Synthesis and evaluation of monoclonal antibody against Plasmodium falciparum merozoite surface antigen 2

ObjectiveTo assess the quality of expressed MSP-2 and also to confirm the immune response against different domains of these proteins.MethodsMice were immunized with a schizont extract to stimulate the immune system to make antibodies against different antigens of the late stage parasite including production of antibodies against different domains of Plasmodium falciparum (P. falciparum) MSP-2. B lymphocytes of immunized mice were extracted from the spleen and the fusion was performed using NS-1 myeloma cells and the hybridoma cells were assayed by ELISA either with a schizont extract or different domains of MSP-2 and/or by IFAT with whole schizont preparation. Fusion of NS-1 and spleen cells was performed. The positive hybrids were cloned and ELISA was applied against different dilutions. The positive clones were transferred to a small tissue culture flask and after developing they were assayed against schizont extract and the different MSP-2 domains. The positive clones were expanded to large (75 cm²) flask and cultured under the same conditions, checking them using both ELISA and IFAT and the positive cells were frozen as soon as possible.ResultsA total number of 7 fusions including 26 plates (2 496 wells) were performed, of which 1 336 hybrids were produced and the overall efficiency (1 336/2496 × 100) was about 53%. ELISA was performed to detect the positive hybrids against crude schizont extract by which the highest frequency to crude schizont extract was found for the supernatant of the hybrids produced in fusion number 3 (66 out of 315 hybrids). The supernatant of both B5 and F1 hybridoma cells were more positive against domain 2 of the MSP-2 recombinant protein in Western blotting test. Western blotting results also showed that different domains of the MSP-2 recombinant protein and also the MSP-2 of the P. falciparum parasite were recognized by some of the positive clones and also immune sera.ConclusionsBringing together all the results of this study it has been confirmed that some clones have recognized both schizont extract and different domains of the MSP-2 recombinant protein and therefore confirming the quality of the MSP-2 domains.     

Granulocyte‐macrophage colony‐stimulating factor,a potent adjuvant for polarization to Th‐17 pattern: an experience on HIV‐1 vaccine model

Cytokines are mediators for polarization of immune response in vaccines. Studies show that co‐immunization of DNA vaccines with granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) can increase immune responses. Here, experimental mice were immunized with HIV‐1_{tat/pol/gag/env} DNA vaccine with GM‐CSF and boosted with recombinant vaccine. Lymphocyte proliferation with Brdu and CTL activity, IL‐4, IFN‐γ, IL‐17 cytokines, total antibody, and IgG1 and IgG2a isotypes were assessed with ELISA. Results show that GM‐CSF as adjuvant in DNA immunization significantly increased lymphocyte proliferation and IFN‐γ cytokines, but CTL response was tiny increased. Also GM‐CSF as adjuvant decreased IL‐4 cytokine vs mere vaccine group. IL‐17 in the group that immunized with mixture of DNA vaccine/GM‐CSF was significantly increased vs DNA vaccine group. Result of total antibody shows that GM‐CSF increased antibody response in which both IgG1 and IgG2a increased. Overall, results confirmed the beneficial effect of GM‐CSF as adjuvant to increase vaccine immunogenicity. The hallmark result of this study was to increase IL‐17 cytokine with DNA vaccine/GM‐CSF immunized group. This study for the first time provides the evidence of the potency of GM‐CSF in the induction of IL‐17 in response to a vaccine, which is important for control of infection such as HIV‐1.     

Nursing profession in Iran: An overview of opportunities and challenges

Aim:  Iran's health-care system has witnessed profound changes in the last decades. Despite its progress, the system has currently faced many challenges in one of the important subsystems, nursing. The present review article aimed to present an overview of the opportunities and challenges of the Iranian nursing system, based on recent literature.
Methods:  A broad search of the English and Persian-language literature was carried out, incorporating both electronic and manual components from 1999 to 2009. The results of the investigations among the searched literature are summarized.
Results:  The major challenges are nursing shortages, job dissatisfaction, poor social position of nurses, the gap between theory and practice, lack of community-based nursing care, lack of an appropriate student recruiting system, and shortages in the nursing educational curriculums.
Conclusion:  The authors believe that media, political and public support play a pivotal role in improving the image of nursing in society, increasing motivation among Iranian nurses, and promoting the sociocultural climate and the welfare of nurses, which will result in higher levels of quality of care as well as greater patient satisfaction.     

