Theses supervised by Prof. Dr. Hacı Ömer Ateş
25 theses · Tokat Gaziosmanpaşa University
COVID-19 hastalarında ACE gen polimorfizminin belirlenmesi
The causal factor behind the respiratory illness COVID-19 is the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). This virus poses one of the most formidable challenges to human health in recent centuries, given its high morbidity and mortality rates. In March 2020, the international proclamation by the World Health Organization designated the worldwide dissemination of COVID-19 as a pandemic. The SARS-CoV-2 virus demonstrates an increased proclivity for binding to the angiotensin-converting enzyme-2 (ACE2) receptor. ACE2 and angiotensin-converting enzyme-1 (ACE1) are integral components of the renin-angiotensin-aldosterone system (RAAS) and play a pivotal role in preserving local tissue homeostasis. In the current research, we investigated the ACE I/D (Insertion/Deletion) polymorphism in a total of 141 participants, the sample set of 100 COVID-19 patients and 41 healthy controls employing the polymerase chain reaction technique. The research examines the potential link between ACE genetic variations and the intensity of symptoms caused by SARS-CoV-2. This study aimed to evaluate the association of angiotensin-converting enzyme (ACE1) gene Insertion/Deletion (I/D) polymorphism (rs1799752) with the COVID-19 severity. The mean age of the participants was 49.5 years (ranging from 17 to 94). The data analysis revealed that the p-value to be 0.989 when comparing ACE gene polymorphism in the patient cohort to the control group, hence statistically non-significant. A potential limitation of this study was the relatively small sample size. In summary, no discernable correlation was detected between the I and D allele polymorphism in ACE gene and the severity of symptoms in SARS-CoV-2 patients within the population studied. Keywords: ACE gene, gene polymorphism, Covid-19, angiotensin converting enzyme
COVID-19 hastalarında MTHFR gen polı̇morfı̇zmı̇nı̇n belı̇rlenmesı̇
The Covid-19 pandemic denotes an exceptionally transmissible illness originating from the SARS-CoV-2 virus, a recent inclusion within the coronavirus lineage. It was first identified in December 2019 in Wuhan, a city located in the Hubei province of China. The rapid spread of this illness across international borders led the World Health Organization (WHO) to officially declare it a global pandemic in March 2020. In the scope of this academic research endeavor, our main objective was to examine the effects of genetic variations within the methylenetetrahydrofolate reductase (MTHFR) gene C677T in individuals affected by COVID-19. A cohort comprising 140 participants including 99 Covid-19 patients and 41 healthy control was procured from Tokat Government Hospital, subsequently transported to the Faculty of Medicine, and subjected to comprehensive analysis within the laboratory facilities of the Department of Medical Biology. The investigative procedures involved DNA isolation, employing a series of methodologies, including Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) techniques. The obtained results were subsequently visualized through Gel Electrophoresis, facilitating the identification of genetic variants. The primary objective of this analytical endeavor was to discern potential correlations between the Methylenetetrahydrofolate Reductase (MTHFR) gene C/T polymorphism (rs1801133) with the severity of Covid-19 manifestations with a p-value less than 0.05. Significantly, a discernible proportion of our results, precisely 42.86%, demonstrated statistical significance in the comparisons of genotypes between the Control group and patients, as well as between mild and inpatient cases in Turkish population. The calculated p-values for these respective genotype comparisons were 0.0154, 0.0222, and 0.0204.
COVID-19'da protrombı̇n polı̇morfı̇zmı̇nı̇n belı̇rlenmesı̇
The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has led to global health, societal, and economic concerns. The virus attaches to ACE2 receptors in various tissues, initiating illness. A heightened immune response plays a crucial role, and severe cases may involve a cytokine storm. The exact pathophysiological mechanisms and the impact of genetic polymorphism on disease progression remain unclear. Clinical manifestations vary, with thrombotic events linked to the genetic polymorphism Factor II G20210A in prothrombin, a key coagulation component. The aim of this study was to investigate the correlation between the G20210A polymorphism (rs1799963) of the prothrombin gene and the development of severe symptoms in individuals infected with SARS-CoV-2. A cohort comprising 100 COVID-19 patients was enrolled for this investigation, and genotyping of the prothrombin G20210A polymorphism was performed using Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) analysis. The frequency of the G20210A polymorphism was compared among the three groups, and associations with clinical outcomes were examined. The results of our study revealed that all patients exhibited the GG genotype. This observation implies that, within this particular patient cohort, the presence of the prothrombin G20210A polymorphism is not associated with the severity of COVID-19.
