Search

Introduction: The assessment of the clonal spread of methicillin resistant Staphylococcus aureus (MRSA) in nosocomial and community-acquired infections through characterization of the isolates is critical for tracking the evolution of the epidemics, implementing effective control measures, and preventing future outbreaks of MRSA. In this context, it is aimed with this study to determine the clonal relationships between the S. aureus isolates obtained from the patients receiving treatment in the intensive-care units of a state hospital in Turkey. Methodology: A total of 80 MRSA isolates obtained from the patients receiving treatment in three different intensive-care units were analyzed for their antibiotic susceptibilities, pulsed-field gel electrophoresis profiles, and multilocus sequence types. Results: The dendrogram of the pulsed-field gel electrophoresis profiles revealed two major pulsed-field gel electrophoresis pulsotypes: A and B, which were further divided into two (A1 and A2) and four (B1-B4) subgroups, respectively. Multilocus sequence typeanalysis indicated that all isolates belonged to a single MRSA clone, sequence type 239. No significant difference was found between the antibiotic sensitivity profiles of strains isolated from different intensive-care units. All of the strains were sensitive to linezolid and vancomycin. Conclusions: It was concluded that the MRSA strains isolated from the patients receiving treatment at the intensive-care units of the hospital constituted two major pulse-field types and belonged to the ST239 lineage, one of the most extensively distributed MRSA lineages throughout the world.

Tonsil tissue is a very important component of the human immunity system, contributing to the functioning of the cellular and humoral defence system, especially in childhood. The endoplasmic reticulum (ER) is an organelle that has a very important function in the balanced functioning of cells, in which the accumulation of a cellular protein called ER stress occurs in case of dysfunction. ER stress influences the pathogenesis of many diseases and immune system functions. We aimed to investigate the relation between the diseases of tonsil tissue and ER stress response to elucidate the mechanisms of diseases related with the immune system. A prospective study was conducted in 46 children aged between 2 and 16 years who underwent tonsillectomy for chronic tonsillitis or tonsillar hypertrophy. Tonsil tissue was separated into two groups according to their size and evaluated in terms of ER stress markers and apoptosis markers by Real-time PCR and Western blot analysis. The ΔCT levels of ER stress markers (ATF4, ATF6, CHOP, GRP78, EIF2AK3, ERN1, GRP94) were greater in children with chronic tonsillitis (p

Amaç: Ailevi Akdeniz Ateşi (AAA) otoinflamatuar bir hastalıktır ve çeşitli klinik bulgular olarak kendini gösterebilir. Kemokinler, inflamatuar sürecin önemli unsurlarıdır. MCP-1 ve onun reseptörü CCR2, FMF'de kritik roller oynayabilen monositler/makrofajlar için ana kemokinlerdir. Bundan dolayı MEFV gen fonksiyonunu etkileyebilecek MCP-1 (A-2518G) ve CCR2 (G190A) polimorfizmlerinin ve MCP-1 ekspresyon düzeyinin araştırılması amaçlanmıştır.Gereç ve Yöntem: FMF'li hastalar Tel-Hashomer kriterlerine göre belirlendi. Elde edilen kan örneklerinden DNA ve RNA izole edildi. Genotipleme analizi, PCR-RFLP tekniği ile yapıldı. Ayrıca Real-time PCR yöntemi ile ekspresyon analizleri yapıldı. Elde edilen sonuçlar istatistiksel olarak değerlendirildi.Bulgular: Çalışmaya toplam 229 birey (125 erkek ve 104 kadın) dahil edildi. Bunlardan 120 kişide FMF kliniği bulunurken, 107 kişide yoktu. Kalan iki kişi şüpheli klinik duruma sahipti. Çalışmaya alınan bireyler MEFV genotiplemesine göre değerlendirildiğinde ise 75 birey homozigot mutant, 77 birey Heterozigot saptanırken 77 birey ise MEFV geninde mutasyon taşımıyordu. Yapılan analizde Hem FMF kliniği hem de MEFV genotipleri ile MCP-1 (A-2518G) ve CCR2 (G190A) genotipleri arasında anlamlı bir ilişki bulunmadı. Ekspresyon analizinde, FMF kliniği olan hastalarda olmayanlara göre MCP-1 ekspresyonu artmış olarak saptandı. Ayrıca heterozigot MEFV grubunda mutasyonu olmayanlara göre MCP-1 ekspresyonu artmış olarak saptandı, Dahası homozigot MEFV grubunda MCP-1 ekspresyonu en yüksek düzeydeydi. Ek olarak, MCP-1 (A-2518G) genotiplendirmesine göre, MCP-1 ekspresyonu, Wild type gruba kıyasla hem homozigot hem de heterozigot gruplarda yükselmiştir.Sonuç: FMF hastalığında MCP-1 ekspresyonu artmış olup, bu durum FMF hastaları arasındaki klinik farklılıkları açıklayabilir. MEFV mutasyonları, MCP-1 transkripsiyonunu artırarak inflamasyonu şiddetlendirebilir. MCP-1(A-2518G) mutasyonlu hastalarda MCP-1 ekspresyonu artar, bu da FMF kliniğini ağırlaştırır.Anahtar Kelimeler: Ailevi Akdeniz Ateşi, MCP-1, CCR2, Expresyon analizi, Background: Familial Mediterranean Fever (FMF) is an autoinflammatory disease and may express as various clinical findings. Chemokines are crucial elements of the inflammatory process. MCP-1 and its’ receptor CCR2 are the main chemokines for monocytes/macrophages that may play critical roles in FMF. Thus, it was aimed to investigate the MCP-1 (A-2518G) and CCR2 (G190A) polymorphisms and MCP-1 expression level, which may affect MEFV gene function. Material and Method: Patients with FMF were identified according to the Tel-Hashomer criteria. DNA and RNA were isolated from the obtained blood samples. Genotyping analysis was performed by PCR-RFLP technique. In addition, expression analyzes were performed by Real-time PCR method. The obtained results were evaluated statistically. Results: A total of 229 individuals (125 male and 104 female) were included in the study. While 120 individuals had FMF clinic, and 107 individuals did not have. The remaining two individuals had suspicious clinical status. In addition, while 75 individuals were homozygous mutants, 77 individuals were heterozygous mutants, and 77 individuals did not carry mutation in the MEFV gene. No significant relationship was found in between both FMF clinic and MEFV genotypes, and MCP-1 (A-2518G) and CCR2 (G190A) genotypes. In the expression analysis, MCP-1 expression increased in patients with FMF clinic compared to those without. In addition, MCP-1 expression was increased in the heterozygous MEFV group compared to those without mutation, moreover, the expression level was highest in homozygous MEFV group. In addition, according to the MCP-1 (A-2518G) genotyping, MCP-1 expression elevated in the homozygous as well as the heterozygous groups, compared to the Wild type group.Conclusion: MCP-1 expression is increased in FMF disease, which may explain the clinical differences between FMF patients. MEFV mutations may exacerbate inflammation by increasing MCP-1 transcription. MCP-1 expression is increased in patients with MCP-1(A-2518G) mutations, which aggravates FMF clinic. MCP-1 expression may be assessed as a marker in suspicious cases. Keywords: Familial Mediterranean Fever, MCP-1, CCR2, expression

