Kursat O Yaykasli | Okayama University, Japan (original) (raw)

Papers by Kursat O Yaykasli

Although rarely seen, Amanita phalloides var. alba, a variety of A. phalloides type mushrooms , c... more Although rarely seen, Amanita phalloides var. alba, a variety of A. phalloides type mushrooms , causes mushroom poisoning resulting in death. Since it is frequently confused with some edible mushrooms due to its white colored cap and macroscopic appearance, it becomes important in toxicological terms. Knowledge of the toxin amount contained in this mushroom type is invaluable in the treatment of cases involving poisoning. In this study, we examined the toxin levels of various parts of the A. phalloides var. alba mushroom growing Duzce region of Turkey. Toxin analyses were carried out for A. phalloides var. alba, which were collected from the forests Duzce region of Turkey in 2011, as a whole and also separately in its spore, pileus, gills, stipe and volva parts. The alpha amanitin, beta amanitin, gamma amanitin, phalloidin and phallacidine analyses of the mushrooms were carried out using the RP-HPLC method. A genetic analysis of the mushroom showed that it had similar genetic characteristics as A. phalloides and was a variety of it. The lowest toxins quantity was detected in spores, volva and stipe among all parts of the mushroom. The maximum amount of amatoxins was measured in the gills. The pileus also contained a high amount of amatoxins. Generally, amatoxins and phallotoxin concentrations were lower as compared to A. phal-loides, but interestingly all toxins other than gamma toxin were higher in the spores of A. phalloides var. alba. The amount of toxin in all of its parts had sufficient concentrations to cause death. With this study, the amatoxin and phallotoxin concentrations in A. phalloides var. alba mushroom and in its parts have been revealed in detail for the first time.

Acne, a chronic inflammatory skin disease, can be seen at any age but it most often occurs in ado... more Acne, a chronic inflammatory skin disease, can be seen at any age but it most often occurs in adolescents and young people. Several factors, including increased sebum production, abnormal cornification of the piloseba-ceous units, proliferation of Propionibacterium acne, and extracellular matrix (ECM) remodeling, are thought to be associated with the pathogenesis of the acne. The remodeling of the ECM is regulated by a balance between matrix metalloproteinases (MMPs) and their inhibitors called tissue inhibitors of metalloproteinases (TIMPs). The current study investigated the potential association between MMP-2 (-1306 C/T) and TIMP-2 (-418 G/C) polymor-phisms and the risk for acne in a Turkish population. The study was conducted with 85 subjects who presented to the Dermatology Department of Duzce University Hospital. DNA was isolated from 2 ml of peripheral blood taken from each subject, and their genotypes were analyzed with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The CC, CT, and TT genotypes for MMP

Background: Psoriasis is a T cell-mediated immune disease in which various cytokines, primarily t... more Background: Psoriasis is a T cell-mediated immune disease in which various cytokines, primarily tumor necrosis factor-a (TNF-a), are complexly involved. Mannose-binding lectin (MBL) gene polymorphisms decrease MBL serum levels, thereby increasing the synthesis of proinflammatory cytokines such as TNF-a.
Objectives: This trial was designed to evaluate the role of the MBL2 codon 54 polymorphism in the pathogenesis of psoriasis.
Methods: Fifty patients diagnosed with psoriasis vulgaris and 53 healthy subjects were included in the trial. The polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) method was applied to determine the MBL2 codon 54 polymorphism. Genotypes were determined according to the bands formed in agarose electrophoresis gels. For the statistical analysis, the level of significance was set at P < 0.05.
Results: A total of 33 (66.0%) of the 50 psoriasis patients were detected to have A/A genotype and 17 (34.0%) had B/B genotype. Of the control subjects, 44 (83.0%) had A/A genotype and nine (17.0%) had B/B genotype. There was a statistically significant difference
between the groups (P = 0.047). The analysis of allele frequencies revealed A allele prevalences to be 79 (79.0%) and 95 (89.6%), and B allele prevalences to be 21 (21.0%) and 11 (10.4%), in the patient and control groups, respectively. A statistically significant difference between allele frequencies was detected (P = 0.031).
Conclusions: This study suggests that the MBL2 codon 54 polymorphism may have an association with psoriasis in the Turkish population.