Böbrek kanserinde WAVE11 gen ekspresyon düzeyinin analizi
Cancer is characterized by uncontrolled cell proliferation and is clearly the most destructive groups of human pathology. Renal cancer was previously thought to exist as a single illness, but it is currently recognized as several distinct cancer types that develop from kidneys, any one of them vary from histological features, disease progression, treatment responses, and genetic basis factors. Several of risk factors influenced renal cancer, involving lifestyle factors, medical condition factors, environmental factors, as well as genetic factors. The WASP family member 1 (WASF1) gene codes for WASP verprolin-homologous protein1 (WAVE1) which is an essential actin cytoskeleton control protein that is highly abundant in the brain. Particularly, WAVE1 is one of the subunits of the WAVE regulatory complex (WRC) that permits activation of the complex of the actin-related protein 2/3 (ARP 2/3) and results in inducing the polymerization of actin that is important in diverse cell events like cellular migration and cellular adhesion. In this paper, we looked at the WAVE1 (also known as WASF1) expression levels within renal cancer. For our work, 11 tissue samples of renal cancer as well as 11 healthy renal tissue samples from the same patient were evaluated. The renal cancer tissues were composed of 6 samples at stage I, 4 samples at stage III, and 1 sample at stage II. To analyze the WAVE1 gene expression level in renal cancer and healthy tissues of any samples, we employed the quantitative real-time polymerase chain reaction (qRT-PCR) method. The findings of this study exhibited that the level of WAVE1 gene was overexpressed in renal cancer tissue samples compared with healthy tissues (Fold change=5.08), which was statistically significant (p = 0.030). As a result, with all of these data, our research findings recommend that the WAVE1 gene does have a significant role in renal cancer progress and growth. Key Words: Actin Cytoskeleton, Kidney Cancer, qRT-PCR, WAVE1.
Böbrek kanserinde WAVE3 gen ekspresyonunun analizi
, xi + 60 pages Cancer is a disease distinguished by uncontrollable cell proliferation that can develop in almost any organ or tissue, such as the kidney. Kidney cancer is a complex illness. Multiple distinct subtypes of kidney cancer develop in the kidney's cells; these subtypes vary in their histology, clinical manifestations, therapeutic responses, and underlying genetic causes. The Wiskott-Aldrich syndrome protein family verprolin homologous protein 3 (WAVE3) gene, whose official name is WASP family member3 (WASF3), belongs to the WASP-WAVE family and is an actin cytoskeleton remodeling protein. WAVE3 is required for many regulatory functions in cells, including organizing cell shape, regulating actin-filament polymerization of the cell membrane, motility, migration, and cell metastasis. Furthermore, it is associated with cellular proliferation and cytokinesis. Nonetheless, WAVE3 has a major function in cancer cell invasion and metastasis. Thus, it acts as a metastasis-enhancing protein. This present investigation on the Turkish population is the first to examine the relationship between the WAVE3 gene and kidney cancer. The aim of present study was to determine the expression level of the WAVE3 gene in kidney cancer tissues relative to healthy tissues. The study involved 11 kidney cancer tissues and 11 healthy tissues from the same participants. The gene's expression level was measured by the quantitative real time polymerase chain reaction technique (qRT-PCR). According to the findings of the present study, there is no difference in WAVE3 gene expression in kidney cancerous tissues when compared with healthy tissues (fold change = 0.93), and regarding this finding, statistically, no significant correlation was found (p = 0.87). Accordingly, further studies with large samples are needed to analyze and clearly understand the WAVE3 expression level relationship to kidney cancer. Key Words: Kidney cancer, Real-Time PCR, WAVE 3.
Böbrek kanserinde actr2 gen ekspresyonunun belirlenmesi
Kidney cancer, particularly renal cell carcinoma (RCC), poses a significant health challenge globally, with its incidence steadily rising. Understanding the molecular mechanisms underlying its pathogenesis is crucial for developing targeted therapies. Among the multitude of genes implicated in kidney cancer, the Actin-Related Protein 2 (ACTR2) gene has lately received attention for its potential role in tumor development and disease furtherance. ACTR2 is an important member of the actin-related protein (ARP) group that participates in several cellular activities like cytoskeletal dynamics, intracellular transport, and gene expression control. The study aim to explore the functional implications of ACTR2 dysregulation in kidney cancer, highlighting its role in tumor cell proliferation, invasion, and resistance to conventional therapies. The study used quantitative Real-time polymerase chain reaction (qRT-PCR) to analyse gene expression rates in 11 cancer and 11 normal tissue samples from the kidney of the same patient. The research indicated a 1.206-fold elevation in ACTR2 gene expression in kidney cancer tissue samples relative to normal kidney tissues; however, this increase did not reach statistical significance (p = 0.234). Accordingly, we believe that further studies with large samples are needed to analyze and understand the ACTR2 expression level relationship to kidney cancer.
Böbrek kanserinde ARPC1A gen ekspresyonunun belirlenmesi
Renal cell carcinoma (RCC), in particular, is an important medical issue that is becoming more commonplace worldwide. Developing targeted therapeutics requires an understanding of the molecular pathways behind the disease's etiology. The ARPC1A gene is one of many genes linked to kidney cancer; however it has lately come to light due to its possible involvement in carcinogenesis and the advancement of the disease. Protein linked to actin the actin-related protein (ARP) family, which includes Subunit1A, is also known by the shortened form ARPC1A. This family of proteins is essential to many cellular functions, like the control of gene expression, intracellular transport, and cytoskeletal dynamics. The ARPC1A gene, located on chromosome 7q22.1, encodes the ARPC1A protein, a crucial component of the ARP2/3 complex. In order to enable cytoskeletal rearrangements that support cell motility, shape maintenance, and membrane dynamics—activities that include endocytosis, cell-cell adhesion, and cell motility—the ARP2/3 complex is a crucial regulator of actin filament nucleation and branching. This research analyzed gene expression rate samples in 11 effected cells and 11 control cells from the kidney of the same patient using quantitative reverse transcription PCR. In comparison to normal kidneys, the study found that kidney cancer tissue samples had 1.14 -fold higher ARPC1A gene expression; however, this increase is not statistically significant (p = 0.75). This abstract discusses the role of the ARPC1A gene in kidney cancer.
