Detection of genomic instability in hypospadias patients by random amplified polymorphic DNApolymerase chain reaction (RAPD-PCR) method

Hypospadias is a urogenital malformation, and it is a common inborn disorder in male individuals. The etiology of hypospadias is still unsolved. The present study is aimed to identify the genetic instability in hypospadias patients. Random amplified polymorphic DNA (RAPD), a polymerase chain reaction (PCR) based technique, was adopted using ten random primers in twelve cases and twelve controls. The primer detectability on genomic instability in 12 samples ranged from 25% with primer OPA-01 to 66% with OPA-08. Case 2 showed the highest genomic instability (80%). The lowest genomic instabamility was (10%) case 6. The results determined numbers of genomic instabilities among hypospadias patients. In addition, the RAPD-PCR technique is a powerful tool for detection of genomic instability in hypospadias patients. Further larger studies are needed, which include low and high grade of patients to: 1) Obtain RAPD markers useful for hypospadias early diagnosis; 2) investigate different genes directly involved in the etiology of hypospadias; 3) analyze chromosomal instability among hypospadia patients.Key words: Hypospadias, random amplified polymorphic DNA (RAPD), genomic instability

Evaluation of the genetic effects of the in vitro antimicrobial activities of Rhazya stricta leaf extract using molecular techniques and scanning electron microscope

Rhazya stricta plants have always played a major role in the treatment of human and animal diseases and it has main role in the folk medicine. The aim of this study was to explore the potential antimicrobial activities of the aqueous leaves extract of R. stricta on Gram-negative and Gram-positive food-borne bacteria and evaluate the antimicrobial effect at the molecular level. The results indicate that the aqueous leaves extract of R. stricta exhibited the antimicrobial activity against tested microorganisms. A clear, but significantly smaller, inhibition zones were formed after the treatment of two Gram-negative bacteria (Escherichia coli and Aeromonas hydrophila) and one Gram-positive bacteria (Staphylococcus aureus) with the aqueous leaves extract of R. stricta (50 mg) comparing with those formed after the treatment with streptomycin (15 mg). Moreover, the results obtained after the treatments of bacterial strains with elevated concentrations of aqueous extracts of the wild plant of R. stricta leaves reveled that the extract has potent lethal activities as the growth turbidity decreased as the concentration or time of exposure increased. In addition, the observation by the scanning electron microscope showed that cells of the bacterial strains were damaged after the treatment with plant extracts. The noticed antimicrobial effect was explored at the molecular level, using restriction fragment length polymorphism (RFLP) analysis of the plasmid DNA and random amplification of polymorphic DNA (RAPD) analysis of the genomic DNA extracted from the control (untreated) and R. stricta leaf extract-treated bacterial strains. The results demonstrate polymorphic band pattern for most treated microbes compared with the wild type (untreated) strain. Concerning gene expression under the same conditions, total protein contents of the three treated bacteria showed significantly gradual increase in all of the treatment doses compared to control. In addition, the SDS-PAGE of the bacterial cellular proteins resulted in the induction of some protein bands under the treatment conditions. All these results strongly point out the mutagenicity, lethal and antimicrobial effect of the leaves extract of R. stricta. The results indicate the possibility of using the leaves extract of R. stricta as a source of antibacterial compounds for treatment of infections caused by multi-drug resistant (MDR) bacterial pathogens.Keywords: Medicinal plants, Rhazya stricta, antimicrobial, mutagenicity, RAPD, RFLP, SEM, E. coli, S. aureus, A. hydrophilaAfrican Journal of Biotechnology Vol. 12(21), pp. 3171-318

Mobile phone as potential reservoirs of bacterial pathogens

Mobile phones are increasingly used by professionals, university staffs and health care personnel for communication. These can harbor various potential pathogens. This study evaluates and identifies the bacterial contamination rate of mobile phones in the university setting that are in frequent contact with faculty members, personnel, students and/or physicians and nurses in the university clinic. A total of 101 mobile phones belonging to different categories working in various departments at Taif University, KSA were screened for microorganisms’ contamination. Out of the total 101 mobile phones, growth was obtained in 78 (77.2%) mobile phones; 70 (89.7%) from staffs, personnel, students and 8 (10.3%) from clinical workers. Staphylococcus spp and Bacillus spp were the most commonly isolated organisms. Coagulase negative Staphylococcus was the most frequently isolated; 60 (27.12%). The efficacy of decontamination with 70% isopropyl alcohol was found to be 71.3%, as only 29 mobile phones showed growth after decontamination. It was found that around 61.5% of the mobile phones of health care workers at university clinic were contaminated and thus acted as a potential source of nosocomial infections. According to morphological, physiological characteristics, APi profiles and sequencing of 16S-rRNA gene, the selected eight isolates were identified as Bacillus pumilus, Bacillus cereus, Staphylococcus aureus, Staphylococcus hominis, Staphylococcus succinus, Staphylococcus xylosus and Staphylococcus saprophyticus. Based on random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), 32 unique RAPD fragments were identified among the selected isolates. Such unique fragments could be considered as specific markers and might be utilized in tracking the bacterial isolates.Key words: Mobile phones, contamination, pathogen carriers, coagulase negative staphylococci, Bacillus species, 16S-rRNA, random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR)

Genetic diversity and DNA fingerprint study in tomato (Solanum lycopersicum L) cultivars grown in Egypt using simple sequence repeats (SSR) markers

A collection of ten cultivars of tomato grown in Egypt were screened with 20 simple sequence repeat (SSR) primers in order to determine genetic identities, genetic diversity and genetic relationships among these  cultivars. On an average, 38 alleles were amplified using SSR primers with scorable fragment sizes ranging  from approximately 75 to 275 bp. 23 alleles were polymorphic thus revealing 60.5% of polymorphism. The  genetic similarity estimated according to SSR data was scaled between 17.6 and 93.2%, suggesting the  potential of SSR markers in discriminating among plants of close or distant genetic backgrounds. Unweighted pair group method with arithmetic mean (UPGMA) clustering grouped the cultivars into two groups where the  two Egyptian cultivars Edkawy and Giza 80 were clustered in different group. In addition, clustering was found  consistent with the known information regarding growth habit. The genetic distance information obtained in  this study might be useful to breeder for planning crosses among these cultivars.Key words: Tomato cultivars, diversity, Simple sequence repeats (SSR), Egypt