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Studies on the Use of Hbv Epitope Chimeras

Thesis Info

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Author

Malik, Imran Riaz

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2014

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/2514/1/3063S.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727356976

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Hepatitis B virus (HBV) is a common cause of viral hepatitis with significant health complications including cirrhosis and hepatocellular carcinoma. Major advances have been made in the treatment of Hepatitis B recent years. However, much remains to be accomplished because current antiviral drugs do not eradicate infection. The currently licensed hepatitis B vaccines, consisting of recombinant hepatitis B surface antigen (HBsAg) and alum, are highly effective and induce protective antibody titers in 95% of vaccinated individuals after 3 immunizations before onset of HBV infection. Present studies were aimed to develop chimeric vaccine (proteins) that may stimulate B- and T- cell responses and generate enhance immune response to treat the HBV patients after infection. In this regard, five HBV chimeric vaccine constructs were designed and developed using the HBcAg gene encoding amino acids 1-78 and amino acids 80-144. Amino acid- position 79 where deleted and replaced with preS epitopes sequences. All five chimeric constructs (pIJMcsc-1, pIJMcsc-2, pIJMcsc-3. pIJMcsc-4 and pIJMcsc-5) contained HBcAg as a carrier molecule for the epitopes of preS regions primed HBcAg- specific antibodies response. These constructs (chimeric plasmids) were transformed into E.coli to see the protein product on SDS-PAGE. The expressed proteins (csc-1, csc-2, csc-3. csc-4 and csc-5) were purified based on affinity chromatography using His tag at the C terminal of expressed proteins. The purified chimeric proteins were further subjected to Enzyme linked Immunosorbent assay (ELISA) and western blotting assays using secondary anti- rabbit AP conjugated antibody and secondary anti-mouse AP-conjugated antibody respectively. All five chimeric proteins showed positive antibody response. In vivo study was performed for chimeric protein (csc-5) only. This chimeric protein (csc-5) was evaluated in C57BL/6J mice for activation of humoral and cellular immune responses, primed both HBcAg- specific T cells and antibodies to preS1. IXResult obtained indicated that the csc-5 protein can induce HBV-specific antibodies and T cells, two functions that well complement the activity of antiviral compounds. Thus, the csc-5 protein may be a future component in therapy for chronic HBV infections where the host is gaining a sustained control of the HBV infection and also a viable approach to develop an effective bi-functional therapeutic vaccine as an add-on for treatment of chronic HBV.
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خواجہ محمد علی شاہ رحمانی

خواجہ محمد علی شاہ رحمانی
افسوس ہے کہ گزشتہ جون میں’’برہان‘‘کے خاص مضمون نگار اوررفیق خواجہ محمد علی شاہ صاحب رحمانی کی سہارنپور میں وفات ہوگئی۔مرحوم کی صحت عرصے سے ٹھیک نہیں تھی اس کے باوجود علمی ریسرچ کے کاموں میں لگے رہتے تھے۔ دارالعلوم دیوبند اور مظاہرالعلوم سہارنپور کے ممتازفاضل تھے۔السنۂ مشرقیہ کی بھی بہت سی ڈگریاں ان کے پاس تھیں۔اکابر دیوبند خاص طورپر حضرت الاستاذ علامہ سید محمد انورشاہ صاحب کشمیریؒ اورحضرت مولانا شبیر احمد صاحب عثمانی مرحوم سے والہانہ تعلق رکھتے تھے اوران حضرات کی علمی خصوصیات پران کی گہری نظر تھی۔ پختہ استعداد، واضح العقیدہ مسلمان تھے۔طبیعت میں قناعت اور خودداری کاجوہر نمایاں تھا۔پوری زندگی گوشہ نشینی اور خدمت علم میں بسر کردی۔ جس روزان کی رحلت ہوئی اسی روز کچھ دیر کے بعد ان کے والد ماجد کاسانحہ ارتحال پیش آگیا۔ان کے والد صاحب خانقاہ رحمانی سہارنپور کے سجادہ نشین تھے اور عوام ان سے فیض حاصل کرتے تھے۔رحمھما اﷲ رحمۃ واسعۃ۔
[ستمبر۱۹۶۷ء]

 

The Life and Achievements of Hyder Bux Jatoi, With an Special Reference to His Social and Political Contributions

Comrade Hyder Bux Jatoi, also known as ‘Baba e Sindh’, was the most prominent leader of the farmer societies of pre-independence Pakistan. Although he was a bureaucrat turned into a farmer activist, but later he played a very important role in the social and political settings of the country at that time. Most of the Pakistani people remember him as a sign of change and renovation in the agricultural history of Pakistan. This was a great effort to credit the front-runners and their struggle; it is very prominent among the laborers and landless leaders of the world. He devoted his entire life to set peace up for the struggle of land ownership rights to dispossessed farmers, and highlighted the cause, to be noticed by the notables. The vision of Mr. Jatoi is still reflected in many situations when initiatives are taken by the various governments of the world to provide the masses with the basic requirements of development and peace.

