Search or add a thesis

Advanced Search (Beta)
Home > Preparation of Polysaccharide-Protein Conjugates of Vi Negative Salmonella Typhi As Potential Candidates for Vaccines

Preparation of Polysaccharide-Protein Conjugates of Vi Negative Salmonella Typhi As Potential Candidates for Vaccines

Thesis Info

Access Option

External Link

Author

Salman, Muhammad

Program

PhD

Institute

Pakistan Institute of Engineering and Applied Sciences

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/handle/123456789/1139

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726997364

Asian Research Index Whatsapp Chanel
Asian Research Index Whatsapp Chanel

Join our Whatsapp Channel to get regular updates.

Similar


Salmonella enterica serovar Typhi (S. Typhi) is the cause of typhoid fever in humans. Although typhoid fever has almost vanished from developed countries, it is still a major cause of morbidity and mortality in the developing world. Its global annual incidence has increased from 21 million cases to 26.9 million cases during the period 2000 to 2010. Typhoid fever is curable with high dose of several antimicrobials but the treatment is time consuming and expensive. Vaccines are the eventual source of prevention from typhoid fever. At present, licensed typhoid vaccines are based on Vi polysaccharides. These vaccines are not useful for children of less than 2 years of age and are ineffective against the infections caused by Vi negative S. Typhi strains. O-Specific polysaccharides (OSP) from lipopolysaccharides (LPS) are universally present in both Vi positive and Vi negative S. Typhi and their conjugation with a potential immunogenic protein can make them useful for children below 2 years of age and to elicit humoral as well as cellular immune response. Therefore, we planned to prepare polysaccharide-protein conjugates of Vi negative S. Typhi as potential vaccine candidates. Initially, we purified OSP from the extracted LPS of a local Vi negative S. Typhi isolate and checked its antigenicity against polyclonal mice antisera. To conjugate a carrier protein with the purified OSP, we used the recombinant exoprotein A of Pseudomonas aeruginosa (rEPA) and human serum albumin (HSA). For production of recombinant rEPA, full length rEPA gene was cloned in expression vector. Protein expression was optimized using different isopropyl-β-D- thiogalactoside (IPTG) concentrations, various temperatures and post-induction time. The expressed protein was purified on Ni-NTA chromatography column from soluble fraction as well as from inclusion bodies. His-tag was removed from rEPA using tobacco etch virus (TEV) protease. Polyacrylamide gel and Western blot procedures were performed to check the purity of the protein. We synthesized four glycoconjugate vaccine candidates of Vi negative S. Typhi OSP using sodium cyanoborohydride (reductive amination) and 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP). Each of the conjugate was injected to mice on day 1, day 21 and day 42. The immune response (IgG) against prepared conjugates were determined using enzyme-linked immunosorbent assay (ELISA) and the results were interpreted by one way ANOVA using all conjugate groups vs. OSP control group. The conjugate 2 elicited significantly high (P = 0.0001) antibody titer, while the titers produced by conjugate 1 (P = 0.037), conjugate 3 (P = 0.302) and conjugate 4 (P = 0.416) were not significantly higher than that of the control group. We conclude that the conjugate 2 (OSP-rEPA) has the potential to be evaluated further and recommended for the clinical trials.
Loading...
Loading...

Similar Books

Loading...

Similar Chapters

Loading...

Similar News

Loading...

Similar Articles

Loading...

Similar Article Headings

Loading...

جو گل سانوں کھا گئی اے

جو گل سانوں کھا گئی اے
ربی چیتے آ گئی اے

خورے موت کرے گی کیہ
جند تے حال ونجا گئی اے

راہ نہ چھڈی جیون دی
اِنج تقدیر ہرا گئی اے

توں کنج ہور دا ہویا ایں
ایہہ گل اندروں کھا گئی اے

سوہنیا ربا! بس کر دے
رہ کیہ ہور سزا گئی اے؟

A Phenomenological Study of the Lived Experiences of Tagumpay National High School Teachers Involved in Online Learning Action Cell Session

