Analizi
Mihriban ÖZEN1,2, Mete YILMAZ2
1Uludağ Üniversitesi Fen Edebiyat Fakültesi Biyoloji Bölümü, Bursa
2Bursa Teknik Üniversitesi, Doğa Bilimleri, Mimarlık ve Mühendislik Fakültesi, Biyomühendislik Bölümü, Bursa
Sunum: Sözlü Özet
Mikrosistinler, tatlı sularda en yaygın görülen siyanotoksinlerdir. Protein fosfataz 1A ve 2A aktivitesini inhibe ederek hepatotoksik etki göster- mektedirler. Bu toksinlerin sulardaki varlığı, hayvan ve insan sağlığı açısından önem teşkil edip hastalıklara ve hatta ölümlere neden olabilmek- tedir. Mikrosistinler en yaygın olarak Microcystis, Anabaena, Planktothrix, Oscillatoria ve Nostoc taksonları tarafından üretilmektedir. Ayrıca bu toksinler, besin zinciri vasıtasıyla taşınarak organizmalarda biyomagnifikasyon göstermekte ve o bölgedeki halk sağlığını tehdit etmektedir. Bu çalışma mikrosistin sentetaz A (mcyA) geninin Uluabat Gölü’nde tespit edilmesini ve potansiyel mikrosistin üreticilerinin varlığını göstermeyi amaçlamıştır. Arazi örnekleri, siyanobakteri populasyonunun en bol bulunduğu 2015-2016 yıllarının yaz aylarında 4 farklı istasyondan toplanmış- tır. Plankton çekimi örneklerinin genomik DNA’ları ticari bir kit ile izole edilmiştir. McyA geninin 950-970 baz çifti uzunluğundaki fragmenti, MAIF-MAIR primerleri kullanılarak PZR ile amplifiye edilmiştir. 47 arazi örneğinden 39’unda mcyA geni tespit edilmiştir. PZR ürünleri ticari bir kitle klonlanmış, plazmid saflaştırmasını takiben ürünler dizilenmiştir. Filogenetik analizler Mega 6 yazılımı kullanılarak yapılmıştır. PZR ürün di- zilerinin yine aynı gölden izole edilen bir Microcystis aeruginosa suşuna ait mcyA dizisine benzer olduğu görülmüştür. Uluabat arazi örneklerinde mcyA geninin varlığı, özellikle yaz aylarında gölde mikrosistin üreticilerinin olduğunu göstermiştir. Bölge halkı geçimini balıkçılık ile sağladığı ve sulama amaçlı gölden su çektiği için Uluabat gölü’nün toksik siyanobakteriler açısından sürekli takip edilmesi gerekmektedir.
Anahtar Kelimeler: McyA geni, PZR, Uluabat Gölü
Teşekkür: Bu çalışma TÜBİTAK Projesi 114Y641 ile desteklenmiştir.
PCR Detection, Cloning and Phylogenetic Analysis of Microcystin Synthetase A Gene (McyA) in Lake
Uluabat
Abstract
Microcystins are the most common cyanotoxins in freshwaters. They exhibit hepatotoxic effects by inhibiting protein phosphatases 1A and 2Ain mammals. The presence of these toxins in surface waters is important for animal and human health and can cause illnesses and even deaths. Micro- cystins are most commonly produced by species of Microcystis, Anabaena, Planktothrix, Oscillatoria and Nostoc. Additionally, these toxins may biomagnify through the food chain and threaten the health of humans and animals in that area. This study aimed to detect microcystin synthetase A (mcyA) gene in samples collected from Lake Uluabat and to show the presence of potential microcystin producers. Field samples were collected from four different study sites during summer months of 2015 and 2016 when cyanobacteria populations are most abundant. Genomic DNA of plankton net samples were isolated with a commercial kit. A 950-970 base pair length fragment of the mcyA gene was amplified by PCR using MAIF-MAIR primers. Out of the 48 environmental samples, mcyA gene was detected in 39 samples. PCR products were cloned and plasmids were purified followed by sequencing of plasmids. Phylogenetic analyses were performed in Mega software, version 6. Sequences were similar to the mcyA sequence of a Microcystis aeruginosa strain isolated from the same lake. Presence of mcyA gene in Uluabat field samples pointed out the presence of microcystin producers in the lake especially during the summer months. Fisheries and agriculture are the main sources of income for villagers around the lake, where they draw water from the lake for irrigation purposes. Therefore continuous monitoring of toxic cyanobacteria and cyanotoxins in Lake Uluabat is warranted.
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Preparation and Characterization of Drug-Loaded Gelatin Cryogels
Suzan Melis YAMAN, Nimet BÖLGEN KARAGÜLLE
Mersin University, Engineering Faculty, Chemical Engineering Department, Çiftlikköy Campus, Yenişehir/MERSİN
Sunum: Sözlü Özet
Tissue engineering aims to improve or restore damaged tissues by using cells, scaffolds and some biochemical factors. In tissue engineering, one of the most important concepts is the scaffold because it has a key role in keeping up and promoting the growth the cells. These scaffolds have been considered to be used as a drug delivery system recently. Cryogels are supermacroporous interconnected porous networks which form at subzero temperature by gelation of polymeric precursors. Gelatin is a natural, biodegradable and biocompatible polymer. Simvastatin has repor- ted to promote osteoblastic activity depending on its local concentration. The aim of this study is to investigate the release of simvastatin from macroporous gelatin cryogels to be used as scaffolds for bone regeneration. Macroporous gelatin cryogels with different glutaraldehyde concent- rations were synthesized by the cryogelation method. The cryogelation and crosslinking reaction occurred in the cryostat at -16°C for 24 hours. Simvastatin was used in two forms (hydrophilic and hydrophobic) and these forms were loaded in the cryogels by incorporating or by embedding. As the glutaraldehyde concentration increased gradually, swelling ratio of cryogels decreased. As the glutaraldehyde concentration increased the pores size of the cryogels desreased, which was demonstrated by SEM analysis. Release profile of simvastatin was observed by ultraviolet-visible spectrophotometry. Release rate of simvastatin from cryogels depended on cryogel composition and form (hydrophilic and hydrophobic) of the loaded simvastatin.
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