Derya Biriken - Academia.edu (original) (raw)
Papers by Derya Biriken
Ankara Üniversitesi tıp fakültesi mecmuası, Apr 1, 2019
Objectives: In this study, it was aimed to evaluate appropriate model of application time and dos... more Objectives: In this study, it was aimed to evaluate appropriate model of application time and dosage of Phorbol-12-Myristate-13-Acetate (PMA) to get activated macrophages from monocytic cell line THP-1 cells. The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated. Materials and Methods: THP-1 cells were produced as suspension as 2x10 5 cells/mL in 6-well plates. Afterwards, THP-1 monocyte cells were treated with PMA of 10, 20, 40, 60 ng/mL concentrations and incubated for 24, 48 and 72 hours. Differentiation of suspended THP-1 cells into adherent macrophage-like cells was confirmed by tissue-culture microscope by observing cell adherence to cell culture flasks and phenotypic changes. Cell viability was determined by trypan blue staining. Results and Conclusion: The best results for phenotypic changes and cell viability for THP-1 cells were obtained with 20 ng/mL PMA at 48h incubation time and the cell viability was detected as 92.2%. The administration of 40 ng/mL PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72h application. These findings show that the differentiation of THP-1 monocytes into macrophages needs to be optimized in order to reflect the real physiologic conditions of macrophage models used in in vitro studies. These findings suggest that THP-1 cells are well differentiated by 20 ng/mL PMA at 48h incubation.
Türkiye klinikleri tıp bilimleri dergisi, 2013
Mikrobiyoloji Bulteni, 2004
The aim of this study was to investigate the presence of Herpes simplex virus type 1, in the spec... more The aim of this study was to investigate the presence of Herpes simplex virus type 1, in the specimens from patients clinically prediagnosed as herpetic keratitis or keratoconjunctivitis, by virus isolation and direct immunoperoxidase methods. The samples obtained from a total of 33 patients included ulcerated corneal epithelium scrapings and, swabs from ulcer, corneal epithelium, and lower conjunctivas. For virus isolation, both scraping and swab samples for each patient, were inoculated into monolayered Vero cell lines which have previously cultivated on 24-well plates, and examined for the presence of cytopathic effects typical for HSV-1. At the end of 5-days incubation period, the culture media were discarded, the cells were fixed and stained with peroxidase labeled specific HSV-1 antibodies (direct immunoperoxidase--DIP--method). Simultaneously, the smears were prepared from the samples which were sufficient in amount, and stained with DIP method. As a result, cytopathic effects on cell cultures were observed in 4 (12.1%) of the samples, of which 2 were also positive with DIP method. Following DIP staining of smears prepared from 15 samples, HSV-1 antigen positivity was detected in only 1 of the samples. This sample was negative in cell culture. In contrast, one of the 2 cell culture positive samples yielded negative result in smear examination, while the other could not be examined since the sample was not enough. In conclusion, HSV-1 has been shown as the etiologic agent in 3 (9.1%) of our patients clinically prediagnosed as herpetic keratitis or keratoconjunctivitis, and lesion scrapings were determined as the most appropriate samples for virus isolation.
Mikrobiyoloji Bulteni, Jul 1, 2006
Epithelial cells take part in the stimulation of immune system during the conversion of nonspecif... more Epithelial cells take part in the stimulation of immune system during the conversion of nonspecific immune response to the adaptive one in the mucosal immune system. Epithelial cells also have critical roles in designating immune homeostasis towards immune regulation or tolerance. Its response type is determined according to the dose and structure of the antigen and the way it is presented. In this study, we aimed to evaluate the response of human urinary epithelial cells to different doses of Escherichia coli K12 by means of their bactericidal effect. Urine was collected from 16 healthy volunteers and urinary epithelial cells were prepared. The cells (effector) were stimulated with bacteria (target) in microplates for one hour with different effector/target cell ratios. At the end of the incubation period, the bactericidal effects were calculated and compared with microorganism controls. Mann-Whitney U test was used for statistical analysis. Urinary epithelial cells which were stimulated by E. coli K12 showed dose dependent bactericidal effect, calculated mean value for bactericidal effects were 60.7% at the 1/100.000 effector/target cell ratio and 11.3% 1/1.000.000 effector/target cell ratio, indicating that the bactericidal effect was dose dependent.
PubMed, Apr 1, 2005
The functions of epithelial cells are more than a physical selective barrier for protection from ... more The functions of epithelial cells are more than a physical selective barrier for protection from mucosal pathogens. The epithelial response regulated by host-microorganism interaction may change due to histological and physiological differences between sterile and nonsterile sites. In this study we examined the presence of a difference between bactericidal capacities of urinary and oral epithelial cells against different strains of Escherichia coli. Oral epithelial cells were collected from unstimulated saliva and uroepithelial cells were collected from urine of eight healthy volunteers. E. coli K12 and O75 strains and epithelial cells were prepared for the experiment, added to microplates and incubated for one hour. The growth of bacteria was detected by the quantitative colony counting method. Antibacterial effects of the oral and urinary epithelial cells against E. coli K12 and O75 strains were 49.8%, 34.7%, 45.7% and 29.2%, respectively. As a result, it was detected that the antibacterial activity of oral epithelial cells to E. coli K12 and O75 were higher than urinary epithelial cells.