A qualitative phenomenological approach was used in this study to describe the lived experiences of Tagumpay National High School (TNHS) teachers on Online Learning Action Cell (LAC) session. LAC is a school-based professional development for teachers implemented by the Philippine Department of Education (DepEd). Due to teacher’s lack of participation on classroom LAC, a fully-online mode option is explored by offering TNHS teachers Online LAC session using Facebook as a Learning Management System (LMS). To capture the lived experience of teachers, an in-depth interview with a purposive sample of one TNHS teacher is done in the process. The data gathered went through “Hycner’s Explicitation Process” (1999, in Groenewald, 2004) which includes bracketing, delineating, clustering, summarizing and extracting unique themes. Validity and Credibility were accomplished through an intercoder agreement between researchers, Facebook chat records, bracketing, and member checking. Results identified three themes in relation to teacher’s experience of Online LAC session including usefulness, barriers, and preference. Findings revealed the major role of TNHS teacher’s context on how Online LAC is utilized. Recommendations include administrator and expert teacher working with classroom teachers and the inclusion of teachers’ voices as input in the program design, implementation and evaluation stages of Online LAC to better address curriculum needs and facilitate the delivery of high-quality professional development for teachers’ professional growth.

Evaluation of Different Bait Formulations for the Management of Indian Crested Porcupine, Hystrix Indica Kerr

Field trials were conducted in Abbotabad-Balakot tract (Pakistan), during April 2004 to January 2007 to select cheap, locally available and suitable bait formulations for Indian crested porcupine (Hystrix indica) and tested these for their potentials to carry lethal quantities of rodenticides. Results of no-choice, multiple-choice and paired-choice tests revealed that groundnut was preferred, both in the whole and cracked forms, over maize, wheat, millet, rice, gram and oats. All the food items were consumed in significantly higher quantities than in their whole form, except for rice. Consumption of all the food grains significantly increased with increasing length of its exposure to the novel food, indicating a careful nature of the species in selecting new food. No-choice tests revealed that groundnut - maize 1:1 mixture can be cost effective bait base and its consumption was not signiifcantly different from that of pure groundnut. Saccharin (5%) supplemented groundnut – maize (1:1) bait was consumed in significantly higher quantities, while all other aditives (common salt, egg yolk, egg shell powder, fish meal, peanut butter, mineral oil, bone meal, coconut oil and yeast powder) at both 2% and 5% concentrations and saccharin at 2% concentration did not significantly increase the intake of groundnut – maize (1:1) bait. Consumption of bait, offered after 3 days of pre-baiting, significantly declined when poisoned with 3% zinc phophide and such effects were delayed with 2% and 1% zinc phosphide. Groundnut – maize (1:1) poisoned with 2% zinc phosphide resulted in maximum decrease (55%) in porcupine burrow activity. Strychnine alkaloid (2%) added groundnut – maize (1:1) bait was seriously shuned by porcupine and caused minimum reduction (25%) in burrow activity. Coumatetralyl (0.0375%) added bait caused the highest reduction (80%) in burrow activity. Saccharin (5%) supplemented poison baits were more acceptable and caused some 10% higher reduction in burrow activity using allpoison cereal bait formulations. Field trials on fresh food items suggested that guava was preferred over potato, carrot and sweet potato. Peanut butter did not significantly increase consumption of guava. Consumption of zinc phosphide impregnated guava, offered after 3 days of pre- baiting, exhibited a decline and no bait was consumed on 3 rd day of poison baiting. Decline was the most rapid with 3% zinc phosphide and minimum with 1% zinc phosphide. Maximum reduction in burrow activity (55%) was recorded with 2% zinc phosphide, followed by 1% (35%) and 3% (25%). Fumigation of porcupine burrows with aluminium phosphide tablets produced 100% reduction in burrow activity, by applying 8 tablets per burrow, 85%, by 6 tablets and 75% by 4 tablets per burrow. A 100% reduction in burrow activity was recorded by applying 4 tablets of aluminum phosphide per burrow in small, 6 tablets in medium and 8 tablets in large burrows. The present study suggested that initial control of the porcupine can be achieved by applying 2% zinc phosphide using guava or 1:1 groundnut – maize mixture supplemented with 5% saccharin. Coumatetralyl (0.0375%) poisoned bait can give a higher control of porcupine, yet involves a higher labour cost and can used as a second line of action. Fumigation of burrows with aluminum phosphide is though cost effective and can be used in the porcupine management programme, yet is non- specific in its action and hence can be pressed into action under emergent conditions only.