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Home > Phage Therapy for Treatment of Multiple Drug Resistant Organisms

Phage Therapy for Treatment of Multiple Drug Resistant Organisms

Thesis Info

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Author

Golkar, Zhabiz

Program

PhD

Institute

University of Karachi

City

Karachi

Province

Sindh

Country

Pakistan

Thesis Completing Year

2009

Thesis Completion Status

Completed

Subject

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/handle/123456789/1976

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726864161

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In the hunt of phages for Gram-negative multiple drug resistant organisms, we have collected lytic phages. These were characterized by identification of genome nucleic acids and proteins profile of capsid. However present study was proceed with the phages isolated form urine of athlete lady. Microbiological examination of urine sample revealed the presence of E. coli as etiologic agent in urinary tract infection (UTI) patient. Filter sterilized urine produced enornomous amount of phages as indicated by plaque assay on the lawn of two local strains of Pseudomonas aeruginosa, P5 and P6. Electron micrograph of the lysates prepared from urine in Ps5 and Ps6 strains indicated the particles similar to Siphovirideae and Myoviradeae respectively. Absence of Pseudomonas in urine sample during active disease and absence of phages in convalescent patient’s urine was significant features that refer the association of these phages to E. coli. Urea induced E. coli lysate was used to raise lysates in P5 and P6 strain of Ps. aeruginosa called U9P5 and U9P6. Small clear and large clear plaques were exhibited on the lawn of P5 and P6 by these lysates. P5S and PS6 lysates were raised in P5 and P6 from single small plaque each on the lawn of P5 and P6 respectively. Similarly PL5 and PL6 lysates were produced in respective hostfrom single large plaque each. Genomes of these Фs were identified as dsDNA. DNA restriction analysis of all four lysates revealed very similar restriction pattern. Hybridization with DNA probe of PS6 and PL5 has indicated the homology in the DNA from these lysates. Methylated and unmethylated CpG motifs were identified in genome DNA of these phages. To test the immuno- stimulatory effects mice and rabbits were vaccinated with killed cell of S. typhi with or without fragmented genome DNA of these phages. In addition adjuvant efficacy of CpG motifs was compared with mineral oil. In antisera, types of antibodies were determined by immunoelectrophoresis and Western blotting. ELISA was done for Quantative analysis, have shown IgG immunoglobulin was produced in BALB/c mice and rabbit when fragmented DNA was used as adjuvant. In antisera raised by Ф DNA vaccine, strong and specific immunogenicity has been attributed due to the presence of CpG motifs. Difference in immune response in animals immunized by Ф DNA vaccine was detected that may be associated with the frequency of CpG island in the DNA. Ps. Ф DNA has additional CpG sites, compare to E.coli Ф DNA and this may enhance immunopotentional of its CpG motifs. PS5 phage has shown cross reactivity with HCV antisera. In order to confirm the cross reactivity of phage and antisera, the 100 sera samples from the HCV +ve patients were tested byELISA on coated plates with HCV antigen, phage and Ps. and have exhibited significant cross reaction with phage lysates. In order to determine genomic relation, PCR products raised by HCV specific primers were sequenced and aligned with HCV genome. BLAST, Gene locater and QuickAling software have shown homology with the 5’-UTR, 3’-UTR and NS3 regions of different HCV genotypes. A range of primers pair corresponding to different antibiotic resistance trait were used to see whether this phage is a carrier of any antibiotic resistant trait. ClustalW, BLASTn, BLASTx, ScanProsite and Swiss-Prot analysis of sequenced PCR products have shown that this phage carries VanA cassette implicated to vancomycin resistance. Van S & R primers have shown homology with histidine kinase and DNA-binding response regulators respectively. Interestingly Ps Ф genome was found to contain a composite transposon which was evolved from the combination of IS1216 and 1546 transposons. These transposons were associated with Gram-positive bacteria i.e. Enterococci. VanB gene primers highlighted seven signature sequence on phage genome correspond to Walker B motifs of ABC-transporter proteins of Enterococcus faecalis V583 in addition phage genome has reflected the presence of some functional domain i.e. SAM which is commonly present in eukaryotic proteins. Our studies provide the strong evidence that lytic life cycle of thisphage has therapeutic implications, not only for superficial infection, but effective for systematic infections as well. It is interesting to note that CpG motifs of phage DNA may have potential to enhance efficacy of antimicrobial therapies supported by DNA vaccin
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فخر ا لدین علی احمد