Böbrek hücreli karsınomda ACTR3 gen ekspresyonunun analizi
Renal cell carcinoma (RCC), in particular, is a major health concern that is becoming more commonplace worldwide. Developing targeted therapeutics requires an understanding of the molecular pathways behind the disease's etiology. The Actin-Related Protein 3 (ACTR3) gene has drawn interest among the many genes linked to kidney cancer because of its possible involvement in tumor growth and disease progression. A crucial component of the actin-related protein (ARP3) group, ACTR3 is involved in a number of biological processes including intracellular transport, cytoskeletal dynamics, and gene expression regulation. The purpose of the study is to compare the role of ACTR3 gene expression level between renal cell carcinoma tissue and control renal tissue. Quantitative Real-time polymerase chain reaction (qRT-PCR) was employed in the study to analyze gene expression. The study analyzed gene expression levels by examining ten renal carcinoma tissue samples and ten corresponding control renal tissue samples, both collected from the same group of patients using quantitative Real-time polymerase chain reaction (qRT-PCR). The research demonstrated a fold change with 1.147 in ACTR3 gene expression in kidney cancer tissue samples relative to normal kidney tissues; however, this increase did not meet statistical significance, since (p = 0.578). Therefore, we think that more research using larger sample sizes is necessary to examine and comprehend the connection between kidney cancer and ACTR3 expression levels.
Pterjiumda CRABP2, FABP5 ve PPARD genlerinin rolü
Pterjium, anormal fibrovasküler proliferasyon, matriks remodelizasyonu ve invazyonuyla karakterize edilen kanat şeklinde fibrovasküler bir dokudur. Pterjiumun patogenezi hala bilinmemektedir. Retinoik asit (RA), hücre içerisinde iki farklı lipit bağlayıcı protein olan hücresel retinoik asit bağlayıcı protein 2 (CRABP2) ve yağ asidi bağlayıcı protein 5 (FABP5) tarafından taşınmaktadır. RA sinyalleri tümör baskılayıcı yolak olan RA reseptörü (RAR) ve tümör ilerletici yolak olan proliferatör aktive edici reseptör β/δ (PPAR β/δ) aracılı yolak olmak üzere iki farklı yol izler. RA sinyallerinde görülen bozukluklar Pterjiumun gelişimine ve/veya nüksüne katkıda bulunabilir. Çalışmamızın amacı primer ve nüks pterjium dokusunda CRABP2, FABP5 ve PPARD genlerinin mRNA ve protein ekspresyonunu ve CRABP2/FABP5 oranını belirlemektir. Çalışmamızda cerrahi operasyon sırasında 30 hastadan pterjium ve sağlıklı konjonktiva dokusu elde edildi. Elde edilen pterjium ve normal konjonktiva dokularında CRABP2, FABP5 ve PPARD'nin mRNA ve protein ekspresyonları gerçek zamanlı PZR ve western blot yöntemleri kullanılarak değerlendirildi. Primer pterjium ve normal konjonktiva dokuları karşılaştırıldığında CRABP2 ve FABP5 genlerinin ekspresyonu değişmemiştir (1,03±0,22- fold change, p=0,895; 1,02±0,19 p=0,917, sırasıyla). Nüks pterjium ve normal konjonktiva dokuları karşılaştırıldığında ise CRABP2 ve FABP5 genlerinin ekspresyonu azalmıştır (0,71±0,39- fold change, p=0,577; 0,88±0,43 p=0,577, sırasıyla). Hem primer hem de nüks pterjium dokusu normal konjonktiva dokusuyla karşılaştırıldığında PPARD'nin ekspresyonu değişmemiştir (1,08±0,28-kat değişikliği, p=0,784; 1,10±0,71 p=0,095, sırasıyla). Pterjium etiyopatogenezinin anlaşılması, erken aşamalarda pterjium gelişiminin engellenmesinde daha umut verici tedavilerin bulunmasına yardımcı olacaktır. Anahtar Kelimeler: CRABP2, CRABP2/FABP5 oranı, FABP5, PPAR β/δ, retinoik asit sinyal yolağı
Bir yüzey yapışma proteini olan e-kaderinin pterjiyum gelişimindeki rolünün araştırılması
Gözde et büyümesi olarak ifade edilen pterjiyum, kornea üzerinde konjonktivanın kontrolsüz bir şekilde büyüyerek ilerlemesiyle sonuçlanan yaygın bir göz hastalığıdır. Özellikle anormal çoğalmaya sebep olarak fibrovasküler dokunun epitel hücrelerinde görülen bozukluklar gösterilmiştir. Epitelyal kaderin olarak da bilinen E-kaderin, hücreler arası adezyona aracılık etmenin ötesinde hücre tanıma, sınır oluşumu ve korunması, koordineli hücre hareketleri de dâhil pekçok biyolojik süreçte anahtar rol oynamaktadır. E-kaderin ekspresyonun azalması, hücre adezyonun kaybına, tümör büyümesi ve proliferasyonuna neden olarak metastaza yol açtığı bilinmektedir. Bu çalışmada, tümör oluşumuna benzer davranışlar gösteren pterjiyum ile bir tümör supresör gen olan E-kaderinin ilişkisi araştırılmıştır. Çalışmada 36 hastaya ait pterjiyum dokusunda E-kaderinin geninin promotör metilasyonu metilasyona spesifik PZR yöntemi ile, ekspresyon düzeylerindeki farklılık kantitatif revers transkriptaz polimeraz zincir reaksiyonu qRT-PZR yöntemi ile ve protein ifade farklılıkları western blot yöntemi ile belirlenmiştir. Elde edilen verilere göre, kontrol grubuna göre pterjiyumlu dokularda eE-kaderin geni metilasyon ve ekspresyon düzeyleri istatistiksel olarak anlamlı bulunmuştur (p<0,005). E-kaderin genindeki metillenme ve ekspresyon ifadelerindeki azalışın pterjiyum oluşumu ve gelişimiyle ilişkili olabileceği düşünülmektedir.
