Person: AKBAŞ, FAHRİ
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Publication Metadata only Molecular cloning and characterization of NAD(+) dependent isocitrate dehydrogenase enzyme from Shewanella putrefaciens(2021-10-01T00:00:00Z) Fahri, Akbas; Metin, Demirel; Ahmet, Ozaydin; Sahabettin, ŞAHBETTİN; AKBAŞ, FAHRİ; DEMİREL, METİN; SELEK, ŞAHABETTİNIsocitrate dehydrogenase (IDH) is a fundamental enzyme for carbon metabolism in the Krebs cycle. This enzyme is required for oxidation-reduction reactions in both eukaryotic and prokaryotic cells and plays a critical role in their growth and pathogenesis. In this study, we cloned the gene encoding NAD(+) dependent isocitrate dehydrogenase from Shewanella putrefaciens. The expression of recombinant protein was induced with 0.5 mM of IPTG. His-tagged IDH overexpressed in E. coli was purified and characterized. The expressed IDH enzyme was purified in an active soluble form. The molecular weight of the enzyme was confirmed with Western blotting. High sequence homology was observed with IDH sequences of other Shewanella strains and remarkable sequence homology was found with other bacteria reported in the database.Publication Metadata only Lucilia sericata Larval Secretions Stimulating Wound Healing Effects on Rat Dermal Fibroblast Cells(2020-09-01T00:00:00Z) AKBAŞ, FAHRİ; Ozaydin, Ahmet; Polat, Erdal; Onaran, Ilhan; AKBAŞ, FAHRİThe extract from larval Lucilia sericata is used clinically to promote wound healing and tissue regeneration. However, its effect and underlying mechanisms on fibroblast cells, which are involved in the wound healing process, are still poorly understood. This study aimed to examine the effects of larval secretions on dermal fibroblast activity and gene expression and to evaluate the wound healing potential of their major components. Primary rat fibroblasts were cultured and treated with larval secretions. Following the treatment, the cells were used to extract RNA for gene profiling. In addition, migration to the injury site was studied with the scratch healing assay. Our results showed that larval secretion accelerated the migration of the fibroblasts compared to the control cells and that several mRNAs were differentially expressed during a period of 72 h incubation. Additionally, we analyzed the chemical composition of larval secretions and showed that fumaric acid, ferulic acid, and p-coumaric acid, which were selected and identified for their major components, enhanced the migration of the fibroblasts. Therefore, these results indicate that L. sericata larval secretions could modulate the mRNA expression of some wound healing-related genes of the fibroblasts and contain the effective components for wound healing.Publication Open Access The Effects of Lucilia sericata Larval Secretions on The Expressions of MicroRNAs that are Suggested to be Related with Wound Healing in Experimental Diabetic Rat Wound Model(2020-01-01T00:00:00Z) Kilinc, Oyku; Arkan, Hulya; AKBAŞ, FAHRİ; Polat, Erdal; Tuncdemir, Matem; Onaran, Ilhan; Ozaydin, Ahmet; AKBAŞ, FAHRİObjective: Normal wound healing is achieved by a cascade of many cellular activities. This process is affected by some of the metabolic diseases like Diabetes Mellitus (DM). DM causes bad prognosis and is one of the major contributors to chronic wound healing problems. Recently, Lucilia sericata larvae are used for wound healing as they are very effective agents in wound healing process. It-s still unclear that how the larvae affect the molecular mechanisms and signaling pathways of chronic wound healing. MicroRNAs (miRNAs) can induce gene expression in post-transcriptional mechanisms. In this study, our aim was to determine whether the larvae secretions could change the expression patterns of selected miRNAs on the diabetic microenvironment.Publication Metadata only Cloning, expression and characterization of recombinant CagA protein of Helicobacter pylori using monoclonal antibodies: Its potential in diagnostics.(2020-09-14T00:00:00Z) Salih, BA; Karakus, C; Ulupinar, Z; Akbas, FAHRİ; Uslu, M; Yazici, D; Bolek, BK; Bayyurt, N; Turkay, C; AKBAŞ, FAHRİPublication Metadata only Iron alters Ca<sup>2+</sup> homeostasis in doxorubicin-resistant K562 cells.