آہ! جناب فخرالدین علی احمد صاحب صدر جمہوریہ ہند
معارف کے اس شمارہ کی طباعت ہوچکی تھی کہ یکایک ریڈیو سے صدر جمہوریہ ہند کی المناک رحلت کی خبر ملی، ملک اس قدر جلد ان کی دائمی جدائی کے لیے تیار نہ تھا، فلک کا کیا بگڑتا جو نہ وہ مرتے کوئی دن اور، پہلے ڈاکٹر ذاکر حسین مرحوم اور اب جناب فخرالدین علی احمد کی وفات راشٹرپتی بھون ہی میں ہوئی، دونوں کی صدارت کے ساتھ،
پیچھے پیچھے وہ دبے پاؤں قضا بھی آئی
وہ جاچکے، جب ان کی سوانح عمری لکھی جائے گی تو وہ ایک پرجوش مجاہد آزادی، قابل فخر محب وطن، کامیاب بیرسٹر، آسام کے معزز ایڈوکیٹ جنرل، اسی ریاست کی حکومت کے قابل اعتماد وزیر خزانہ، پھر ملک کی لوک سبھا کے ہر دلعزیز ممبر، اقوام متحدہ کے ہندوستانی وفد کے بڑے لائق رکن، مرکزی حکومت کے مختلف محکموں کے بہت ہی کارگزار وزیر سیکولرزم کے بہترین نمائندہ، قومی یکجہتی کے زریں نشان اور آخر میں جمہوریہ ہند کے محبوب صدر کی حیثیت سے برابر یاد کئے جائیں گے، ہندوستان کی خارزار سیاست میں داخل ہوکر کسی مسلمان رہنما کا کامیاب ہونا آسان نہیں، کچھ مسلمان قائد ایسے ہوئے جو مسلمانوں میں تو مقبول تھے لیکن ہندوؤں میں اچھی نظروں سے نہیں دیکھے گئے اور کچھ مسلمان لیڈر ایسے بھی گذرے جو ہندوؤں میں تو محبوب لیکن مسلمانوں میں غیر محبوب رہے، جناب فخرالدین علی احمد صاحب کا نمایاں وصف یہ تھا کہ وہ دونوں حلقوں میں عزت کی نگاہوں سے دیکھے گئے، ان کے کسی اخباری بیان کسی عمل کی نشاندہی نہیں کی جاسکتی ہے، جس سے ہندو خوش اور مسلمان ناخوش یا مسلمان خوش اور ہندو ناخوش ہوئے۔
ان کی علم نوازی کی یادوں کی بھی مشعل روشن رہے گی، ۱۹۶۹؁ء میں غالب کی صدسالہ برسی پورے ملک میں ان...

اختلاف الدلالات للكلمات المشتركة بين العربية والأردية وأثره في تعليم اللغة العربية

Arabic language is a family member of Semitic languages whereas Urdu is the member of Indo-European Languages. The Arabic language though is not from the same language family but amazingly it provides much of its share through alphabets, words with its meanings and pronunciation. These features of both languages have provoked to study it under the contrastive linguistics through semantic study of commonly used words. This research is a semantic study of commonly used words in both languages of different language family along with the applied linguistics in Language teaching. There are large numbers of Arabic words that are used in Urdu language and there are significant numbers of words that are used in different meanings, this change in meaning led to change in semantic field. This research paper also study the effect of semantic change of these words on Arabic Language teaching to the Pakistani students whose native language is Urdu. This study will also reveal the reasons of errors during language learning with the help of semantic study if commonly used words.

Molecular Studies on Major Potyviruses Infecting Tomato and Chilli Crops in Pakistan

Tomato and chilli are economically important crops of Pakistan. Both of these are necessary part of our daily diet in the form of vegetables, salad and other culinary uses. Among biotic factors, the viruses are considered as substantial limiting factors reducing yield and deteriorating quality and quantity of tomato and chilli crops. The potyviruses like Chilli veinal mottle virus (ChiVMV), Potato virus Y (PVY), Pepper veinal mottle virus (PVMV), Pepper vein banding virus (PVBV) and Tobacco etch virus (TEV) have been reported as tremendous threats to the production of these crops worldwide including Pakistan. Management of plant viruses depends upon rapid and specific detection and identification, which is mostly facilitated by molecular techniques like PCR/RT-PCR/NA hybridization. The Molecular data on native Pakistani solanacious potyviruses was scare and their genetic variability was unknown. So the present research was conducted to determine the incidence, distribution and genetic variability based on molecular characterization of Pakistani solanaceous Potyviruses. A total of 2423 tomato and chilli leaf samples from 16 tomato and chilli growing districts viz; Badin, Thatta, Umerkot, Hyderabad, Tandu Allayar, Bahawalpur, Multan, Lodhran, Muzafargarh, Faisalabad, Vehari, Rawalpindi, Attock, Chakwal, Sialkot, Sheikupura, Karak and Loralai were collected in 2013-2014, of which only 920 samples were positive for potyviruses by PTA ELISA. Of these positive samples, the ChiVMV, PVY and ChiRSV were detected in 526, 323 and 71 samples respectively. The overall incidence of Potyvirus, ChiVMV, PVY and ChiRSV was recorded as 38%, 21.7%, 13.33% and 0.3% respectively. 20 The co-infections of ChiVMV and PVY was detected in 29 tomato samples and 31 chilli samples. While, co-infection of ChiVMV and ChiRSV and of ChiRSV and PVY in 18 and 24 chilli samples respectively. The highest incidence was recorded from Lodhran, Multan, Bahawalpur and Rawalpindi districts and the lowest from Karak, Loralai, Sheikhupura and Mansehra districts. The sequences of five ChiVMV, three PVY and 2 ChiRSV isolates were submitted to genbank. The Pakistani isolates of ChiVMV shared nucleotide identities of 90-97.5% with each other and 82.4-90.5% with other ChiVMV isolates. The nucleotide sequences of PVY isolates were 98.699.1% to each other and 98.2-99% with other PVY isolates. The ChiRSV isolates were 98.4% identical to each other, while they shared 92.6-98.1% identities with other ChiRSV isolates. The Pakistani isolates of ChiVMV clustered and shared maximum nucleotide identities with Indian isolates, while PVY and ChiRSV with chines isolates.