Background: Psoriasis is a chronic inflammatory disease of uncertain pathogenesis. Omentin is a n... more Background: Psoriasis is a chronic inflammatory disease of uncertain pathogenesis. Omentin is a new adipokine with anti-inflammatory properties; however, the relationship between psoriasis and omentin has not been fully established yet.
Objectives: This study was designed to evaluate the relationship between psoriasis and omentin serum levels and Val109Asp polymorphism in exon 4 of the omentin gene.
Methods: Forty-nine patients with plaque-type psoriasis and 39 healthy subjects were included in the study. Omentin concentrations were determined by using enzyme-linked immunosorbent assay. Val109Asp polymorphism in exon 4 of the omentin gene was assessed by the polymerase chain reaction–restriction fragment length polymorphism
method. Genotypes were determined according to the bands formed in agarose electrophoresis gels. In the statistical analysis, the level of significance was set at P < 0.05.
Results: The serum omentin levels of the patients with psoriasis (354.2 152.0) were found to be significantly lower than those in the control group (488.7 190.3) (P = 0.001). A moderate level negative correlation was determined between serum omentin level and body mass index and waist circumference. No significant differences were observed
between the patient and control groups in terms of the genotype and allele frequency of Val109Asp polymorphism in exon 4 of the omentin gene (P > 0.05).
Conclusions: Omentin serum levels were determined to be low in patients with psoriasis. No significant difference was found regarding Val109Asp polymorphism of the omentin gene. To the best of our knowledge, our study is the first clinical study to examine the relationship between psoriasis and omentin in terms of serum and genomic levels.

The fungus Amanita phalloides is known to contain two main groups of toxins: amanitins and phallo... more The fungus Amanita phalloides is known to contain two main groups of toxins: amanitins and phallotoxins. The amanitins group effectively blocks the RNA polymerase II enzyme found in eukaryotic cells. As alpha amanitin has a lethal effect on the majority of eukaryotic cells, it can be valuable as an antiparasitic or antifungal drug. It can be used externally against ectoparasites. It is critical that percutaneous applications of the alpha amanitin toxin are not harmful to the recipient. In this study, the absorption and the toxicity of percutaneous and intraperitoneal (ip) applications of 1 mg/kg alpha amanitin to mice were compared. Potential skin, liver and kidney toxicities were investigated through pathological examination. HPLC analysis was used to determine the amount of the toxin. No toxicity or toxin were found in the skin, liver, or kidneys of the mice in the control group. Interestingly, the percutaneous application group also showed no toxicity, and the toxin was not present in this group. After 24 h, Councilman-like bodies and pyknotic cells were observed in the mice in which alpha amanitin was applied intraperitoneally, demonstrating the presence of toxicity. Peak levels of alpha amanitin (mg/mL) in the liver, kidney, and blood in the ip application group were measured at 3.3 (6 h), 0.2 (6 h) and 1.2 (1 h), respectively. The results demonstrated that the toxin was not absorbed through the skin of the mice and that the percutaneous application of alpha amanitin did not have any toxic effects. Thus, alpha amanitin may be administered percutaneously for therapeutic purposes.

