The preparation and characterization of solid phase micro extraction fiber for pesticide analysis

Çevre ve tarım açısından büyük önem arzeden pestisitlerin, formülasyonlarda, karmaşık yapılı gıda ve çevre örneklerinde analizi genellikle yüksek verimli, seçimli ve duyarlı yöntemlerin geliştirilmesini gerektirir. Tepe boşluğu katı faza mikro özütleme (HS-SPME) tekniği klasik örnek hazırlama tekniklerine göre birçok üstünlüğe sahiptir. Bu çalışmada pahalı ticari fiberlere alternatif oluşturmak ve özel seçimlilik kazandırmak üzere laboratuar yapımı fiber geliştirilmesi amaçlanmıştır. Fiber oluşturmada elektrokimyasal polimerizasyon tekniği ile pirol monomeri çelik tel üzerine kaplanmış ve HS-SPME koşullarına kullanımı araştırılmıştır. Adsorpsiyon sıcaklığı ve süresi, desorpsiyon sıcaklığı, karıştırma hızı ve tuz miktarı gibi denel parametreler optimize edilmiş ve sırasıyla 70oC ve 45 dk. , 200oC, 600 rpm ve 10 g/L olarak saptanmıştır. Bu tez kapsamında seçilen Chlorpyrifos (CP), Penconazole (PNZ), Procymidone (PRC), Brompropylate (BRP) ve Lambda-Cyhalothrin (LMD) pestisit türleri için kalibrasyon grafikleri oluşturulmuştur. Çalışmada doğruluğun artırılabilmesi için iç standart olarak aldrin(ALD) kullanılmıştır. Bu yöntemin uygulaması Türk sarap örneklerine yapıldığında, makul gerikazanım değerleri hesaplanmıştır

Conversion of specific lncRNAs to biomarkers in exhaled breath condensate samples of patients with advanced stage non-small-cell lung cancer

Objectives: Lung cancer (LC) is one of the most prevalent cancers with the highest fatality rate worldwide. Long noncoding RNAs (lncRNAs) are being considered potential new molecular targets for early diagnosis, follow-up, and individual treatment decisions in LC. Therefore, this study evaluated whether lncRNA expression levels obtained from exhaled breath condensate (EBC) samples play a role in the occurrence of metastasis in the diagnosis and follow-up of patients with advanced lung adenocarcinoma (LA).Methods: A total of 40 patients with advanced primary LA and 20 healthy controls participated in the study. EBC samples were collected from patients (during diagnosis and follow-up) and healthy individuals for molecular analysis. Liquid biopsy samples were also randomly obtained from 10 patients with LA and 10 healthy people. The expression of lncRNA genes, such as MALAT1, HOTAIR, PVT1, NEAT1, ANRIL, and SPRY4-IT1 was analyzed using cfRNA extracted from all clinical samples.Results: In the diagnosis and follow-up of patients with LA, lncRNA HOTAIR (5-fold), PVT1 (7.9-fold), and NEAT1 (12.8-fold), PVT1 (6.8-fold), MALAT1 (8.4-fold) expression levels were significantly higher than those in healthy controls, respectively. Additionally, the distinct lncRNA expression profiles identified in EBC samples imply that decreased ANRIL-NEAT1 and increased ANRIL gene expression levels can be used as biomarkers to predict the development of bone and lung metastases, respectively.Conclusion: EBC is an innovative, easily reproducible approach for predicting the development of metastases, molecular diagnosis, and follow-up of LC. EBC has shown potential in elucidating the molecular structure of LC, monitoring changes, and discovering novel biomarkers.The research project received support from the Scientific and Technological Research Council of Turkey (TUBITAK-121S932). The article processing charges were funded by the Presidency of Strategy and Budget of the Republic of Turkey (2019K12-149080).Scientific and Technological Research Council of Turkey [TUBITAK-121S932]; Presidency of Strategy and Budget of the Republic of Turkey [2019K12-149080

* Concordance in molecular genetic analysis of tumour tissue, plasma, and exhaled breath condensate samples from lung cancer patients

Yalcin, Femin/0000-0003-0602-9392; Tetik Vardarli, Asli/0000-0001-9890-3256; Aldag, Ceyda/0000-0003-3130-4739; Pelit, Levent/0000-0001-8090-703XWOS: 000528569000001PubMed: 32031993Aim. Lung adenocarcinoma is characterized by poor prognosis and short survival rates. Therefore, tools to identify the tumoural molecular structure and guide effective diagnosis and therapy decisions are essential. Surgical biopsies are highly invasive and not conducive for patient follow-up. To better understand disease prognosis, novel non-invasive analytic methods are needed. the aim of the present study is to identify the genetic mutations in formalin-fixed paraffin-embedded (FFPE) tissue, plasma, and exhaled breath condensate (EBC) samples by next-generation sequencing and evaluate their utility in the diagnosis and follow-up of patients with lung adenocarcinoma. Method. FFPE, plasma, and EBC samples were collected from 12 lung adenocarcinoma patients before treatment. DNA was extracted from the specimens using an Invitrogen PureLink Genomic DNA Kit according to the manufacturer's instructions. Amplicon-based sequencing was performed using Ion AmpliSeq Colon and Lung Cancer Research Panel v2. Results. Genetic alterations were detected in all FFPE, plasma, and EBC specimens. the mutations in PIK3CA, MET, PTEN, SMAD4, and FGFR2 genes were highly correlated in six patients. Somatic and novel mutations detected in tissue and EBC samples were highly correlated in one additional patient. the EGFR p.L858R and KRAS p.G12C driver mutations were found in both the FFPE tissue specimens and the corresponding EBC samples of the lung adenocarcinoma patients. Conclusion. the driver mutations were detected in EBC samples from lung adenocarcinoma patients. the analysis of EBC samples represents a promising non-invasive method to detect mutations in lung cancer and guide diagnosis and follow-up.Scientific and Technological Research Council of TurkeyTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TUBITAK-1003-216S591, 216S435]The research project was supported by the Scientific and Technological Research Council of Turkey (TUBITAK-1003-216S591 and 216S435)