While forming the stable IVT mRNA molecule, high concentration and purity plasmid DNA must be obtained to ligase the ORF antigen sequence initially copied from the plasmid DNA with the UTR regions. In this study, in the stage of creating the mRNA molecule, which is the first step of the COVID-19 mRNA vaccine, comparison and optimization of the pDNA containing the ORF target antigen sequence were performed as a result of isolation with alkaline lysis method and commercial kit. Plasmid DNA bacteria containing the target antigen ORF sequence were grown under appropriate conditions. Plasmid DNA was isolated by commercial kit and alkaline lysis method from bacterial cultures stopped at different OD600 nm values (0.02-0.05, 0.05-0.1, 0.1-0.2, 0.2-0.3, 0.3-0.4, 0.4-0.5). After the obtained pDNAs were visualized on an agarose gel, their purity and concentration were measured by spectroscopic measurement. After the stab culture is resuscitated in SOC medium, bands are formed in a single form after isolation with the kit, and in multiple forms (linear, supercoiled, circular) after pDNA isolation by alkaline lysis method. The ideal OD600 nm for both methods was 0.3-0.4. As a result of isolation with the kit, higher purity on the contrary low concentration pDNA was obtained. The ideal OD600 nm value is a critical parameter that affects the concentration and purity of pDNA. The alkaline lysis method is a cheap and powerful technique that can be used as an alternative for mRNA vaccine development compared to kit isolation., Kararlı IVT mRNA molekülü oluşturulurken başlangıçta plazmid DNA’sından kopyalanan ORF antijen sekansının UTR bölgeleri ile ligaze edilebilmesi için yüksek konsantrasyon ve saflıkta plazmid DNA'nın elde edilmesi gerekmektedir. Bu çalışmada, COVID-19 mRNA aşısının ilk basamağı olan mRNA molekülünün oluşturulması adımında, ORF hedef antijen sekansını içeren pDNA’nın alkalen lizis method ve ticari kit ile izolasyonu sonucundaki kıyaslama ve optimizasyonu yapılmıştır. Hedef antijen ORF sekansını içeren plazmid DNA bakterisi uygun üreme koşullarında çoğaltılmıştır. Farklı OD600 nm değerlerinde (0.02-0.05, 0.05-0.1, 0.1-0.2, 0.2-0.3, 0.3-0.4, 0.4-0.5) durdurulan bakteri kültürlerinden ticari kit ve alkalen lizis yöntemi ile plazmid DNA izolasyonu yapılmıştır. Elde edilen pDNA’lar bir agaroz jelde görüntülendikten sonra, spektroskopik ölçüme tabi tutularak saflığı ve konsantrasyonu ölçülmüştür. Stab kültür SOC medium içerisinde canlandırıldıktan sonra, kit ile izolasyon sonrasında tek formda, alkalen lizis metodu ile pDNA izolasyonu sonrasında birden çok formda (lineer, süper kıvrımlı, sirküler) bant oluşur. Her iki yöntem için de ideal OD600 nm 0.3-0.4 olarak bulunmuştur. Kit ile izolasyon sonucunda daha yüksek saflıkta ancak düşük konsantrasyonda pDNA elde edilmiştir. İdeal OD600 nm değeri, pDNA'nın konsantrasyonunu ve saflığını etkileyen önemli bir parametredir. Alkalen lizis yöntemi, kit izolasyonuna kıyasla mRNA aşı geliştirmeye alternatif olarak kullanılabilecek ucuz ve güçlü bir tekniktir.

