Ten isolates belonging to Proteus spp. were collected and obtained from Department of Biology, College of Science, the University of Baghdad. The diagnosis was done by polymerase chain reaction (PCR) technique using 16S rRNA gene and urease C gene. All isolates (100%) were sensitive to meropenem, imipenem, ciprofloxacin, gentamycin, amoxicillin, clavulanic acid and levofloxacin. These isolates also showed 60% sensitivity to cefixime and nitrofurantoin. However, both species of P. mirabilis and P. vulgaris showed the lowest sensitivity when treated with tetracycline (60%) and amikacin (20%). Cephalothin had a variable effect on the species under study as P. mirabilis isolates were 100% sensitive in comparison with the 80% sensitivity of P. vulgaris isolates. The antibacterial activities of Ag and TiO 2 nanoparticles (NPs) were investigated. The minimum inhibitory concentration (MIC) value of Ag NPs against both species isolates was 10 mg/mL, while the MIC value of TiO 2 NPs was 14 mg/mL against P. mirabilis and 15 mg/mL against P. vulgaris isolates. P. mirabilis isolates showed larger swarming diameter than P. vulgaris, but this motility phenomenon of P. vulgaris was arrested rapidly after incubation with sub-MIC of TiO 2 NPs and Ag NPs comparatively with control. All isolates showed shifting to down-regulation in the fliL gene expression under the effect of the NPs using TiO 2 NPs and Ag NPs. In conclusion, down-regulation of the fliL gene expression is directly linked to the inhibition of swarming movement of Proteus species. We encourage using these inhibitors (after tests to ensure minimal toxicity to human) in combination with antibiotics to ensure bactericidal /bacteriostatic effect to treat Proteus infections.
Aluminum oxide nanoparticles were prepared and characterized with a wide variety of applications. This study aimed at the preparation of AlO 2 nanoparticles by the liquid particles treated with laser (1064 nm), immersed in deionized water at varied laser energy (350 J/cm 2 ) and diverse numbers of 300 pulse. For antimicrobial, antioxidants, animal's model, the prepared nanoparticles were confirmed by using ultraviolet-visible spectroscopy (UV-Vis), X-ray diffraction (XRD) and scanning electronic microscope (SEM). The antimicrobial activity of synthesized Al 2 O 3 nanoparticles was tested using well diffusion method against three types of bacteria: Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The best concentration of aluminum oxide nanoparticles was 100 mg/mL, which exhibited a higher antibacterial activity against both gram-negative and -positive bacteria. In addition, the results showed that the gram-negative bacteria were more resistible than gram-positive bacteria. The results also indicated that the aluminum oxide nanoparticles possessed antioxidant effects, as investigated by DPPH method. Finally, the wound healing activity of the aluminum oxide nanoparticles was investigated, and the results showed that the aluminum oxide nanoparticles stimulated the wound healing activity by using animal's model. The results presented in this study demonstrated that the aluminum oxide nanoparticles had biomedical applications could be applied in biomedical uses in the future.
Introduction and Aim: Cancers are a complex group of genetic illnesses that develop through multistep, mutagenic processes which can invade or spread throughout the body. Recent advances in cancer treatment involve oncolytic viruses to infect and destroy cancer cells. The Newcastle disease virus (NDV), an oncolytic virus has shown to have anti-cancer effects either directly by lysing cancer cells or indirectly by activating the immune system. The green fluorescent protein (GFP) has been widely used in studying the anti-tumor activity of oncolytic viruses. This study aimed to study the anticancer effect of a recombinant rNDV-GFP clone on NCI-H727 lung carcinoma cell line in vitro. Materials and Methods: The GFP gene was inserted to a NDV strain to create a recombinant NDV (rNDV- GFP) using reverse genetics technology. The MTT assay was used in evaluating the oncolytic effect of rNDV- GFP on the lung carcinoma NCI-H727 cells. Light and fluorescent microscopy was used to study the cytopathic effects of rNDV-GFP. Results: MTT assay showed that rNDV-GPF inhibited the NCI-H727 tumor cell death in a time-dependent manner. A significant inhibitory effect (78.3%) for rNDV-GPF on cancer cells was observed at 96h in comparison to rNDV (22.7%) and the cytotoxicity rate was directly proportional to the MOI used. Microscopic studies showed rNDV-GPF to induce cytopathic effect post 24 h of infection. Conclusion: The GFP-expressing recombinant NDV strains exhibited encouraging results in terms of tumor growth inhibition. Our research set the groundwork for employing recombinant NDV as an anticancer viral vector.
Introduction and Aim: The antinuclear antibodies (ANA) are unwanted molecules which bind and destroy certain structures within the nucleus. Immunofluorescence is a powerful technique that utilizes fluorescent-labeled antibodies to detect specific target antigens. The aim of this study was to detect the anti-testicular antibody among infertile males in Baghdad city and determine the most common type of infertility. Materials and Methods: The study involved 73 male (53 infertile and 20 non-infertile) volunteers, at Kamal Al-Samarrai Teaching Hospital in Baghdad, Iraq. Serum collected from the study subjects was tested for steroid-cell antibodies (STC-Ab), anti-nuclear antibody (ANA) and anti-testicular antibodies (ATCA) by Indirect Immunofluorescence assay (IIFA). Data obtained was subjected to statistical analysis using the SPSS program. Results: In the current study 52.9% of infertile men tested positive for testicular antibodies compared to the control group. The highest rate of testicular cell antibodies was observed in the serum of infertile patients aged between 30-39 years and the lowest in patients aged 50-59 years. The marriage duration among those showing the highest percentage of antibodies against testicular cells was 1-10 years. Study for the association of male infertility type to those positive for ATCA showed almost all (85.7%) patients with necrozoospermia to be positive for ATCA. This was followed by males with azoospermia (50%) and oligospermia (46.9%). The patients were negative for Addison’s disease while a few (28.6%) were positive for lupus erythematosus.
