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Preparation of Multiple Congugates Lips-Protein for Immunization Against Typhoidal Diseases

Thesis Info

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Author

Amir Ali

Program

PhD

Institute

Quaid-I-Azam University

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2009

Thesis Completion Status

Completed

Subject

Applied Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/342

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726996728

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Classically Salmonella enterica serovar Typhi (S. Typhi) is associated with typhoid, a major health problem in developing countries. However, in recent years Salmonella enterica serovar Paratyphi A (S. Paratyphi A) and Vi negative variants of S. Typhi have emerged rapidly. For development of multiple protein-polysaccharide conjugates with potential to be used as vaccines, we needed to have a first hand knowledge of local distribution of these pathogens. For this purpose, we developed a nested multiplex PCR targeting 5 different genes for differential diagnosis of typhoidal pathogens which has been optimized to be directly applicable on clinical blood samples. Out of 42 multiplex PCR positive blood samples, 26, 9, and 2 were Vi-positive S. Typhi, Vi-negative S. Typhi and S. Paratyphi A, respectively and 5 patients were found to have mixed infection. Seventeen patients grew Salmonella from blood culture and the remaining 25 were positive in the Salmonella specific PCR. Tests with several common pathogens confirmed the specificity of the assay. We conclude that the proposed multiplex PCR is rapid, sensitive and specific for the diagnosis of typhoidal pathogens directly from blood samples. Detection of Vi negative strains of S. Typhi and S. Paratyphi A among typhoidal patients suggested the need to work on vaccines which are not based on Vi polysaccharide only. Therefore, O-specific polysaccharides (OSP) purified from lipopolysaccharides (LPS) of S. Typhi and S. Paratyphi A were conjugated with diphtheria toxoid (DT) using adipic acid dihydrazide (ADH) as linker, and evaluated for immunogenicity in mice. Use of DT as carrier protein for Salmonellae has not been reported before. S. Typhi OSP-AH-DT conjugate 1 and 2 elicited significantly higher IgG anti-LPS ELISA titer (P = 0.0241 and 0.0245 respectively) than polysaccharide alone. The injection schedule-B (three injections with 4-weeks interval) was found better than schedule-A (three injections with 2-weeks interval). The conjugate of S. Paratyphi A OSP with DT without linker molecule did not elicit sufficient immune response to be used as conjugate vaccine candidate while antibody response against S. Paratyphi A OSP-A H-DT c onjugate was found significantly higher (P = 0.0446) than polysaccharide alone.
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کردے رہو نت کسب کمال

کر دے رہو نت کسب کمال
کھائو مت پرایا مال
امت دے رکھوالے بن کے
آئے آپ سخی لجپال
بے فیضے یاراں نے رل کے
کیتا سانوں آن کنگال
منہ شریک دا رتا ہووے
اپنا کریے چاٹاں نال
چادر ویکھ کے پیر پھیلایئے
رکھیے اپنا آپ سنبھال
روشن چن ستارے بھانویں
گھر دا رکھیے دیوا بال
جیہڑا ہتھاں نال کمائو
اوہو سمجھو رزق حلال
اپنی کہن سیانے بھلیا
’’کوا چلیا ہنس دی چال‘‘
لوکی پہنچے چن دے اتے
توں حنیف نہ بدلیں حال

Trash Rush – Educational Mobile Game For Android Devices

As population grows faster, the rise of garbage collection follows.  Massive information dissemination of waste management has been done by local representatives to educate people but despite the effort exerted still mismanagement of waste exist that caused serious environmental consequences to nature and human beings.  This condition ignites the researchers to develop a technology-based means to help disseminate proper disposal of garbage through the use of mobile phones.  Trash Rush educational mobile game application utilized Lua Scripting Language for game features, system behavior and synchronizes the phone sensor to the system. The Lua tool is decoded to JSON codes and sent to PHP MySQL in the webserver for storage. For tilting motion and orientation of the phone, the researchers employ the accelerometer sensor as the main navigation controller.   For sound and background effects, the Reactable Application is utilized.  The Box2D game engine is used to analyze and interpret the images and graphic effects while the Box2D API is utilized for object movement and animation. The Motion Parallax effect is also used to give more realistic effects in movements of the objects and the graphical background of the game. All these components are extracted through the APK file of Corona SDK to build and publish the app.  Upon reaching to the deployment stage of iterative model, it was found out that the system became viral to grade levels in the campus because of its story and time attack mechanism.

Ecological Modulation of Phytosterol of Selected Food Commodities to Control Hyperlipidemia