A novel coronavirus (SARS-CoV-2) outbreak, responsible for a pneumonia-associated respiratory disorder (COVID-19), has started in early December 2019 in Wuhan, China, and has rapidly spread around the world. Rapid and accurate diagnostic testing plays a crucial role in tackling the COVID-19 pandemic. In this study, it was aimed to compare 5 primer sets designed to amplify different regions for the detection of SARS-CoV-2 and to perform sequence analysis. Conventional RT-PCR was carried out using primers targeting different regions of the virus genome including ORF1ab, Envelope (E), RNA-dependent RNA polymerase (RdRp), Spike (S) and Nucleocapsid (N) genes for the diagnosis of COVID-19. DNA sequence of ORF1ab gene from each sample were compared with the DNA sequence data of SARS-CoV-2 stored in the GenBank and ORF1ab phylogenetic tree was constructed. The amplicon sizes of ORF1ab, S, E, N and RdRp genes were 588 bp, 440 bp, 145 bp, 323 bp and 196 bp, respectively. The SARS-CoV-2 RNA was detected from 74% of total samples from RdRp gene, 87% for N gene, 74% for S gene, 61% for E gene and 82% for ORF1ab region. The ORF1ab sequences of SARS-CoV-2 from 82 patients were had 100% identity to the sequence of Wuhan isolate and among themselves. The phylogenetic analysis revealed that all isolates formed a cluster. The results of this study suggest that the N region is the best for SARS-CoV-2 identification.

Objectives Appropriate antibiotic therapy and prevention of cross-contamination are the most important subjects in tuberculosis (TB) control. The aim of this study was to investigate the major phylogenetic clades and transmission rate of multidrug-resistant (MDR) Mycobacterium tuberculosis isolates (n = 200) from patients with TB in Sivas and Konya Provinces of Turkey. Methods The phylogenetic relationship among the isolates was investigated by spoligotyping method. In addition, the 24-locus mycobacterial interspersed repetitive unit–variable-number tandem repeat (MIRU-VNTR) typing method was used to reveal cross-contamination. Results Spoligotyping revealed 13 different spoligotypes. A total of 188 strains (94.0%) were included in the cluster. The most prominent spoligofamily was the T family (43.0% of strains), followed by LAM (26.0%), H (8.0%), X and S (both 6.0%) and U (5.0%). Also, 12 strains (6.0%) belonged to the Beijing profile. MIRU-VNTR results showed 176 (88.0%) different genotypes among the isolates. In total, 24 strains (12.0%) were in the cluster. Conclusions According to spoligotyping, there is a heterogeneous M. tuberculosis population in Turkey. MIRU-VNTR results showed that cross-contamination observed between MDR M. tuberculosis isolates in Turkey is controllable.

Amac: Palatogenez, fetal yasamin erken evrelerinde ortaya cikan metabolik bir olaydir. Raporlar, hipoksinin damak olusumunda ve yukselmesinde ve ayrica dudaklarin fuzyonunda kritik bir durum oldugunu gostermektedir. Cevresel faktorlerin yani sira, genetik faktorlerin hipokside onemli rol oynadigi bilinmektedir. Hucrelerin hipoksiye yol acan oksijen basincindaki degisikliklere cevap verebilme yetenegi, hipoksi ile induklenebilir faktorler (HIF'ler) olarak bilinen bir transkripsiyon faktoru ailesinin aktivasyonuna baglidir. Bu calisma, yarik dudak/damakli cocuklar ve bu cocuklarin annelerinde hipoksi ile induklenebilir faktor 1, alfa alt birimi (HIF1A)'nin iki polimorfizminin dagilimini belirlemek icin yapilmistir. Yontemler: HIF1A'nin (Pro582Ser ve Ala588Thr) iki polimorfik yapisi, calisma grubu olan yarik dudak/damakli cocuklar ve annelerinde ve kontrol grubu cocuklar ve annelerinde polimeraz zincir reaksiyonu fragment uzunlugu polimorfizmi (PCR-RFLP) yontemi kullanilarak incelendi. DNA fragmentleri, kesim isleminden sonar agaroz jelde goruntulendi. Bulgular: Polimorfik yapilardan Ala588Thr karsilastirmasinda, yarik dudak/damakli cocuklar ve anneler ile kontrol grubu cocuklar ve anneleri arasinda fark gozlenmemistir. Pro582Ser karsilastirmasinda ise; anne ve cocuk genotiplerinin kontrol gruplariyla karsilastirilmasinda farkliliklar gorulmustur (P=0.034 ve P=0.023, sirasiyla). Alell karsilastirmalarinda, yarik dudak/damakli cocuklarin anneleri ile kontrol grubu anneleri arasinda fark gozlenmistir (P=0.001). Cocuklardaki allel karsilastirmalarinda da, annelerdeki kadar yuksek olmasa da fark gorulmustur (P=0.026). Sonuc: HIF1A proteinindeki Prolin aminoasitlerinin degisimine neden polimorfizmler, HIF1A proteininin aktivitesini veya omrunu etkiliyor ve yarik dudak/damak olusunda rol aliyor olabilir.

There is an increasing interest about the role of miRNAs in the pathogenesis of bipolar disorder (BD). In this study, we aimed to examine the role of miRNAs as potential diagnostic and clinical biomarkers in BD.Fifteen miRNAs in plasmas obtained from BD patients (n = 66) and from the healthy control group (n = 66) were analyzed by a qPCR test. Clinical variables including lithium treatment response were assessed with various test batteries. The correlation of the miRNA levels with the clinical variables and scale scores was examined. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed using the DIANA-miRPath v.3.0 software to identify the possible target genes.The miR-132, miR-134, miR-152, miR-607, miR-633, and miR-652 levels were significantly increased, whereas the miR-15b and miR-155 levels were found to be significantly decreased in the patient group compared to the controls. The miR-15b-5p and miR-155-5p levels and increases in the miR-134-5p and miR-652-3p levels were calculated to have 83.3 % sensitivity and 78.8 % specificity in determining the risk of BD. miR-155-5p was associated with the disease burden and severity. Fatty acid biosynthesis and metabolism, viral carcinogenesis, the EBV infection, and extracellular matrix and adhesion pathways were highlighted as target pathways.We can conclude that miRNAs may play a role in the pathophysiology of BD through various biological pathways and that miRNAs may be used as a screening test to distinguish bipolar patients from healthy controls. Our findings will provide a basis for long-term follow-up studies with larger samples.