Mikrobiyoloji Bulteni, Jul 1, 2009
Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactio... more Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactions and releasing proinflammatory mediators. In this study, we aimed to illuminate the underlying mechanisms of contribution of macrophages and epithelial cells in the struggle against pathogens. Therefore, Streptococcus pyogenes and Escherichia coli activated Caco-2 cells and THP-1 macrophage-like cells were investigated for their bactericidal activities and nitric oxide (NO) production when they were alone, in contact with eachother and when their contact was blocked by continuing exposure of soluble mediators. Caco-2 epithelial cells and THP-1 macrophage cells were used as effector cells and S. pyogenes or E. coli as target cells and were incubated at an effector cell/target cell ratio of 1/1 in duplication in 24-well plates. After 5 hours incubation, supernatants were collected from each well for growth inhibition assay and were inoculated onto blood agar plates for S. pyogenes and eosin methylene blue agar plates for E. coli. Following overnight incubation at 37 degrees C, bactericidal effect rate was calculated by counting the colony forming units. The supernatants were also collected after 5 and 24 hours incubation for measurement of NO production by using Griess reagent. Bactericidal effect of Caco-2 cells alone against S. pyogenes and E. coli were found 21.9% and 36.2%, when seperated from THP-1 cells via an insert was 31.8% and 30.5%, and when cell-cell contact was established with THP-1 cells was 24.4% and 55.7%, respectively. Bactericidal effect of THP-1 cells alone against S. pyogenes and E. coli was 27.7% and 63.9%, when seperated from Caco-2 cells via an insert was %27.5 and 43.6%, and when cell-cell contact was established with Caco-2 cells was 24.4% and 55.7%, respectively. As a result, we found that Caco-2 epithelial cells and THP-1 macrophage cells had an antibacterial effect against S. pyogenes and E. coli (p < 0.05), and this effect was higher in macrophage cells than epithelial cells. NO levels in epithelial and/or macrophage cell culture supernatants collected after exposure to S. pyogenes and E. coli were significantly higher for S. pyogenes at 5 hours incubation and for E. coli at 24 hours incubation (p < 0.05). Morever, it can be concluded that macrophages played a more active role than epithelial cells in bactericidal effect and NO response. Besides, epithelial cells and macrophages activated each other more when they were in contact than when they were alone or when their contact was blocked.
Mikrobiyoloji Bulteni, Jul 16, 2021
MİKROBİYOLOJİ BÜLTENİ Biriken D, Arıbal Ayral P, Yazıhan N. dığı bulunmuştur. Vit D3 hiperglisemi... more MİKROBİYOLOJİ BÜLTENİ Biriken D, Arıbal Ayral P, Yazıhan N. dığı bulunmuştur. Vit D3 hiperglisemik şartlarda TNF-α, IL-10 ve MK sekresyonlarını düzenlemektedir. MK ve TNF-α seviyelerinin NF-κB ve IL-10 ile ilişkili olduğu tespit edilmiştir. Çalışmada, elde edilen sonuçlar Vit D3'ün NF-κB ve sitokin/kemokin benzeri molekül MK supresyonu ve proenflamatuvar/antienflamatuvar sitokin dengesini düzenlemesiyle immün modülasyonda rol oynayabileceğini göstermiştir. LPS uyarımı ile monositlerden proenflamatuvar sitokinler salındığı; Vit D3'ün, antienflamatuvar sitokin IL-10 seviyelerini arttırarak ve NF-κB seviyelerini baskılayarak enflamasyon üstünde baskılayıcı etki yaptığı görülmektedir. Vit D3'ün farklı şartlar altında etki mekanizmasının ayrıntılı olarak incelenmesi gerekmektedir.
Ankara Üniversitesi tıp fakültesi mecmuası, Mar 1, 2019
In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA t... more In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA to get activated macrophages from monocytic cell line THP-1 cells. The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated. Materials and Methods: THP-1 monocytic cells (2x105 cells/ml) were treated with PMA of 10, 20, 40, 60 ng/ml concentrations and incubated for 24 h, 48 h and 72 h to induce differentiation into adherent macrophage-like cells. Differentiation was confirmed by observation of cell adherence and phenotypic changes using phase contrast microscopy. Cell viability was determined by trypan blue staining. Results: Results and conclusion: The best results were obtained with 20 ng/ml PMA at 48 h incubation and the cell viability was 92.2 % at this concentration. The administration of 40 ng/ml PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72 h application. These results indicate that the differentiation of THP-1 monocytes to macrophages requires optimization to ensure that this invitro macrophage model more precisely reflects real physiologic conditions. These findings suggests that THP-1 cells are well differentiated by 20 ng/ml PMA at 48 h incubation.
Keratitis ve Keratokonjunktivitis Olgularinda Herpes Simplex Virusunun Hucre Kulturu izolasyonu v... more Keratitis ve Keratokonjunktivitis Olgularinda Herpes Simplex Virusunun Hucre Kulturu izolasyonu ve Direkt Immunoperoksidaz Teknigi ile Gosterilmesi Bu calismada cesitli goz kliniklerinden HSV keratitis veya keratokonjunktivitis on tanisi ile laboratuvara gonderilen orneklerde, hucre kulturu iriokulasyonu yontemi ile HSV izolasyonunu takiben immunoperoksidaz (IP) yontemi ile identifikasyonu ve tiplendirilmesi, ayrica hazirlanan direkt frotilerde direkt immunoperoksidaz (DIP) yon temi ile HSV antijen tespiti ve tiplendirilmesi arastirilmistir. Toplam 33 olguya ait kornea epitel kazintisi ile ulser, kornea epiteli veya alt konjunktivadan alinan swablar HSV-1 antijeni yonunden enfekte Vero hucre kulturunde incelenmistir. Buna gore klinik olarak HSV keratit veya keratokonjunktivit on tanisi ile orneklenen 33 olgunun 4 (%12.1) tanesinde virus izolasyonu (VI) gerceUestMlmistir. Yapilan DIPT sonucunda hucre kulturlerinde epe (sitopatik etki) sergileyen 4 adet olgu ya ait materyallerden sadece 2 tanesinde HSV-1 tanisi yapilirken, epe pozitif diger 2 ornekte spesifik boyanma tespit edilememistir. Yayma frotilerde ise DIP yontemi ile boyanmasi sonucunda bu materyallerin %6.6'sinda antijen saptanmistir. Keratitis veya keratokonjunktivitis vakalarinda %15.1 nispetinde viral etiyoloji tespit edilmis olup HSV olarak identifiye edilen olgu sayisi 3 (%9.1) olarak bulunmustur. Bu sonuc ile hasta grubu icinde yapilan virus tespitlerinin %60'ini HSV olusturmustur. Ay rica izolasyon materyali olarak en uygun ornegin lezyonlardan