Pterjiumda WASP geninin analizi
Analysis WASP Gene Expression in Pterygium. Tokat Gaziosmanpaşa University, Health Sciences Institute, Department of Medical Biology, MSc. Thesis, Tokat. Pterygium is a benign growth of the conjunctive tissue that covers the sclera that is proliferative and causes intrusive ocular surface and wide matrix remodeling. The WASP gene is located on the p arm (short arm) of the X chromosome at locations Xp11.4→ p11.21 and encodes a 502 amino acid sequence. The WASP gene comprises 12 exons, and the protein has a molecular weight of about 65 kD. The WASP gene has multiple distinct domains, including a GTPase, GBD, PH domain, cofilin, and verprolin homology domain, SH3 binding motifs with a proline-rich locus, and the C-terminal acidic region, which plays a role in actin cytoskeleton remodeling by beginning Arp2/3. The WASP gene appears to play a function in signal transduction as well as cytoskeletal remodeling. Actin restructuring, platelet formation/removal, cell-cell contact, actin polymerization, cell trafficking and motility, immunological synapse arrangement, and neutrophil formation/release are all linked to WASP. The goal of this research was to see if the WASP gene was expressed in pterygium.Tissues were removed throughout surgery with the patient's agreement and knowledge. By collecting cDNA from the tissue levels established by real-time PCR, we employ the WASP gene expression technique. WASP gene expression levels were determined to be statistically significant higher in tissues with pterygium compared to the control group (p < 0.005), based on our findings that the increase in WASP gene expression is hypothesized to be linked to the formation and development of pterygium. According to qRT-PCR data, WASP gene expression is pterygium tissue compared to healthy conjunctive tissue (1.9100.367-foldchange, p=0.0001).
Pterjiyumda ARPC1B gen ekspresyonunun analizi
Pterygium, or flesh growth in the eye, is a common eye illness that causes the conjunctiva on the cornea to expand out of control. The most common cause of epithelial tissue abnormalities is long-term UV radiation exposure from the sun. Although the precise etiology of the pterygium disease is unknown, additional elements such as dry weather, dust, and a vaporization of the tear film, and other genetic factors, Despite the fact that the exact cause of pterygium illness is uncertain, there seems to be a correlation between outdoor work and the illness's development. Subunit 1B of the Actin Related Protein 2/3 Complex the name of the ARPC1B gene. The human Arp2/3 protein complex has seven subunits, and this gene codes for one of them. Sensory rhodopsin-2 protein is a component that belongs to the SOP2 family. The goal of this research was to see how the ARPC1B gene affected pterygium illness. Tissue was extracted during surgery with the subjects' agreement and consent. We employed the technique expression of ARPC1B gene by extracting cDNA synthesis from group of tissues levels assessed by Real-Time PCR, and the diagnosis of pterygium was validated (polymerase chain reaction). According to the findings of qRT-PCR, there was no discernible change. in ARPC1B gene levels of expression in pterygium and normal conjunctival tissues (p>0.005).
Pterygiumda actin ile ilgili protein 2/3 kompleks alt bira 2 gen ekspresyonunun analizi
The most important aspects of diagnosing a disease are determining its cause and then attempting to find a treatment. Pterygium is a disorder that isn't completely understood yet and it's described as a growth disorder by some and a degenerative disorder by others. However, several studies have been conducted in this field, and certain factors for this disease have been identified, including UV, virus, growth factors, apoptosis, ECM, MMP, and interleukins. Uncontrolled proliferation of conjunctiva cells onto the cornea, metaplasia, and certain reports of malignant traits have been documented. Furthermore, the ARPC-2 gene has been discovered to play a role in the progression and induction of a certain type of cancer, which is one of the Arp 2/3 complex's subunits that is involved in actin assembly and therefore sustains and maintains cellular integrity, the objective of this paper is to determine how ARPC-2 gene expression relates to pterygium development and formation. The quantitative reverse transcription PCR method was used to determine the rate of gene expression in 18 samples with pterygium and 18 normal conjunctiva of the same eye. The gathered data showed that the expression of ARPC-2 in pterygium has up-regulated (3.24±1,75) compared to normal conjunctiva, although there was not a significant difference between the two groups (p = 0.209). In conclusion the up-regulation of ARPC-2 gene is thought that it may contribute to pterygium development and formation.
