Significant increase in ZNF304 and decrease in CXCR4 gene expressions may alter anoikis in prostate cancer

Amaç: Prostat kanser hücre hattı (DU145) ve prostat normal epitel hücre hatları (RWPE) arasında anoikis mekanizmasını arttıracak veya inhibe edebilecek genlerin analizini yapmak ve kanser gelişiminde olası rolünü incelemek. Gereç ve Yöntem: İnsan prostat epitel hücre hattı (RWPE) ve prostat kanseri hücre hatları (DU-145) Amerikan Tip Kültür Koleksiyonu (ATCC)’den temin edildi. Hücre hatlarının çoğaltılmasında ve sürdürülmesinde RPMI 1640 (Biological Industries) besi ortamı kullanıldı. Transkriptom analizi için RNA izolasyonu yapılarak, kütüphane oluşturuldu, kütüphanenin kantitasyonunun ardından NextSeq500 (illumina) ile sekanslama yapıldı. Dizileme, haritalandırma, bağıl gen ifadeleri ölçümleri gibi biyoinformatik analizler Genomics Workbench v 8 (Qiagen) yazılımı kullanılarak GRCh38 referans sekansı ile yapılmıştır. Bulgular: RWPE Normal prostat epitel hücre kültürleri ile DU145 prostat kanser hücreleri karşılaştırıldığı zaman DU-145 prostat kanser hücre kültürlerinde, ZNF304, PYCARD ve Notch3 gen expresyonlarında anlamlı bir artış (p<0,05) görülürken, CXCR4, Pak3, SerpınB1 gen ekspresyonlarında anlamlı bir azalma (p<0,05) görülmüştür. Sonuç: DU145 prostat kanseri hücre hattında anoikis ile ilişkili önemli gen ekpresyonlarında artış ve azalma gözlemledik. Değişime bağlı olarak hücrelerin anoikisden kaçarak metastatik özellik kazanabileceğini düşündük.Aim: To analyze genes that may increase or inhibit the anoikis mechanism between prostate cancer cell line and prostate normal epithelial cell line and examine the possible role of cancer in cancer development. Materials and Methods: Human prostate epithelial cell line (RWPE) and prostate cancer cell line (DU145) were acquired from ATCC. Both cell lines were maintanied in RPMI 1640 (Biological Industries) medium. Total RNA were isolated and fragmented. Adapters were ligated to prepare RNA library for whole trasncriptome experiments. Statistics and bioinformatics analysis including mapping, clustering, sequencing were done by using Genomics Workbench v 8. (Qiagen) software. Results: As we compared the normal prostate ephitalial cells (RWPE) and prosatate cancer cells (DU 145); ZNF304,PYCARD ve NOTCH3 were significantly (p<0.05) up-regulated in DU145 cells, on the other hand, CXCR4, PAK3, SERPINB1 genes were significantly down-regulated. Conclusion: We found that there are significant differential gene expressions in DU-145 cells which may lead to metastatic state via evasion of anoikis process

Design and characterization of nanocarriers loaded with Levofloxacin for enhanced antimicrobial activity; physicochemical properties, in vitro release and oral acute toxicity

Inorganic and carbon based nanomaterials are widely used against several diseases, such as cancer, autoimmune diseases as well as fungi and bacteria colonization. In this work, Santa Barbara Amorphous mesoporous silica (SBA), Halloysite Nanotubes (HNTs) and Multiwalled Carbon Nanotubes (CNTs) were loaded with fluoroquinolone Levofloxacin (LVF) to be applied as antimicrobial agents. The prepared via adsorption nanocarriers were characterized by Fourier-Transformed Spectroscopy, Scanning Electron Microscopy as well as High Pressure liquid Chromatography. In vitro release studies were carried out using Simulated Body Fluid at 37o C and data analyzed by various kinetic models showing slow dissolution over 12-24 hours. Antimicrobial studies showed improved antibacterial activity against Escherichia coli, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus, and Staphylococcus epidermidis compared to neat nanomaterials. CNTs were found to be the most promising candidates for LVF delivery and they were chosen to be further studied for their acute oral toxicity and histopathological examination using C57/Black mice. Histological examination depicted that drug loading did not affect mice organs morphology as well as hepatocyte degeneration, central vein degeneration and parenchymal necrosis scores. To conclude, the prepared nanomaterials present significant characteristics and can act as antimicrobial drug carriers; CNTs found to be safe candidates when orally fed to mice