alman kazinti oldugu saptanmistir. Ancak virus izolasyon sansim yukseltmek icin suratle inokulasyonu zorun ludur. Herpes simplex virusunun olusturdugu goz enfeksiyonlari, stromal keratiti izleyen korneal yapisal hasar, hastaligin kronik seyri, tedaviye karsi gorulen direnc ve bunlarin sonucu olusan gorme kaybi nedeniyle goz kliniklerinde onemli bir problem olarak ortaya cikmaktadir. Etkin antiviral tedavinin bulunmasi virusun tanisina deger kazandirmakta dir. Ayrica laboratuvar tanisi konmadan baslanilan antiherpetik tedaviler, baska bir viral etiyoloji soz konusu olan olgularda gereksiz ve masrafli bir uygulama olarak karsimiza cikmaktadir. Etken surme preparatlarda DIP ile hizla tespit edilebilir ancak kazinan huc re miktarinin yeterli olmasi gerekir. Sonuc olarak HSV'nin hizli ve dogru tespitinde, duyarli hucre sistemlerine materyalin inokulasyonu ve takiben DIPT ile dogrulamasinin daha pratik, guvenilir ve tercih edilebilir oldugu kanisina varilmistir.Abstract Detection of Herpes Simplex Virus by Cell Culture and Direct Immunoperoxidase in cases with Keratitis and Keratoconjunctivitis In this study, specimens obtained from patients suspected of having HSV keratitis or keratoconjunctivitis were examined for identification and typing of HSV by immunoperoxidase (IP) following isolation of virus by inoculation of cell cultures and for detection and typing of HSV antigen by direct immunoperoxidase on direct smears. Specimens obtained from overall 33 patients by scraping of ulcerated corneal epithelium or swabbing from corneal epithelium or lower conjunctiva were examined for HSV-1 antigen in infected Vero cell cultures. Virus isolation was performed in 4 of 33 cases suspected of having HSV keratitis or keratoconjunctivitis (12.1%). DIPT showed that only 2 of 4 cases with cpe positivity on cell culture had HSV infection, but specific staining was not observed in the specimens of the remaining two cases. Following direct IP staining of smears, HSV antigen was detected in 6.6% of these specimens. 15.1% of cases with keratitis or keratoconjunctivitis were found to have a virus infection whereas 9.1% (3 cases) of cases had infection with HSV. As a result, HSV accounted for 60% of virus infections. The most appropriate material for virus isolation was found to be scraping of lesions. In order to increase frequency of virus isolation, specimens should be inoculated as quickly as possible. Ocular infections with HSV are of great importance in opthalmology clinics in that they are chronic, resistant to treatment and cause blindness due to damage to corneal structure. Detection of virus has a great value for effective antiviral therapy. The causative agent can be detected rapidly by DIPT on smears, but the number of epithelial cells obtained by scraping must be adequate. In addition, antiherpetic treatment initiated before detection of virus are ineffective and costly. In conclusion, following inoculation of specimen into sensitive cell cultures, DIPT is a practical and reliable method for rapid and accurate detection of HSV.
Journal of Medicinal Chemistry
Ankara Üniversitesi Veteriner Fakültesi Dergisi
Musculoskeletal injuries as a kind of trauma that the human body is exposed to, adversely affect ... more Musculoskeletal injuries as a kind of trauma that the human body is exposed to, adversely affect the quality of life and workforce of individuals due to restriction of movement function. This study aimed to evaluate the effects of dose-dependent ascorbic acid (AA) administration on the repair process after gastrocnemius muscle injury in rats. In this study, 5-month-old 66 male Wistar Albino rats were used and rats were randomly divided into 6 groups of 11 each [control, muscle injury, healthy (with 5 mg/10 mg/kg/day AA-treated group), injury (with 5 mg/10 mg/kg/day AA-treated group)]. A linear incision was made in the gastrocnemius muscle of thirty-three animals included in the muscle injury groups. AA (5-10 mg/kg/day) was administered to the four groups intraperitoneally just after surgery once a day. Animals were sacrificed twenty-one days later. Blood and tissue samples were used for cytokine, collagen, and histological measurements. It was found that a dose of 5 mg/kg/day AA adm...
Mikrobiyoloji Bulteni, 2006
Epithelial cells (EC) have an important role in the constitution of both innate and acquired immu... more Epithelial cells (EC) have an important role in the constitution of both innate and acquired immune responses. The aim of this study was to investigate the alterations of bactericidal effects and cytokine production patterns of human oral epithelial cells (OEC) against different doses of Streptococcus pyogenes. For this purpose, OEC have been stimulated with S. pyogenes with an effector/target (E/T) cell ratio of 1/1, 1/100, 1/1.000 and 1/10.000, and bactericidal effects and interleukin (IL)-6, IL-8 and IL-10 levels were detected in the first and sixth hours of incubation. The mean rates of bactericidal effect detected in the first and sixth hours were 38.7% and 54.5%, respectively. The bactericidal effects observed at 1/1 E/T cell ratio in the first hour, and at 1/1 and 1/100 E/T cell ratio in the the sixth hour were found significantly higher then the other cells ratios (p<0.05). Time- and dose-depended differences were detected in the cytokine responses of OEC for different S. pyogenes concentrations. IL-6 levels produced by stimulated OEC were found higher, and IL-8 levels were found lower then the levels which were produced by unstimulated OEC (p<0.05, p<0.001, respectively) in the first hour, while there were no change in IL-10 levels after stimulation with different bacterial concentrations (p>0.05). At the sixth hour there were no differences in the IL-6 levels produced by stimulated and unstimulated cells, while the levels of IL-8 produced by stimulated cells were found lower then the levels produced by unstimulated cells in the E/T cell ratio of 1/100, 1/1000 and 1/10.000 (p<0.05, p<0.01, p<0.01, respectively). Nevertheless IL-10 levels in the E/T cell ratio of 1/100 and 1/1.000 were statistically higher then the levels produced by unstimulated cells (p<0.05, p<0.05). As a result OEC stimulated with S. pyogenes showed dose dependent manner in bactericidal effect and cytokine production. It is suggested that epithelial cells stimulation with different doses of antigen contributed to the immune system activation or tolerance.
