World Journal of Experimental Biosciences
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Antibacterial Effect of Flavonoids Extracted from Seeds of Silybum marianum against Common Pathogenic Bacteria
Get PDFThis research was carried out to evaluate the antibacterial effect of Silybum marianum flavonoids extracted from seeds on pathogenic bacteria (Staphylococcus saprophyticus, Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae) and detect the bioactive compounds found in the seed extract. Different methods were used to tech the aims of the study. Agar well diffusion method and mixed up the extract with agar media were used to evaluate the antibacterial activity of the flavonoids extracted from seeds of S. marianum. The chemical analysis to seeds showed that it contained several antibicrobial compounds such as terpenoids, flavonoids and tannins. The cold alcoholic extract of milk thistle did not appear significant antibacterial activity against the bacterial isolates when agar well diffusion method was used while mixing the seed extracts with agar media showed antibacterial activity (1500-2900 μg/ml) against S. sprophyticus, E. coli, K. pneumoniae and S. aureus.
 
Evaluation of anti-Helicobacter pylori antibodies level in sera of patients with chronic hepatitis B
Get PDFThe relationship between chronic hepatitis B virus and Helicobacter pylori infection was evaluated to determine, seventy five patients with chronic hepatitis B infection (8-70 years) were investigated. The results were compared with the results of 50 healthy volunteers. Anti-H. pylori antibodies IgA and IgG were measured by Indirect fluorescent antibody test (IFAT) in sera of patients and healthy groups. The percentage of anti-H.pylori IgA antibodies (26.67%) were significantly (P<0.01) higher than healthy control group. While, no significant difference was found between the percentages of anti-H. pylori IgG antibodies (48 %) in patient sera and these kind of antibodies in sera of healthy control group (P > 0.05). The present results indicated that the acute infection with H. pylori may be correlated with severity of chronic hepatitis B infection.
 
Immunization of Male Rabbits with Pseudomonas aeruginosa Increases Spermatogenesis
Get PDFThe impact of immunity on spermatogenesis is documented in the literature. This study highlighted the influence of Pseudomonas aeruginosa hyper-immunization on male rabbit testicular spermatogenesis as the effect of immune response on sex hormones changes. Morphometric criteria were used to evaluate spermatogenesis. The level of hormone was estimated by enzyme-linked immunosorbent assay (ELISA). We found that male rabbits vaccinated with the bacteria showed a highly significant (P<0.001) decreased in spermatogonia cell number (11.46±8.11 in test vs 45.19±10.76 in control). In addition there was a significant increase (P<0.001) in spermatocytes cell number represented by pachytene and zygotene cells in vaccinated animal as compared to control (88.5±8.11 in test vs 54.14±10.13 in control). In concomitance with these changes, there was a significant elevation in level of testosterone in immunized groups (3.55±3.86 in test vs 0.98±0.8 in control). While, no significant change was observed in progesterone (2.00±0.80 in test vs 1.83±0.79 in control) and estradiol (21.42±14.02 in test vs 19.31±3.72 in control) levels. The histopathological changes included detachment of spermatogonia in the test group was observed. The increase in spermatogenesis and the changes in hormones level as well as histopathological changes suggest that the immune mechanisms impact spermatogenesis in males rabbits.
 
Molecular detection of blaCTX-m_2 in Proteus mirabilis
Get PDFFrom 152 different clinical samples collected from patients in Baghdad, 20 isolates of Proteus mirabilis were identified by bacteriological and biochemical assays and confirmed by vitek 2 identification system. It was found that 20 (66.6%) isolates were identified as P. mirabilis and 10 (33.3%) isolates were P. vulgaris. Susceptibility of P. mirabilis to cefotaxime was done, Notable, 13 (65%) P. mirabilis isolates were resistant to cefotaxime, whereas, 6 (30%) P. mirabilis isolates were extended spectrum beta-lactamases producers. MIC value of cefotaxime was estimated to 7 isolates (512 μg/ml), and the MIC was 4096 μg/ml for other isolates, One isolate had MIC equal to 128 μg/ml while, another isolate showed MIC equal to 8192 μg/ml to cefotaxime. DNA was extracted from 8 P. mirabilis isolates that was resistant to cefotaxime and 2 isolates were sensitive to cefotaxime. The blaCTX-M-2 gene was detected by polymerase chain reaction (PCR). The 8 cefotaxime resistant P. mirabilis isolates and one cefotaxime sensitive isolate carried blaCTX-M-2 gene, while the cefotaxime-sensitive isolate did not harbored the gene. Furthermore, this study confirmed that blaCTX-M-2 gene was carried on chromosomal DNA, by extraction the DNA from 10 P. mirabilis isolates having blaCTX-M-2 gene. DNA profile showed chromosomal DNA band only. Curing of blaCTX-M-2 gene was done, 4 cefotaxime-resistant P. mirabilis isolates were treated with ethidium bromide as a chemical curing agent in different concentrations. The results confirmed that blaCTX-M-2 gene was carried on chromosomal DNA.
 