Gastrointestinal Cancers (GICs) are the most important causes of mortality and morbidity in indus... more Gastrointestinal Cancers (GICs) are the most important causes of mortality and morbidity in industrialized world. Sister chroma-tid exchange (SCE), as an index of chromosomal instability, involves cancer. The aim of this study is to determine whether SCE frequency is a heritable factor for GIC or not. The study groups consisted of 15 gastrointestinal carcinoma patients, 13 patient relatives and 15 healthy subjects as the control group. After collection of 2 ml peripheral blood, lymphocytes were cultured for 3 days and sister chromatid exchange (SCE), mitotic index, and replication index were analyzed. SCE was significantly increased (p<0.01) in patients (16.06±22.37) and in their relatives (5.23±2.64) compared with controls (3.51±1.58). There was no significant difference between patients' relatives and control group in terms of the incidence of SCE frequency. Mitotic index was significantly decreased (p<0.05, p<0.01) in patients (5.4±3.13) compared with healthy relatives (7.15±2.15) and controls (9.00±2.26). Replication index was also significantly lower (p<0.01) in patients (1.39±0.35) and in their relatives (1.7±0.21) compared with controls (2.04±1.13). The results of this study indicate that SCE is a heritable factor for GICs. Increased SCE reflects genomic instability, which is an important factor in carcinogenesis. Although the most putative factors causing genomic instability are epigenetics marks, further studies in combination with epigenetic modifications are needed using more subjects. Birinci Derecede Akrabalarda Gastrointestinal Kanserli Hastalarda Artmış Kromatid Değişimler ÖZET Gastrointestinal kanserler, sanayileşmiş ülkelerdeki mortalite ve morbiditenin en önemli sebeplerinden biridir. Kromozomal kararsızlığın ölçüsü olan kardeş kromatid değişimi (KKD) kanser etiyolojisinde yer almaktadır. Bu çalışmanın amacı, kardeş kro-matid değişim sıklığının gastrointestinal kanserde kalıtsal bir faktör olup olmadığının araştırılmasıdır. Çalışma grupları 15 gas-trointestinal kanser hastası ve 13 hasta yakını, kontrol grubu ise 15 sağlıklı bireyden oluşmaktadır. Olgulardan 2 ml periferik kan alındıktan sonra lenfositler 3 gün süreyle kültüre edildikten sonra, kardeş kromatid değişimi, mitotik indeks ve replikasyon indeksi analiz edildi. Kardeş kromatid değişimi açısından hasta (16,06±22,37) ve hasta yakınları (5,23±2,64) grubunda kontrol grubuna (3,51±1,58) göre istatistiksel olarak anlamlı bir artış gözlemdi (p<0,01). Hasta grubunun (5,4±3,13) mitotik indeksi hasta yakınları (7,15±2,15) ve kontrol grubuna (9,00±2,26) göre anlamlı derecede düşük çıkmıştır (p<0,05, p<0,01). Benzer olarak hasta grubunun replikasyon indeksi (1,39±0,35), hasta yakınları (1,7±0,21) ve kontrol grubuna (2,04±1,13) göre anlamlı derecede düşük çıkmıştır (p<0,01). Elde edilen sonuçlar kardeş kromatid değişiminin gastrointestinal kanserler için kalıtsal bir faktör olduğunu göstermektedir. Artan kardeş kromatid değişimi, karsinogenezde etken olduğu bilinen genomik kararsızlığın ölçüsüdür. Fakat ge-nomik kararsızlığın başlıca sebepleri arasında epigenetik değişiklikler olduğundan, epigenetik değişikliklerle kombine daha ileri araştırmaların yapılması gerekmektedir.

Aim: Prostate cancer is the most commonly diagnosed malignancy and the second most common cause o... more Aim: Prostate cancer is the most commonly diagnosed malignancy and the second most common cause of cancer deaths in the
Western male population. Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) modulate the remodeling of the
extracellular matrix (ECM). The imbalance between MMPs and TIMPs may lead to an emergence of pathological processes such as
cancer. In this study, the association between TIMP-2 (–418 G/C) and MMP-2 (–1306 C/T) polymorphisms and prostate cancer in the
Turkish population was investigated.
Materials and methods: Sixty-one prostate cancer patients and 46 healthy subjects were included in the study. DNA was isolated from 2
mL of peripheral blood taken from subjects, and genotypes were analyzed by the polymerase chain reaction-restriction fragment length
polymorphism method.
Results: The TIMP-2 –418 (GC) genotype was found in 15 cases (32.6%) in the control group and in 9 cases (14.8%) in the patients
group, and statistical significance was determined (P = 0.037, OR = 0.346). The MMP-2 –1306 (CT) genotype was found 2.17 times more
in the patient group than in the control group (P = 0.149, OR = 2.17).
Conclusion: Our results show that the TIMP-2 –418 (GC) genotype had a putative protective effect against prostate cancer.