Journal of Medicinal Chemistry
Journal of Ankara University Faculty of Medicine, 2019
In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA t... more In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA to get activated macrophages from monocytic cell line THP-1 cells. The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated. Materials and Methods: THP-1 monocytic cells (2x105 cells/ml) were treated with PMA of 10, 20, 40, 60 ng/ml concentrations and incubated for 24 h, 48 h and 72 h to induce differentiation into adherent macrophage-like cells. Differentiation was confirmed by observation of cell adherence and phenotypic changes using phase contrast microscopy. Cell viability was determined by trypan blue staining. Results: Results and conclusion: The best results were obtained with 20 ng/ml PMA at 48 h incubation and the cell viability was 92.2 % at this concentration. The administration of 40 ng/ml PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72 h application. These results indicate that the differentiation of THP-1 monocytes to macrophages requires optimization to ensure that this invitro macrophage model more precisely reflects real physiologic conditions. These findings suggests that THP-1 cells are well differentiated by 20 ng/ml PMA at 48 h incubation.
Acta orthopaedica et traumatologica turcica, 2018
The aims of this study were 1) to identify the level of inflammatory biomarkers interleukin (IL)-... more The aims of this study were 1) to identify the level of inflammatory biomarkers interleukin (IL)-1α, IL-1β, IL-6, IL-8, IL-17, C-reactive protein (CRP), granulocyte colony-stimulating factor (GCSF), ferritin, and tumor necrosis factor (TNF)-α in serum and synovial fluid samples of patients who underwent revision arthroplasty surgery; 2) to establish the relationship between serum and synovial fluid levels; 3) to determine if any of the 11 genetic polymorphisms of TNFα, IL-1, IL-6, IL-8, IL-17, and GCSF on the encoding genes was associated with periprosthetic joint infection (PJI). Synovial fluid and serum was collected from 88 patients who underwent revision arthroplasty surgery. The Musculoskeletal Infection Society definition was used to classify these patients into 2 groups: 36 PJIs and 52 aseptic failures. Synovial fluid and serum samples were tested for 9 biomarkers using a micro enzyme-linked immunosorbent assay. Genetic polymorphisms were evaluated with polymerase chain react...
Mikrobiyoloji Bulteni, 2017
Mononükleer fagositer hücreler ve epitel hücreleri, doğal bağışıklığın başlatılması ve düzenlenme... more Mononükleer fagositer hücreler ve epitel hücreleri, doğal bağışıklığın başlatılması ve düzenlenmesinde etki gösteren hücrelerdir. Hem mekanik olarak hem de sitokin salınımı ile etkileşerek mukozal bağışık yanıtta etkin rol oynamaktadırlar. Bunun yanında defensinler, çeşitli hücrelerden salınarak ilk hat bağışık yanıtta etkili olduğu düşünülen mikrobisidal peptitlerdir. Toxoplasma gondii'ye karşı oluşan hücresel bağışıklıkta IL-12 ve IL-10, sırasıyla pro-inflamatuvar ve anti-inflamatuvar özellikleri ile konakta immünopatoloji oluşmadan enfeksiyonun kontrolünü sağlayan iki sitokindir. Bu çalışmada, Caco-2 (insan kolon epidermal adenokarsinom hücresi) ve THP-1 (insan lösemi monosit hücresi) hücre dizilerinin birlikte veya tek başlarına, canlı T.gondii takizoitleri ile doğrudan veya araya teması engelleyecek bir filtre konarak karşılaştırılması sonucunda, hücrelerden IL-12 ve IL-10 salınımında ve insan beta-defensin-3 (hBD-3) ekspresyonunda meydana gelebilecek değişikliklerin in vitro koşullarda gözlenmesi amaçlanmıştır. Deney kuyucuklarındaki hücrelere RH suşu takizoitlerinin eklenmesinden 24 saat sonra toplanan üst sıvılarda insan IL-12 ve IL-10 sitokinlerinin saptanması için ticari ELISA kitleri (Invitrogen) üreticinin talimatları
Mikrobiyoloji bülteni, 2009
Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactio... more Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactions and releasing proinflammatory mediators. In this study, we aimed to illuminate the underlying mechanisms of contribution of macrophages and epithelial cells in the struggle against pathogens. Therefore, Streptococcus pyogenes and Escherichia coli activated Caco-2 cells and THP-1 macrophage-like cells were investigated for their bactericidal activities and nitric oxide (NO) production when they were alone, in contact with eachother and when their contact was blocked by continuing exposure of soluble mediators. Caco-2 epithelial cells and THP-1 macrophage cells were used as effector cells and S. pyogenes or E. coli as target cells and were incubated at an effector cell/target cell ratio of 1/1 in duplication in 24-well plates. After 5 hours incubation, supernatants were collected from each well for growth inhibition assay and were inoculated onto blood agar plates for S. pyogenes and eo...
Turkiye Klinikleri Journal of Medical Sciences, 2013
Bratislavske lekarske listy, 2021
BACKGROUND AND OBJECTIVES Epithelial cells and macrophages play major roles in modulating the sta... more BACKGROUND AND OBJECTIVES Epithelial cells and macrophages play major roles in modulating the state of inlammatory response regulated by intracellular signaling pathways. The aim of this study was to characterize changes in cell proliferation, apoptosis and inflammation related intracellular signalling pathways MAPKs and NF-κB in Caco-2 epithelial cells and THP-1 macrophage-like monocytes contacted and filter-seperated co-cultures in the presence of LPS stimulation. METHODS We assessed the apoptosis and inflammation by measuring caspase-3 activity, TNF-α and IL-10 cytokines, total and phosphorylated forms of intracellular signalling pathway molecules p53, JNK, Jun, ERK, p38, NF-κB p65 and IkB. RESULTS The contacted co-culture of Caco-2 and THP-1 cells represented higher levels of JNK, jun and p38 MAPK pathway proteins associated with cell proliferation, whereas apoptosis related molecule caspase-3 and p53 increased in the filter-separated co-culture condition. Also, the contacted co...
