Administrative Information المعلومات االدارية Project Title - عنوان المشروع A Novel Strategy for Efficient Monitoring of Diabetic Nephropathy Disease and Hypoalbuminemia by Cooperative Binding of a Fluorescent Probe in a Suprabiomolecular Host–Protein Assembly Principal Investigator - الباحث الرئيسي رقم الهاتف العنوان االلكتروني Telephone e-mail 3985 dp03@aub.edu.lb العنوان Address Dept. Of Chemistry, American University of Beirut, Beirut, Lebanon الوظيفية المؤسسة Post Institution Assistant American Professor University of Beirut االسم Name Digambara Patra Co-Workers - الباحثون المشاركون العنوان االلكتروني e-mail 24 months المؤسسة Institution االسم Name : Duration -المدة التعاقدية للمشروع Scientific Information علمية ّ المعلومات ال Objectives - الهدف The specific aims of the proposed project were: 1) Physical and Optical characterization of fluorescent probes and PAHs in various solvent media, micelle, and membrane. 2) Study on interaction of different fluorescent probes and PAHs with albumins 3) Investigating different fluorescent guest molecules e.g. PAHs in various cucurbit[n]urils host. 4) Cooperative binding study of albumins with various cucurbit[n]urils incorporating different guest molecules. 5) Development of analytical technique based on above study and applying synchronous fluorescence scan spectroscopy and life time technique for efficient monitoring of diabetic nephropathy disease and hypoalbuminemia by directly measuring albumin 6) Application of above mentioned technique towards real sample analysis Achievements -أالنجازات المحققة Physical and Optical characterization of fluorescent probes and PAHs in various solvent media, micelle, and membrane which was successfully achieved during the project work. Study on interaction of different fluorescent probes and PAHs with albumins which was achieved during this period. Investigating different fluorescent guest molecules e.g. PAHs in various cucurbit[n]urils host that was successfully carried out during this period. Cooperative binding study of albumins with various cucurbit[n]urils incorporating different guest molecules, this aim was well investigated during the project work. Development of analytical technique based on above study and applying synchronous fluorescence scan spectroscopy and life time technique for efficient monitoring of diabetic nephropathy disease and hypoalbuminemia by directly measuring albumin, though project aimed at both lifetime and synchronous fluorescence scan spectroscopy as analytical tools, our results failed to correlate with fluorescence lifetime. However, a very sensitive method for albumin determination was developed based on synchronous fluorescence spectroscopy scan during this period, which has been published in Biosensors and Bioelectronics. Perspectives - آفاق البحث The synchronous fluorescence biosensor has high sensitivity for the determination of albumin, particularly for BSA and -globunin, when chrysene is used as a fluorescence probe and -CD, CB6 or CB7 as host molecule. Appearance of separate SFS peak in the lower (blue) wavelength region upon binding of chrysene with albumins can easily indentify albumin present in the sample. Thus, unlike conventional fluorescence emission or excitation spectral measurement, SFS becomes a unique tool to identify the presence of albumin in the biological sample. Cooperative binding of chrysene in a supramolecular host–protein assembly improves the interference from coexisting foreign metal ions. SFS measurement could be done much faster than conventional separation based method for protein estimation and fluorescence process gives higher sensitivity which could be further improved by confining the probe molecule in a host system. Therefore, apart from reducing interference, applying the proposed method higher sensitivity, simplicity and rapidity could be achieved. Further studies in this field will open up the way to applications of cooperative binding of probe in a supramoleccular host-bioanalyte assembly in biosensor and analytical biochemistry. Publications & Communications - المنشورات والمساهمات في المؤتمرات 1. Digambara Patra*, Synchronous fluorescence based biosensor for albumin determination by cooperative binding of fluorescence probe in a supra-biomolecular host–protein assembly, Biosensors and Bioelectronics, 25, 1149–1154 (2010) 2. Brigitte Wex, Fadi M. Jradi, Digambara Patra, Bilal R. Kaafarani, End-capping of conjugates thiophene aromatic systems, 66, 8778 - 8784 (2010) 2. Digambara Patra*, Arlin Jose Amali, Rohit Kumar Rana*, Preparation and photophysics of HPTS based nanoparticle-assempled microcapsules, Journal of Material Chemistry, 19, 40174021 (2009) 3. Jad A. Degheili, Rasha M. Moustafa, Digambara Patra*, Bilal R. Kaafarani, Effect of chain length on photophysical properties of pyrene based molecules substituted with extended chains, The Journal of Physical Chemistry A,113, 1244 – 1249 (2009) 4. Rasha M. Moustafa, Jad A. Degheili, Digambara Patra*, Bilal R. Kaafarani, Synthesis and detailed photophysical studies of pyrene based molecules substituted with extended chains, The Journal of Physical Chemistry A, 113, 1235 – 1243 (2009) 5. Digambara Patra*, Tarek H. Ghaddar, Application of synchronous fluorescence scan spectroscopy for size dependent simultaneous analysis of CdTe nanocrystals and their mixtures, Talanta,77, 1549 – 1554 (2009) Abstract - موجز عن نتائج البحث In diabetic nephropathy disease, albumin proteins can cross the kideny of the patient and can be detected in urine sample. All patients with kidney disease, diabetes, or hypertension and metabolic syndrome should be screened for albuminuria. Recently developed methodologies using high-performance liquid chromatography (HPLC) are sensitive and specific when compared with older methods of detection and may prove very useful for earlier identification of high-risk patients. This is important since studies have shown that albuminuria levels below the microalbuminuria range, determined by conventional methodologies in uncomplicated essential hypertensive men, are associated with an adverse cardiovascular and metabolic risk profile. Fluorescence is fast, highly sensitive and selective, thus, fluorescence based method would not only improve the sensitivity and selectivity for albumin detection but also unlike HPLC it does not need a time consuming pre-separation procedure opening up the possibility for fast optical sensor development. Here our goal is to develop an efficeint methodology for monitoring albumin by cooperative binding of a fluorescent probe in a supramolecular hostprotein assembly. Fluorescent properties of probe molecules through host guest assemplies or associations with proteins enable biomedical sensor applications and improve the factors governing molecular recongination. Macrocyclic hosts impose a structural confinement of the included chromophoric guest molecules, increase their thermal and photochemical stability, and isolate them from the bulk water, which together improves their radiative properties. Cucurbit[n]uril is a synthetic pumpkin-shaped cation receptor and have recently attracted much attention because of their remarkably strong binding of organic dyes, protonated alkyl and aryl amines, and metal ions. Cucurbit[7]uril (CB7), in particular, is known for its exceptional thermal and photochemical stabilization of fluorescent dyes. CB7 shows a strong affinity for protonated amines. The goal of the present study is therefore to modify the radiative property of certain fluorescent probe like polycyclic aromatic hydrocarbons (PAHs) and other fluorescent probes by introducing a combination of supramolecular (using cucurbit[n]urils as a macrocyclic host) and biomolecular (using HAS/BSA as a protein) interactions and apply them as an analytical tool for albumin monitoring.