Genetic transformation in antibiotic resistant escherichia coli O157:H7

Five hundred stool samples were collected from patients with diarrhea [infants and children under ten years of age] admitted to the Pediatric and Maternity Hospital in Erbil City from March 2007 to September 2007. The samples were cultured on different culture media and according to the colony morphology, biochemical reactions and by the use of API 20E system, 35 [7%] were diagnosed as E.coli I, 8 [1.6%] E.coli II, 17 [3.4%] E.coli III, 22 [4.4%] E.coli IV, 8 [1.6%] Shigella dysenteriae, 16 [3.2%] Salmonella arizonae, 12 [2.4%] Salmonella typhi and 6 [1.2%] Vibrio cholerae. In addition, cases of Entamoeba histolytica 175 [35%], Giardia lamblia 102 [20.4%] and Hymenolepis nana 2 [2.4%] were identified. No infectious agents were found in 75 [15%] of the samples. 22 [4.4%] of the samples had mixed infections. The sensitivity of E.coli O157:H7 isolate to different antibiotics was performed. There was a variation in the resistance ranging from 8.5-90%. The determination of the site of genes responsible for the antibiotic resistance in E.coli O157:H7 was performed using the genetic transformation method for E.coli DH5 alpha laboratory strain with the DNA that is absent from the highly resistant strains, E.coli O157:H7 4 and E. coli O157:H7 6. The transformation process succeeded when using the plasmid DNA for strain 4 and failed when using strain 6. It was evident that the genes responsible for resistance to the following antibiotics were located on the plasmid DNA: amoxicillin, amoxiclav, ampicillin, cephalexine, cefixime, cefotaxime, doxycyclin, gentamycin, nalidixic acid, nitrofurantoin, rifampicin, streptomycin and tetracycline. Whereas the genes responsible for the following antibiotic resistance were located on the chromosome: amikacin, erythromycin, chloramphenicol, ciprofloxacin, tobramycin and trimethoprim

Immunostaining method to explore cellular immunity against toxoplasma gondii in Iraqi women with a history of abortion

To explore cellular immunity against Toxoplasma gondii in Iraqi women with a history of abortion using the immunostaining method. Sixty women who had abortion were selected for this study. The presence of anti-Toxoplasma antibodies in their sera was confirmed via ELISA testing. Heparinized blood and serum were collected. The women were divided into three groups according to the presence of specific anti-Toxoplasma antibodies in their serum. These groups included: women with IgG [18], women with IgM [14] and women with both IgG and IgM [12]. The other 16 women had no antibodies against Toxoplasma. Twenty-four healthy-looking women had been selected as controls. Those that revealed any antibody titer against Toxoplasma were excluded from the study. The isolated peripheral blood leukocytes [PBL] were tested for cell surface markers [CD3, CD4, CD8 and CD71] using monoclonal antibodies against T-cell subsets. All the surface makers revealed significantly higher percentages when compared to the healthy controls. The results indicate a strong cellular immune response during toxoplasmosis

Possible cellular expression of IFN-gamma in women with abortion infected with toxoplasma gondii