Ankara Üniversitesi tıp fakültesi mecmuası, Apr 1, 2019
Objectives: In this study, it was aimed to evaluate appropriate model of application time and dos... more Objectives: In this study, it was aimed to evaluate appropriate model of application time and dosage of Phorbol-12-Myristate-13-Acetate (PMA) to get activated macrophages from monocytic cell line THP-1 cells. The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated. Materials and Methods: THP-1 cells were produced as suspension as 2x10 5 cells/mL in 6-well plates. Afterwards, THP-1 monocyte cells were treated with PMA of 10, 20, 40, 60 ng/mL concentrations and incubated for 24, 48 and 72 hours. Differentiation of suspended THP-1 cells into adherent macrophage-like cells was confirmed by tissue-culture microscope by observing cell adherence to cell culture flasks and phenotypic changes. Cell viability was determined by trypan blue staining. Results and Conclusion: The best results for phenotypic changes and cell viability for THP-1 cells were obtained with 20 ng/mL PMA at 48h incubation time and the cell viability was detected as 92.2%. The administration of 40 ng/mL PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72h application. These findings show that the differentiation of THP-1 monocytes into macrophages needs to be optimized in order to reflect the real physiologic conditions of macrophage models used in in vitro studies. These findings suggest that THP-1 cells are well differentiated by 20 ng/mL PMA at 48h incubation.
Türkiye klinikleri tıp bilimleri dergisi, 2013
Mikrobiyoloji Bulteni, 2004
The aim of this study was to investigate the presence of Herpes simplex virus type 1, in the spec... more The aim of this study was to investigate the presence of Herpes simplex virus type 1, in the specimens from patients clinically prediagnosed as herpetic keratitis or keratoconjunctivitis, by virus isolation and direct immunoperoxidase methods. The samples obtained from a total of 33 patients included ulcerated corneal epithelium scrapings and, swabs from ulcer, corneal epithelium, and lower conjunctivas. For virus isolation, both scraping and swab samples for each patient, were inoculated into monolayered Vero cell lines which have previously cultivated on 24-well plates, and examined for the presence of cytopathic effects typical for HSV-1. At the end of 5-days incubation period, the culture media were discarded, the cells were fixed and stained with peroxidase labeled specific HSV-1 antibodies (direct immunoperoxidase--DIP--method). Simultaneously, the smears were prepared from the samples which were sufficient in amount, and stained with DIP method. As a result, cytopathic effects on cell cultures were observed in 4 (12.1%) of the samples, of which 2 were also positive with DIP method. Following DIP staining of smears prepared from 15 samples, HSV-1 antigen positivity was detected in only 1 of the samples. This sample was negative in cell culture. In contrast, one of the 2 cell culture positive samples yielded negative result in smear examination, while the other could not be examined since the sample was not enough. In conclusion, HSV-1 has been shown as the etiologic agent in 3 (9.1%) of our patients clinically prediagnosed as herpetic keratitis or keratoconjunctivitis, and lesion scrapings were determined as the most appropriate samples for virus isolation.
Mikrobiyoloji Bulteni, Jul 1, 2006
Epithelial cells take part in the stimulation of immune system during the conversion of nonspecif... more Epithelial cells take part in the stimulation of immune system during the conversion of nonspecific immune response to the adaptive one in the mucosal immune system. Epithelial cells also have critical roles in designating immune homeostasis towards immune regulation or tolerance. Its response type is determined according to the dose and structure of the antigen and the way it is presented. In this study, we aimed to evaluate the response of human urinary epithelial cells to different doses of Escherichia coli K12 by means of their bactericidal effect. Urine was collected from 16 healthy volunteers and urinary epithelial cells were prepared. The cells (effector) were stimulated with bacteria (target) in microplates for one hour with different effector/target cell ratios. At the end of the incubation period, the bactericidal effects were calculated and compared with microorganism controls. Mann-Whitney U test was used for statistical analysis. Urinary epithelial cells which were stimulated by E. coli K12 showed dose dependent bactericidal effect, calculated mean value for bactericidal effects were 60.7% at the 1/100.000 effector/target cell ratio and 11.3% 1/1.000.000 effector/target cell ratio, indicating that the bactericidal effect was dose dependent.
PubMed, Apr 1, 2005
The functions of epithelial cells are more than a physical selective barrier for protection from ... more The functions of epithelial cells are more than a physical selective barrier for protection from mucosal pathogens. The epithelial response regulated by host-microorganism interaction may change due to histological and physiological differences between sterile and nonsterile sites. In this study we examined the presence of a difference between bactericidal capacities of urinary and oral epithelial cells against different strains of Escherichia coli. Oral epithelial cells were collected from unstimulated saliva and uroepithelial cells were collected from urine of eight healthy volunteers. E. coli K12 and O75 strains and epithelial cells were prepared for the experiment, added to microplates and incubated for one hour. The growth of bacteria was detected by the quantitative colony counting method. Antibacterial effects of the oral and urinary epithelial cells against E. coli K12 and O75 strains were 49.8%, 34.7%, 45.7% and 29.2%, respectively. As a result, it was detected that the antibacterial activity of oral epithelial cells to E. coli K12 and O75 were higher than urinary epithelial cells.
