Multi Drug Resistant K. pneumoniae is considered as a serious health concern worldwide. Infections caused by K. pneumoniae are of prime importance among elderly, neonates and immunocompromised patients in hospital settings, however, this pathogen is also credited with various community-acquired diseases in which sepsis and pneumonia are blatant ones. Fluoroquinolone antibiotics are broad-spectrum and potent antimicrobial agents frequently opted by the clinicians to manage various infectious diseases. Several mechanisms like modifying enzymes, efflux pumps, target site mutation and target protective proteins are responsible for resistance against these antimicrobial agents. Emergence of fluoroquinolone-resistant bacteria is elevating with a worrying rate. In Pakistan, this important subject has been neglected so far, as little data is available about molecular insights of MDR K. pneumoniae especially with reference to Fluoroquinolone, which is rapidly being used as the drug of choice to treat bacterial infections caused by the Gram-negative bacteria, in different clinical and health care settings. Henceforth, present study was designed with the manifesto to study the molecular insights and transcriptional analysis of K. pneumoniae against the Fluoroquinolones. Clinical samples (n = 889) including urine (UTI Patients n = 278), Tracheal aspirates (n = 222), Wounds (n = 174) and Blood (n = 215) were collected from hospital settings. Isolation and identification of K. pneumoniae was done with the help of conventional microbiological procedures. Characterization of K. pneumoniae was done based on phenotypic and genetic characteristics. Phenotypic characterization was done based on cultural and morphological properties. Moreover, biochemical characterization was completed by employing the biochemically characterized samples to API® 20E strip (bioMérieux, France). Polymerase Chain Reaction (PCR) was then performed to target a portion of 16S rDNA for molecular identification of K. pneumoniae. Both, Disc-Diffusion Assay and Broth Micro-Dilution Assay were performed to study the antibiotic susceptibility profiling of K. pneumoniae isolates against various antibiotics including β-lactams and Penicillin, Quinolones, Cephalosporins, Tetracyclines, Aminoglycosides, Sulpha drugs and others like Chloramphenicol and Tigecycline. Minimum Inhibitory Concentration (MIC) of the drug Ciprofloxacin against K. pneumoniae was determined by broth dilution method. Afterwards, molecular insights of Fluoroquinolone resistance were studied by targeting various genes of chromosomal origin, responsible for resistance which include Quinolone Resistant Determining Regions i.e. QRDRs (gyrA & gyrB) and then from the plasmid origin, Plasmid Mediated Quinolone Resistant determinants i.e. PMQRs (qnrA, qnrB & qnrS). Lastly, transcriptional analysis of K. pneumoniae was observed under Ciprofloxacin stress in time depended manner using qPCR by targeting genes like gyrA, lexA, eu-Tu, mutS, dnaK, ftsZ and clpB. Findings of the present study revealed out that among 889 clinical samples that were collected from various sample sources, 200 (22.49 %) were confirmed as K. pneumoniae. Whereas, distribution of K. pneumoniae from confirmed (200) isolates was as follow Urine (UTI patients); 86 (43%), Tracheal aspirates; 51 (25.5 %), Purulent wounds; 33 (16.5 %) and Blood; 30 (15 %). Isolates of K. pneumoniae has shown resistance to almost all the used antibiotics (19) in the study except for Cefotaxime and Tigecycline. Additionally, significant resistance was found against Fluoroquinolones. Out of 200 confirmed isolates of K. pneumoniae 179 (89.5 %) showed resistance to Ciprofloxacin. MIC of 64 µg/ml was recorded for 42 % isolates. Total 81 (45.5 %) out of 179 isolates were found to harbor qnrB gene. Sequence analysis of qnrB revealed that it is qnrB1 variant and has been assigned the Accession number MF953600. The gyrA was found with a strong mutation for Ciprofloxacin resistance i.e. Ser83→Ile. Out of 179 ciprofloxacin resistant isolates in 98 (54.75 %) gyrA gene was present whereas 41 (22.9 %) isolates harbor qnrB1 gene. In 40 (22.5 %) isolates both gyrA and qnrB1 genes were present. Transcriptional analysis of K. pneumoniae isolates has revealed that basal gene expression is altered against Ciprofloxacin stress in a time depended manner. With the finding of current study, it was concluded that MDR K. pneumoniae is an urgent public health pathogen in Pakistan. It has been found that it is resistant to Ciprofloxacin which is considered as one of the most important and frequently used antibiotics in the country. Indigenous K. pneumoniae has fluoroquinolone resistance due to qnrB1 variant of PMQR and mutation in gyrA gene of QRDRs. In addition, Ciprofloxacin has influenced the gene expression of indigenous K. pneumoniae. Mounting antibiotic resistance and emerging MDR superbugs like K. pneumoniae pose a significant threat worldwide especially in developing countries, and it could only be contained by devising comprehensive studies and strategies to control the menace of infectious disease caused by resistant pathogens.