Preparation and characterization of sodium dodecyl sulfate doped polypyrrole solid phase micro extraction fiber and its application to endocrine disruptor pesticide analysis

WOS: 000320085300015PubMed ID: 23669608A robust in house solid-phase micro extraction (SPME) surface has been developed for the headspace (HS)-SPME determination of endocrine disruptor pesticides, namely, Chlorpyrifos, Penconazole, Procymidone, Bromopropylate and Lambda-Cyhalothrin in wine sample by using sodium dodecylsulfate doped polypyrrole SPME fiber. Pyrrole monomer was electrochemically polymerized on a stainless steel wire in laboratory conditions in virtue of diminishing the cost and enhancing the analyte retention on its surface to exert better selectivity and hence the developed polymerized surface could offer to analyst to exploit it as a fiber in headspace SPME analysis. The parameters, mainly, adsorption temperature and time, desorption temperature, stirring rate and salt amount were optimized to be as 70 degrees C and 45 min, 200 degrees C, 600 rpm and 10 g L-1, respectively. Limit of detection was estimated in the range of 0.073-1.659 ng mL(-1) for the pesticides studied. The developed method was applied in to red wine sample with acceptable recovery values (92-107%) which were obtained for these selected pesticides. (C) 2013 Elsevier B.V. All rights reserved.Ege UniversityEge University [2010/Fen/033]The authors thank to the Ege University for financial support (Project 2010/Fen/033)

Preparation and Characterization of a Novel Solid-Phase Microextraction Material for Application to the Determination of Pesticides

9th Aegean Analytical Chemistry Days (AACD) -- SEP 29-OCT 03, 2014 -- Chios, GREECEWOS: 000373911300004The preparation and characterization of a novel solid-phase microextraction fiber is reported with application to the determination of pesticides in fruit juice. The fiber was fabricated by electrochemically coating a stainless steel wire with a thin polymeric film of 4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl) benzenamine. The procedure was initiated in 10mL of acetonitrile containing 5.5mg of monomer, 0.1mol NaClO4, and 0.1mol LiClO4 by cycling the potential between -0.5 and 1.2V with a scan rate of 100mV/sec. The morphology of the fiber surface was examined by scanning electron microscopy and its stability was characterized by thermal gravimetric analysis. The fiber was exposed to headspace extraction of bromopropylate, chlorpyrifos, lambda-cyhalothrin, penconazole, and procymidone prior to the analysis by gas chromatography with an electron capture detector. Operational parameters affecting the extraction efficiency, adsorption and desorption times and temperature, and stirring rate were screened using a Plackett-Burman Design. Emerging parameters were further optimized via Central Composite Design that were 20min at 64 degrees C for adsorption and 4.4min at 250 degrees C for desorption. Solution parameters were optimized to be 5.0mL of sample in pH 2.0 Britton-Robinson buffer containing 0.1mg/L NaCl to promote the volatilization of the analytes. The limits of detection were at the ng/mL level for the pesticides. The fiber was used as a selective and sensitive tool for the trace determination of these pesticides in grape juice

cfDNA in exhaled breath condensate (EBC) and contamination by ambient air: toward volatile biopsies

###EgeUn###Exhaled breath is a source of volatile and nonvolatile biomarkers in the body that can be accessed non-invasively and used for monitoring. The collection of lung secretions by conventional methods such as bronchoalveolar lavage, induced sputum collection, and core biopsies is limited by the invasive nature of these methods. Non-invasive collection of exhaled breath condensate (EBC) provides fluid samples that are representative of airway lining fluids. Various volatile and nonvolatile biomarkers can be detected in volatile condensates, such as H2O2, nitric oxide, lipid mediators, cytokines, chemokines, DNA, and microRNAs. Studies have examined cell-freeDNA(cfDNA) in plasma samples from non-small-cell lung cancer patients, offering to new insights and fostering development of the liquid biopsy. However, few studies have examined cfDNA in EBC samples. This study examined whether EBC is an appropriate source of cfDNA using housekeeping-gene-specific primer probes and quantitative real-time polymerase chain reaction in healthy subjects. Ambient (room) air is contaminated with DNA, so caution is needed. Preliminary studies indicated that volatile biopsies are becoming an important diagnostic tool in lung cancer.Department of Scientific Research Projects of Dokuz Eylul University [KB.SAG.049]; Scientific and Technological Research Council of TurkeyTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [216S435, 216S591]Informed consent was obtained from all research subjects. The study was approved by the Ethics Committee of Ege University and was supported by grants from the Department of Scientific Research Projects of Dokuz Eylul University (Project no: 2017.KB.SAG.049) and the Scientific and Technological Research Council of Turkey (Project no: 216S435 and 216S591)