Pterjiumda ACTR-3 gen ifadesinin analizi
Pterygium is a disorder of the ocular surface defined by uncontrolled cell proliferation, fibrovascular growth that crosses the cornea of the eye and disruption of cell shape and morphology, moreover ACTR-3 is a gene that encodes for one of the major subunits of ARP2/3 complex that help regulate actin assembling in which it gives cell its shape and motility. The purpose of this study is whether there's a link between ACTR-3 gene expression and pterygium development.The study included 18 pterygium patients and 18 volunteers with normal conjuctiva.The expression of the ACTR-3 gene in each group of individuals was evaluated using a quantitative reverse transcription PCR technique.. The result of the study was statistically significant (P=0.015) and there was an increase in ACTR-3 gene (13.62±4.91) in pterygium tissues compare to normal conjuctiva. We concluded that this increase in expression of ACTR-3 gene may have role in pterygium formation.
Pterjiyum tanısı almış hastalarda COL1A1 geninin analizi
Pterjiyum, göz kapağı aralığına yerleşen üçgen şekilli, genellikle bilateral, nazal tarafta bulbar konjoktivadan orjin alan dejeneratif, anormal fibrovasküler dokunun kornea üzerine ilerlemesi olarak tanımlanmaktadır. Kollajen, hareket sisteminin yapı taşlarını, özellikle kemik, kıkırdak, lif ve eklemleri oluşturan proteindir. Bu protein birbiri üzerine sarılmış üç alfa zincirinden meydana gelmektedir. 28 tane değişik tipi tanımlanmış olup, tip I, tip II şeklinde isimlendirilmektedir. Tip I kollajen ECM'de en fazla bulunan kollajen tipi olarak bilinmektedir. Kollajen tip I alfa 1 (COL1A1) geni ise, iki alfa 1 zinciri ve bir alfa 2 zincirinden oluşan tip I kollajenin pro-alfa 1 zincirlerini kodlamaktadır. COL1A1 geni 17q21.33 pozisyonunda kromozom 17 üzerinde yer almakta ve 51 ekzon içermektedir. Pterjiyum ile ilgili birçok çalışma olmasına rağmen, hastalığın patogenezi tam olarak açıklanamamıştır. Pterjiyum patogenezinin hücresel proliferasyon ve anti-apoptozis ile karakterize edilmesi, pterjiyumun dejeneratif bir hastalıktan ziyade bir tümör anoloğu olduğunu düşündürmektedir. Kollajen familyasının üyelerinin çeşitli doku tiplerinde karsinojenezde rol oynadığına dair kanıtlar bulunmaktadır. COL1A1 gen mutasyonlarının patojenitesinin birçok hastalıkla ilişkili olduğu söylenmektedir ve COL1A1'nin anormal ekspresyonun bazı kanserlerde rol oynadığı bildirilmiştir. Pterjiyum tanısı almış 27 olgunun pterjiyum ve normal konjonktiva dokusu çalışmaya alındı. Pterjiyum dokusunda COL1A1 geninin RNA düzeyinde ekspresyonu için qRT-PCR analizi yapıldı. Çalışma sonucunda elde edilen verilere göre normal konjonktiva dokusuna kıyasla pterjiyum dokusunda COL1A1 geni ekspresyon düzeyinin arttığı fakat bu artışın istatistiksel olarak anlamlı olmadığı bulunmuştur (p>0,05). Anahtar Kelimeler: COL1A1, Pterjiyum, qRT-PCR
WASL(N-WASP) gen ekspresyonunun pterjiyumdaki etkisi
Pterygium is a benign growth of the conjunctive tissue that covers the sclera with a high proliferation property which causes intrusive ocular surface and wide matrix remodeling. WASL which also known as n-Wasp (Neural Wiskott-Aldrich syndrome protein) is a nucleation-promoting factor that promotes the formation of branching actin filaments. The WASL gene is located on the q arm (long arm) of chromosome 7, which locate q31.32 and encodes a 505 amino acid sequence. The WASL gene comprises 11 exons, and the encoded protein has a molecular weight of about 54.84 kD. The WASL gene has several important molecular function, including a actin binding, GTPase regulator activity. SH3 binding motifs with a proline-rich locus, and the C-terminal acidic region, which plays a role in actin cytoskeleton remodeling by stimulating Arp2/3. The WASL gene appears to play a function in signal transduction as well as cytoskeletal remodeling. Regulation of cortical actin filament rearrangement in response to external stimuli is mediated through the Arp2/3 complex, which links small GTPases (e.g., Rac and Cdc42) with actin polymerization. The aim of this study was to see if the expression of the WASL gene changed in pterygium versus healthy cataract tissue. Tissues were removed with the patient's consent and awareness throughout the procedure. We use the WASL gene expression approach to harvest cDNA from tissue levels determined by real-time PCR. WASL gene expression levels were determined to be statistically significant higher in tissues with pterygium compared to the control group (p < 0.05), based on our findings that the increase in WASL gene expression is hypothesized to be linked to the formation and development of pterygium. According to qRT-PCR data, WASL gene expression is pterygium tissue compared to healthy conjunctive tissue (1.320±0,254-foldchange, p=0.001).