Wherever person goes, he finds unless it finds advertisement tools in various shapes and sizes. These ads, which have become an important pattern in the life of traders are indispensable for them than in contemporary reality. But now companies are specializing in the production of these ads and incentives, taking advantage of all the modern means of magazines and broadcasting video and audio formats and even mobile internet services. Each of these companies have their own philosophy of it, some of which consider the legitimate controls in advertisements but other are to make money regardless of the appropriateness of these ads or following the controls of legitimacy and this research aims at highlighting the most important of these Islamic perspective controls which must be adhered when designing these commercials ads. Islam regulates the trade system and provides a sound guideline to its followers. It has forbidden all the malpractices being exercised in business and its advertisement. The research talks about the commercial advertisements, their types, aims and the opinions of Islamic experts about them. Besides, the research proves that advertisement is permissible according to shariah.
Shisham (Dalbergia sissoo) is an important tree of Pakistan and is widely grown in different areas of the country mainly for furniture, timber and fuel. Dieback is a serious threat to this tree and has caused huge damage not only in Pakistan but also in India, Nepal and Bangladesh. A nursery experiment of sexually (seedlings) and asexually (cuttings) propagated D. sissoo was conducted during 2009 and 2010. Both seedlings and cuttings were inoculated with the most commonly found fungi in the dieback affected trees i.e Fusarium solani, Botryodiplodia theobromae, Curvularia lunata and Ganoderma lucidum. Overall highest disease incidence was observed in plants inoculated with F. solani (31.39%). In seedlings F. solani caused 46.18% disease while in cuttings it was only 16.61%. No disease was recorded in controlled conditions. A significant (P<0.05) correlation of seedlings and cuttings was observed with climatic variables. Good association of seedlings (r= 0.734) and cuttings (r=0.629) was observed with maximum temperature. Disease predictive models of seedlings and cuttings were developed with climatic variables; Y = -58.3 +7.58x 1 +0.0054x 2 -1.14x 3 +2.47x 4 -1.09x 5 R 2 = 0.62 (2 years data) Where Y= Disease in cuttings, x 1 = Rainfall, x 2 = Relative humidity, x 3 = Minimum temperature, x 4 = Maximum temperature and x 5 = Wind velocity R 2 =0.48 (2 years data) Y = -134 +15x 1 +0.158 x 2 -2.32 x 3 +5.27 x 4 -3.70x 5 Where Y= Disease in seedlings, x 1 = Rainfall, x 2 = Relative humidity, x 3 = Minimum temperature, x 4 = Maximum temperature and x 5 = Wind velocity A comprehensive survey of shisham was carried out in four agro-ecological zones (Sandy deserts, Northern irrigated plains, Barani areas and Sulaiman piedmont) of Punjab province. For survey eleven districts were selected being the most productive and shisham frequenting districts from the above said zones. Trees were divided into age classes and on the basis of disease severity into healthy, partially affected and fully affected trees. Age class one (1-20 years old trees) was observed as the healthiest while age class three (above 40 years) was found to be the most affected in all zones and districts. Maximum number of healthy trees was observed in age class one of Barani areas (92.16%) Maximum number of partially affected trees was seen in Sulaiman piedmont (22.89%) and less number was in Barani areas. Maximum number of dead or fully affected trees was found in class three of Northern irrigated palins (23.60%) and only 4.68% dead trees were recorded in age class one of Barani areas. In selected districts maximum number of healthy trees was in class one of Rawalpindi district (95%) and maximum partially affected trees were in age class three in Dera Ghazi Khan (22.89%). Maximum number of dead trees (33%) was found in age class three of Toba Tek Singh. Rawalpindi was the least affected district with only 5 % dead trees in class one. Water table between 15-20 feet of depth was considered as the most suitable depth where less number of dead trees were recorded in all age classes. Significant (P<0.05) correlation was observed between age class, relative humidity, minimum temperature, maximum temperature and wind velocity. Correlation was non significant with rainfall and water table. Good association was observed between tree age and dieback disease in Healthy (r=0.626), partially affected (r=0.539) and fully affected trees (r=0.613). A disease predictive model based on two years disease survey data was developed: FA = 2.51 + 4.15 x 1 + 5.25 x 2 + 0.00866 x 3 - 0.113 x 4 - 0.0611 x 5 - 0.0115 x 6 - 0.0092 x 7 + 0.541 x8 R 2 = 0.89 Where FA= Fully affected, x 1 = Age, x 2 = Year, x 3 = Water table, x 4 = Rainfall, x 5 = Relative humidity, x 6 = Minimum temperature, x 7 = Maximum temperature, x 8 = Wind velocity