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Transcript of RESEARCH ETHICS
Supervised By: Dr. M. Arfat Yameen
Presented By: Sana Mushtaq
Attachment host cell Colonization
Pathogenecity is based on:
Exchange of Genetic Information
and MRSA are resistant to Beta-Lactam, Macrolides, Aminoglycosides, & quinolones.
Emerging vancomycin resistant
Community associated & Nosocomial infections
Major cause of epidemic infant diarrhea is O:H serotypes of
an emerging threat to patients
GIT commensal- found in every healthy individual
In US, 73480 patients are infected with
Designing new drug regimen
Adopting alternative drug delivery systems
New drug moiety
Aims & Objectives
To evaluate the enhanced antibacterial activity of antibiotic loaded chitosan nanoparticles against resistant strains of G +ive and G -ive bacteria
Isolation & identification of resistant strains of MRSA &
Evaluation of antimicrobial susceptibility
Synthesis & characterization of blank & drug loaded nanoparticles
Evaluation of synergistic antibacterial acitivity
Biochemical & Molecular identification
Growth on selective/differential media
Mannitol salt agar-
Sheep blood agar-
DNA extraction & PCR
A (270 bp) &
A (533 bp) gene detection
16S rRNA (1400 bp) &
A (884 bp) gene detection
Antimicrobial susceptibility testing
Disc diffusion test
Kirby Bauer Method
Methods of CNPs preparation
Ionic gelation method
Chitosan- low molecular weight deacetylated chitin poly D- glucosamine (Sigma Aldrich)
Sodium tripolyphosphate 85% Techical grade (Sigma Aldrich)
Ceftriaxone sodium- 83.2% potency (Cirin phm.)
Irregular and spherical nanoparticles
Antimicrobial susceptibility testing with blank and Ceftriaxone loaded CNPs
Antimicrobial susceptibility testing
Evolution of mankind lead towards evaluation of diseases
- The Gram +ive
- The Gram -ive
causes blood, soft tissue, and skin infections.
MRSA causes nosocomial infections
One third of individuals are
80,000 patients per year are infected with MRSA
Three major classes
Extra intestinal pathogenic strains
Colonization on host mucosa Evade host defense system Multiplication
Pathogenecity is based on
are resistant against beta lactams & qiunolones.
Emergence of rapid resistance against third and fourth generation cephalosporins & flouroquinolones .
(Aries de Sousa & de Lencastre, 2004)
(Gordon & Lowy, 2008)
(Novick, 2000; Decker, 2008; Gordon & Lowy, 2008; Huang & Platt, 2003; Cookson et. al., 2007)
(Levine & Edelman, 1984; States et. al., 2005; Allocati et. al., 2013)
(Russo & Johnson, 2000; Salayers & Whitt, 2002; Levine & Edelman, 1984)
Targeted drug delivery
Nanoparticles- Novel carrier system
Drug incorrporated synthetic & natural polymer nanopartcle
Chitosan- an example of natural polymer
Catalase & Coagulase tests
MRSA- Catalase & Coagulase +ive
TSI, Citrate, & Indole tests
- TSI & Indole +ive & Citrate -ive
Synthesis of Chitosan nanoparticles (CNPs)
Different ratios of Chitosan:TPP:Ceftriaxone
Characterization of CNPs
Nanoparticle size analysis
blank- 0.31 & Cef-CNPs- 0.462
(Zeta ZF, Malrem UK)
Morphology of CNPs
MIC with ceftriaxone
Entrapment efficiency & Loading capacity
Questions & Answers
In-vitro study revealed that chitosan blank nanoparticles possess their own antibacterial activity against resistant strains of both Gram positive and Gram negative bacteria
Loading of Ceftriaxone in CNPs gave the enhanced antibacterial acitivity.
(Greenwood & Kendall, 1999; Hanaor et. al., 2012)
Dr. Nisar ur Rehman
HOD/Chairman, Dept. of pharmacy, CIIT, Atd.
Dr. M. Arfat Yameen
Assistant prof., Dept. of pharmacy, CIIT, Atd.
Dr. Faiz Rabbani
Assistant prof., Dept. of Chemistry, CIIT, Atd.
Dr. M. Arfan
Assistant prof., School of Natural Schiences, NUST, Isd.
Evaluation of enhanced antibacterial effects of antibiotics against resistant microbes by In-vivo studies.
Targeted antibiotic loaded polymeric nanoparticles against resistant microbes using mAbs.
Toxicological studies of nanoparticles for their safe use.