Norofibromatozis tip I (NF1), 17. kromozomda norofibromin protein kodlayan genin mutasyonuna yol acan karmasik bir hastaliktir. NF1 otozomal dominant bir hastaliktir. Bireylerde NF1 prevalansi yaklasik 1: 2500 ila 1: 3500'tur. Erkekler ve kadinlarda esit olarak gorulur. Bu calismada cok sayida hiperpigmente deri makulu, birden fazla cafe-au-lait lekesi, aksiller cillenmesi, optik gliomu, yuzlerce yumusak deri norofibromu ve her iki gozun irisinde lisch nodulleri bulunan 27 yasinda bir kadin hasta sunulmustur. Klinik ozelliklere gore, norofibromatozis tip 1'den suphelenilen hastadan, NF1 geninin dizi analizi yapildi ve NF1 geninde bir heterozigot mutasyon c.980 T> G (p.L327R) tespit edildi. Bu mutasyon literaturde daha once rapor edilmemistir.

WOS: 000484847800013, The purpose of this study was to determine the phenotypic antibiotic susceptibility patterns, extended-spectrum beta-lactamase (ESBL) genes and genotypic profiles of ESBL-positive Escherichia coli strains isolated from urine samples obtained from outpatients with urinary tract infection in Turkey. A total of 120 E. coli strains during 2017, 2018, and 2019 (40 patients per year) were examined for antibiotic susceptibility patterns by disc diffusion method, for ESBL genes using PCR and sequencing and for molecular typing by pulsed-field gel electrophoresis (PFGE) method. The isolates were evaluated for their sensitivity to 21 different antibiotics. Four different antimicrobial resistance patterns were determined according to antibiotic susceptibility status of the isolates. The beta-lactamase genes detected in the isolates were CTX-M-15 + OXA-1 (n = 14), CTX-M-15 (n = 24), TEM-1 + CTX-M-15 (n = 52), TEM1 + SHV-12 (n = 6), SHV-12 1 (n = 6), TEM-1 + CTX-M-1 (n = 6), TEM-1 + CTX-M-16 (n = 6) and TEM-1 + CTX-M-9 (n = 6). The CTX-M-15 was the most prevalent ESBL enzyme in the isolates. As a result of PFGE analysis performed by XbaI enzyme restriction process, one major PFGE profile and three main groups (Group I-I-III) were observed. While antibiotic resistance profiles of the strains showed four groups (RI-RII-RIII-RIV), PFGE band profiles showed a major group (90% similarity ratio). High ESBL production and decreased susceptibility to broad-spectrum cephalosporins were observed in E. coli strains. In addition, PFGE analysis showed high clonal similarity among E. coli isolates.

WOS:000623839300001 PMID: 33645410 The anti-proliferative effects of 5-methylquinolinium (5MQ) of nicotinamide N-methyltransferase (NNMT) have not been previously investigated on a cervical cancer cell line. NNMT is a metabolic enzyme that is correlated with tumour progression and metastasis. 5MQ is a small molecule inhibitor of NNMT. 0.1-500 mu M of 5MQ was tested on the HeLa epithelial cervical cancer cell line. Cell viability was assessed with the MTT test. TWIST, ZEB1, SERPIN1, SIRT1, CD16, mRNA and various protein expression levels were analysed with Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and Western Blotting, respectively. 5MQ significantly inhibited HeLa cell proliferation in a concentration and time-dependent manner. Increased cell shrinkage, loss of cellular adhesions and apoptotic bodies were observed in HeLa cells after 5MQ treatment. Following treatment with 5MQ, ZEB1, SIRT1, CD16 mRNA levels were increased while TWIST and SERPIN1 mRNA levels were reduced. Expressions of oncogenic proteins phospho-Akt and SIRT1 were decreased. 5MQ can effectively inhibit HeLa cell proliferation without apparently affecting HEK-293 cell proliferation. IMPACT STATEMENT What is already known on this subject? NNMT is a cytosolic enzyme involved in tumour progression, metastasis and treatment resistance. It was overexpressed in many human malignancies. 5-amino-1-methylquinolinium (5MQ) is a novel small molecule inhibitor of NNMT that has shown promising results in the treatment of obesity and in senescent muscle regeneration. 5MQ has not been tested on the HeLa cervical cancer cell line, previously. What do the results of this study add? In this study, 5MQ was tested on the HeLa cervical cancer cell line for the first time and the molecular changes associated with 5MQ treatment were analysed. 5MQ demonstrated significant anti-proliferative activity on HeLa cells, which displayed morphological signs of apoptosis. Treatment of HeLa cells with 5MQ led to an increase in ZEB1, SIRT1 mRNA while TWIST mRNA was decreased. Phospho-Akt and Sirtuin1 protein expressions were decreased. What are the implications of these findings for clinical practice and/or further research? 5MQ can effectively inhibit HeLa cell proliferation without apparently affecting HEK-293 cell proliferation. 5MQ treatment was associated with a decrease in the expression of phospho-Akt and Sirtuin1 proteins, both of which have been reported to maintain tumour progression. 5MQ can further be investigated and modified for anti-cancer therapy.