(2020-03-05T00:00:00Z) Yalcintepe, L; Erdag, D; Akbas, FAHRİ; Kucukkaya, B; AKBAŞ, FAHRİPublication Open Access The utility of serum microRNA-93 and microRNA-191 levels for determination of injury severity in adults with multiple blunt trauma(2020-12-01T00:00:00Z) Sogut, Ozgur; Metiner, Merve; Kaplan, Onur; Calık, Mustafa; Cakmak, Sumeyye; Umit, Tugba Betul; Ergenc, Huseyin; Akbas, Fahri; Süsgün, Seda; AKBAŞ, FAHRİ; SÜSGÜN, SEDABACKGROUND: Various scoring systems have been developed to determine the trauma severity and prognosis of patients following multiple blunt trauma (MBT). However, these scoring systems do not provide exactly the desired severity assessment. In recent years, serum concentration of many specific miRNAs, especially for head trauma, has been shown to play an important role in determining the diagnosis, severity and prognosis of injury. To date, however, no studies have investigated serum microRNAs in patients with MBT. Thus, this study measured the expression of miRNA-93 and -191 in the serum of adults with MBT and examined the correlations of Injury Severity Score (ISS) and Revised Trauma Score (RTS) values with serum miRNA-93 and -191 levels in these patients with the aim of predicting trauma severity based on the miRNA levels.METHODS: This prospective case–control study enrolled 50 consecutive adults with MBT and age- and sex-matched 60 healthy controls. The patients were divided into ISS >16 (group 1, major or severe trauma) and ISS ≤16 (group 2, minor or mild-moderate trauma) groups. Serum miRNA-93 and -191 levels were assessed using quantitative real-time reverse transcription-PCR. We evaluated whether the miRNAs were differentially expressed in major and minor MBT patients and determined their utility for assessing the severity of injury.RESULTS: The mean serum miRNA-93 and -191 levels were significantly elevated in the patients compared to the controls and were higher in patients with ISS >16 compared to those with ISS ≤16, although the difference was not significant. In the patients with multitrauma, ISS was significantly, negative and weak correlated with serum miRNA-191 level (rho = –0.320, p = 0.023) but not with the serum miRNA-93 level. No optimal cutoff for the serum miRNA-93 level was found with respect to trauma severity (AUC 0.617, [0.455–0.779]). However, an optimal cutoff value for serum miRNA-191 was identified, with values <1.94 indicating severe trauma (AUC 0.668 [0.511–0.826]; 65.6% sensitivity, 77.8% specificity).CONCLUSION: miRNA-191 and -93 levels were significantly upregulated in multitrauma patients compared to controls. The level of miRNA-191 in conjunction with ISS, but not that of miRNA-93, may be a useful biomarker for determining injury severity in patients with multitrauma.Publication Open Access Fibromiyalji Sendromu ile Bağırsak Mikrobiyotası Arasındaki İlişkinin Araştırılması(2021-03-01T00:00:00Z) Albayrak, Büşra; Süsgün, Seda; Küçükakkaş, Okan; Akbaş, Fahri; Yabacı, Ayşegül; Özçelik, Semra; SÜSGÜN, SEDA; KÜÇÜKAKKAŞ, OKAN; AKBAŞ, FAHRİ; YABACI TAK, AYŞEGÜL; ÖZÇELİK, SEMRAFibromiyalji sendromu (FMS), yetişkin popülasyonda bildirilen prevalansı %3-10 olan, kronik yaygın ağrının sık görülen formlarından biridir. Tipik ağrının klinik görünümü ve ilişkili somatik ve psikolojik semptomların varlığı tanıya temel oluşturur. FMS sinir sistemi disfonksiyonu ile ilişkilidir ve nörotransmitterler fibromiyalji için onaylanmış bir dizi ilacın hedefi olarak rol oynamaktadır. Bununla birlikte, FMS’de altta yatan mekanizmalar kesin olarak henüz bilinmemekle birlikte birçok hipotez ortaya konmuştur. Fibromiyalji ve irritabl bağırsak sendromu (IBS) arasındaki ilişki göz önüne alındığında, değişen bağırsak mikrobiyomu fibromiyalji ile ilişkili olabilir. Bu çalışmada, FMS tanılı hastalarda sağlıklı kontrollere göre değişen bağırsak mikrobiyom düzeylerinin araştırılması amaçlanmıştır. Mikrobiyom çalışması için FMS tanılı 54 hastadan ve 36 sağlıklı kişiden oluşan kohorttan fekal örnekler toplanmıştır. Kontrol grubunda herhangi bir mental ve/veya fiziksel hastalığı olanlar çalışma dışında bırakılmıştır. FMS’li hasta grubu, -American College of Rheumatology (ACR)- 2010 tanı kriterlerine bağlı kalınarak belirlenmiştir. Fekal örnekler, kullanılana kadar -80°C’de muhafaza edilmiş ve buz üzerinde çözülmüş; her ekstraksiyon için 0.3 g feçes tartılmıştır. DNA izolasyonu, ticari kit ile üreticinin protokolüne uygun olarak gerçekleştirilmiştir. Örnekler, gerçek zamanlı polimeraz zincir yöntemi ile Bacteroidetes, Firmicutes, Enterobacter, Lactobacillus, Streptococcus ve Bifidobacterium’a özgül primerler ile 16S rRNA gen amplifikasyonu yapılarak karşılaştırılmıştır. Sonuçlara göre, hasta grubunda Bacteroidetes ve Bifidobacterium istatistiksel olarak anlamlı bir şekilde artarken (p< 0.05), Firmicutes’in azaldığı saptanmıştır (p< 0.001). Enterobacter, Streptococcus ve Lactobacillus için istatistiksel olarak anlamlı sonuç bulunamamıştır (p> 0.05). Bakteriler