Pterjiyumda ACTR2 gen ekspresyonu
Pterygium is a benign, uncontrolled growth of conjunctival tissue extending over the sclera. Pterygium is characterized by inflammatory, proliferative, invasive ocular surface and extensive matrix remodeling. Pterygium is a benign growth of conjunctival tissue lining the sclera that is proliferative and causes invasive ocular surface and extensive matrix remodeling. the ACTR2 gene is located on the p arm (short arm) of autosomes (body chromosome(s)) at locations 2p14 encoding the 399 amino acid sequence. The ACTR2 gene consists of 10 exons and consists of 3798 base pairs. ACTR2 encodes Actin-Associated Protein 2 (ARP2), is involved in the formation of branched actin networks in the cytoplasm, important for cell shape. It provides force for cell motility and takes part in cell proliferation. loss of ACTR2 or the presence of abnormal transcripts can result in aberrant cellular proliferation by the activity of ACTR2. In this study, it was aimed to determine the ACTR2 gene expression, which may cause abnormal benign tissue growth from the conjunctiva (mucous membrane covering the eye) to the cornea. It depends on cell adhesion, motility, and deformability resulting from quantitative changes and rearrangement of the actin-related protein. Our aim in this study is to determine the expression level of ACTR2 gene in pterygium tissue. qRT-PCR analysis was performed for expression of ACTR2 gene at RNA level in pterygium tissue. According to the data obtained at the end of the study, ACTR2 gene expression levels were found to be statistically significantly higher in tissues with pterygium compared to the control group (p < 0.005) (p=0.005).
Pterjium gelişiminde WASF3'ün rolünün araştirilmasi
Pterygium is a dysplastic Triangular-shaped and fleshy fibro vascular structure. In general pterygium color appears as pearl white to pink in the eyes. Pterygium grows up across the limbus toward cornea as sub epithelial ingrowth of degenerative bulbar conjunctival tissue. Previously, it has been mentioned by the studies that pathology of pterygium correlated with several causes which are known as risk factors. These factors consist of environmental, genetic, immunological, geographical, viral and aspectic inflammation factors. WASF3 belongs to the Wiskot-Aldrich syndrome proteins WASP/WAVE family of actin-binding protein that play essential roles in regulating cell morphology, actin polymerization, cytoskeleton remodeling, cell motility and invasion.it regulates many aspects of cancer progression, cell invasion, cell migration and metastasis, as well as remodeling of cytoskeleton. WASF3 was classified as a key driver of tumor progression and metastasis in cancers from several origins as well. The aim of this study was to investigate the effect of WASF3 on the behaviors of pterygium disease and to evaluate its clinical impact. In this framework, we assessed the association of WASF3, a familiar actin-binding protein and analogous to tumor suppressor gene with pterygium which exhibits tumor-like behavior. In our study We analyzed the expression level of WASF3 gene in pterygium tissue belonging to 27 patients was determined by qRT PCR method. According to the obtained data, the expression levels of the WASF3 gene in tissues with pterygium were statistically significant compared to the control group (p <0.005). It is thought that the decrease in the expression of the WASF3 gene may be related to the formation and development of pterygium. Key Words: Pterygium, Real-Time PCR, WAVE 3
ARPC1A geninin periodontitis gelişimindeki rolünün araştırılması
Periodontitis konak immun yanıtı sonucunda oluşan; yumuşak ve sert doku yıkımı ile karakterize inflamatuar bir hastalıktır. Periodontitis, zamanında tedavi edilmediğinde, dişleri destekleyen periodontal bağın bağlanma kaybına, diş kaybına ve birleşme epitelinin apikal göçüne neden olan, kronik yapısı nedeniyle tekrarlama riski yüksek bir hastalıktır. Hücre göçü, gelişim, doku onarımı ve bağışıklık tepkileri gibi temel fizyolojik süreçler için gereklidir. Bağışıklık ve hematolojik hücrelerde aktin hücre iskeletinin koordineli olarak yeniden düzenlenmesinin önemi, bu sürecin bozulduğu insanlarda çeşitli hastalıklarla vurgulanmaktadır. Birçok hastalık hücre iskeleti ve proteinlerdeki anormalliklerle ilişkilendirilmiştir. Periodontitisin böyle bir hücre iskeleti hastalığı olduğu düşünülmektedir. ARPC1A, aktin ile ilgili protein ARP2/3 kompleks ailesinin bir üyesidir. ARPC1A geni, hücrelerde aktin polimerizasyonunun kontrolünde yer alan insan Arp2/3 kompleksinin p41 alt birimini kodlar. ARPC1A'ın hücre göçünde rol aldığına dair bir çok literatür çalışması mevcuttur. Bu çalışmada, ARPC1A geni aktin hücre iskeleti düzenleyicisinin, bir inflamatuar hastalık olan perodontitisde ilişkisi araştırılmıştır. Periodontitis nedeniyle ileri diş destek doku kaybı oluşumuna bağlı diş çekimi için başvuran periodontitis tanısı almış 20 hastanın, diş çekim endikasyonu bulunan dişler çekildikten sonra, yumuşak doku düzeltmesi gereksinimi olan bölgelerden periodontitisli yapışık diş eti doku örnekleri elde edilmiştir. Kontrol grubu doku örnekleri için aynı hastadan rezektif gingival cerrahi endikasyon varlığında sağlıklı diş eti örneği alınmıştır. Periodontitis dokusunda ARPC1A geninin mRNA düzeyinde ekspresyonu için RT-PCR analizi yapıldı. Çalışma sonucunda elde edilen verilere göre periodontitis dokusunda ARPC1A geninin ekspresyon seviyesi normal dokuyla karşılaştırıldığında periodontitis dokusunda artış olduğu fakat bu artışın (p=0,433, değeri p>0.05), istatistiksel olarak anlamlı olmadığı saptanmıştır.