Objective: Coronary artery disease (CAD) is the most important morbidity and mortality disease in... more Objective: Coronary artery disease (CAD) is the most important morbidity and mortality disease in the world. It is also one of the leading causes of death in Turkey. Omentin, a recently found adipocytokine, is reported to regulate insulin sensitivity. It has anti-inflammatory properties and is inversely associated with CAD. Omentin gene polymorphism in patients with CAD has not been studied yet. The aim of this study is to investigate the relationship between omentin Val109Asp polymorphism and CAD. Methods: This is an observational study on genetic association. 157 consecutive patients who had undergone coronary angiography were included in the study. Seventy-five of them had CAD and the rest serves the control group. Val109Asp polymorphism was analyzed and compared. Chi-square test was used in comparison of genotype frequencies, whereas ANOVA and chi-square tests were used in comparison of clinical characteristics according to the genotypes. Results: There was no significant difference between CAD patients and control subjects regarding omentin Val109Asp polymorphism. However, a 2.5 fold increase in Val/Val (homozygous mutant) genotype was detected in patients with CAD. The OR (80% Cl) for Val/Val genotype was 3.46 (1.14-10.49). Conclusion: Although no significant difference was detected regarding omentin Val109Asp polymorphism, Val/Val genotype frequency was found to be more in patient group than control group. In conclusion, it may be speculated that Val/Val genotype increases the tendency for CAD, but this experiment should done with larger population to clarify this issue. (Anadolu Kardiyol Derg 2014; 14: 511-4)

Background: Colon cancer is one of the most common cancers worldwide. Apoptosis is a necessary ph... more Background: Colon cancer is one of the most common cancers worldwide. Apoptosis is a necessary physiological process for cell elimination which is very important both cellular homeostasis and cell proliferation and differantiation. Dysregulation can lead to uncontrolled cell growth and tumor development. Survivin, a member of the IAP family, plays a key role in promotion of cell proliferation as well as inhibition of apoptosis in cancer cells. The aim of this study was to investigate whether specific genetic polymorphisms of survivin could be associated with colon cancer development and progression in a Turkish population. Our study is the first to our knowledge to investigate the relationship between colon cancer risk and survivin gene polymorphisms.
Materials and Methods: The relation between colon cancer and survivin -31 G/C (rs9904341), -241 C/T (rs17878467) and -625 C/G (rs8073069) polymorphism in promotor site of survivin gene associated with apoptosis was investigated using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results:
Individuals with -31C allele and CC genotype were found to have a higher risk of developing colon cancer (OR=13.4, p=0.01). The -241 CT genotype considerably increased the risk of colon cancer (OR=12.0, p=0.0001). However, there was no significant varaition of the survivin -625 C/G polymorphism among colon cancer patients and controls in our study. Conclusions: This study provides the first evidence that survivin -31 G/C and -241 C/T SNP significantly contribute to the risk of colon cancer in the Turkish population.

In carbon monoxide (CO) poisoning, CO affects the oxygen-carrying capacity of the hemoglobin mole... more In carbon monoxide (CO) poisoning, CO affects the oxygen-carrying capacity of the hemoglobin molecule. Nucleolar-organizing regions (NORs) are genetic loci on chromosomes that are composed of ribosomal DNA and proteins. NORs can be stained with silver. A total of 18 rats were exposed to CO in three different concentrations (1000, 3000, and 5000 ppm) with 6 rats as controls. The animals were euthanized 7 days after CO intoxication. Lung tissues were taken, embedded in paraffin blocks, and sectioned at 5 mm thickness. Argyrophilic nucleolar-organizing region (AgNOR) staining was carried out. One hundred nuclei per individual were evaluated, and total AgNOR number per total nuclear number and total AgNOR area per nuclear area (TAA/NA) for each nucleus were analyzed. The CO exposure groups had significantly higher TAA/NA values and AgNOR numbers than the control group (p < 0.05). Although the differences between 1000 ppm and the other two CO-exposed groups were meaningful (p < 0.05) in the TAA/NA values, there were no differences among the CO exposure groups for the AgNOR number (p > 0.05). The increase in TAA/NA value depends on the increase in the CO exposure. Significant correlations between both the AgNOR values and histopatholo-gical scoring methods were found. Therefore, AgNOR staining method may be used as an indirect indicator for evaluating the degree of cell damage rate.

Most of the fatal cases of mushroom poisoning are caused by Amanita phalloides. The amount of tox... more Most of the fatal cases of mushroom poisoning are caused by Amanita phalloides. The amount of toxin in mushroom varies according to climate and environmental conditions. The aim of this study is to measure a-, band nd g-amanitin with phalloidin and phallacidin toxin concentrations. Six pieces of A. phalloides mushrooms were gathered from a wooded area of Düzce, Turkey, on November 23, 2011. The mushrooms were broken into pieces as spores, mycelium, pileus, gills, stipe, and volva. a-, band nd g-Amanitin with phalloidin and phallacidin were analyzed using reversed-phase high-performance liquid chromatography. As a mobile phase, 50 mM ammonium acetate þ acetonitrile (90 þ 10, v/v) was used with a flow rate of 1 mL/min. C18 reverse phase column (150 Â 4.6 mm; 5 mm particle) was used. The least amount of g-amanitin toxins was found at the myce-lium. The other toxins found to be in the least amount turned out to be the ones at the spores. The maximum amounts of amatoxins and phallotoxin were found at gills and pileus, respectively. In this study, the amount of toxin in the spores of A. phalloides was published for the first time, and this study is pioneering to deal with the amount of toxin in mushrooms grown in Turkey.

