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    Biofortified Whey/Deglycosylated Whey and Chickpea Protein Matrices: Functional Enrichment by Black Mulberry Polyphenols
    (Springer, 2022) Özleyen, Adem; Çınar, Zeynep Özlem; Karav, Sercan; Bayraktar, Ayşe; Arslan, Ayşenur; Kayılı, H. Mehmet; Salih, Bekir; Boyuneğmez Tümer, Tuğba
    Morus nigra L. (black mulberry-BM) is a promising nutraceutical fruit containing biologically active polyphenols like anthocyanins, proanthocyanidins, catechins, and stilbenes, with well-established anti-inflammatory, antidiabetic, anti-obesity, and anticancer biofunctions. However, these health-promoting properties in raw fruit are greatly masked due to the presence of soluble and insoluble carbohydrates in excess amounts restricting daily intake of the required dose to achieve targeted effects. In the current study, different protein sources (defatted whey and chickpea flours) were optimized through different conditions to capture polyphenols from BM juice while diminishing its glucose content. To optimize polyphenol-protein interactions, various pHs (3.7, 4.2, and 4.7), matrix concentrations (20, 50, and 80 g protein/L), and incubation times (5, 20, and 45 min) were tested. In the present work, optimized BM polyphenol enriched whey matrix inhibited pro-inflammatory mediators and promoted Nrf-2 dependent cytoprotective enzyme expressions in lipopolysaccharide (LPS) induced macrophages at low doses. In addition, whey proteins were also subjected to an enzymatic deglycosylation process by using recently identified EndoBI-1 enzyme for the specific cleavage of N-glycan core in all glycan types including high mannoses, hybrids as well as complex glycans found on defatted whey proteins. After this process, the polyphenol sorption capacity of deglycosylated whey proteins was found to be significantly higher (37%) than the capacity of non-treated normal whey protein under optimized conditions. In conclusion, deglycosylation of protein matrices could be a novel strategy for efficient sorption/concentration of polyphenols from fruits and vegetables, however, more detailed studies are needed to understand this effect.
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    Correction: Distinguishing Turkish pine honey from multi-foral honey through MALDI-MS-based N-glycomics and machine learning
    (Springer, 2024) Masri, Saad; Aksoy, Sena; Duman, Hatice; Karav, Sercan; Kayılı, Hacı Mehmet; Salih, Bekir
    [Anstract Not Available]
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    Distinguishing Turkish pine honey from multi-floral honey through MALDI-MS-based N-glycomics and machine learning
    (Springer, 2024) Masri, Saad; Aksoy, Sena; Duman, Hatice; Karav, Sercan; Kayılı, Hacı Mehmet; Salih, Bekir
    Honey, a multifaceted blend of sugars, amino acids, vitamins, proteins, and minerals, exhibits compositional variability dependent upon the floral source. While previous studies have attempted to categorize honey, the use of glycomic profiles for honey classification remains an unexplored avenue. This investigation seeks to establish a methodology for distinguishing honey types, specifically multi-floral and pine honey, employing mass spectrometry-based glycomic analysis in tandem with machine learning. In this search, seven samples of pine honey and eight samples of multi-floral honey were obtained from diverse regions of Turkey. Subsequently, the proteins within these honey samples were extracted, and glycans were enzymatically released. The released glycans were labeled with 2-aminobenzoic acid (2-AA) and subjected to analysis via matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The glycan profiles of pine and multi-floral honey were determined through these analytical procedures, revealing a total of 76 distinct N-glycan structures. Among these, 13 N-glycan profiles consistently established at high levels across experimental replicates and were incorporated in subsequent analyses. Following the quantification of individual glycan abundances, statistically significant differences in glycan profiles were determined. Notably, N-glycans Hex5HexNAc2, Hex4HexNAc3, and Hex5HexNAc3 displayed considerable differences. Using the 13 N-glycan profiles, an accuracy rate of 93.5% was obtained from machine learning analysis, which increased to 100% when incorporating the identified significantly changed glycans. The most productive models were identified as subspace and fine k-nearest neighbors (KNN). The findings underscore the potential of mass spectrometry-based glycomics in conjunction with machine learning as a robust tool for precise honey type classification and its prospective utility in quality control and honey product authentication.
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    ESI-IM-MS characterization of cyclodextrin complexes and their chemically cross-linked alpha (α-), beta (β-) and gamma (γ-) cyclodextrin particles as promising drug delivery materials with improved bioavailability
    (Elsevier, 2023) Yılmaz, Aynur Sanem; Öztürk, Serhat; Salih, Bekir; Ayyala, Ramesh S.; Şahiner, Nurettin
    Cyclodextrins (CDs) are natural cyclic oligosaccharides with a relatively hydrophobic cavity and a hydrophilic outer surface. In this study, alpha (alpha-), beta (beta-) and gamma (gamma-) CD particles were prepared by directly using alpha-, beta-, and gamma-CDs as monomeric units and divinyl sulfone (DVS) as a crosslinker in a single-step via reverse micelle microemulsion crosslinking technique. Particles of p(alpha-CD), p(beta-CD), and p(gamma-CD) were perfectly spherical in sub 10 mu m size ranges. The prepared p(CD) particles at 1.0 mg/mL concentrations were found biocompatible with > 95 % cell viability against L929 fibroblasts. Furthermore, p(alpha-CD) and p(beta-CD) particles were found non-hemolytic with < 2 % hemolysis ratios, whereas p(gamma-CD) particles were found to be slightly hemolytic with its 2.1 +/- 0.4 % hemolysis ratio at 1.0 mg/mL concentration. Furthermore, a toxic compound, Bisphenol A (BPA) and a highly antioxidant polyphenol, curcumin (CUR) complexation with alpha-, beta-, and gamma-CD molecules was investigated via Electrospray-Ion Mobility-Mass Spectrometry (ESI-IM-MS) and tandem mass spectrometry (MS/MS) analysis. It was determined that the most stable noncovalent complex was in the case of beta-CD, but the complex stoichiometry was changed by the hydrophobic nature of the guest molecules. In addition, BPA and CUR were separately loaded into prepared p(CD) particles as active agents. The drug loading and release studies showed that p (CD) particles possess governable loading and releasing profiles.