Modeling the growth of Salmonella typhimurium under the effect of Zataria multiflora essential oil, pH, and temperature by artificial neural networks

Salmonella is the most important cause of bacterial food-borne disease outbreaks in the world. In this study, we have trained and validated artificial neural networks (ANNs) to predict the combined effect of Zataria multiflora essential oil (EO), pH, and temperature on the probability percentage of growth initiation (log P%) of Salmonella. Z. multiflora was collected in the Fars province of Iran. Lyophilized cultures of Salmonella typhimurium ATCC 25923 was used in this study. This design included four levels of EO (0.0, 0.015, 0.03, and 0.06%), three levels of pH (5.5,6, and 7.3), three storage temperatures (35, 25, and 15°C), and repeated observations (18 times) for growth in brain heart infusion broth for up to 43 days. We have designed a standard and the so-called feed-forward ANN, including four input neurons, eight neuron in hidden layer, and one output neuron to predict the combined effect of Z. multiflora EO, pH, and temperature on the probability percentage of growth initiation (log P%) of S.typhimurium. The mean and standard deviation of ANN and real outputs were −2.9771 ± 2.43 and −2.9722 ± 2.39, respectively. The mean differences (and 95% CIs) between the ANN and real outputs were 0.0049 (0.0009–0.0089). Result showed better prediction compare to the previous study(R = 0.998).     

Prevalence of intestinal parasitic pathogens among HIV-positive individuals in Iran

Parasites are important enteric pathogens among patients with human immunodeficiency virus (HIV) infection. There have been very few reports on the prevalence of intestinal parasites among such patients in Iran. To determine the prevalence of intestinal parasites among HIV-positive individuals, we collected single stool samples and analyzed them for detection of various intestinal parasites from 206 HIV-positive individuals with different immune status visited in different medical centers in Iran. The data were tested for statistical significance with chi(2) and Mann-Whitney U tests. The overall prevalence of intestinal parasites was 18.4% (95%CI: 13.7, 24.3). More specifically, the following parasites were identified: Giardia lamblia (7.3%), Blastocystis hominis (4.4%), Entamoeba coli (3.9%), and Cryptosporidium parvum (1.5%). Other parasites observed included Strongyloides stercoralis and Hymenolepis nana in two cases and Dicrocoelium dendriticum in one. Of the 38 patients who tested positive for intestinal parasites, 15 (39.2%) had diarrhea. Intestinal parasites were significantly more common among patients with diarrhea than those without (P < 0.001). Further, CD4 counts were significantly lower among individuals with diarrhea than those without (P < 0.001). This study highlights the importance of testing for intestinal parasites among Iranian HIV-positive patients, especially those with low immunity presenting with diarrhea.     

Histopathologic Findings of Pneumatocele in a Patient with Hyper-IgE syndrome, compatible with cystic adenomatoid malformation

Hyper-immunoglobulin E syndrome is a rare primary immunodeficiency disease characterized by recurrent abscess formation, respiratory tract infections and very high titers of serum IgE associated with peculiar face and skeletal features. We report a seven-year old girl presenting with persistent productive cough and history of chronic eczematoid facial lesions since infancy and two episodes of hospitalizations due to pneumonia and perianal abscess. Additionally, in physical examination finger tip clubbing, laxity of joints and crackles in both lungs were detected. Immunologic work up revealed markedly raised IgE level and eosinophilia. The patient was diagnosed as hyper IgE syndrome based on his clinical and laboratory findings. Chest X-ray revealed multiple large cystic lesions in left lung which were confirmed by spiral CT-scan. Pneumonectomy specimen examination showed cystic adenomatoid malformation, characterized by the presence of various cysts lined by epithelium in different sizes. There are few reports of cystic adenomatoid malformation in children. To our best known, this is the first report of cystic adenomatoid malformation in a child with hyper IgE syndrome. Early diagnosis and surgical therapy are helpful in prevention of repeated infections in these patients.