International Journal of Applied Biology and Forensics

Investigation of Biochemical Changes in Saliva and Blood of Diabetic Patients

Abdul Razzaq, Muhammad Sajid and Nasir Iqbal

Institute of Molecular Biology and Biotechnology (IMBB), The University of Lahore, Lahore, Pakistan.

Correspondence: arazzaq1122@gmail.com

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Abstract

Diabetes mellitus is group of metabolic disorders characterized by an inability to produce sufficient insulin or to responds insulin that results high blood glucose level. Type-1 diabetes mellitus considered as an autoimmune disease and required life-long treatment with exogenous insulin. Blood glucose levels along with serum insulin level required for diagnosis. In present study serum and salivary biochemical changes in type-1 diabetes mellitus patients were analyzed to evaluate non-invasive diagnostic markers. For this purpose, serum calcium, phosphate, total protein, alkaline phosphatase and salivary calcium, phosphate and alpha amylase were analyzed. Overall high serum calcium levels were found in diabetic patients. Serum phosphate levels were found lower in diabetic patients as compare to normal individuals. Serum alkaline phosphatase and total protein levels were also significantly higher in diabetic patients. Salivary calcium levels increased significantly in diabetic patients as compare to normal individuals. While the salivary phosphate and salivary alpha amylase levels significantly decreased in type-1 diabetic patients. It is suggested that saliva is absolute surrogate of blood that can be used to diagnose and evaluate the progression of type-1 diabetes mellitus and as non-invasive diagnostic body fluid.

Keywords: Diabetes Mellitus, Exogenous Insulin, Autoimmune, Non-Invasive Diagnostic Markers, Surrogate.

Extraction, Purification and Characterization of Proteolytic Enzyme from Fig (Ficus carica) and Kachri (Cucumis trigonus)

Haseeb Akram Sindhu, Abu Saeed Hashmi, Faiza Masood and Muhammad Sajid

Institute of Biochemistry and Biotechnology (IBBT), UVAS, Lahore, Pakistan.

Correspondence: sirhaseebakram@gmail.com

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Abstract

Fig (Ficus carica) and Kachri (Cucumis trigonus) contain high concentration of proteolytic enzymes. In current research work, purification and characterization of papain (a proteolytic enzyme) from Kachri (Cucumistrigonus) and Fig (Ficus carica) was carried out. Kachri and Fig was crushed separately in a food blender byusing 0.1M (PO4)-3 buffer of pH 8.00, distilled water and 70% ethanol. Liquid mixture obtained was centrifuged and supernatant used as enzyme source. Purification of papain was carried out by Ammonium Sulphate precipitation and dialysis followed by Gel filtration by Sephadex G-50. Then characterization of papain such as protein estimation, determination of proteolytic activity (International Unit) of enzyme and SDS-PAGE analysis were performed to determined molecular weight. Finally, the yield and proteolytic activity of papain was measured and compared with the commercial products. Crude preparation of enzyme has a wide specificity due to the presence of various proteinase and peptidase isozymes. These enzymes being present in natural fruits were free from any toxic effects and consequently can be used in food and pharmaceutical industries.

Key words: Kachri, Fig, Papain, SDS-PAGE, Sephadex G-50, Isozymes