Aim: To explore whether chrysin has any protective effect against the deadly effect of alpha aman... more Aim: To explore whether chrysin has any protective effect against the deadly effect of alpha amanitin on hepatocytes that is one of the major toxins in the structure of Amanita phalloides, which is considered the most important mushroom responsible for fatal mushroom poisonings.
Materials and methods: For this study, alpha amanitin was purified from A phalloides mushroom using the preparative HPLC (high performance liquid chromatography) method as described in the literature. Four hours after administering alpha amanitin in a concentration of 10 μg/mL on the cells in a hepatocyte cell line (C3A), silibinin and chrysin were administered in various concentrations. The MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] test was used to determine cell viability.
Results: The alpha amanitin was obtained in high purity from the mushrooms and was found to decrease cell viability down
to a level of 63% after 48 hours due to its toxic effect on the liver cells in the cell culture. In the groups given silibinin and chrysin,
both substances were seen to reduce the toxicity caused by alpha amanitin.
Conclusion: Chrysin may play a role in decreasing the tissue damage caused by toxins and lowering the mortality rates in fatal mushroom poisonings associated with alpha amanitin.

We aimed to obtain gamma amanitin with high purity through a purification process and compare tox... more We aimed to obtain gamma amanitin with high purity through a purification process and compare toxic effects of alpha, beta, and gamma amanitin. Specific concentrations of the toxins (25, 10, 1, and 0.1 mg/mL) were applied to the C3A human hepatocytes. A MTT test was performed to determine the level of toxicity. Alpha amanitin showed a higher toxicity in 48 h while the lowest toxicity was observed in beta amanitin. The toxicity level of gamma amanitin was found between the alpha and beta amanitin toxicity. Our method is suitable for obtaining
gamma amanitin with high purity (499%) as well as for obtaining alpha amanitin and beta amanitin. Gamma amanitin has been shown to have equal responsibility for toxicity as alpha amanitin in amanita poisoning.

Purpose: Chronic inflammation is an important etiological factor in the development of arthritic ... more Purpose: Chronic inflammation is an important etiological
factor in the development of arthritic diseases. Several factors
contribute to aggregation of chronic inflammation, including
the presence of excess adipose tissue.
Methods: The putative induction mechanisms of ADAMTS-5
by resistin were investigated in normal primary human articular
articular
chondrocytes. Expression levels of the ADAMTS-5 gene
were determined at several resistin doses and durations.
Results: Human chondrocytes were activated and associated
with upregulated ADAMTS-5 gene expression after exposure
to resistin (also known as adipose tissue-specific secretary factor,
ADSF). Release of ADAMTS-5 leads to joint cartilage
degradation, a key event in the development of arthritic diseases
rheumatoid arthritis (RA) and osteoarthritis (OA). Activation
of chondrocytes was associated with upregulated NF-κB
protein levels in a time-dependent fashion. Co-incubation of
human chondrocytes with JNK and p38 inhibitors lead to abrogated
levels of NF-κB, indicating that these MAPKs are important
in the activation of chondrocytes after stimulation
with resistin. Similarly, ADAMTS-5 expression levels were
abrogated when co-incubated with p38, NF-κB, JNK, MEK
and PI3K inhibitors. Our results demonstrate that resistin,
released from adipose tissue, may be involved in the
development of RA and OA in obese patients through degradation
of joint cartilage via ADAMTS-5 released from activated
chondrocytes.