Prevalence of neural tube defect (NTD) has reduced after folic acid intake. However; which mechanisms are effective in NTD are not known exactly. In this study; due to the possible effects on hypoxic pathway and embryonic development, particularly on extracellular matrix components, Hif-1α Pro582Ser and TGF-β3 IVS5+104 A/G SfaN1 polymorphisms were studied by PCR-RFLP method both on children with NTDs and mothers. Statistical differences were seen for Hif-1α and TGF-β3 IVS5+104 A/G SfaN1 polymorphisms in children with NTDs but no difference was seen in mothers. Both genes are effective on many pathways and our results suggest that regulation of extracellular matrix components of children during fetal life is important in neural tube defects formation. The results of this study show that Hif-1α Pro582Ser and TGF-β3 IVS5+104 A/G SfaN1 polymorphisms may play a role in NTDs.

Vancomycin-resistant enterococcus (VRE) is a global threat to public health. Knowledge about the occurrence of vanA-carrying enterococci in broiler and environmental samples is important as antibiotic resistance can be transferred to human bacteria. The aim of this study was to investigate the presence of VRE in broiler cloacal and environmental (house) samples and to genotype the isolates. In this study, 350 swabs were collected from broiler farms. All samples were plated onto enterococcus selective agar containing 6 mg/L vancomycin and 64 mg/L ceftazidime. Minimum inhibitory concentration (MIC) values were determined for vancomycin and teicoplanin. Vancomycin-resistant Enterococcus faecium (VREfm) was isolated from 6 out of 300 (2%) broiler cloacal samples and 13 out of 50 (26%) house samples. All E. faecium isolates had vanA genes. All VREfm isolates (19 isolates) were confirmed to be 95% similar to each other. In conclusion, although 20 years have passed since the ban on avoparcin in Turkey, the present study shows that VREfm isolates are still present in broiler production and especially in broiler houses, and most importantly, a major VREfm clone was isolated from broiler cloacal and house samples.

Tunicamycin is an inhibitor of glycosylation that disturbs protein folding machinery in eukaryotic cells. Tunicamycin causes accumulation of unfolded proteins in cell endoplasmic reticulum (ER) and induces ER stress. ER stress is an essential mechanism for cellular homeostasis has role in cell death via reprogramming of protein processing, regulation of autophagy and apoptosis. In this study we show effect of tunicamycin on subpopulation and invasion of CD44+/CD24- MCF7 breast cancer stem cells. CD44+/CD24- cells were isolated from MCF7 cell line by fluorescence activated cell sorting (FACS) and treated with tunicamycin. ER stress was monitored by evaluation of X-box binding protein 1(XBP-1) mRNA splicing, cleaved activating transcription factor 6 (ATF6) nuclear translocation and CCAAT/enhancer-binding protein homologous protein (CHOP) expression. CD44+/CD24- subpopulation was analyzed using flow cytometry. Invasion was investigated by scratch assay, trypan blue staining, 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) proliferation and in vitro migration assays. Increased level of spliced XBP-1, ATF6 nuclear translocation and CHOP protein expression were detected in CD44+/CD24- and original MCF7 cells treated with tunicamycin. Also, a significant decline in CD44+/CD24- cell subpopulation was determined in the cells treated with tunicamycin. The results also showed inhibited invasion, increased cell death, suppressed proliferation and reduced migration in the CD44+/CD24- and CD44+/CD24- rich MCF7 cell culture, under effect of tunicamycin. Our results indicate that CD44+/CD24- phenotype MCF7 cells are susceptible to tunicamycin. The results showed that tunicamycin-induced ER stress suppresses CD44+/CD24- phenotype cell subpopulation and in vitro invasion and accelerates tumorosphore formation. These results suggest that tunicamycin-induced ER stress inhibits CD44+/CD24- phenotype MCF7 breast cancer stem cells. We conclude that using ER-targeting chemicals like tunicamycin is an interesting approach to target breast cancer stem cells inside tumor.

WOS: 000473333700003, …

Prevalence of neural tube defect (NTD) has reduced after folic acid intake. However; which mechanisms are effective in NTD are not known exactly. In this study; due to the possible effects on hypoxic pathway and embryonic development, particularly on extracellular matrix components, Hif-1α Pro582Ser and TGF-β3 IVS5+104 A/G SfaN1 polymorphisms were studied by PCR-RFLP method both on children with NTDs and mothers. Statistical differences were seen for Hif-1α and TGF-β3 IVS5+104 A/G SfaN1 polymorphisms in children with NTDs but no difference was seen in mothers. Both genes are effective on many pathways and our results suggest that regulation of extracellular matrix components of children during fetal life is important in neural tube defects formation. The results of this study show that Hif-1α Pro582Ser and TGF-β3 IVS5+104 A/G SfaN1 polymorphisms may play a role in NTDs.

WOS: 000469890500001, PubMed: 30853450, Many reports have been published on the suspected vertical transmission of Encephalitozoon cuniculi; however, prior to 2003, these reports were based on circumstantial evidence, such as histopathological, immunohistochemical, or serological diagnosis of the infection. In 2003, vertical transmission of the parasite was confirmed by detection of E. cuniculi DNA in fetuses with the nested polymerase chain reaction (PCR) technique. However, the passage of the parasite to eyes of fetus during the intrauterine stage still requires verification. In the current study, natively infected with parasite spores female rabbits were mated with non-infected males. All resulting offspring that died before ten postpartum days were investigated using molecular techniques to confirm the intrauterine transmission of the parasite to the offspring' eyes. In total, 119 DNA samples from rabbit offspring tissues were collected from blood, kidney, brain, eye (both eyes were used as single samples), lung, placenta, liver and heart were used for PCR. Parasitic DNA in the eyes of offspring was detected (54%) 6 of 11 naturally seropositive mother rabbits. PCR results were found to be positive for the eyes of 63% (19/30) of the offsprings from seropositive rabbits. Therefore, mother rabbits naturally infected with E. cuniculi showed the molecular presence of the parasite in their offspring' eyes. Sequence analysis confirmed the partial DNA sequence data of E. cuniculi and blast analysis identified the agent as genotype I. These results confirm transmission of E. cuniculi to rabbit offspring' eyes in the intrauterine period. This is the first molecular evidence to show ocular transmission of the infection via an intrauterine route in rabbits.