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    Identification and comparison of N-glycome profiles from common dietary protein sources
    (Elsevier, 2025) Bolino, Matthew; Avcı, İzzet; Kayılı, Hacı Mehmet; Duman, Hatice; Salih, Bekir; Karav, Sercan; Frese, Steven A.
    The N-glycomes of bovine whey, egg white, pea, and soy protein isolates are described here. N-glycans from four protein isolates were analyzed by HILIC high performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry (HILIC-FLD-QTOF-MS/MS). In total, 33 N-glycans from bovine whey and egg white and 10 N-glycans from soy and pea glycoproteins were identified. The type of N-glycans per glycoprotein source were attributable to differences in biosynthetic glycosylation pathways. Animal glycoprotein sources favored a combination of complex and hybrid glycan configurations, while the plant proteins were dominated by oligomannosidic N-glycans. Bovine whey glycoprotein isolate contained the most diverse N-glycans by monosaccharide composition as well as structure, while plant sources such as pea and soy glycoprotein isolates contained an overlap of oligomannosidic N-glycans. The results suggest N-glycan structure and composition is dependent on the host organism which are driven by the differences in N-glycan biosynthetic pathways.
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    Superporous poly(β-Cyclodextrin) cryogels as promising materials for simultaneous delivery of both hydrophilic and hydrophobic drugs
    (Pergamon-Elsevier Science Ltd, 2022) Ari, Betül; Demirci, Şahin; Ayyala, Ramesh S.; Salih, Bekir; Şahiner, Nurettin
    Here, one step, simple preparation of superporous p(beta-cyclodextrin) (p(beta-CD)) cryogels in the presence of various ratios of crosslinker, divinyl sulfone (DVS) e.g., 100, 150, 200% mole with respect to the mole ratio of 6 hydroxyl groups on beta-CD unit under cryogenic conditions were reported for the first time. The swelling properties, and hydrolytic degradation of p(beta-CD) cryogels was directly related to the used crosslinker ratio. It was observed that the p(beta-CD) cryogels showed higher swelling% in DMSO and DMF than in DI water. The prepared p(beta-CD)-1 cryogel that was 100% crosslinked exhibited 14 +/- 3.7%, 34 +/- 4.8%, and 45 +/- 6.2% weight losses within 20 days in pH 5.4, 7.4, and 9.0 buffer solutions, respectively. An acceptable hemolysis index of < 5% and a blood coagulation index, >89% were observed for the p(beta-CD) cryogels at 1 mg/mL concentrations. Additionally, the potential useability of p(beta-CD) cryogels as in vitro drug delivery systems for both hydrophilic vancomycin, and hydrophobic tetracycline as model drugs were individually illustrated at pH 7.4 in PBS. The cumulative drug release from p(beta-CD) cryogels, in sustained release profiles with 76.7 +/- 9.0 mg/g of vancomycin (89.9 +/- 10.5% of the loaded amount) and 146.5 +/- 19.4 mg/g tetracycline g (83.1 +/- 6.3% of the loaded amount) were attained in 6 and 54 h, respectively at pH 7.4 in PBS. Most importantly, p(beta-CD) cryogels exhibited simultaneous loading and releasing the ability for both hydrophilic vancomycin, and hydrophobic tetracycline drugs by concurrently releasing 37.0 +/- 2.7 mg/g (23.4 +/- 1.8% of the loaded amount), and 36.3 +/- 1.3 mg/g (23.6 +/- 0.8% of the loaded amount), respectively at pH 7.4 (PBS) in 10 h.
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    Synthesis, characterization and thermal study of some tetradentate Schiff base transition metal complexes
    (Springer, 2009) Doğan, Fatih; Ulusoy, Mahmut; Özturk, Ömer F.; Kaya, İsmet; Salih, Bekir
    Several mononuclear Co(II), Ni(II), Cu(II), and Fe(II) complexes of tetradentate salpren-type diimine, obtained from 3,5-di-tert-butyl-2-hydroxybenzaldehyde and 1,3-diaminopropane have been prepared and characterized by analytical, spectroscopic (FT-IR, UV-VIS) techniques, magnetic susceptibility measurements and thermogravimetric analyses (TG). The thermodynamic and thermal properties of complexes have been investigated. For further characterization Direct Insertion Probe-Mass Spectrometry (DIP-MS) was used and the fragmentation pattern and also stability of the ions were evaluated. The characterization of the end products of the decomposition was achieved by X-ray diffraction. The thermal stabilities of metal complexes of N,N'-bis(3,5-di-t-butylsalicylidene)-1,3-propanediamine ligand (L) were found as Ni(II) > Cu(II) > Co(II) > Fe(II).

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