Mikrobiyoloji Bulteni, Jul 1, 2009
Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactio... more Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactions and releasing proinflammatory mediators. In this study, we aimed to illuminate the underlying mechanisms of contribution of macrophages and epithelial cells in the struggle against pathogens. Therefore, Streptococcus pyogenes and Escherichia coli activated Caco-2 cells and THP-1 macrophage-like cells were investigated for their bactericidal activities and nitric oxide (NO) production when they were alone, in contact with eachother and when their contact was blocked by continuing exposure of soluble mediators. Caco-2 epithelial cells and THP-1 macrophage cells were used as effector cells and S. pyogenes or E. coli as target cells and were incubated at an effector cell/target cell ratio of 1/1 in duplication in 24-well plates. After 5 hours incubation, supernatants were collected from each well for growth inhibition assay and were inoculated onto blood agar plates for S. pyogenes and eosin methylene blue agar plates for E. coli. Following overnight incubation at 37 degrees C, bactericidal effect rate was calculated by counting the colony forming units. The supernatants were also collected after 5 and 24 hours incubation for measurement of NO production by using Griess reagent. Bactericidal effect of Caco-2 cells alone against S. pyogenes and E. coli were found 21.9% and 36.2%, when seperated from THP-1 cells via an insert was 31.8% and 30.5%, and when cell-cell contact was established with THP-1 cells was 24.4% and 55.7%, respectively. Bactericidal effect of THP-1 cells alone against S. pyogenes and E. coli was 27.7% and 63.9%, when seperated from Caco-2 cells via an insert was %27.5 and 43.6%, and when cell-cell contact was established with Caco-2 cells was 24.4% and 55.7%, respectively. As a result, we found that Caco-2 epithelial cells and THP-1 macrophage cells had an antibacterial effect against S. pyogenes and E. coli (p &amp;amp;amp;amp;amp;amp;amp;lt; 0.05), and this effect was higher in macrophage cells than epithelial cells. NO levels in epithelial and/or macrophage cell culture supernatants collected after exposure to S. pyogenes and E. coli were significantly higher for S. pyogenes at 5 hours incubation and for E. coli at 24 hours incubation (p &amp;amp;amp;amp;amp;amp;amp;lt; 0.05). Morever, it can be concluded that macrophages played a more active role than epithelial cells in bactericidal effect and NO response. Besides, epithelial cells and macrophages activated each other more when they were in contact than when they were alone or when their contact was blocked.
Mikrobiyoloji Bulteni, Jul 16, 2021
MİKROBİYOLOJİ BÜLTENİ Biriken D, Arıbal Ayral P, Yazıhan N. dığı bulunmuştur. Vit D3 hiperglisemi... more MİKROBİYOLOJİ BÜLTENİ Biriken D, Arıbal Ayral P, Yazıhan N. dığı bulunmuştur. Vit D3 hiperglisemik şartlarda TNF-α, IL-10 ve MK sekresyonlarını düzenlemektedir. MK ve TNF-α seviyelerinin NF-κB ve IL-10 ile ilişkili olduğu tespit edilmiştir. Çalışmada, elde edilen sonuçlar Vit D3'ün NF-κB ve sitokin/kemokin benzeri molekül MK supresyonu ve proenflamatuvar/antienflamatuvar sitokin dengesini düzenlemesiyle immün modülasyonda rol oynayabileceğini göstermiştir. LPS uyarımı ile monositlerden proenflamatuvar sitokinler salındığı; Vit D3'ün, antienflamatuvar sitokin IL-10 seviyelerini arttırarak ve NF-κB seviyelerini baskılayarak enflamasyon üstünde baskılayıcı etki yaptığı görülmektedir. Vit D3'ün farklı şartlar altında etki mekanizmasının ayrıntılı olarak incelenmesi gerekmektedir.
Ankara Üniversitesi tıp fakültesi mecmuası, Mar 1, 2019
In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA t... more In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA to get activated macrophages from monocytic cell line THP-1 cells. The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated. Materials and Methods: THP-1 monocytic cells (2x105 cells/ml) were treated with PMA of 10, 20, 40, 60 ng/ml concentrations and incubated for 24 h, 48 h and 72 h to induce differentiation into adherent macrophage-like cells. Differentiation was confirmed by observation of cell adherence and phenotypic changes using phase contrast microscopy. Cell viability was determined by trypan blue staining. Results: Results and conclusion: The best results were obtained with 20 ng/ml PMA at 48 h incubation and the cell viability was 92.2 % at this concentration. The administration of 40 ng/ml PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72 h application. These results indicate that the differentiation of THP-1 monocytes to macrophages requires optimization to ensure that this invitro macrophage model more precisely reflects real physiologic conditions. These findings suggests that THP-1 cells are well differentiated by 20 ng/ml PMA at 48 h incubation.
Keratitis ve Keratokonjunktivitis Olgularinda Herpes Simplex Virusunun Hucre Kulturu izolasyonu v... more Keratitis ve Keratokonjunktivitis Olgularinda Herpes Simplex Virusunun Hucre Kulturu izolasyonu ve Direkt Immunoperoksidaz Teknigi ile Gosterilmesi Bu calismada cesitli goz kliniklerinden HSV keratitis veya keratokonjunktivitis on tanisi ile laboratuvara gonderilen orneklerde, hucre kulturu iriokulasyonu yontemi ile HSV izolasyonunu takiben immunoperoksidaz (IP) yontemi ile identifikasyonu ve tiplendirilmesi, ayrica hazirlanan direkt frotilerde direkt immunoperoksidaz (DIP) yon temi ile HSV antijen tespiti ve tiplendirilmesi arastirilmistir. Toplam 33 olguya ait kornea epitel kazintisi ile ulser, kornea epiteli veya alt konjunktivadan alinan swablar HSV-1 antijeni yonunden enfekte Vero hucre kulturunde incelenmistir. Buna gore klinik olarak HSV keratit veya keratokonjunktivit on tanisi ile orneklenen 33 olgunun 4 (%12.1) tanesinde virus izolasyonu (VI) gerceUestMlmistir. Yapilan DIPT sonucunda hucre kulturlerinde epe (sitopatik etki) sergileyen 4 adet olgu ya ait materyallerden sadece 2 tanesinde HSV-1 tanisi yapilirken, epe pozitif diger 2 ornekte spesifik boyanma tespit edilememistir. Yayma frotilerde ise DIP yontemi ile boyanmasi sonucunda bu materyallerin %6.6'sinda antijen saptanmistir. Keratitis veya keratokonjunktivitis vakalarinda %15.1 nispetinde viral etiyoloji tespit edilmis olup HSV olarak identifiye edilen olgu sayisi 3 (%9.1) olarak bulunmustur. Bu sonuc ile hasta grubu icinde yapilan virus tespitlerinin %60'ini HSV olusturmustur. Ay rica izolasyon materyali olarak