ELISA Sisteminde Protein-İnorganik Hibrit Konjugatın Kullanılabilirliği

yokIn the Enzyme-linked immunosorbent assay method (ELISA), it is important to increase the immobilization efficiency of the functional molecules on the microplate and increase the signal-to-noise ratio in order to detect the target molecules with high sensitivity and selectivity. For this purpose, antibody functionalized materials can generate remarkable signal amplification with high enzyme capacity by using conventional immobilization methods. Recently, hybrid structures containing protein/enzyme and Cu3(PO4)2, which is a different enzyme immobilization method, have higher catalytic activity compared to the free form of protein-containing molecules as they have hierarchical microstructures and form large active surface areas. In this study; using protein-inorganic hybrid structure synthesis method, hybrid functionalized conjugate systems with enzyme, antibody and Cu3(PO4)2 were synthesized all together and characterization of the resulting structures was performed by SEM, EDX, XRD and FTIR analysis. When the findings obtained from the researches were evaluated, the hybrid nano structure showed 183.5 EU/mg peroxidase activity and free HRP enzyme showed 59.01 EU/mg activity. The performance of the ELISA system was measured using hybrid conjugate constructs prepared using various TNF-alpha specific antibodies at a -1 concentration of 5-1000 g mL . The performance of hybrid conjugate structure containing multiple organic molecules in ELISA system was higher than other structures. This method is a highly practical method that can replace enzyme-labeled antibody method in ELISA.yo

An investigation of acute effects at various doses of malathion on glucose homeostasis and insulin resistance in rat liver, pancreas and serum

Objective. This study investigates acute effects of various doses of Malathion on glucose homeostasis and insulin resistance in rat. Methods. Rats were randomly divided into four groups of 6 animals each. Corn oil was given orally to Group 1. Group 2, Group 3, and Group 4 received malathion dissolved in corn oil via oral administration at the doses of 100, 200 and 400 mg/kg, respectively. 24 hours later the rats were sacrificed. Results. Acute administration of Malathion led to a decrease in serum butryl cholinesterase (BChE) levels at all doses tested. It also caused a significant increase in serum advanced glycation end products (AGEs), insulin, and TNF-α levels at all doses. Moreover, Malathion administration raised the liver ALT, AST and LDH, TNF-α, and glycogen levels in a dose dependent manner. It also led to a remarkable increase in pancreatic insulin levels at all doses. Conclusions. Acute administrations of Malathion affect glucose homeostasis in a dose dependent manner through its effects on the liver, serum, and pancreas

The Investıgatıon Of Malathıon s Effect On Oxıdant And Antıoxıdant systems, Inflammatory Marker And Some Metabolıc Bıomolecules In rat Serum

Çalışmamızda tarımda pestisit olarak kullanılan bir organofosfat olan Malathion un akut (24 saat) farklı doz uygulamarıyla Wistar albino rat serum parametrelerine etkileri incelenmiştir. Bu amaçla her grupta 6 tane olmak üzere kontrol (Grup1) ve 100 mg/kg (Grup 2), 200 mg/kg (Grup 3), 400 mg/kg (Grup 4) doz grupları oluşturularak seçilen serum parametreleri üzerindeki değişimler araştırılmıştır. Serum malondialdehit (MDA), glukoz, glukoz-6-fosfat dehidrojenaz (G6PD), laktat dehidrojenaz (LDH), amilaz, alkalen fosfataz (ALP), kreatinin ve üre (BUN) seviyelerinde, gruplar arasında istatistiksel olarak farkın anlamlılığı bulunamamıştır. Serum aspartat aminotransferaz (AST) düzeylerinde Grup 3 ve Grup 4, Grup 1 ve Grup 2 ye göre anlamlı olarak artmıştır. Serum alanin aminotransferase (ALT) düzeylerinde Grup 3, Grup 1 e göre anlamlı bir artış bulunmaktadır. Kreatin kinaz (CK) düzeylerinde Grup 3 ve Grup 4 ile Grup 1 ve Grup 2 ye göre bir artış gözlenmiştir. Lipaz kontrol grubu dışındaki tüm doz gruplarında inhibe olmuştur. Total oksidan durum (TOS) düzeyinin Grup 4 te, Grup 1 ve Grup 2 ye göre arttığı bulunmuştur. Serum ileri glikolizasyon son ürünleri (AGEs), insulin ve tümor nekroz faktör-alfa (TNF-α ) Grup 4 te Grup 1 e göre artış gözlenmiştir. Korelasyon analizlerine bakıldığında Serum AGEs ve Serum TOS düzeyleri arasında pozitif bir korelasyon bulunmuştur. AGEs ile TNF- α düzeyleri aralarında pozitif korelasyon olduğu tespit edilmiştir. TNF- α ile TOS, TNF- α ile AST arasında pozitif bir korelasyon gözlenmişken, TNF-α ve ALP arasında negatif bir korelasyon tespit edilmiştir. Ayrıca ALP ile AST ve ALP ile CK arasında da kuvvetli negatif bir korelasyon olduğu bulunmuştur. Glukoz ile G6PD arasında ve TOS ile insülin arasında da pozitif bir korelasyon olduğu gözlenmiştir. İnsulin ile AGEs arasında da pozitif bir korelasyon olduğu tespit edilmiştir. Sonuçlarımıza göre Malathionun akut artan doz uygulamalarında, serum parametreleri incelendiğinde oksidan-antioksidan sistem, doku hasarı, inflamasyon ve bazı metabolik olayları etkilediği ve bu sebeple insan-toplum-çevre sağlığı açısından doz uygulamalarında gerekli eğitim ve önlemlerin alınması gerektiği sonucu ortaya çıkmaktadır.In this study, we investigated the acute effects of different doses of Malathion, which is an organophosphate used as a pesticide in agriculture, on serum parameters of Wistar albino rat. For this purpose, we set 4 groups containing 6 animals (control (Group 1), 100 mg/kg (Group 2), 200 mg/kg (Group 3), 400 mg/kg (Group 4)) and the change of serum parameters was analyzed. We found that there was no significant change in serum malondialdehyde (MDA), glucose, glucose-6- phosphate dehydrogenase (G6PD), lactate dehydrogenase (LDH), amylase, alkalen phosphatase (ALP) creatinine and urea levels among the Groups. Serum aspartate aminotransferase (AST) levels of Group 3, and Group 4 were significantly higher than Group 1 and Group 2. The serum alanine aminotransferase (ALT) significantly increased in Group 3 compare to Group 1. Creatine kinase (CK) increased in Group 3 and Group 4 compare to Group 1 and Group 2. Serum lipase activity has lose all dose groups (Group 2, Group 3 and Group 4). Serum total oxidant status (TOS) levels increased in Group 4 compare to Group 1 and Group 2. Serum advanced glycation end products (AGEs), Insulin and tumor necrosis factor-alpha (TNF- α) levels were increased in Group 4 compare to Group 1. In correlation analysis, we found that thare is a positive correlation between serum TOS levels and AGEs levels. We observed that there is a strong positive correlation between AGEs levels and TNF- α levels in serum. Also there is positive correlation between TNF- α and TOS levels and TNF- α and AST levels. On the other hand negative correlation has found between TNF- α and ALP levels in serum. Additionally, negative correlation has observed between ALP and AST levels and ALP and CK levels in serum. There is strong positive correlation has found between glucose and G6PD levels and TOS and insulin levels. Also we observed that positive correlation between serum AGEs and insulin levels. According to our result, In acute ascending dose of Malathion application, when serum parameters examined, it effects oxidant-antioxidant status, tissue damage, inflammation and any other metabolic status, so for this purpose we consider that necessary precautions and educations should be taken for human, public and environment heal