A novel approach for skin infections: Controlled release topical mats of poly (lactic acid)/poly(ethylene succinate) blends containing Voriconazole

WOS: 000438500700010The oral and injectable formulations of Voriconazole (VRZ), a known antifungal agent with low solubility, seem to cause severe side effects. Consequently, topical application of VRZ could be advantageous for skin fungal infections. In this study, VRZ embedded in a polymeric matrix composed of biocompatible poly(lactic acid) (PLA) and poly(ethylene succinate) (PESu). The mats were prepared via solvent evaporation and fully characterized by Fourier-Transformed Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Scanning Electron Microscopy (SEM), in vitro hydrolysis and release studies. The prepared blends defined as immiscible by DSC and SEM while FTIR spectroscopy did not disclose noticeable interactions between the polymers. It was found that hydrolysis was improved by increasing PESu content into the blend. VRZ loaded blends spectra exhibit slight differentiation compared to neat blends while the absence of VRZ melting peak, as DSC illustrated, indicated drug amorphization. Lastly, in vitro release studies depicted a controlled release pattern dependent on mats' hydrolysis degree. An improved antifungal activity of mats was detected by disc diffusion method against various microorganisms. Ex vivo studies of VRZ did not determine high permeation while histopathology results using mice were profitable. The irritation experiments displayed that the mats did not induce any skin irritation

Arum macalatum bitkisinin yara iyileştirici aktivitesi

Objective: In this study, the antioxidant properties of Arum maculatum plant were evaluated. This study reported for the first time the wound healing activity of the methanol extract of A. maculatum fruits. This study aimed to assess and determine the possible pharmacological activities of A. maculatum and evaluate its potential to act as a wound care plant. Methods: The antioxidant and antimicrobial activities of A. maculatum were investigated using excisional in vivo and in vitro wound healing mouse models. A total of 32 Balb-c mice were used, which were equally, divided into four groups: saline control group, control group, A. maculatum group, and Centella asiatica extract group. Treatment applications were performed topically once per day. Wound area narrowing, wound healing percentage, and epithelialization time were analyzed. Results: A. maculatum application supported the healing process in in vivo and in vitro wound models. A. maculatum contributed to the healing process by promoting granulation tissue formation, epidermal regeneration, and angiogenesis. Conclusions: Wound healing is a complex and well-organized process that requires communication between cells. The antioxidant and antimicrobial activities of A. maculatum extract have been determined by current studies. A. maculatum extract may provide significant benefits in promoting the wound healing process.Amaç: Bu araştırmada antioksidan özelliklerini değerlendirmek için Arum maculatum bitkisi seçilmiştir. Bildiğimiz kadarıyla A. maculatum meyvelerinin metanol özünün yara iyileştirici aktivitesi ilk kez bu çalışmada rapor edilmiştir. Bu çalışma, A. maculatum’un olası farmakolojik aktivitelerini belirlemek, değerlendirmek ve bir yara tedavi edici bitki olarak etki gösterme potansiyelini değerlendirmek içindi. Yöntemler: A. maculatum’un antioksidan ve antimikrobiyal aktiviteleri, farelerde eksizyonel in vivo ve in vitro yara iyileşme modelleri kulanılarak araştırılmıştır. Toplamda 32 Balb-c fare kullanılmış olup salin kontrol grubu, kontrol grubu, A. maculatum uygulanan grup ve Centella asiatica özütü uygulanan grup olmak üzere 4 gruba ayrılmıştır. Tedavi uygulamaları günde bir kez topikal olarak gerçekleştirilmiştir. Skar alanı hacminde gerçekleşen değişim, yara iyileşme yüzdesi ve epitelizasyon süresi analiz edilmiştir. Bulgular: A. maculatum uygulaması in vivo ve in vitro yara modelinde iyileşme sürecini desteklemiştir. A. maculatum, granülasyon dokusunu artırarak iyileşme sürecine katkıda bulunmuş, epidermal rejenerasyonu ve anjiyogenezi artırmıştır. Sonuçlar: Yara iyileşmesi, hücreler arası iletişimi gerektiren karmaşık ve iyi organize edilmiş bir süreçtir. Mevcut çalışmalar doğrultusunda antioksidan ve antimikrobiyal aktivitesi belirlenmiş olan A. maculatum özü, yara iyileşme sürecinin desteklenmesinde önemli bir fayda sağlayabilir