Hypoglycemic Effect of 24-Methylencycloartan-3-one Isolated from Prosopis juliflora Pods in Alloxan Induced Diabetic Rabbits
Get PDFProsopis juliflora is herbal plant uses in different countries to treat diabetes mellitus. In current study, oil extract was extracted from P. juliflora pods. Qualitative preliminary tests showed the extract contained terpenoids. The oil compound was isolated purified and identified by thin layer chromatography (TLC), gas chromatography-mass (GC-MS), nuclear magnetic resonance (H1-NMR and C13-NMR) and infra red (IR) spectroscopy. It was found that oils extract contain 24-Methylencycloartan-3-one. Hypoglycemic effect for this compound was checked in alloxan induced fasted diabetic rabbits. The dose of 0.3 gm/kg of terpenoidic at different time intervals (2,4,6 and 24 h) reduced blood glucose significantly with values 321.42, 294.60, 251.40 and 172.30 mg /100 ml, respectively. The toxicity study has shown that the 24-Methyl cycloartan compound has no toxic effect on red blood cells; therefore we suggest that this compound can be successfully and safely use to treat diabetes mellitus instead of insulin
 
Phytoremediation of Cadmium in river water by Ceratophyllum demersum
Get PDFThe current study included an assessment of the ability of Ceratophyllum demersum Phytoremediation of cadmium in accumulative concentration by measuring the heavy metal in the tissues of the plant to check the ability of C. demersum to reduce or eliminate the harmful and toxic effect, which resulting from this type of pollutants in the most important ecosystem. In present study the plant was grown in natural conditions for two months and treated to two concentrations of cadmium (3.714, 4.952 mg/l). The samples of water and a certain weight of the plant were taken before and after treating with cadmium chloride in order to estimate the level of cadmium in these samples before and after treatment. The residual amount of cadmium in the water and also in the plant has also been calculated at different time intervals (3, 6 and 9 days). The present study showed that cadmium accumulated rapidly in the plant and increased dramatically with time, thus the highest level of cadmium was observed at day nine. C. demersum showed high ability to remove cadmium and was at the ninth day of the experiment and that was 1.53, and 1.089 mg /l) when exposed to 3.714, 4.952 mg /l of cadmium respectively. The current study proved strongly the ability of C. demersum to eradicate the cadmium in ecosystem.
Histological study in liver of albino mice post exposing to shisha smoke
Get PDFThe effect of shisha smoking on liver parenchyma tissues were evaluated in (120) male adult mice. The animals were divided into four groups according to kind of exposed smoke (cigarette, shisha and mixture of cigarette and shisha). The mice exposed to fresh air considered as control group. A special inhalation chamber designed locally was used to expose the animals to different kinds of smokes. Exposure to cigarette smoke was done for 5 min/day, while exposure to shisha smoke was done for 15 min /day for 4, 8, 12 weeks. The tissues of control and exposed groups were processed for histological study. The results showed different histological changes such as hepatocytes degeneration, congestion, sinusoid dilation and infiltration of inflammatory cells in liver parenchyma tissues. These changes observed clearly when the period of exposure increased especially in groups exposed to shisha smoke and shisha plus cigarette smoke for 12 weeks. The results showed that shisha smoke causes mild damage in liver parenchyma tissue and this damage increased concomitantly with the increase of duration of exposure.
 
Analysis of genetic diversity among pomegranates cultivated in Diyala, Iraq using RAPD-PCR method
Get PDFThis study attempts to identify genetic diversity among four genotypes of pomegranates cultivated in Diyala city, Iraq and find the genetic polymorphism among them by using DNA markers generated by polymerase chain reaction (PCR). Total genomic DNA of genotypes studied was extracted from dry young leaves by using cetyltrimethyl ammonium bromide (CTAB) procedure. Molecular analysis was performed by using twelve random markers in random amplified polymorphic DNA (RAPD-PCR) technique. The analyses based on five primers OPH – 08, OPH – 13, OPH – 18, RI – 3 and RI – 16, which used in amplification and polymorphism for the four cultivated genotypes studied. The genetic polymorphisms value of each primer was determined and ranged between 31 to 100%; primers OPBA–3 and OPBB–4 produced the highest percent of genetic polymorphism compared with primer OPH–13. The genetic diversity and the isolation of the four cultivated genotypes of pomegranates were confirmed obviously by RAPD-PCR technique.
Evaluation the Uricase Produced from Different Clinical isolates of Pseudomonas aeruginosa by Plate Assay Methods
Get PDFOut of 150 clinical samples, 43 isolates of P. aeruginosa were identified based on biochemical tests, API 20 NE, and VITEK 2 system. The results showed that 38 (88.37%) isolates were uricase producers while only 5 (11.62%) isolates were uricase non producers, based on uricolytic activity on agar medium using two methods, qualitative method by using agar plate medium which contains many ingredients (uric acid, dextrose, yeast extract and NaCl), the results obtained by this method indicated by the presence of clear zone around streaking line on an agar plate assay. The another method is semi quantitative, in which agar plate medium contains 0.5%w/v uric acid as the only nutrition source and showed diameter of clear zone around well on an agar plate. The diameter of clear zone around wells ranged between 12-26 mm. In conclusion, both methods gave same results for screening uricase producers, but semi quantitative method gave more accurate results because its medium contain only uric acid as a source of carbon or nitrogen or both which makes this method more specific and sensitive than the other one.
Transformation of Saccharomyces cerevisiae by pET plasmid using lithium acetate
Get PDFThe recombinant pET 16b was propagated and amplified in E. coli using heat shock method, then plated and incubated on luria agar containing ampicillin (100μg/ml) for 18 hours at 37ºC. One colony was picked and mini-culture was made. Transformed E. coli was cultured on luria broth containing ampicillin. After incubation 18 h at 37˚C the recombinant plasmid was extracted using QIAprep Spin Miniprep Kit from transformed E. coli and transferred to Saccharomyces cerevisiae using lithium acetate/SS carrier DNA/PEG. The positive transformed clones were grown on selective media lacking of tryptophan. The plasmid was extracted using the same extraction kit with some modification and the concentration of obtained DNA was 70.8 ng/μl measured by Nanodrop. In this study, the recombinant pET 16b that amplified in E. coli was transformed in S. cerevisiae by lithium acetate method.