Summary Introduction Trichosporon asahii is the most common species that causes trichosporonosis. Materials and methods In the present study, a collection of 68 T. asahii strains recovered from hospitalized patients urine samples between 2011 and 2016 was examined. T. asahii strains were identified by sequencing the intergenic spacer 1 region (IGS1) and genotyped. In addition, proteinase, phospholipase, esterase, haemolytic activity, and biofilm formation of a total of T. asahii strains were investigated. Results The predominant genotype was 1 (79.3%) and followed by 5 (8%), 3 (6.9%), 6 (3.4%), 4 (1.1%), 9 (1.1%). In none of the 68 strains, proteinase and phospholipase activities could be detected, while all were found to be esterase positive. Biofilm production and hemolytic activity were detected in 23.5 and 97% respectively. Discussion Our results indicated that six genotypes were (1, 5, 3, 6, 4, 9) present among T. asahii strains and no property was found to associate with a genotype, in terms of virulence factors.

Melatonin is a hormone of the pineal gland that has a wide range of biological effects such as antioxidant, anti-inflammatory, and anti-tumor activity. Previous studies have shown that melatonin also affects survival, proliferation, and apoptosis of the cells. In this study, we investigated the effect of melatonin on apoptosis, self-renewal, and differentiation. For this purpose, MCF-7 and HEK293 cells were subjected to melatonin treatment. Expression of genes related to apoptosis (Bax and Bcl2) and self-renewal and differentiation (Oct4, Sox2, and Nanog) analyzed after the sorting of cancer stem cells from MCF-7 cells. Results showed that the effect of melatonin is dependent on the melatonin concentration and treatment periods. Melatonin treatment decreased the cell proliferation rate of MCF-7 in contrast to HEK293. Also, this treatment increased apoptosis in MCF-7 cells and decreased in HEK293 cells. Gene expression of Nanog was decreased and Sox2 was increased in both cell groups after the melatonin treatment. Expression of Oct4 was decreased in MCF-7 cells and increased in HEK293 cells. We determined that melatonin decreases apoptosis and differentiation of stem cells in normal HEK293 stem cells, but increases apoptosis and differentiation in the MCF-7 cancer stem cells.

Lung cancer is among the leading causes of cancer-related deaths, and thusmonitoring, treatment and course of the disease has become a major area of research following diagnosis of the disease. The present study aimed to evaluate the role of HIF pathway in development of non-small-cell lung cancer (NSCLC) in patients diagnosed with NSCLC using their tumor and non-tumor tissue specimens. Both tumor and non-tumor tissue specimens were collected from 10 patients with NSCLC.They were placed in Trizol, and stored at - 80 C ° until processing. RNA was isolated from these tissue specimens using the Trizol/chloroform method. Then, the RNA was converted to cDNA by RT PCR. Amplified cDNA was used for gene expression via specific probes and primers. The mean age of patients (9 male, 1 female)was 62.5 years. The HIF (Hypoxia-Inducible Factor)-1 gene expression was 3 times higher in tumor tissuesthan in non-humor tissues. In relation to this pathway, there was an increased gene expression of 3.72forVEGF (Vascular Endothelial Growth Factor), 3.11forMYC (Avian Myelocytomatosis Viral Oncogene Homolog), 2.28 forTGFA (Transforming Growth Factor) and 2.28 for VEGFR-1 (Vascular Endothelial Growth Factor Receptor-1). The increased HIF expression in the present study results in increased hypoxia in the microenvironment of tumor tissue, while induction of HIF by hypoxia results in expression of genes regulated by HIF (VGF, VEGFR-1,MYC, TGFA). Our results indicate that hypoxia and hypoxia-induced pathways play a role in development and metastasis of tumor in patients with NSCLC. Keywords: Non-small-cell lung cancer, gene expression, HIF pathway.

Thromboangiitis obliterans (TAO), which is also known as Buerger’s disease, is a disease characterized by segmental inflammation together with vasoactive phenomena in the arteries and veins of the extremities. In the present study, the relationship between angiotensin-converting enzyme (ACE) insertion/deletion (I/D) gene mutations, which is known to be involved in vasodysfunction, and TAO was determined. Because of this relationship, it was aimed to determine whether the distribution of angiotensin-converting enzyme (ACE) insertion / deletion (I / D) gene mutations in patients with Thromboangiitis obliterans (TAO) is different from healthy subjects in the control group. Eighty patients who were previously diagnosed with TAO were included in the study. The patients were determined based on Olin’s criteria. The control group consisted of 88 healthy volunteers. DNA isolation was performed by venous blood from the patients in both groups. The DNAs were amplified via polymerase chain reaction (PCR) using appropriate primers, the amplicons were eluted by running in 1% agarose gel, and defined via staining. In addition to the signifi cant difference between the patient and control groups in terms of age and smoking due to the selection criteria (Olin’s criteria), a statistically significant difference was also determined between the groups in terms of the distribution of the ACE I/D gene polymorphism. The significant difference between groups with respect to the ACE I/D polymorphism, together with the smokinglike effects of ACE on the cardiovascular system, reveal a consequence which should not be neglected.