ELISA SİSTEMİNDE PROTEİN-İNORGANİK HİBRİT KONJUGATIN KULLANILABİLİRLİĞİ

yokIn the Enzyme-linked immunosorbent assay method (ELISA), it is important to increase the immobilization efficiency of the functional molecules on the microplate and increase the signal-to-noise ratio in order to detect the target molecules with high sensitivity and selectivity. For this purpose, antibody functionalized materials can generate remarkable signal amplification with high enzyme capacity by using conventional immobilization methods. Recently, hybrid structures containing protein/enzyme and Cu3(PO4)2, which is a different enzyme immobilization method, have higher catalytic activity compared to the free form of protein-containing molecules as they have hierarchical microstructures and form large active surface areas. In this study; using protein-inorganic hybrid structure synthesis method, hybrid functionalized conjugate systems with enzyme, antibody and Cu3(PO4)2 were synthesized all together and characterization of the resulting structures was performed by SEM, EDX, XRD and FTIR analysis. When the findings obtained from the researches were evaluated, the hybrid nano structure showed 183.5 EU/mg peroxidase activity and free HRP enzyme showed 59.01 EU/mg activity. The performance of the ELISA system was measured using hybrid conjugate constructs prepared using various TNF-alpha specific antibodies at a -1 concentration of 5-1000 g mL . The performance of hybrid conjugate structure containing multiple organic molecules in ELISA system was higher than other structures. This method is a highly practical method that can replace enzyme-labeled antibody method in ELISA.yo

Effects of organic‑inorganic hybrid nanoflowers’ framework on hemocytes and enzymatic responses of the model organism, Galleria mellonella (Lepidoptera: Pyralidae)

Organic–inorganic hybrid nanofowers have gained considerable attention over the last years due to their morphologies, enhanced enzymatic activities, and relatively high stability. The framework of these hybrid nanostructures has been associ ated with the formation of Cu3(PO4)2.3H2O (pure copper phosphate nanofowers). In this study, we investigated the formation of pure copper phosphate nanofowers (p-CPnfs), and their efects on hemocytes and enzymatic responses in Galleria mel lonella (Lepidoptera: Pyralidae) as a model organism. Therefore, the efects of p-CPnfs on G. mellonella hemocytes and on the activities of hemolymph acetylcholine esterase (AChE), superoxide dismutase (SOD), and catalase (CAT) enzymes were evaluated after a 24-h p-CPnfs treatment in the G. mellonella larvae. Diferent concentrations of p-CPnfs (100, 500, 1000, 2000, 6000 ppm) were cautiously force-fed to the fourth instar larvae through the mouth of the insect by means of a micro fne insulin syringe without causing any physical damage. 1000, 2000, and 6000 ppm of p-CPnfs signifcantly increased the total hemocyte counts (THCs) of the G. mellonella larvae. 500, 1000, 2000, and 6000 ppm of p-CPnfs increased the SOD activity of the larvae. 1000, 2000, and 6000 ppm of p-CPnfs increased the CAT activity in the hemolymph of the larvae. Unlike the results seen in SOD and CAT enzyme activities, all concentrations of p-CPnfs decreased AChE activity in the larval hemolymph. In addition to these results, LC50 value was found to be 8410 ppm for the force-fed larvae at the end of the 24-h period.Nevsehir Haci Bektas Veli University Scientifc Research Projects Coordination Unit