promoter mutations possibly related to HCC, we have analyzed sera samples collected from 61 patients with chronic hepatitis B. We have detected TI653 mutation in 1 of 61 (1,63%), A1896 mutation in 10 of 61 (16,39%), and T1762 - A1764 dual mutation in 4 of 61 (6,55%). T1653 and T1762- A1764 dual mutations were suggested significantly related to HCC in earlier reported studies. Our findings demonstrate that HBx gene and PreC promoter mutations related to HCC are present in our region and prospective clinical chord studies would be useful for better patient management and of early diagnosis of possible HCC cases.

Evaluation of Plasma miR-17-5p, miR-24-3p and miRNA-223-3p Profile of Hepatitis C Virus-Infected Patients after Treatment with Direct-Acting Antivirals.

The expression of miR-223-3p, miR-17-5p, and miR-24-3p was evaluated in hepatitis C virus (HCV) patient serum samples, collected before DAA treatment and after a sustained virological response (SVR). Fifty HCV patients were stratified based on their liver damage stages into three different subgroups (21 with chronic hepatitis-CH, 15 with cirrhosis, and 14 with hepatocellular carcinoma-HCC). Considering the entire HCV population, the miRNA expression levels were significantly downregulated after the SVR compared to pre-treatment ones (p < 0.05). Stratifying the patients based on liver damage, the post-SVR values of the three miRNAs were significantly downregulated compared to the pre-treatment levels for both cirrhosis and HCC patients. No significant differences emerged from the analysis of the CH group. To our knowledge, this is the first study to detail the behavior of miR-223-3p, miR-17-5p, and miR-24-3p levels in patients with HCV-related CH, cirrhosis, and HCC after DAA therapy. Our findings show that HCV-infected patients have different miRNA profiles before and after treatment with DAAs, strongly suggesting that miRNAs may be involved in the pathogenesis of HCV-related damage. In this respect, the correlation observed among the three studied miRNAs could imply that they share common pathways by which they contribute the progression of HCV-induced chronic liver damage.

[Determination of hepatitis B virus genotypes in chronic hepatitis B patients in Mersin province, Turkey]. / Mersin Ilinde Kronik Hepatit B Hastalarinda Hepatit B Virusu Genotiplerinin Belirlenmesi.

Currently, ten genotypes (A-J) of hepatitis B virus (HBV) are identified based on the nucleic acid sequence heterogeneity, and these genotypes have been shown to have distinct geographic distribution. Reports of previous studies indicated that the genotype D is the predominant type among hepatitis B patients in different regions of Turkey, however there is no data for HBV genotypes to date from Mersin region. The aim of this study was to investigate the HBV genotypes by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in chronic hepatitis B patients in Mersin province (located in the Mediterranean region of Turkey). A total of 54 serum samples were obtained from the chronic hepatitis B patients (33 male, 21 female; mean age: 40.05 years) followed-up at Gastroenterology Clinic of Mersin University Hospital. Patients had detectable HBV-DNA levels in their serum samples, and they were under antiviral therapy for at least one year. Genotyping of HBV was performed by RFLP analysis with the use of AvaII and MboI restriction enzymes after amplification of pre-S gene region by PCR. Confirmation of selected 18 cases was carried out with direct DNA sequencing. The genotypes were determined by phylogenetic comparison with 43 reference NCBI (National Center for Biotechnology Information) HBV sequences. Genotype determination was not successful in seven cases; since three of them were negative in preS-PCR, three of them yielded non-specific bands, and one of them exhibited a deleted PCR product, at the 300 bp level that was shorter than expected. Four different restriction patterns were determined in PCR-RFLP analysis of the remaining 47 samples. One of these patterns which was AvaII [-]/MboI [306/89/51], was clearly discriminated in 72.3% (34/47) of the samples as genotype D. Genotype discrimination of three patterns could not be done properly and these patterns were AvaII [- ]/MboI [357/306/89/51] (7/47, 14.9%), AvaII [300/146]/MboI [306/89/51] (5/47, 10.7%), and AvaII [- ]/MboI [357/89/---] (1/47, 2.1%). Phylogenetic comparison of HBV sequences demonstrated that all patterns in our cases were clustered in NCBI genotype D sequences. Patterns of AvaII [300/146]/MboI [306/89/51] and AvaII [-]/ MboI [357/89/---] and deleted sample were recognized as pre-S gene variants of HBV isolates. Our data indicated that the predominant HBV type was genotype D as commonly seen in Turkey and other Mediterranean countries. The results of this study also showed that the genotype uniformity and pre-S gene variants within the HBV isolates could be crucial in terms of understanding the molecular epidemiology of HBV circulating in the Mediterranean region of Turkey.

Possible relation between expression of circulating microRNA and plasma cytokine levels in cases of pulmonary tuberculosis.

INTRODUCTION: Tuberculosis (TB) is a life-threatening infection and early diagnosis is critical for treatment and prevention of transmission. There is evidence of correlation between miRNA expression and cytokine regulation during TB infection. The aim of this study was to determine the relationship between expression levels of miRNAs in plasma and cytokine levels as a potential biomarker for genetic predisposition and/or early diagnosis of TB infection. METHODOLOGY: The expression levels of 86 miRNAs were examined in plasma samples of 44 TB patients and 44 healthy controls by qRT-PCR using BioMarkTM 96.96 Dynamic Array (Fluidigm Corporation, South San Francisco, CA, USA) system. The levels of plasma TNF-α, IFN-γ, IL-1ß, IL-4, IL-6, IL-8, IL-10, and IL-12/P40 were examined with ELISA. RESULTS: We identified dysregulation of 18 miRNAs which included upregulation of miR-1, miR-7-5p, miR-9-5p, miR-10a-5p, miR-10b-5p, miR-100-5p, miR-106b-5p, miR-128-3p, miR-133a-3p, miR-143-3p, miR-193a-5p, miR-200b-3p, miR-205-5p, miR-210-3p, and miR-296-5p, and downregulation of miR-15b-5p, miR-16-5p, and miR-25-3p in plasma samples of patients with pulmonary TB (p < 0.05). A significant correlation between the expression levels of miR-1, miR-7-5p, miR-9-5p, miR-10a-5p, miR-10b-5p, miR-15b-5p, miR-100-5p, miR-143-3p, miR-193a-5p, miR-200b-3p, miR-210-3p and cytokine levels of TNF-α, IFN-γ, IL-1ß, IL-8 and IL-10 was identified (p < 0.05). CONCLUSIONS: We demonstrated that altered expression levels of plasma miRNAs consistent with immunological response have the potential to serve as non-invasive biomarkers for early diagnosis of pulmonary TB. Additional investigations with larger sample sizes will be required to confirm our findings and to determine if miRNAs can be possible targets for TB management strategies.