p53 is a transcription factor that regulates multiple cellular processes that are also important in cellular fates such as cell cycle arrest or programmed cell death. Induction of growth arrest or cell death by p53 prevents the repli­cation of damaged DNA and proliferation of genetically abnormal cells. Therefore, inactivation of p53 by muta­tion or deletion is also important in ensuring the cellular homeostasis. However, studies showed that p53 deficient mice and cells such as Saos-2 cells are maintaining their life. This situation suggests that p53-related proteins might compensate the functions of p53 in p53 deficient organisms. The identification of two p53-related proteins, p63 and p73 revealed the transcription of p53 responsive genes in p53 deficient organisms. Both p63 and p73 pro­teins have high homology with the p53 protein and share some of the functions of p53. In contrast to p53, p63 and p73 rarely mutated in human cancers. Here we studied to summarize the current information about the p53 and other p53-related proteins, p63 and p73 that are included into the p53 gene family., p53, hücre akıbetinin belirlenmesinde önemli rol oynayan hücre döngüsünün durdurulması veya programlı hücre ölümüne yönlendirilmesi gibi hücresel birçok yolağın dü­zenlenmesinde görev alan bir transkripsiyon faktörüdür. p53 aracılığı ile programlı hücre ölümünün başlatılması veya hücre çoğalmasının durdurulması, hasarlı DNA rep­likasyonunu ve genetik olarak normal olmayan hücrelerin çoğalmasını engellemektedir. Bu nedenle p53 işlevlerinin mutasyon ya da delesyon ile engellenmesi hücre home­ostasisin sağlanmasında da ayrıca önemlidir. Bununla birlikte, çalışmalar p53\'ten yoksun farelerin ve Saos-2 gibi hücrelerin yaşamlarını devam ettirebildiklerini göstermek­tedir. Bu durum, p53\'ten yoksun olan organizmalarda p53 işlevini karşılayan p53-ilişkili proteinlerin olabileceğini dü­şündürmüştür. p53 ilişkili p63 ve p73 proteinlerinin keşfi, p53\'ten yoksun hücrelerde p53 bağımlı genlerin transkrip­siyonlarına bir açıklama getirmiştir. p63 ve p73 proteinle­rinin her ikisi de p53 proteini ile yüksek oranda benzerlik göstermekte ve p53\'ün bazı işlevlerini de paylaşmaktadır­lar. p53\'ten farklı olarak p63 ve p73 insan kanserlerinde nadir olarak mutasyona uğramıştır. Burada p53 hakkında­ki güncel bilgilerle beraber p53-ilişkili proteinler olan p63 ve p73 hakkındaki bilgileri de özetlemeye çalıştık.

Homocysteine levels have been investigated in patients with obstructive sleep apnoea syndrome (OSAS), a syndrome associated with a high level of comorbid cardiovascular disease (CVD). While significant increases in homocysteine levels have been observed in OSAS patients with CVD, no increases have been noted in OSAS patients without CVD. This study was designed to investigate the methylenetetrahydrofolate reductase ( MTHFR) gene, which is essential for homocysteine metabolism and has been shown to have a causal role in the development of CVD. Eighty subjects, 30 diagnosed with OSAS by polysomnography and 50 controls (healthy volunteers with no symptoms of OSAS) were enrolled. Two mutations in the MTHFR gene were investigated using polymerase chain reactions and restriction fragment length polymorphisms. No significant differences were found in mean age, body mass index, homocysteine levels, or MTHFR allele or genotype distributions between patient and control groups.

Citation Yenicesu GI, Cetin M, Ozdemir O, Cetin A, Ozen F, Yenicesu C, Yildiz C, Kocak N. A prospective case–control study analyzes 12 thrombophilic gene mutations in Turkish couples with recurrent pregnancy loss. Am J Reprod Immunol 2010; 63: 126–136 Problem Recurrent pregnancy loss (RPL) is a heterogeneous disorder. The contribution of specific thrombophilic genes to the pathophysiology of RPL has remained controversial. We evaluated the prevalences of 12 thrombophilic gene mutations among homogenous Caucasian couples with RPL and fertiles. Method of study This was a prospective case–control study evaluating 272 women with RPL and 152 of their male partners, and a control group of 56 fertile couples. We investigated mutations including FV Leiden, factor V H1299R, factor II prothrombin G20210A, F XIII V34L, β-fibrinogen −455G>A, plasminogen activator inhibitor-1, GPIIIa L33P (HPA-1 a/b L33P), MTHFR C677T, MTHFR A1298C, ACE I/D, Apo B R3500Q, and Apo E. Results Overall, heterozygous mutations of FV Leiden, FXIII V34L, GPIIIa L33P, Apo E4, and prothrombin G20210A and homozygous mutations of PAI-1and MTHFR C677T were associated with RPL. There was no meaningful association between RPL and other studied genes. Conclusion In contrast to the other mutations and polymorphisms, FV Leiden, FXIII V34L, GPIIIa L33P, Apo E, prothrombin G20210A, PAI-1 and MTHFR C677T gene mutations may help to identify the couples at risk for recurrent pregnancy loss.

WOS: 000263540800026, Familial Mediterranean fever (FMF) is the most frequent hereditary inflammatory disease characterized by self-limited recurrent attacks of fever and serositis. The aim of the current study is to determine the frequency of the mutations in 365 suspected FMF patients and to reveal whether there is a correlation between genotype and phenotype of these patients. All patients were clinically examined according to Tell-Hashomer FMF criteria and were screened genetically in terms of common 12 Mediterranean fever gene (MEFV) mutations. Various point mutations were detected in 270 (74%) patients. The most frequent mutation was M694V (26.85% of the alleles) and was followed by E148Q (15.55%), M680I (G/C) (9.62%) and V726A (7.96%). Patients who bear M694V homozygous mutation had most severe disease phenotype and high risk for amyloidosis (P = 0.04). Our results indicate that Sivas population has a wide range of heterozygous mutated carriers of MEFV gene and there is a high frequency of E148Q allele when compared to the other Mediterranean groups.