en uygun ornegin lezyonlardan alman kazinti oldugu saptanmistir. Ancak virus izolasyon sansim yukseltmek icin suratle inokulasyonu zorun ludur. Herpes simplex virusunun olusturdugu goz enfeksiyonlari, stromal keratiti izleyen korneal yapisal hasar, hastaligin kronik seyri, tedaviye karsi gorulen direnc ve bunlarin sonucu olusan gorme kaybi nedeniyle goz kliniklerinde onemli bir problem olarak ortaya cikmaktadir. Etkin antiviral tedavinin bulunmasi virusun tanisina deger kazandirmakta dir. Ayrica laboratuvar tanisi konmadan baslanilan antiherpetik tedaviler, baska bir viral etiyoloji soz konusu olan olgularda gereksiz ve masrafli bir uygulama olarak karsimiza cikmaktadir. Etken surme preparatlarda DIP ile hizla tespit edilebilir ancak kazinan huc re miktarinin yeterli olmasi gerekir. Sonuc olarak HSV'nin hizli ve dogru tespitinde, duyarli hucre sistemlerine materyalin inokulasyonu ve takiben DIPT ile dogrulamasinin daha pratik, guvenilir ve tercih edilebilir oldugu kanisina varilmistir.Abstract Detection of Herpes Simplex Virus by Cell Culture and Direct Immunoperoxidase in cases with Keratitis and Keratoconjunctivitis In this study, specimens obtained from patients suspected of having HSV keratitis or keratoconjunctivitis were examined for identification and typing of HSV by immunoperoxidase (IP) following isolation of virus by inoculation of cell cultures and for detection and typing of HSV antigen by direct immunoperoxidase on direct smears. Specimens obtained from overall 33 patients by scraping of ulcerated corneal epithelium or swabbing from corneal epithelium or lower conjunctiva were examined for HSV-1 antigen in infected Vero cell cultures. Virus isolation was performed in 4 of 33 cases suspected of having HSV keratitis or keratoconjunctivitis (12.1%). DIPT showed that only 2 of 4 cases with cpe positivity on cell culture had HSV infection, but specific staining was not observed in the specimens of the remaining two cases. Following direct IP staining of smears, HSV antigen was detected in 6.6% of these specimens. 15.1% of cases with keratitis or keratoconjunctivitis were found to have a virus infection whereas 9.1% (3 cases) of cases had infection with HSV. As a result, HSV accounted for 60% of virus infections. The most appropriate material for virus isolation was found to be scraping of lesions. In order to increase frequency of virus isolation, specimens should be inoculated as quickly as possible. Ocular infections with HSV are of great importance in opthalmology clinics in that they are chronic, resistant to treatment and cause blindness due to damage to corneal structure. Detection of virus has a great value for effective antiviral therapy. The causative agent can be detected rapidly by DIPT on smears, but the number of epithelial cells obtained by scraping must be adequate. In addition, antiherpetic treatment initiated before detection of virus are ineffective and costly. In conclusion, following inoculation of specimen into sensitive cell cultures, DIPT is a practical and reliable method for rapid and accurate detection of HSV.
Journal of Medicinal Chemistry
Ankara Üniversitesi Veteriner Fakültesi Dergisi
Musculoskeletal injuries as a kind of trauma that the human body is exposed to, adversely affect ... more Musculoskeletal injuries as a kind of trauma that the human body is exposed to, adversely affect the quality of life and workforce of individuals due to restriction of movement function. This study aimed to evaluate the effects of dose-dependent ascorbic acid (AA) administration on the repair process after gastrocnemius muscle injury in rats. In this study, 5-month-old 66 male Wistar Albino rats were used and rats were randomly divided into 6 groups of 11 each [control, muscle injury, healthy (with 5 mg/10 mg/kg/day AA-treated group), injury (with 5 mg/10 mg/kg/day AA-treated group)]. A linear incision was made in the gastrocnemius muscle of thirty-three animals included in the muscle injury groups. AA (5-10 mg/kg/day) was administered to the four groups intraperitoneally just after surgery once a day. Animals were sacrificed twenty-one days later. Blood and tissue samples were used for cytokine, collagen, and histological measurements. It was found that a dose of 5 mg/kg/day AA adm...
Mikrobiyoloji Bulteni, 2006
Epithelial cells (EC) have an important role in the constitution of both innate and acquired immu... more Epithelial cells (EC) have an important role in the constitution of both innate and acquired immune responses. The aim of this study was to investigate the alterations of bactericidal effects and cytokine production patterns of human oral epithelial cells (OEC) against different doses of Streptococcus pyogenes. For this purpose, OEC have been stimulated with S. pyogenes with an effector/target (E/T) cell ratio of 1/1, 1/100, 1/1.000 and 1/10.000, and bactericidal effects and interleukin (IL)-6, IL-8 and IL-10 levels were detected in the first and sixth hours of incubation. The mean rates of bactericidal effect detected in the first and sixth hours were 38.7% and 54.5%, respectively. The bactericidal effects observed at 1/1 E/T cell ratio in the first hour, and at 1/1 and 1/100 E/T cell ratio in the the sixth hour were found significantly higher then the other cells ratios (p<0.05). Time- and dose-depended differences were detected in the cytokine responses of OEC for different S. pyogenes concentrations. IL-6 levels produced by stimulated OEC were found higher, and IL-8 levels were found lower then the levels which were produced by unstimulated OEC (p<0.05, p<0.001, respectively) in the first hour, while there were no change in IL-10 levels after stimulation with different bacterial concentrations (p>0.05). At the sixth hour there were no differences in the IL-6 levels produced by stimulated and unstimulated cells, while the levels of IL-8 produced by stimulated cells were found lower then the levels produced by unstimulated cells in the E/T cell ratio of 1/100, 1/1000 and 1/10.000 (p<0.05, p<0.01, p<0.01, respectively). Nevertheless IL-10 levels in the E/T cell ratio of 1/100 and 1/1.000 were statistically higher then the levels produced by unstimulated cells (p<0.05, p<0.05). As a result OEC stimulated with S. pyogenes showed dose dependent manner in bactericidal effect and cytokine production. It is suggested that epithelial cells stimulation with different doses of antigen contributed to the immune system activation or tolerance.