Context: Pituitary stalk interruption syndrome (PSIS) is a rare, congenital anomaly of the pituit... more Context: Pituitary stalk interruption syndrome (PSIS) is a rare, congenital anomaly of the pituitary gland characterized by pituitary gland insufficiency, thin or discontinuous pituitary stalk, anterior pituitary hypoplasia, and ectopic positioning of the posterior pituitary gland (neurohypophysis). The clinical presentation of patients with PSIS varies from isolated growth hormone (GH) deficiency to combined pituitary insufficiency and accompanying extrapituitary findings. Mutations in HESX1, LHX4, OTX2, SOX3, and PROKR2 have been associated with PSIS in less than 5% of cases; thus, the underlying genetic etiology for the vast majority of cases remains to be determined. Objective: We applied whole-exome sequencing (WES) to a consanguineous family with two affected siblings who have pituitary gland insufficiency and radiographic findings of hypoplastic (thin) pituitary gland, empty sella, ectopic neurohypophysis, and interrupted pitiutary stalk— characteristic clinical diagnostic findings of PSIS. Design and Participants: WES was applied to two affected and one unaffected siblings. Results: WES of two affected and one unaffected sibling revealed a unique homozygous missense mutation in GPR161, which encodes the orphan G protein– coupled receptor 161, a protein responsible for transducing extracellular signals across the plasma membrane into the cell. Conclusion: Mutations of GPR161 may be implicated as a potential novel cause of PSIS. (J Clin Endocrinol Metab 100: E140 –E147, 2015)

The Knee, 2015

Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative a... more Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative activity in non-haematopoietic tissues. There is insufficient knowledge about the role of EPO activity in tendon healing. Therefore, we investigated the effect of EPO treatment on healing in rat patellar tendons. One hundred and twenty-six, four-month-old male Sprague-Dawley rats were randomly assigned to three experimental groups: 1, no treatment; 2, treatment with isotonic saline (NaCl) and 3, treatment with EPO. Each group was randomly subdivided into two groups for sacrifice at three (1a, 2a, 3a) or sixweeks (1b, 2b, 3b). Complete incision of the left patellar tendon from the distal patellar pole was performed. We applied body casts for 20days after the incised edges of the patellar tendon were brought together with a surgical technique. Both legs were harvested and specimens from each group underwent histological, biomechanical, and protein mRNA expression analyses. There were statistically significant differences in the ultimate breaking force between the EPO group and others at both weeks three and six (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05); significant differences in fibroblast proliferation, capillary vessel formation, and local inflammation were found between groups 1a and 3a, and 2a and 3a (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). There were statistical differences between 1a, 3a and 2a, 3a for Col III, TGF-β1, and VEGF and between 1b, 3b and 2b, 3b for Col I, Col III, TGF-β1, and VEGF mRNA expressions. EPO had an additive effect with surgery on the injured tendon healing process in rats compared to the control groups biomechanically, histopathologically and with tissue protein mRNA expression. This is the first experimental study to analyze the relationship between EPO treatment and the patellar tendon repair process by biomechanical, histopathological, and tendon tissue mRNA expression methodologies.

ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1 is an inﬂammatory‑ indu... more ADAMTS1 (a disintegrin and metalloproteinase with thrombospondin motifs 1 is an inﬂammatory‑ induced gene. We have previously reported that ADAMTS1 was strongly but transiently expressed in the infarcted heart. In this study we investigated whether a 3ʼ‑untranslated region (UTR aﬀects the mRNA stability of this gene. When stimulated with tissue necrosis factor (TNF‑ the expression level of ADAMTS1 mRNA rapidly

ADAMTS9 is a member of the disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) ... more ADAMTS9 is a member of the disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) genes, with aggrecan-degrading activity. The nuclear factor of activated T cells (NFAT) proteins are a family of transcription factors related to Rel/NFκB family whose activation is controlled by calcineurin. The inhibition of aggrecan degradation likely plays a protective role in the prevention of cartilage collagen breakdown. Recently, an NFAT inhibitor peptide, VIVIT, was developed based on the conserved calcineurin docking site of the NFAT family. The presence of putative NFAT binding elements (NBE) in the ADAMTS9 promoter region prompted an investigation of whether NFAT is one of the activators for ADAMTS9 upon IL-1 stimulation. To test the possibility regarding whether NFAT mediates the induction of ADAMTS9, real-time PCR was done using inhibitors for NFAT, FK506 and 11R-VIVIT. ADAMTS9 expression was decreased to nearly 40-50% by 1 μM of 11R-VIVIT. In addition, western blott was d...