The bacterial isolates were obtained from Al-Kindi Hospital were diagnosed by the Vitek-2 system and re confirm by 16srRNA gene as S. aurous, the results were shown 20 isolates (66.7%) out of 30 isolates were positive to protease production. All bacterial isolates (100%) were sensitive to Gentamicin and Levofloxacin. but resistant (100%) to aztreonam. The best temperature for enzyme production from bacteria was 37 °C, and the best pH for enzyme production was 7. Partial purification of the bacterial enzyme (protease) was carried out using short steps included ammonium sulfate 65% saturation, ion exchange using DEAE- cellulose column and then applied on gel filtration chromatography using Sephadex G-200 column. The enzymatic activity was determined for each purification step. The specific fold and yield of the purified enzyme were 5.91 and 38.3 % respectively. The molecular weight of the purified enzyme was 37 kDa , it was determined by SDS-PAGE. After being exposed to high concentrations of the protease enzyme (800-1000 µg/ml), the proliferation of a breast cancer cell line (MCF-7) was seen to be suppressed, however the inhibitory effect gradually diminished as the concentration of the enzyme decreased. 200–400 µg/ml is the recommended concentration.
Thousands of people were infected with Vibrio cholerae during the outbreak in Iraq in 2007-2009. Vibrio cholerae was shown to be variable in its content of virulence determinants and in its antibiotic sensitivity. This study was designed to isolate and characterize clinical and environmental V. cholerae isolates and to determine antibiotic sensitivity, enzyme and toxin production, and the presence of virulence genes. Eighty clinical and five environmental bacterial isolates were collected and diagnosed by subjecting them to microscopic, biochemical, serological, and molecular analysis. The results revealed that 55% of clinical isolates belonged to the Inaba serotype, 32.5% to the Ogawa serotypes, and 12.5% to the Non-O1 serotype. All environmental V. cholerae isolates belonged to the Non-O1 serotype. All environmental isolates were sensitive to all examined antimicrobial agents, while all clinical isolates showed a high sensitivity (100%) to ampicillin, gentamicin, cephalothin, tetracycline, erythromycin, and ciprofloxacin, and a high resistance (97.5%) to co-trimoxazole, nalidixic acid, and chloramphenicol. It was found that all V. cholerae (O1) isolates were resistant to the Vibrio static O129 and all Non-O1 V. cholerae isolates were sensitive to the Vibrio static O129. All clinical and environmental isolates produced hemolysin (100%) and lecithinase (100%), while they showed various production rates of protease (90% of clinical and 60% of environmental) and lipase (50% of clinical and 20% of environmental). The ompW gene was amplified in all the clinical and environmental V. cholerae isolates, but not in other related and nonrelated bacteria. Multiplex PCR analysis showed that the toxR gene was amplified in all clinical and environmental isolates, while ctxA, ctxB, tcpA genes were amplified only in clinical (O1) isolates. This study indicates the differences in the production of some enzymes and toxins and in the content of virulence genes between clinical and environmental isolates in Iraq during the outbreak (2007-2009).
Introduction and Aim: Brucellosis is an important zoonotic disease caused by Brucella spp. which is an intracellular gram-negative bacterium. Brucella melitensis lacks the "traditional" virulence factors such as exotoxins or cytolysins, but is capable of persisting intracellularly and evading the immune system. This study aims to identify B. melitensis using PCR and discover genes associated with its severity for early detection and therapy. Materials and Methods: Ten ml of unclotted blood sample was collected from each patient (n=100) suspected to be infected with brucellosis. The Castaneda technique was used to inoculate blood samples onto Brucella Basel agar with a selective supplement and tryptone soy broth in a diphasic flask. Biochemical tests were used in identifying the isolated colonies. B. melitensis isolates were further confirmed by the polymerase chain reaction (PCR) technique using, primers targeting a specific region (IS711 gene) of the genome. Multiplex PCR was used to determine the four virulence related genes (lps B, mgtA, omp25, CBG) in all positive samples. Results: Brucella melitensis was detected in 9% (9/100) of the blood samples. Among the virulence factors, LpsB and mgtA, were detected in all the isolates while, the genes omp25 and CBG were detected in 66.6% and 55.5% of the isolates, respectively. Conclusion: Brucellosis could be diagnosed rapidly using molecular techniques. PCR technique could also be used in identifying the Brucella virulence related genes lpsB, mgtA, CBG, and omp25 that are crucial to the bacterium's pathogenicity in the intracellular environment.
Introduction and Aim: Thyroid eye illness is widely recognized as one of the most significant concerns confronting the medical profession today. The thyroid-related ophthalmopathy also known as Graves’ ophthalmopathy is an autoimmune disorder that cannot be reversed and hence, research pertaining to the identification of novel markers that can explain both the prognosis and the recovery of the condition is needed. In this study, we aimed to investigate the IL-38 gene expression levels among Graves’ ophthalmopathy patients in Iraq. Methodology: The TSH levels were measured with an enzyme-linked immunosorbent assay (ELISA). The level of IL-38 gene expression in patient blood samples and normal healthy controls was measured qualitatively using RT-PCR. Results: Patients who were diagnosed with Graves’ ophthalmopathy were observed to have abnormally low levels of thyrotropin (TSH) in their serum. In these patients, the interleukin IL-38 gene expression was observed to be significantly greater compared to healthy controls (P<0.01), the tendency of which continued even after one year of therapy with anti-thyroid drugs. Conclusion: The findings of this study indicate that the IL-38 transcript is important in the autoimmune response. The identification of IL-38 expression levels could contribute in the early clinical diagnosis and treatment of thyroid eye disease
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