Cancer stem cells, their microenvironment and anoikis

Tumors are composed of a variety of cancer cells, all of which contribute to tumor heterogeneity. Among these populations of cells, cancer stem cells (CSCs) have an important role in the initiation and progression of cancer. CSCs are maintained extrinsically within the tumor microenvironment, which contains both cellular and physical factors. As a barrier to metastases, cells normally undergo apoptosis (cell death process) after they lose contact with their extracellular matrix or neighboring cells. This cell death process has been termed “anoikis.” The tumor cells that acquire malignant potential develop mechanisms to resist anoikis. The tumor microenvironment has been acknowledged to contribute to anoikis resistance of bystander cancer cells via modulation of the matrix stiffness, enhancement of oxidative stress, production of pro-survival soluble factors, trigger of epithelial-mesenchymal transition (EMT), and self-renewal ability, thus leading to metabolic deregulations of cancer cells. In this article, we review the significance of perivascular cells, extracellular matrix, tumor stiffness, and hypoxia in the regulation of CSC plasticity and anoikis resistance. With a better understanding of the CSC interaction with its niche and anoikis resistance, it is possible to identify potential therapeutic targets for the development of more effective treatments against cancer

MDAH-2774 insan over kanseri üç boyutlu hücre kültüründe farklı ilaç etkilerinin nitrik oksit sentaz değişiklikleri ile incelenmesi

Objectives: The purpose of our study is to examine the effects of the various medicines at the three dimensional MDAH-2774 human ovary cancer culture with cell proliferation and nitric oxide synthase changing. Methods: Using of medicines at the two dimensional cultures jointly or severally was examined by immunohystochemical method in terms of cell increasing at the 24th, 48th, 72nd and 96th hours and in terms of activities of excitable (inducible) nitric oxide synthase and endothelial nitric oxide synthase at the tree dimensional cultures. Results: Using of these medicines in Paracetamol, gemsitabine and Gemsitabine + Paracetamol and Gemsitabine + Resveratrol combination groups, reduced the number of the cells at all hours according to the control (p;lt;0,05). When performed immunohystochemical analyses were individually compared with the cells of MDAH-2774 or medicine combination, it indicated that caused increasing at inducible nitric oxide synthase and endothelial nitric oxide synthase immunoreactivity. Immunoreactivity increased markedly at the 96th hour in comparison with the 24th hour. Conclusion: Using of the medicines one by one or in combination, reduced the cell proliferation at all hour time zones and increased inducible nitric oxide synthase and endothelial nitric oxide synthase activity.Amaç: MDAH-2774 insan over kanseri iki ve üç boyutlu hücre kültüründe farklı ilaç etkilerinin hücre proliferasyonu ve nitrik oksit sentaz değişiklikleri ile incelenmesidir. Yöntem: İki boyutlu kültürlerde ilaçların ayrı ayrı ve birlikte kullanımları 24, 48, 72 ve 96. saatlerde hücre çoğalması açısından, Üç boyutlu kültürlerde 24 ve 96. saatlerde uyarılabilir (indüklenebilir) nitrik oksit sentaz ve endotelial nitrik oksit sentaz aktivitesi açısından immünohistokimyasal yöntemle incelendi. Bulgular: Parasetamol, Gemsitabine ve Gemsitabine + Parasetamol ve Gemsitabine + Resveratrol kombinasyon gruplarında bu ilaçların kullanılması kontrole göre tüm saatlerde hücre sayısını anlamlı bir şekilde azalttı (p 0,05). Yapılan immunohistokimyasal analizler Gemsitabine’in MDAH-2774 hücreleri ile tek başına veya ilaç kombinasyonu ile karşılaştırıldığında nitrik oksit sentaz ve endotelial nitrik oksit sentaz immunoreaktivitesinde artışa neden olduğunu göstermiştir. İmmunoreaktivite 96 saatte, 24 saate göre belirgin olarak artmıştır. Sonuç: İlaçların tek tek ve kombinasyon halinde kullanımı tüm saat dilimlerinde hücre proliferasyonunu azaltmış, nitrik oksit sentaz ve endotelial nitrik oksit sentaz aktivitesini artırmıştır