The Knee, 2015

Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative a... more Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative activity in non-haematopoietic tissues. There is insufficient knowledge about the role of EPO activity in tendon healing. Therefore, we investigated the effect of EPO treatment on healing in rat patellar tendons. One hundred and twenty-six, four-month-old male Sprague-Dawley rats were randomly assigned to three experimental groups: 1, no treatment; 2, treatment with isotonic saline (NaCl) and 3, treatment with EPO. Each group was randomly subdivided into two groups for sacrifice at three (1a, 2a, 3a) or sixweeks (1b, 2b, 3b). Complete incision of the left patellar tendon from the distal patellar pole was performed. We applied body casts for 20days after the incised edges of the patellar tendon were brought together with a surgical technique. Both legs were harvested and specimens from each group underwent histological, biomechanical, and protein mRNA expression analyses. There were statistically significant differences in the ultimate breaking force between the EPO group and others at both weeks three and six (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05); significant differences in fibroblast proliferation, capillary vessel formation, and local inflammation were found between groups 1a and 3a, and 2a and 3a (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). There were statistical differences between 1a, 3a and 2a, 3a for Col III, TGF-β1, and VEGF and between 1b, 3b and 2b, 3b for Col I, Col III, TGF-β1, and VEGF mRNA expressions. EPO had an additive effect with surgery on the injured tendon healing process in rats compared to the control groups biomechanically, histopathologically and with tissue protein mRNA expression. This is the first experimental study to analyze the relationship between EPO treatment and the patellar tendon repair process by biomechanical, histopathological, and tendon tissue mRNA expression methodologies.

The Knee, 2015

Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative a... more Erythropoietin (EPO), regulating erythropoiesis, is used to provide protective and regenerative activity in non-haematopoietic tissues. There is insufficient knowledge about the role of EPO activity in tendon healing. Therefore, we investigated the effect of EPO treatment on healing in rat patellar tendons. One hundred and twenty-six, four-month-old male Sprague-Dawley rats were randomly assigned to three experimental groups: 1, no treatment; 2, treatment with isotonic saline (NaCl) and 3, treatment with EPO. Each group was randomly subdivided into two groups for sacrifice at three (1a, 2a, 3a) or sixweeks (1b, 2b, 3b). Complete incision of the left patellar tendon from the distal patellar pole was performed. We applied body casts for 20days after the incised edges of the patellar tendon were brought together with a surgical technique. Both legs were harvested and specimens from each group underwent histological, biomechanical, and protein mRNA expression analyses. There were statistically significant differences in the ultimate breaking force between the EPO group and others at both weeks three and six (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05); significant differences in fibroblast proliferation, capillary vessel formation, and local inflammation were found between groups 1a and 3a, and 2a and 3a (p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.05). There were statistical differences between 1a, 3a and 2a, 3a for Col III, TGF-β1, and VEGF and between 1b, 3b and 2b, 3b for Col I, Col III, TGF-β1, and VEGF mRNA expressions. EPO had an additive effect with surgery on the injured tendon healing process in rats compared to the control groups biomechanically, histopathologically and with tissue protein mRNA expression. This is the first experimental study to analyze the relationship between EPO treatment and the patellar tendon repair process by biomechanical, histopathological, and tendon tissue mRNA expression methodologies.

Idiopathic Pulmonary Fibrosis (IPF) is a disease that is characterized by the deposition of an ex... more Idiopathic Pulmonary Fibrosis (IPF) is a disease that is characterized by the deposition of an excessive degree of myofibroblast cells and extracellular matrix components in the lower respiratory tract and lung interstitium. Median survival is 3 years after initial diagnosis. Prevalence rate varies from 14 to 43 per 100 000 people. Today, it is thought that recurrent epithelial damage and aberrant wound healing are the basis of IPF pathogenesis, resulting in the accumulation of fibroblasts in the lung. In addition, coagulation, apoptosis, angiogenesis pathway disorders, oxidative damage, and most recently epithelial–mesenchymal transition (EMT) are implicated in the pathogenesis of this disease. Major aim of our study is to show the fundamental role of osteopontin, Twist and Wnt5a genes in IPF pathogenesis which these genes have been implicated by several studies in EMT and to show whether the suppression of these genes could be effective for IPF treatment examined. First of all, we treated A549 cell line with different dose of TGFβ to create EMT. We showed that after TGFβ treatment, Ecadherin expression is markedly decreased, on the contrary, Vimentin expression increased showing EMT. Then, changes in the expression of genes responsible for EMT formation and fibrosis examined at the level of mRNA and protein performing siRNA knockdown for Osteopontin, Twist and Wnt5a gene transcripts in lung alveolar cell lines.

