Isolation, identification and analysis of antibacterial activity of soil streptomycetes isolates from north Jordan

A total of 90 different Streptomyces isolates were recovered from 36 soil samples and assessed for their antibacterial activity. Nine isolates were identified by the absence of an aerial mycelium. The rest were grouped into six colour series, namely grey, white, yellow, green, red and polymorphic colours (pink, orange or violet) with total numbers of 29, 18, 14, 8, 3 and 9, respectively. The isolates (68%) showed a reverse side culture pigmentation, 30% produced melanin and 25% produced other soluble pigments. Isolates (48%) were characterized by flexuous spore chains, 21% with spiral and 10% for each of the rectus and retinaculum apertum arrangement. The antibiotic activity against a wide range of bacteria was exhibited by 54% of the isolates which were effective against Bacillus subtilis (57%), Staphylococcus aureus (47%), Escherichia coli (24%), Klebsiella spp (16%), and Shigella spp (12%). The lowest activity (8%) was exhibited against Pseudomonas spp and Salmonella spp. The antibacterial activity of the isolates was divided into four groups according to the diameter of the inhibition zone produced. Groups 3 and 4 with larger inhibition zones indicated their potential as a possible source of novel antibiotics.     

Molecular identification of Salmonella isolates from poultry and meat productsin Irbid City,Jordan

The sensitivity and accuracy of molecular diagnosis of Salmonella from meat and poultry products using polymerase chain reaction (PCR) was compared with conventional microbiological methods. A total of 212 samples representing the most frequently used fresh and frozen meat and poultry products (whole, cut, ground, and processed) were collected from different locations within the city of Irbid. DNA was extracted directly from each food sample and amplified using Salmonella-specific primers. Samples were also analysed using conventional microbiological methods for the presence of Salmonella spp. Results showed that Salmonella was detected in 185 samples out of 212 (87%) by PCR technique, while 172 (81%) samples were detected Salmonella positive by conventional microbiological methods. On the other hand, 27 (12.7%) samples were negative by PCR and 40 (18.8%) samples were negative by conventional microbiological methods. PCR assay proved to be an effective method for Salmonella detection in meat and poultry products with high specificity and sensitivity and more importantly a less time-consuming procedure. Using PCR, Salmonella spp. detection could be achieved within 24–36 h compared to 3–8 days for the conventional microbiological methods.     

Plant Hormones Isolated from “Katahdin” Potato Plant Tissues and the Influence of Photoperiod and Temperature on Their Levels in Relation to Tuber Induction

Qualitative and quantitative analyses were carried out on vegetative tissues of potato (Solanum tuberosum cv. “Katahdin”) in search of natural products thought to play a role in tuber induction. Tissues were obtained from plants initially grown in a growth chamber under noninducing conditions (30°C day and 28°C night with an 18-h photoperiod), and then half of the plants were moved to inducing chambers (28°C day and 13°C night with a 10-h photoperiod) for 10 days prior to tissue harvest. Plants from each chamber were then harvested at 2-day intervals for 10 days, separated into above- and belowground portions, and the lyophilized tissues were extracted and subjected to rigorous purification and separation using high-performance liquid chromatography. This was followed by identification and quantification using combined gas chromatography-mass spectrometry. Compounds isolated and identified included gibberellic acid; cytokinins cis-zeatin riboside, trans-zeatin, trans-zeatin riboside, and isopentenyladenine; and jasmonates jasmonic acid, tuberonic acid and its methyl ester, methyl 7-isocucurbate, and 9,10-dihydromethyljasmonate. Methyl 7-isocucurbate and 9,10-dihydromethyljasmonate were detected for the first time in potato tissue as endogenous compounds. Cytokinin and jasmonate levels generally increased under inducing conditions, whereas gibberellic acid levels declined progressively during the 10-day sampling period. Only gibberellic acid, jasmonic acid, and cis-zeatin riboside levels were significantly influenced by induction.     

Polymorphisms of arylamine N-acetyltransferase2 and risk of lung and colorectal cancer

The arylamine N-acetyltransferase 2 (NAT2) enzymes detoxify a wide range of naturally occurring xenobiotics including carcinogens and drugs. Point mutations in the NAT2 gene result in the variant alleles M1 (NAT2 *5A), M2 (NAT2*6A), M3 (NAT2*7) and M4 (NAT2 *14A) from the wild-type WT (NAT2 *4) allele. The current study was aimed at screening genetic polymorphisms of NAT2 gene in 49 lung cancer patients, 54 colorectal cancer patients and 99 cancer-free controls, using PCR-RFLP. There were significant differences in allele frequencies between lung cancer patients and controls in the WT, M2 and M3 alleles (p < 0.05). However, only M2 and M3 allele frequencies were different between colorectal cancer patients and controls (p < 0.05). There was a marginal significant difference in the distribution of rapid and slow acetylator genotypes between lung cancer patients and controls (p = 0.06 and p = 0.05, respectively), but not between colorectal cancer patients and controls (p = 1.0 and p = 0.95, respectively). Risk of lung cancer development was found to be lower in slow acetylators [odds ratio (OR): 0.51, 95% confidence interval (95% CI): 0.25, 1.02, p-value = 0.07]. No effect was observed in case of colorectal cancer. Our results showed that NAT2 genotypes and phenotypes might be involved in lung cancer but not colorectal cancer susceptibility in Jordan.     