Periodontitiste WAVE1 gen ekspresyonunun analizi
Periodontitis is a multifactorial inflammatory autoimmune disorder characterized by disruption of the periodontium. It is a result of the interaction between host inflammatory response and supra- and sub-gingival biofilms produced by periodontopathic bacteria. The condition depends on multiple interacting risk factors, including: genetic, microbial, environmental, and immunological factors, as well as race, gender, and age. To date, specific genes directly involved in periodontitis have not been described. WASP verprolin-homologous protein 1 (WAVE1) is one of the essential actin-cytoskeleton rearrangement proteins that is primarily expressed in neuronal cells. It is a very crucial molecule in the regulation of tumor growth, invasion and metastasis. The role of WAVE1 protein in the control of actin polymerization by activation of the ARP 2/3 complex seems to be important in the formation of actin-based membrane protrusions that are observed in cell migration. The current study is the first to investigate the relationship between periodontitis and WAVE1. In this study, 18 periodontitis patients (10 male and 8 female) were recruited in order to analyze the mRNA expression of WAVE1 gene in periodontitis tissues. They had a mean age of 53.1 years (34-65). The quantitative real time PCR (qPCR) method was used to evaluate the gene expression level in periodontitis and healthy tissues of each patient. The analyzed data showed that WAVE1 was overexpressed in periodontitis tissues when compared to healthy control tissues and the difference was statistically significant (p = 0.0442). One of the limitations of this study was that it had a small sample size. In conclusion, WAVE1 does have a significant role in the development and progression of periodontitis.
Investigation of the role of WASP which is cytoskeletal protein in the development of periodontitis
Periodontitis is an inflammatory autoimmune disease with multifactorial etiology, which affects the supporting tissues of the teeth, is characterized by deepening of periodontal pockets, loss of connective tissue attachment and alveolar bone loss. Inflammation and immune system defects have important roles in immune-mediated inflammatory diseases such as periodontitis. Actin cytoskeleton; it is involved in the cellular process essential for normal immune function, including movement, intercellular interactions, endocytosis, cytokinesis, signal transduction, and maintenance of cell morphology. WASP (Wiskott-Aldrich syndrome protein) is a cytoskeletal regulatory protein known to be involved in biological processes such as cell migration, cell signaling, increased inflammatory cytokines and excessive activation of inflammation. Studies show that changes in WASP expression level cause defects in immune and inflammatory response formation. For this reason, it is thought that WASP may be a risk factor for the development of periodontitis. The aim of our study was to determine the mRNA and protein expression levels of the WASP gene in periodontitis and healthy gingival tissues. In our study, 20 periodontitis and 20 healthy gingival tissues from the same patients were included. Gene expression levels were determined by SYBR green based PCR and protein expression levels were determined by western blot analysis. According to our study results, it was found that the expression level of the WASP gene in periodontitis tissue was statistically significantly increased compared to the control tissue, while the protein level was significantly decreased. Our results suggest that WASP may be effective in the onset and development of periodontitis. Moreover, understanding the inflammatory/immune mechanisms in periodontitis may be useful for diagnosis, prevention and treatment of periodontitis in its early stages. Keywords: Actin Cytoskeleton, Immunity, Inflammation, Periodontitis, WASP
Pterjiumda konneksin 43 gen ekspresyonunun ve protein düzeyinin analizi
Pterjium, limbustan korneaya doğru konjonktivanın kanat şeklinde anormal, epitelyal ve fibrovasküler büyümesidir. Aynı zamanda, proliferatif, invaziv ve oldukça vaskülarize bir doku olduğu da söylenebilir. Pterjium, kanser benzeri özelliklere sahiptir. Bu özellikler, hücre proliferasyonu, invazyonu ve rezeksiyon sonrası tekrarlama şekilde sıralanabilir. Hastalığın en yaygın olarak bilinen faktörü ultraviyole radyasyon olmasına rağmen, patogenezi hala açık değildir. Son yıllardaki çalışmalar pterijiumlarda moleküler değişikliklerin tümör hücrelerine benzer bir mekanizmaya sahip olabileceğini desteklemektedir. Konneksinlerin hücre döngüsünün ve apoptozun düzenlenmesinde rol alan proteinlerle direkt etkileşim halinde olduğu ve bu yolla hücre büyümesini düzenleyebildiği yönünde çalışmalar mevcuttur. Konneksinler (CX), hücreler arası bağlantı birimlerinden olan gap junction kanallarının oluşumundan sorumlu olarak işlev görürler. Bu bağlantılar hücreler arasında iyonların ikincil mesajcıların ve küçük sinyal moleküllerinin doğrudan değişiminde, doku homeostazında, büyüme, farklılaşma ve gelişmenin düzenlenmesinde önemli rol oynarlar. Neredeyse her memeli hücre tipinde görülen bu bağlantı bölgelerinin en önemli özelliği hücreler arası iletişimi sağlamalarıdır. Çalışmada, 26 pterjiumlu olgunun gözüne ait pterjium ve normal konjonktiva dokularında Konneksin 43 (CX43) gen ifade düzeyi Gerçek Zamanlı Polimeraz Zincir Reaksiyonu (RT-PZR) yöntemi ile CX43 protein miktarı ise Western Blot yöntemi kullanılarak araştırılmıştır. Sonuçlarımıza göre, pterjium ve normal konjonktiva dokularında CX43 gen ifade düzeyi ve protein miktarı bakımından ististiksel olarak anlamlı bir fark bulunamadı (p>0.05). Pterjiumda, CX43 gen ifadesinin bir risk etmeni olamayacağı sonucuna varılmıştır.