Objective: ADAMTS-1 belonging a disintegrin and metalloproteinase with trombospondin motifs (ADAM... more Objective: ADAMTS-1 belonging a disintegrin and metalloproteinase with trombospondin motifs (ADAMTs) protease family. Besides the aggrecanolytic activity, ADAMTS-1 is involved in angiogenesis. Angiogenesis is a normal physiological process and described as a formation of new blood vessels. However, aberrant regulation of angiogenesis caused several pathological processes including cancer. Even, the etiology of cancer has been not understood yet, the obesity was assumed one of the main factors for several cancers like chondrosarcoma. Recently, a novel adipokine called leptin secreted from adipose tissue was identified. Then, leptin become target to develop new therapy against to obesity associated diseases like cancer. The aim of this study was to investigate the signaling pathways involved in ADAMTS-1 upregulation induced by leptin in human chondrocytes.
Material and Method: Confluent human articular chondrocytes were cultured with serum-free medium for 24 hours. Then the cells were pretreated with SB203580, SP600125, PD98059, QNZ and LY294002 reagents for inhibition of p38, JNK, MEK1, NF-ĸB and PI3-kinase pathways respectively for 30 min. Subsequently, the cells were treated with leptin at 1000ng/mL for 48 h. At the end of the incubation, total RNA was extracted using TriPure reagent, and 2 µg of total RNA was reverse-transcribed. The ADAMTS-1, and β-actin genes expression were analyzed by real-time polymerase chain reaction.
Results and Conclusion: The current study was investigated that leptin increased the ADAMTS-1 gene expression level by mitogen-activated protein kinases (MAPKs) signaling pathways. Vascular endothelial growth factor (VEGF) has angiogenic property, and leptin caused VEGF upregulation has been identified recently. Similarly, the upregulation of ADAMTS-1 gene expression level by leptin and VEGF was demonstrated. Even, the roles of ADAMTS-1 in angiogenesis are under debate, it was clarified that ADAMTS-1 involved in angiogenesis. So, ADAMTS-1 has pivotal importance for obesity related cancer like chondrosarcoma. In this study, the signaling pathways of ADAMTS-1 upregulation caused by leptin were investigated. The elucidation of these pathways may give new idea to develeop new approach for chondrosarcoma therapy. However, further studies are needed.

Visfatin also called NAMPT (Nicotinamide phosphoribosyltransferase) and PBEF1 (pre-B-cell colony-... more Visfatin also called NAMPT (Nicotinamide phosphoribosyltransferase) and PBEF1 (pre-B-cell colony-enhancing factor 1) is adipokine (adipocytokines), and characterized in 1994. Visfatin with inflammatory feature involves in normal physiology and pathological problems. The underlying putative reasons of inflammation in arthritic diseases such as rheumatoid arthritis (RA), osteoarthritis (OA) still needed to explain. Obesity is considered to be risk factor for RA and OA. It was clarified that visfatin expression was increased in obesity and RA. The upregulation mechanisms of Matrix Metalloproteinase-2 and -9 by visfatin in several cell types were shown before. So, catabolic effect of visfatin on articular cartilage makes visfatin central place in the investigation of arthritic diseases. Visfatin caused induction of ADAMTS-5 (aggrecanases-2) was investigated by our group. However, this induction mechanism was not clarified yet. For this aim, human chondrocytes was preincubated with p38 (SB203580) and NF-ĸB (QNZ) inhibitors for 30 min. Then, 500 ng/ml visfatin was added the cell culture medium, and incubated for 6 hours additionally. At the end of the incubation isolated total RNA was reverse transcribed, and the gene expression level of ADAMTS-5 measured using RT-PCR (real-time polymerase chain reaction) method. As a results, visfatin caused ADAMTS-5 upregulation was almost inhibited by p38 and NF-ĸB inhibitors. This result has vital importance to understand the mechanisms of the obesity caused-inflammation in arthritic diseases. In conclusion, the aggrecanase induction pathways may become a target to develop new therapy for arthritic diseases.