ULIXES, unravelling and exploiting Mediterranean Sea microbial diversity and ecology for xenobiotics’ and pollutants’ clean up

The civilizations in the Mediterranean Sea have deeply changed the local environment, especially with the extraction of subsurface oil and gas, their refinery and transportation. Major environmental impacts are affecting all the sides of the basin with actual and potential natural and socio-economic problems. Events like the recent BP??s oil disaster in the Gulf of Mexico would have a tremendous impact on a close basin like the Mediterranean Sea. The recently EU-funded project ULIXES (http://www.ulixes.unimi.it/) aims to unravel, categorize, catalogue, exploit and manage the microbial diversity available in the Mediterranean Sea for addressing bioremediation of polluted marine sites. The rationale of the project is based on the multiple diverse environmental niches of the Mediterranean Sea and the huge range of microorganisms inhabiting therein. Microbial consortia and their ecology, their components or products are used for designing novel pollutant- and site-tailored bioremediation approaches. ULIXES exploits microbial resource mining by the isolation of novel microorganisms as well as by novel advanced ??meta-omics?? technologies for solving pollution of three major high priority pollutant classes, petroleum hydrocarbons, chlorinated compounds and heavy metals. A network of twelve European and Southern Mediterranean partners is exploring the microbial diversity and ecology associated to a large set of polluted environmental matrices including seashore sands, lagoons, harbors and deep-sea sediments, oil tanker shipwreck sites, as well as coastal and deep sea natural sites where hydrocarbon seepages occur. The mined collections are exploited for developing novel bioremediation processes to be tested in ex situ and in situ field bioremediation trials.     

Intratumoral Delivery of Paclitaxel in Solid Tumor from Biodegradable Hyaluronan Nanoparticle Formulations

In the current study, novel paclitaxel-loaded cross-linked hyaluronan nanoparticles were engineered for the local delivery of paclitaxel as a prototype drug for cancer therapy. The nanoparticles were prepared using a desolvation method with polymer cross-linking. In vitro cytotoxicity studies demonstrated that less than 75% of the MDA-MB-231 and ZR-75-1 breast cancer cells were viable after 2-day exposure to paclitaxel-loaded hyaluronan nanoparticles or free paclitaxel, regardless of the dose. These results suggest that hyaluronan nanoparticles maintain the pharmacological activity of paclitaxel and efficiently deliver it to the cells. Furthermore, in vivo administration of the drug-loaded nanoparticles via direct intratumoral injection to 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumor in female rats was studied. The paclitaxel-loaded nanoparticles treated group showed effective inhibition of tumor growth in all treated rats. Interestingly, there was one case of complete remission of tumor nodule and two cases of persistent reduction of tumor size that was observed on subsequent days. In the case of free paclitaxel-treated group, the mean tumor volume increased almost linearly (R ² = 0.93) with time to a size that was 4.9-fold larger than the baseline volume at 57 days post-drug administration. Intratumoral administration of paclitaxel-loaded hyaluronan nanoparticles could be a promising treatment modality for solid mammary tumors.     

Patterns of testicular cytology in men with primary infertility: any change since the Gulf War?

OBJECTIVE: To evaluate the patterns of testicular cytology in men with primary infertility, to compare the morphologic patterns between the periods 1990-1995, immediately after Gulf War II, and 1997-2001 and to determine whether there is a correlation between hormonal profile, testicular volume and morphologic pattern. STUDY DESIGN: Retrospective study of men with primary infertility. History, complete physical examination, hormonal assay and testicular ultrasound were evaluated. A total of 545 patients had samples for testicular cytology obtained from both testes. The patient's consent was obtained in all cases. Smears were interpreted under light microscopy after treatment with Diff-Quik. A total of 104 healthy, fertile subjects were used for comparison of the hormonal profile and testicular volume. RESULTS: The mean (+/- SD) age was 28.66 +/- 4.36 years and duration of marriage 4.4 +/- 4.36 years. There were 11.2% patients with normal cytology, 55.8% with hypospermatogenesis, 28.4% with testicular atrophy, 2.9% with Sertoli cells only and 1.7% with maturation arrest. A significant increase in hypospermatogenesis and decrease in the Sertoli cell-only pattern were noted in 1997-2001 when compared with 1990-1995. The mean left testicular volume was 10.53 +/- 5.51 mL3 in the infertile group vs. 15.2 +/- 4.97 in the fertile group (p < 0.003); right testicular volume was 10.84 +/- 4.77 vs. 15.15 +/- 5.31 (p < 0.003). The hormonal profile revealed higher luteinizing hormone and follicle-stimulating hormone levels in the infertile group vs. control group (8.53 +/- 9.03 and 16.44 +/- 19.243 vs. 6.98 +/- 4.53 and 7.37 +/- 6.63, respectively [p < 0.001]). Free testosterone was higher in the fertile group (39.69 +/- 12.76 vs. 20.28 +/- 8.5 [p < 0.000]). CONCLUSION: The majority of infertile males in our cohort had hypospermatogenesis; testicular atrophy was the next most common disorder. There was no major change in overall absolute numbers since the Gulf War. Testicular cytology by fine needle aspiration is a safe and well-tolerated complementary investigation for unexplained male infertility.     

Use of RAPD-PCR fingerprinting to detect genetic diversity of soil Streptomyces isolates

Random amplified polymorphic DNA (RAPD) was used for identification and assessment of genetic diversity between isolates of Streptomyces from soil. Genomic DNA from 18 Streptomyces isolates and 2 reference strains were amplified using four different 10-mer primers. Different DNA fingerprinting patterns were obtained for all the isolates. Electrophoretic and cluster analysis of the amplification products revealed incidence of polymorphism among the isolates and none of them was identical to the reference strains although there were some common amplification bands. Two highly divergent groups were determined among the isolates. The results indicate that RAPD is an efficient method for discriminating and studying genetic diversity of Streptomyces isolates.