Pterjiyumda wwox gen ekspresyonunun ve protein seviyesinin analizi
WWOX, yaklaşık 1.1 mb'lık genomik bir bölgeyi kaplayan ve 16q23.3→q24.1'te yerleşik bir tümör baskılayıcı gendir. Bu bölge insanlarda en yaygın ikinci frajil bölge olan FRA16D'yi kapsamaktadır. WWOX geni, 2.2 kb uzunluğunda bir transkriptten ve 414 amino asit'ten oluşan 46 kDa'luk bir protein kodlar. WWOX proteini N-terminalinde iki WW alanından ve C terminalinde ise SDR adlı bir kısa zincirli dehidrojenaz domeininden oluşur. WW alanları, prolin içeren ligandlarla bağlanarak protein-protein etkileşimlerine aracılık etmektedir. WWOX; hücre çoğalması, embriyonik gelişim, metabolizma ve apoptozu düzenleyen birçok sinyal yolağında görev almaktadır. Birçok kanser türünde WWOX'un kaybı veya anormal transkriptlerinin bulunması WWOX'un tümör baskılayıcı aktivitesini göstermektedir. Bu çalışmada, bir tümör analoğu olabileceği düşünülen pterjiyumda WWOX gen ifade ve protein düzeyinin belirlenmesi amaçlandı. Bu amaçla 27 hastaya ait pterjiyum dokusunda WWOX gen ifade değişimi gerçek zamanlı PZR yöntemi ile, WWOX protein değişimi ise Western Blot yöntemi kullanılarak araştırılmıştır. Çalışma sonuçlarımıza göre pterjiyum dokusunda WWOX geni ifade ve protein düzeyinin kontrol dokusuna göre istatistiksel olarak anlamlı bir oranda azaldığı bulunmuştur (p˂0,05). Bu bilgiler ışığında; bir tümör baskılayıcı gen olan WWOX'un, pterjiyum dokusunda ifade ve protein düzeyinin azalması, pterjiyumun kanser analoğu bir doku olabileceği yönündeki iddiaları desteklemektedir.
Yaşa bağlı makula dejenerasyonunda serum interlökin 17 düzeyinin araştırılması
Yaşa bağlı makula dejenerasyonu (YBMD), dünya genelinde yaşlılar arasında geri dönüşümü olmayan görme bozukluğu olup körlüğün önde gelen nedenleri arasında yer almaktadır. YBMD, hem çevresel hem de genetik faktörlerin etkili olduğu multifaktöriyel bir hastalıktır. Genetik faktörlerin YBMD'nin insidansı ve progresyonuna etkisi yapılan çalışmalarda ortaya konulmaktadır. İnterlökinler; hücre içi iletişimi, hücre büyümesini ve farklılaşmasını düzenleyen, immün sistemin gelişiminde ve homeostazında önemli rol oynayan, sitokin ailesinin bir alt kümesi olan protein yapılı moleküllerdir. İnterlökinler sentezlendikleri yere ve yaptıklara görevlere göre çeşitlilik göstermektedirler. İnterlökin 17, Th17 hücreleri tarafından üretilmesine rağmen bunun dışında çok çeşitli hücre tarafından da üretildiği bilinmektedir. Bu çalışmada, serum interlökin 17'nin yaşa bağlı makula dejenerasyonu ile olan ilişkisi değerlendirildi. 100 YBMD li hasta ile 100 sağlıklı kontrol dâhil edilen bu çalışmada ELİSA tekniği kullanıldı. Çalışma sonuçlarımızda, interlökin 17 nin hasta ve kontrol grupları arasındaki ilişki istatistiksel olarak incelendiğinde anlamlı bulunmuştur (p:0,001). Hasta bireylerde cinsiyet farklılığının ise interlökin 17 seviyesini etkilemediği bulunmuştur. Anahtar kelimeler: Yaşa bağlı makula dejenerasyonu, Elisa, İnterlökin 17.