The rhythmicity of life: A review of the circadian clocks

Physiology of the mammalian body has been adapted to diurnal cycles of around 24 h, an evolutionary situation that affects a wide spectrum of biological events including sleep-to-wake transitions, feeding/fasting, body temperature, and hormonal regulations. The patterns of the diurnal cycle occur due to rhythmic oscillations that arise from the suprachiasmatic nucleus of hypothalamus, which also can be defined as the pacemaker of the system. The clock can be defined as a molecular machinery driven by the core clock genes that encode clock proteins in a rhythmic oscillatory fashion maintained by the light/dark cycles of the environment. Although the well-established knowledge refers to the function of the circadian rhythm as maintenance of the normal physiology, growing evidence shows that disruptions in the system usually caused by genetic and/or epigenetic misregulations may have a direct effect to lead major pathological conditions, such as carcinogenesis. This review outlines the main molecular aspects of circadian physiology, and reveals the reasons for and results of the circadian disruptions at different levels. In spite of the fact that more proof is needed for a direct correlation between circadian disruptions and oncogenesis and other pathological events, data obtained from current research supports the role of circadian rhythms in malfunctioning of the normal cellular metabolism

Kanser Kök Hücresi

There has been a growing important in recent years in the role stem cells play in health and disease. In the cancer tissue, this population of long lived cells with extraordinary expansion potential has been called tumor initialing cells or cancer stem cells (CSCs). It ıs thought that CSCs are responsible for these aggressive and reccurent tumors.Research has been increasing in recent years into the application of stem cell biology to clinical medicine, particularly it’s role in the evolution and metastasis tumours.Cancer stem cell may have a specific role in tumour metastasis, and their understanding may provide insight into the development of predictive and prognostic markers, future studies shoul lead to development of CSCs targeted therapies for cancer treatment.Kök hücrelerin sağlıklı organizma ve hastalıklar üzerindeki rolü son yıllarda yapılan çalışmalarda önemli gelişmelere neden olmuştur. Kanser dokusunda, patogenezden sorumlu tutulan artmış büyüme potansiyeline sahip, uzun ömürlü hücre popülasyonu kanser kök hücreleri (KKH) olarak adlandırılırlar. KKH’lerinin kemoterapiye dirençli, saldırgan ve tekrarlayan tümörlerden sorumlu olduğu düşünülmektedir. Klinik tıpta hücre biyolojisine dair bildirimler artmıştır, özellikle tümör metastazı ile gelişmelerde önemli bir rol oynamaktadır. Kanser kök hücreleri tümör metastazında önemli bir role sahip olabilir ve prognostik belirleyiciler anlaşıldığı takdirde gelecekte yapılacak çalışmalar kanserin tedavisi için KKH’lerini hedef alan tedavilerin geliştirilmesine öncülük edecektir