The present study was aimed to improve the phytosterol contents in food commodities. Plant microbe interaction is an efficient and ecofriendly way to induce nutritional or desired contents, plant biochemicals to get improved food crops. The present study addresses the issue of nutritional improvement of food crops to provide healthy food, to the people for global green food security. The plant microbe interactions establish a direct relation with plant throughout its development. This resistance termed as induced systemic resistance (ISR) can be achieved applying non-pathogenic bacterial strains. This phenomenon of ISR has been used to manage barley crop using Acetobacter aceti as a biological inducer. Initially, staple food crops Triticum aestivum (wheat), Cicer arietinum (white chickpea), Cicer arietinum (black chickpea), Hordeum vulgare (barley), Oryza sativa (rice), Zea mays (corn) and Pennisetum glaucum (millet) were screened for their phytosterol contents on the basis of their nutritional quantities e.g. biochemicals, physical texture, vitamin contents. All these staple crops were evaluated through transcriptional analysis of squalene synthase (SS) genes which have a direct relation with increased phytosterol production. Biochemical analyses were performed using standard procedures and it was recorded that food crop varieties exhibit variable levels of pectins, alkaloids, saponins, phenolics, terpenoids, phytosterols and flavonoids. High nutritional and biochemical staple food varieties were screened. Barley (Hordeum vulgare) screened out among other staple food crops showed the maximum amount of phytosterols 0.239 ± 0.04 g/kg and other plant biochemicals which are essential in plant growth. These phytosterols compounds are one of the documented remedies for the treatment of hyperlipidemia. Thus, regular consumption of food with high contents of phytosterols controls lipid absorption most efficiently than other food products. Additionally, its dietary benefits and nutritional facts further support its use as most recommended staple food crop worldwide. Squalene synthase expression analysis including a family of genes i.e. SSA, SS1, SS2 and SS3 was carried out using reverse transcription polymerase chain reaction (RT-PCR) in selected staple food crops yielding elevated expression of the most of these genes. Maximum expression of SSA was recorded in chickpea black 64.3 ± 4.63 ng/5μL, which was closely related to barley and chickpea white with 62.91 ± 4.23 and 60.8 ± 3.98 ng/5μL respectively. Wheat and millet exhibited close expression of SSA gene 54.87 ± 3.86 and 54.79 ± 3.68 ng/5μL respectively. Comparatively, rice showed lesser expression of SSA gene 50.07 ± 3.01 ng/5μL, whereas, corn recorded least expression with 28.92 ± 1.27 ng/5μL. Gene SS1 showed maximum expression in barley along with chickpea black and corn 30.14 ± 1.78, 29.91 ± 1.69 and 29.0 ± 1.03 ng/5μL. SS2 gene recorded its maximum expression in barley 41.7 ± 1.99 ng/5μL, chickpea black 39.8 ± 1.98 ng/5μL and corn 37.9 ± 1.76 ng/5μL. Wheat 37.2 ± 1.69 ng/5μL showed no difference in SS2 gene expression than corn, however, SS2 was significantly less expressive in millet 33.8 ± 1.31 ng/5μL and chickpea white 33.1 ± 1.29 ng/5μL. SS3 gene is mainly responsible for squalene production in wheat 44.21 ± 2.43 ng/5μL. Among other staple food crops barley, 42.84 ± 2.16 ng/5μL and chickpea black 42.39 ± 2.01 ng/5μL recorded a second highest expression of SS3 genes. Following with corn 41.86 ± 1.97 ng/5μL, rice 40.87 ± 1.46 ng/5μL and white chickpea recorded 39.8 ± 1.16 ng/5μL. whereas, millet recorded no SS3 gene expression. Different strains of bacillus i.e. AC1, AC2, AC3…AC8 was analyzed as plant inducers and AC8 was screened out as the best inducer in barley. It induced highest quantities of phtosterols 0.008 ± 0.001 g/kg including other biochemicals (i.e. phenolics, alkaloids and terpenoids). AC8 also showed its maximum activity in SS gene expression analysis. The most expressive gene recorded under AC8 treatment was SSA, which has the maximum role in the up regulation of squalene synthase and phytosterols. SS2 reported second highly induced gene against AC8 treatment. SS3 recorded at third level. SS1 was reported as the least expressive gene. AC8 reported as the most favorable microbial strain which showed the best relationship with barley and reported the maximum genetic expression of SSA. AC7 reported as least significant strain in an expression of gene SS1. The current study revealed that among eight microbial strains AC8 had a maximum potential to increase ascorbic acid, pantothenic acid, pyridoxine, thiamine and riboflavin in barley than other microbial strains. AC8 screened out among other microbial strains on the basis of its high vitamins induction potential. AC3 plus AC6 were reported second in the recorded list although other strains had a chronological reduction in vitamins as AC2 > AC7 > AC4 > AC5 and AC1. Overall temperature variation results revealed that AC8 treated barley showed significant induction of phytosterols 0.009 ± 0.003 g/kg and vitamins at T3 (26.5 ± 1.5 °C). Chromatographic techniques such as preparative thin layer chromatography (PTLC), column chromatography (CC) and gas chromatography mass spectrometry (GCMS) were used to identify bioactive compounds of A. aceti. Identified bioactive compounds were responsible for vitamins and phytosterol induction in barley. A total of 13 bioactive compounds were identified through bioactivity guided assay and were analyzed through principle component analysis. Mainly four chemical compounds i.e., quinolinic acid, pyridoxic acid, p.Aminobenzoate and α-Oxobutanoic acid were evaluated through PCA directly associated with increased vitamin contents. Solvent system chloroform: ethanol (4:1) was used to extract bioactive compounds of A. aceti from its crude metabolites. Selected crops were tested against cholesterol reduction in albino rats. The rats were fed with different selected crops and found that the barley 121.7 ± 6.26 mg/dl reduced maximum blood cholesterol level. The selected crop barley was further treated with eight different bacterial strains to enhance its nutritional values and checked against cholesterol reduction. The AC8 treated barley 119.9 ± 5.19 mg/dl reported a maximum reduction in total cholesterol. Following with AC8 treated barley grown at T3 (26.5 ± 1.5 °C) temperature reported maximum cholesterol reduction with 112.6 ± 4.16 mg/dl. This study on small-scale produced good results in the rhizospheric induction of microbes in barley crop. This association promoted the production of phytosterols, vitamins and other nutrients in barley crop. Such barley crop fortified with nutrients could be used to manage the lipid metabolic syndrome for a healthy life.