This study included one hundred and twenty six women with spontaneous abortion [6 of them with induced abortion]. Venous blood was collected from these women, and serum was obtained for the performance of the ELISA test for the detection of anti-Toxoplasma IgM to indicate the acute T. gondii infection. Paraffin embedded blocks of the trophoblas-tic tissue obtained from each patient were prepared. The trophoblastic tissue was investigated for the presence of T. gondii antigen by using immunohistochemical analysis, using specific monoclonal antibodies for T. gondii. According to the immunohistochemical analysis, the patients were divided into three groups, [group 1] 26 positive for Toxoplasma, [group 2] 26 negative for Toxoplasma, and [group 3] 6 negative for Toxoplasma [induced abortion group]. The results indicated a high frequency of the T. gondii infection among women with abortion, 23 of 120 women [19.17%] have IgM Abs against T. gondii by ELISA test. The use of ELISA test for the detection of anti-Toxoplasma IgM was highly specific [100%] but not highly sensitive [88.46%]. This could explain the use of more sensitive techniques for the detection of T. gondii infection like immunohistochemical analysis. The results of the IHC revealed that 26 of 120 women [21.66%] had Toxoplasma antigen within the trophoblastic tissue. The sensitivity and specificity of IHC were 100%, 96.91%, respectively. The current study showed that there was no significant difference between the mean age of positive and negative groups, while there was a highly significant difference between the mean age of positive and induced abortion groups. Similarly, the mean age between negative and induced abortion groups showed a highly significant difference. In this protocol, the majority of patients within the positive and negative groups were found to have no previous abor-tions, while patients with previous abortions constituted a less percent. Among the induced abortion group, it has been found that all the six cases had no previous abortions. The mean gestational age among the three groups was compared in this study, where it revealed a highly significant dif-ference between the positive and induced abortion groups. The same result was found between the mean gestational age of negative and induced abortion groups. In addition, it revealed the lack of significant difference between the mean gestational age of positive and negative groups. The majority of abortions within the positive group for T. gondii fall in the period of 12 weeks [41%] followed by 8 [15%] and 10 [12%] weeks of gestational age. Levels of IFN-gamma were investigated by using immunohistochemical analysis. The results revealed a highly significant dif-ference regarding the mean percent of IFN-gamma when compared between the positive and negative groups. In this work, t-test revealed a highly significant difference regarding the mean percent of IFN-gamma between positive and induced abortion groups. Similarly, a significant difference was found when the mean percent of IFN- gamma was compared between the negative and induced abortion groups. The results of immunohistochemical analysis of IFN-gamma within positive group showed that there were negative and highly significant correlations with gestational age but not with number of previous abortions. In conclusion, the data of this study strengthen the possibility that IFN-gamma may explain the role of type 1 cytokines in the pathogenicity of abortion in the positive group for T.gondii

Evaluation of B-lymphocyte marker [CD19] and interleukin-4 [IL-4] in women infected with Trichomonas vaginalis

The aim of this investigation was to evaluate the B-lymphocyte subset [CD-19] and interleukin-4 [IL-4] in women injected with Trichomonas vaginalis. Vaginal swabs, washes and blood specimens were collected from 65 women attending outpatient clinic at Al-Kadhimyia Teaching Hospital in Baghdad suffering from vaginal discharge starting from January 2005, to October 2005. Twenty healthy looking age matched women were also included for control studies. Blood was taken to heparinized tubes and serum was separated. Heparinized blood was used for evaluation of the CD marker; CD-19 using the indirect immunostaining technique. The cytokine IL-4 was evaluated in serum and vaginal washes using the ELISA technique. Trichomonas vaginalis was isolated from 25 women with a prevalence rate of 38.5%. The results of CD marker showed significant differences between the infected women and controls. There was a significant increase in IL-4 in the infected women. It was found that this parasite has the ability to stimulate the cell-mediated immunity which eventually led to production of specific immunoglobulin against Trichomonas vaginalis

[Insulin dependent diabetes mellitus and antibodies to glutamic acid decarboxylase]

Insulin dependent diabetes mellitus [IDDM] is an autoimmune disease, the factors that induce the selective destruction of the insulin producing islet cells of the pancreas are unknown. Although autoreactive T-cells rather than autoantibodies are responsible for the destruction of beta-cells, the identification and chararcterization of the autoantibody-autoantigen system in IDOM are of crucial and fundamental importance in prediction and potential immunotherapy of the disease. Currently, Cytoplasmic Islets Cell Antibodies [ICAs Insulin Autoantibodies [IAA], Glutamic Acid Decarboxylase Antibodies [Anti-GADs] and 37-k antigen antibodies are potentially useful hurnoral markers for such prediction. The attempt to identify the earliest events in the autoimmune process suggest that Anti-GAD could be the first and most important autoantigen