WOS: 000393279900008, PubMed: 28058155, Papillophlebitis is known as central retinal vein occlusion seen in young patients. It usually presents as unilateral optic disc edema with cotton wool spots and hemorrhage in the peripapillary region. As it may be due to many autoimmune and inflammatory causes, a thorough systemic evaluation of the patient is warranted. In this case report we describe a bilateral, simultaneous papillophlebitis case thought to be related to hyperhomocysteinemia secondary to C677T polymorphism of methylenetetrahyrofolate reductase enzyme.

Bacterial strain typing is an important process for diagnosis, treatment and epidemiological investigations. Current bacterial strain typing methods may be classified into two main categories: phenotyping and genotyping. Phenotypic characters are the reflection of genetic contents. Genotyping, which refers discrimination of bacterial strains based on their genetic content, has recently become widely used for bacterial strain typing. The methods already used in genotyping of bacteria are quite different from each other. In this review we tried to summarize the basic principles of DNAbased methods used in genotyping of bacteria and describe some important DNA regions that are used in genotyping of bacteria. J Microbiol Infect Dis 2011;1(1):42-46.

WOS: 000298011800002, PubMed: 21897766, Objective: The purpose of this study was to evaluate the relationship between 5,10-methylenetetrahydrofolate reductase (MTHFR) polymorphisms and Attention Deficit Hyperactivity Disorder (ADHD) in a sample of Turkish children. Study Design: MTHFR gene polymorphisms were assessed in 40 patients with ADHD and 30 healty controls. Two mutations in the MTHFR gene were investigated using polymerase chain reactions and restriction fragment length polymorphisms. Results: Although there were no statistically significant differences in genotype distributions of the C677T alleles between the ADHD and the control groups (p=0,678) but the genotypic pattern of the distributions of the A1298C alleles was different between the ADHD patients and the controls (p=0,033). Conclusions: Preliminary data imply a possible relationship between A1298C MTHFR polymorphisms and the ADHD.

Growing evidences indicate that tumor regrowth and invasion refer a small subpopulation of tumor cells called cancer stem cells (CSCs). Autophagy is an essential catabolic process for cell homeostasis that involves in cell survival and death and also development of various human diseases including cancer. Here we aimed to investigate the effect of autphagy on the subpopulation of CD44 C /CD24 � /low phenotype MCF7 and Hep-2 cells. CD44 C /CD24 � /low cells were isolated from MCF7 and Hep-2 cell cultures by using fluorescence-activated cell sorting (FACS). The CD44 C /CD24 � /low and the original cells were separately treated with 20 mM of C2 ceramide for 24 hours in order to stimulation of autophagy. LC3 proteins aggregation was monitored to evaluate autophagy in the cells by using LC3 targeted immunofluorescence staining assay. Subpopulation of CD44 C /CD24 � /low cells in total cultures were investigated by flow cytometry analysis and eventually statistics were carried out. Results showed an enhanced aggregation of LC3 proteins in the cells and also CD44 C /CD24 � /low cells under effect of C2 ceramide. Moreover, the subpopulation ofCD44 C /CD24 � /low cells werefoundsignificantlylower in thecellscultures treated with C2 ceramide in comparison with negative controls. The results elucidated that CD44 C /CD24 � /low MCF7 and Hep-2 cells were susceptible to autophagy stimulatory reagent C2 ceramide. Additionally, our results suggest that CD44 C /CD24 � /low cell subpopulation in the cell cultures is reduced under autophagic condition. Therefore, we suppose that autophagy may has a negative impact on the breast and Larynx cancer stem cells self-renewal potency or expression of CD44 and CD24 cell surface markers, however further studies are needed to reveal the exact mechanism.

Sarkoidoz, nedeni tam olarak bilinmeyen ancak etiyolojisinde genetik, enfeksiyöz ajanlar, mesleksel maruziyet ve çevresel faktörlerin rol oynadığı multisistemik granülomatöz bir hastalıktır. Sarkoidozda ailesel yatkınlık bilinmekle beraber kalıtım şekli tam olarak tanımlanamamıştır. Bu makalede histopatolojik olarak sarkoidoz tanısı koyduğumuz iki kız kardeşi sunduk. Yapılan genetik incele- me sonucunda her iki kardeşte de Human Lökosit Antijen (HLA) HLA A24, B51, Cw07, DRB1 15, DQB1 06 genleri ortak olarak saptandı. Her iki olguya steroid tedavisi başlandı ve takibe alındı. Sonuç olarak her iki kız kardeşte tesbit edilen bulgulara göre sarkoidozda ailesel genetik yatkınlık olabileceği ve sarkoidoz tanısı alan bireylerin ailesinin sarkoidoz yönünden değerlendirilmesi gerektiği kanısına varıldı., Sarcoidosis is a multisistemic, granulomatous disease with an unknown etiology, but genetic factors, infectious agent occupatinal exposure and environmental factors may play a role in etiology. Although familial predisposition is known in sarcoidosis, the type of inharitance is not defined exactly. In this article we reported two sisters with the diagnosis of sarcoidosis confirmed histopathologi- cally. The result of genetic analysis reaveled that were common in human leucocyte antigen (HLA) HLA A24, B51, Cw07, DRB1 15, DQB1 06 genes both of the sisters. Steroid therapy was initiated to both patients and they were followed up. In conclution , according to findings found in both sisters, we thought that genetic predisposition could be seen in sarcoidosis and the family members of the sarcoidosis cases should be evaluated for sarcoidosis.