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Biological Evaluation of Phospholipid Complexes from Extracts of Bistora Amplexicaulis

Thesis Info

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Author

Batool, Salma

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Biochemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/12229/1/Salma%20Batool%20biochemistry%202019%20arid%20rwp%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725625370

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Medicinal plants cater healthcare needs of about 80% of world population particularly in developing countries. Bistorta amplexicaulis belongs to the genus Polygonum (Polygonaceae), and is found in temperate regions of the world. The plant is extensively used as herbal medicine in North Pakistan, India and China and has proven antioxidant and antitumor activities with high phenolic contents. Plant phenolics being quantitatively highest in the extract and are most bioactive, suffer from low cellular uptake due to their hydrophilic nature. The in vivo activity of phenolics is often limited due to stability, low bioavailability and cellular uptake. This translates to using a higher dosage of the extract for effective intracellular concentrations which in turn limits the affectivity and increases side effects. So the researchers are focusing on approaches that can improve the cellular uptake of these compounds to enhance their activity. Keeping in view, the current study was designed for nano-liposomal encapsulation of B. amplexicaulis extracts for increased anti-cancer activity due to increased cellular uptake. Plant extract was prepared using maceration technique, analyzed for its phytochemical constituents using Ultra performance liquid chromatography (UPLC) with C18 columns and screened for its anticancer activity against HCT-116 Human Colon cancer cell lines. Liposomes were prepared using thin film method with DPPC, PEG2000DSPE and Cholesterol in a ratio of 1.85:0.15:1 with 2mg/mL and 4mg/mL ethanolic extract of B. amplexicaulis respectively. Prepared liposomes were characterized using Scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) and Size was determined using dynamic light scattering (DLS) whereas charge was determined using Zeta sizer. Encapsulation efficiency was estimated using spectrophotometer and UPLC. A cytotoxicity comparison was established between encapsulated and un-encapsulated extract through in-vitro MTS cell proliferation assay using Hepatocellular carcinoma cell lines (HepG2) and breast cancer cell lines (MCF-7). Furthermore, encapsulated and un-encapsulated extract was also tested for cytotoxicity to normal Human Umbilical Vein Endothelial Cells. The UPLC analysis identified gallic acid, caffeic acid, chlorogenic acid, catechin and epicatechin in ethanolic rhizome extract. Liposomes encapsulating rhizome extract were characterized having zeta potential of -19.8 mV and -16.9mV, size of 155nm and 143nm with PDI of 0.09 and 0.16 for 2mg/mL and 4mg/mL ethanolic extract formulations respectively. Encapsulation efficiency was in a range of 71-81% for both formulations. The HepG2 cells were found most sensitive against free extract with IC50 of 27mg/mL as compared to MCF-7 with IC50 of 67mg/mL. The rhizome extract loaded liposomes as compared to free extract has shown to improve anticancer activity by 30% against HepG2 cells. However, in normal Human Umbilical Vein Endothelial Cells (HUVEC) the LD50 value of free extract was 13 mg/mL compared to liposomes encapsulated extract having LD50 54.0 mg/mL and LD50 65.9 mg/mL for 2mg/mL and 4mg/mL ethanolic extract formulations respectively. The increased LD50 values of liposomes encapsulated extract against HUVEC cells indicates that its toxicity is decreased to the normal cell whereas liposomal encapsulation of extract has enhanced its anticancer activity. Hence, Nano-liposomal encapsulation could be used as a mean of targeting the B. amplexicaulis rhizome extract to the cancer cells to enhance its anticancer potential avoiding cytotoxicity to the normal cells. To our knowledge this is the first report of the liposomal encapsulation of an extract, enhanced uptake of extract and an improved activity against hepatocellular carcinoma (HepG2).
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گزشتہ ماہ علامہ شیح عبدالعزیز بن عبدﷲ بن باز نے ۸۸ برس کی عمر میں داعی اجل کو لبیک کہا، انا ﷲوانا الیہ راجعون۔ وہ مملکت سعودیہ کے جلیل القدر عالم، مفتی اعظم، علمائے کبار کی سپریم کونسل نیز دارالافتاء اور مجلس بحوث علمیہ کے سربراہ رابطہ عالم اسلامی، الجمع الفقہی الاسلامی اور اس نوع کی متعدد عالمی سطح کی علمی و تحقیقی، دعوتی اور فلاحی انجمنوں اور اداروں کے اساسی رکن تھے۔ دارالمصنفین میں شیخ کی وفات کی خبر بڑے رنج و غم سے سنی گئی اور تغریت کے لیے برقیہ بھی بھیجا گیا۔
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Watermarking of Medical Images for Content Authentication and Copyright Protection

Due to development of latest technologies in the areas of communication and computer networks, present businesses are moving to the digital world for effectiveness, convenience and security. One of the examples is in the area of medical applications, where traditional diagnosis is being replaced by e-diagnosis. This paradigm gave birth to number of applications in healthcare industry like teleconsulting, telesurgery and telediagnosis. All these applications require the exchange of medical images in digital format from one geographical location to another throughout the globe via a cheap and fast network such as Internet. However, digital form of medical image can easily be manipulated through image processing softwares. Insurance companies, hospitals as well as patients might want to modify the medical images for a number of reasons. The tampered images may be used for illegal purposes. The objective of this dissertation is to investigate how to protect a medical images, detect tampered images and recover the original image using the technique of Digital Watermarking. Watermarking tech- nology can be categorized into robust, fragile and semi-fragile watermarking, each serving for different purposes. In order to facilitate sharing and remote handling of medical images, we propose techniques to solve the problem of copyright protection and content authentication using robust and fragile watermarking respectively. In the first part of this dissertation, we design a fragile watermarking system for the exact authentication of medical images. The proposed system authenticates the computed tomography (CT) scan medical images of thorax area against different distortions. The system first enhances the embedding capacity of a CT scan image by isolating the actual lung parenchyma unlike classical watermarking techniques, which use logical square or ellipse for isolating region of interest (ROI). Secondly it embeds watermark only in region of non interest (RONI), thus not compromising the diagnosis value of medical image. The method utilizes the spatial domain watermarking and least significant bit (LSB) replacement method for embedding the watermark. Experimental results reveal that the proposed system detects both legitimate and illegitimate distortions. Robust watermarks are well suited for copyright protection because they stay intact with the image under various manipulations. In the second part of the dissertation, we investigate an approach of robust watermarking for protection of medical images which ensures the security of medical images against geomet- rical distortions. In the proposed method we firstly normalize the medical image using image moments in order to make it invariant against geometric transformations. Secondly, we select the salient regions in the normalized image based on feature points detected by Harris corner detector. Watermarking is performed in Discrete Cosine Transform (DCT) domain of salient regions by adapting block based embedding. The scheme is completely blind, that it does not require the original image to detect the watermark. We thus address the issue of tolerating affine transformations in medical images that is hardly be found in existing schemes of medical image watermarking. In third part of the dissertation, we investigate the complete reversible watermarking system. The proposed method uses integer wavelet transform and companding technique which can embed and recover the secret information as well as can restore the image to its pristine state. The method takes advantage of block based watermarking and iterative optimization of threshold for companding, which avoids histogram pre and post-processing. Consequently, it reduces the associated overhead usually required in most of the reversible watermarking techniques. As a result, it keeps the distortion small between the marked and the original images. Experimental results show that the proposed method outperforms the existing reversible data hiding schemes reported in the literature.