arasındaki ilişki değerlendirildiğinde, Bacteroidetes ile Firmicutes yüzdesi arasında istatistiksel olarak yüksek anlamlı ve negatif korelasyon bulunurken (r= -0.778, p< 0.001), Enterobacter ve Bifidobacterium yüzdesi arasında orta derecede istatistiksel anlamlılık ve pozitif korelasyon gözlenmiştir (r= 0.460, p= 0.005). Sonuçlar, bağırsak mikrobiyotasının fibromiyaljide rol oynayabileceğini göstermektedir. Bağırsaktaki Firmicutes ve Bacteroidetes filumlarının dengesinin bağırsak homeostazı için önemli etkilere sahip olduğu bilinmektedir. Özetle, daha büyük kohortlarda yapılacak büyük ölçekli araştırmaların, bağırsak mikrobiyomu ve FMS arasındaki ilişkiyi anlamada ve olası tedavi seçeneklerini değerlendirmede etkili olacağı açıktır.Publication Open Access Detection of Anti-CagA Antibodies in Sera of Helicobacter pylori-Infected Patients Using an Immunochromatographic Test Strip(2020-03-01T00:00:00Z) Karakus, Cebrail; Ulupinar, Zeynep; AKBAŞ, FAHRİ; Yazici, Duygu; AKBAŞ, FAHRİThe cagA gene of Helicobacter pylori that encodes an immunodominant CagA protein provokes severe mucosal damage and acts as a risk factor for the development of peptic ulcer disease and gastric cancer. Our aim is to develop an immunochromatographic test strip (ICTS) using our previously developed recombinant CagA (rCagA) protein and anti-rCagA monoclonal antibody (Mab) for the detection of anti-CagA antibodies in sera of infected patients. The rCagA was firstly conjugated to gold nanoparticle and placed into the conjugate pad. A nonconjugated rCagA and anti-rCagA Mab (CK-02) were immobilized on the test line and control line, respectively. Biopsy and serum samples from 30 H. pylori-infected patients were used. The presence of cagA gene in biopsy samples was first detected by PCR (Polymerase Chain Reaction), and 22 patients were found positive while 8 were negative. When serum samples were tested by our developed ICTS, 21 were positive for anti-CagA antibodies while 9 were negative. The serum samples were also tested by a commercial ELISA (Enzyme Linked Immunosorbent Assay), and when compared to the ICTS a sensitivity of 95% and a specificity of 100% were obtained. The ICTS can be used for rapid detection of CagA-positive H. pylori infection instead of expensive, time consuming and laborious invasive approaches.Publication Open Access Platelet-rich plasma and platelet-derived lipid factors induce different and similar gene expression responses for selected genes related to wound healing in rat dermal wound environment(2020-12-01T00:00:00Z) AKBAŞ, FAHRİ; Ozdemir, Busra; BAHTİYAR, Nurten; Arkan, Hulya; ONARAN, İlhan; AKBAŞ, FAHRİAlthough platelet-rich plasma (PRP) is the plasma fraction that contains higher levels of platelet-sequestered proteins such as growth factors and chemokines, it is also abundant in bioactive lipids whose role in wound healing has not been well characterized. This study provides a preliminary evaluation for the effect of the lipid component of PRP on selected genes related to wound healing. Sprague-Dawley rats were classified into four groups after induction of full thickness excisional wounds: the lipid fraction (LF) (lipid extract from PRP) group, PRP group, dimethyl sulfoxide group, and sham group. Subsequently, relevant groups were topically treated with test preparations. Healing wounds were collected on 3rd, 7th and 14th days, and expression levels of 12 genes were determined using qPCR. LF treatment-induced gene expression signature distinct from that induced by PRP treatment, although there are some overlaps in LF- and PRP-responsive genes. Differentially expressed all eight genes (Cxcl5, Cxc11, Egfr, Tgfb1, IL10, Tgfa, Mmp1, and Mmp7) to LF response were significantly down-regulated at either 3rd, 7th, or 14th days. Also, the comparison between LF- and PRP-treatment groups showed that the LF significantly decreased expression of Cxcl11, Mmp7, and Tgfa mRNA on day 7 of healing. This study revealed that PRP and its LF induced different and similar gene expression responses of the skin during the repair of full thickness excisional wounds. Identifying mRNA response to LF treatment at whole transcriptome level can be beneficial for comprehensive understanding of the role of platelet-derived lipid factors in wound healing processes.Publication Metadata only The effect of lucilia sericata larvae secretions on healing of osteochondral defects in a rat model(2021-07-29) BOTANLIOĞLU H.; KARAİSMAİLOĞLU B.; AYDINGÖZ Ö.; ÖZŞAHİN M. K.; YETİŞMİŞ S. C.; POLAT E.; ONARAN İ.; RAMADAN S.; TANRIVERDİ G.; DAĞISTANLI F.; et al.; AKBAŞ, FAHRİ