Pharmacokinetic Study of Type-2 Diabetes Drugs in Combination Vildagliptin and Metformin Hydrochloride Using Liquid Chromatographic Technique

Pharmacokinetic studies are performed to examine the absorption, distribution, metabolism and excretion of a drug under investigation in healthy volunteers and/or patients. Data obtained from such studies are useful for the design and conduct of subsequent clinical trials. Such studies also provide useful information for the appropriate and safe conduct of clinical trials and for the evaluation of the mechanism of action in human subjects. They are also necessary for appropriate analysis and evaluation of the efficacy and safety data obtained in clinical trials for new drug development and in postmarketing clinical trials. Outcomes of clinical pharmacokinetic studies are useful for determining the appropriate use of medicines according to patient characteristics, such as disease and genotype of drug-metabolizing enzymes, and for predicting the influence of pharmacokinetic drug interactions. The results can also provide information for therapeutic drug monitoring. Development of a new medicine may be achieved, thereby avoiding unnecessary studies. Recently, a growing body of information obtained from different studies conducted around the globe showed a striking rise in healthcare expenditures and the developing countries are not exception. Previous studies conducted in Pakistan showed that a considerable portion of the healthcare costs goes to medicine price. The present dissertation was the first detailed investigation in the relationship between the patient’s socioeconomic status and physician’s choice on multisource solid oral drugs. Such association had not been fully investigated in developing countries, particularly in Pakistan. My dissertation work will provide the information about important aspect and parameters involved in Pharmacokinetic studies.The purpose of this research was to evaluate Pharmacokinetic parameters of a combination formulation of Vildaglipton and metformin hydrochloride50/1000 mg tablet of an established branded formulation.One set of parallel study design was employed in healthy normal rabbits as well as in Alloxan induced diabetic rabbits.The other set of parallel study design was employed in healthy male human subjects as well as in diabetic patients.The concentrations of Vildagliptonand metformin hydrochloride in blood plasma were analyzed using High Performance Liquid Chromatographic (HPLC) technique. The plasma concentration-time curves were used to obtain the different pharmacokinetic parameters. Six healthy rabbits, six diabetic rabbits, twelve diabetic Pakistani patients and twelve healthy Pakistani subjects were selected to participate in this study. A single dose, open label parallel study design was employed in all study subjects. In this study, a oncedaily dose of metformin HCl 1000 mg and vildagliptin 50 mg was used in human subjects, as this allowed assessment of the full pharmacokinetic profile of metformin HCl and vildagliptin over 24 hours at the maximum individual dose. All the rabbits, weighing between 1.25 to 1.5 kg, were given combined dose of drugs metformin HCl 70 mg per 1.5 kg and vildagliptin 15 mg per 1.5 kg orally. Blood samples were withdrawn at predetermined intervals of 0, 0.25, 0.50, 1, 2, 3, 4, 5, 6, 8, 10 and 12 hours from a marginal ear vein puncture. The concentrations of vildagliptin and metformin HCl in blood plasma were analyzed using High Performance Liquid Chromatographic (HPLC) technique and using developed and validated method. The concentration of drugs in blood plasma were determined using standard calibration curves of metformin HCl and vildagliptin having concentrations 50, 100, 200, 500 and 750 ng/mL for vildagliptin and 250, 500, 1000, 2500 and 3750 ng/mL for metformin HCl. The pharmacokinetic parameters were calculated by using different pharmacokinetic formulae. Data is shown as arithmetic mean±SD. The aim of this work was to develop and validate a simple, rapid, more sensitive, more accurate and precise method for simultaneous determination of metformin HCl and vildagliptin in formulations and blood serum. Furthermore, its applications for studying and comparison of pharmacokinetic parameter in Pakistani population among healthy and diabetic individuals along with healthy and diabetic rabbits were analyzed. No significant difference was observed in pharmacokinetic parameters for healthy and diabetic individuals among the humans and the rabbits. There was no statistically significant difference for the treatment values. In both of groups healthy and diabetic pharmacokinetic parameters were equal in terms of rate and extent of absorption. There was no significant difference of drugs absorption and clearance in healthy and diabetic Pakistani subjects.