Journal of Medicinal Chemistry
Journal of Ankara University Faculty of Medicine, 2019
In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA t... more In this study, it was aimed to evaluate appropriate model of application time and dosage of PMA to get activated macrophages from monocytic cell line THP-1 cells. The differentiation of THP-1 cells into macrophages when stimulated with different dosage and application time of phorbol myristic acid (PMA) was shown morphologically and the effects on cell viability were evaluated. Materials and Methods: THP-1 monocytic cells (2x105 cells/ml) were treated with PMA of 10, 20, 40, 60 ng/ml concentrations and incubated for 24 h, 48 h and 72 h to induce differentiation into adherent macrophage-like cells. Differentiation was confirmed by observation of cell adherence and phenotypic changes using phase contrast microscopy. Cell viability was determined by trypan blue staining. Results: Results and conclusion: The best results were obtained with 20 ng/ml PMA at 48 h incubation and the cell viability was 92.2 % at this concentration. The administration of 40 ng/ml PMA resulted in a significant decrease in cell count compared to 10 and 20 ng/ml PMA administration at the 48h, but no difference was observed between groups after 72 h application. These results indicate that the differentiation of THP-1 monocytes to macrophages requires optimization to ensure that this invitro macrophage model more precisely reflects real physiologic conditions. These findings suggests that THP-1 cells are well differentiated by 20 ng/ml PMA at 48 h incubation.
Acta orthopaedica et traumatologica turcica, 2018
The aims of this study were 1) to identify the level of inflammatory biomarkers interleukin (IL)-... more The aims of this study were 1) to identify the level of inflammatory biomarkers interleukin (IL)-1α, IL-1β, IL-6, IL-8, IL-17, C-reactive protein (CRP), granulocyte colony-stimulating factor (GCSF), ferritin, and tumor necrosis factor (TNF)-α in serum and synovial fluid samples of patients who underwent revision arthroplasty surgery; 2) to establish the relationship between serum and synovial fluid levels; 3) to determine if any of the 11 genetic polymorphisms of TNFα, IL-1, IL-6, IL-8, IL-17, and GCSF on the encoding genes was associated with periprosthetic joint infection (PJI). Synovial fluid and serum was collected from 88 patients who underwent revision arthroplasty surgery. The Musculoskeletal Infection Society definition was used to classify these patients into 2 groups: 36 PJIs and 52 aseptic failures. Synovial fluid and serum samples were tested for 9 biomarkers using a micro enzyme-linked immunosorbent assay. Genetic polymorphisms were evaluated with polymerase chain react...
Mikrobiyoloji Bulteni, 2017
Mononükleer fagositer hücreler ve epitel hücreleri, doğal bağışıklığın başlatılması ve düzenlenme... more Mononükleer fagositer hücreler ve epitel hücreleri, doğal bağışıklığın başlatılması ve düzenlenmesinde etki gösteren hücrelerdir. Hem mekanik olarak hem de sitokin salınımı ile etkileşerek mukozal bağışık yanıtta etkin rol oynamaktadırlar. Bunun yanında defensinler, çeşitli hücrelerden salınarak ilk hat bağışık yanıtta etkili olduğu düşünülen mikrobisidal peptitlerdir. Toxoplasma gondii'ye karşı oluşan hücresel bağışıklıkta IL-12 ve IL-10, sırasıyla pro-inflamatuvar ve anti-inflamatuvar özellikleri ile konakta immünopatoloji oluşmadan enfeksiyonun kontrolünü sağlayan iki sitokindir. Bu çalışmada, Caco-2 (insan kolon epidermal adenokarsinom hücresi) ve THP-1 (insan lösemi monosit hücresi) hücre dizilerinin birlikte veya tek başlarına, canlı T.gondii takizoitleri ile doğrudan veya araya teması engelleyecek bir filtre konarak karşılaştırılması sonucunda, hücrelerden IL-12 ve IL-10 salınımında ve insan beta-defensin-3 (hBD-3) ekspresyonunda meydana gelebilecek değişikliklerin in vitro koşullarda gözlenmesi amaçlanmıştır. Deney kuyucuklarındaki hücrelere RH suşu takizoitlerinin eklenmesinden 24 saat sonra toplanan üst sıvılarda insan IL-12 ve IL-10 sitokinlerinin saptanması için ticari ELISA kitleri (Invitrogen) üreticinin talimatları
Mikrobiyoloji bülteni, 2009
Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactio... more Epithelial cells and macrophages contribute to innate immune responses by cell to cell interactions and releasing proinflammatory mediators. In this study, we aimed to illuminate the underlying mechanisms of contribution of macrophages and epithelial cells in the struggle against pathogens. Therefore, Streptococcus pyogenes and Escherichia coli activated Caco-2 cells and THP-1 macrophage-like cells were investigated for their bactericidal activities and nitric oxide (NO) production when they were alone, in contact with eachother and when their contact was blocked by continuing exposure of soluble mediators. Caco-2 epithelial cells and THP-1 macrophage cells were used as effector cells and S. pyogenes or E. coli as target cells and were incubated at an effector cell/target cell ratio of 1/1 in duplication in 24-well plates. After 5 hours incubation, supernatants were collected from each well for growth inhibition assay and were inoculated onto blood agar plates for S. pyogenes and eo...
Turkiye Klinikleri Journal of Medical Sciences, 2013
Bratislavske lekarske listy, 2021
BACKGROUND AND OBJECTIVES Epithelial cells and macrophages play major roles in modulating the sta... more BACKGROUND AND OBJECTIVES Epithelial cells and macrophages play major roles in modulating the state of inlammatory response regulated by intracellular signaling pathways. The aim of this study was to characterize changes in cell proliferation, apoptosis and inflammation related intracellular signalling pathways MAPKs and NF-κB in Caco-2 epithelial cells and THP-1 macrophage-like monocytes contacted and filter-seperated co-cultures in the presence of LPS stimulation. METHODS We assessed the apoptosis and inflammation by measuring caspase-3 activity, TNF-α and IL-10 cytokines, total and phosphorylated forms of intracellular signalling pathway molecules p53, JNK, Jun, ERK, p38, NF-κB p65 and IkB. RESULTS The contacted co-culture of Caco-2 and THP-1 cells represented higher levels of JNK, jun and p38 MAPK pathway proteins associated with cell proliferation, whereas apoptosis related molecule caspase-3 and p53 increased in the filter-separated co-culture condition. Also, the contacted co...