ORIGINAL_ARTICLE Comparison of Th1 and Th2 Responses in Non-Healing and Healing Patients with Cutaneous Leishmaniasis Background: Cutaneous leishmaniasis is an endemic disease in many regions of Iran, including the city of Mashhad. In recent years, some cases have not responded to Glucantime, the usual treatment for this disease. The cellular immune response caused by T-helper type 1 (Th1) cells has an important role in protection against leishmaniasis, and activation of the T-helper type 2 (Th2) response causes progression of the disease. By analyzing these responses we hope to find a more effective treatment than that currently in use for leishmaniasis patients. Methods: The cellular immune responses in 60 cases of non-healing and healing cutaneous leishmaniasis, and individuals in a control group, were analyzed by measuring cytokines released by peripheral blood mononuclear cells (PBMCs) when stimulated with Leishmania major antigens by Enzyme Linked Immuno Sorbent Assay (ELISA). Results: Subjects from the healing group secreted more interleukin-12 (IL-12) and interferon gamma (IFN-γ) (p<0.05) and less interleukins -4, -5, -10 (IL-4, IL-5, and IL-10) (p<0.005) and -18 (IL-18) (p=0.003) than the non-healing group. Conclusions: The results demonstrate that secretion of cytokines that activate Th2 response including IL-4, IL-5 and IL-10 in non-healing subjects was higher than healing subjects and secretion of cytokines that activate Th1 response including IL-12 and IFN-γ in healing subjects was higher relative to the non-healing subjects. In this study it has been shown that the level of IL-18 progresses disease in non-healing patients when the level of IL-12 gets decreased. http://rbmb.net/article-1-29-en.pdf 2013-04-30 43 48 Cutaneous leishmaniasis Cytokines Glucantime Maryam Shahi Shahi.maryam@gmail.com 1 Ghazvin University of Medical Sciences, Ghazvin, Iran AUTHOR Masoud Mohajery 2 Parasitology & Medical Mycology Department, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR Seyyed Ali Akbar Shamsian 3 Parasitology & Medical Mycology Department, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR Hossein Nahrevanian 4 Parasitology Departmen, Pasteur Institute of Iran, Tehran, Iran. AUTHOR Seyyed Mohammad Javad Yazdanpanah 5 Parasitology & Medical Mycology Department, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Molecular Cloning, Characterization, and Expression of Cuc m 2, a Major Allergen in Cucumis melo Background: Several studies reported the clinical features of IgE-mediated hypersensitivity after ingestion of melon. Melon allergy is a common IgE-mediated fruit allergy in Iran. This prompted us to investigate immunochemical and molecular properties of the major allergen in melon fruit, to compare the IgE-binding capacity of the natural protein with the recombinant allergen, and to determine cross-reactivity of the major allergen with closely-related allergens from other plants displaying clinical cross-reactivity with melon. Methods: Identification and molecular characterization of the major melon allergen were performed using IgE immunoblotting, allergen-specific ELISA, affinity-based purifications, cross-inhibition assays, cloning, and expression of the allergen in Escherichia coli. Results: Melon profilin was identified and isolated as a major IgE-binding component and designated as Cuc m 2. Sequencing corresponding cDNA revealed an open reading frame of 363 bp coding for 131 amino acid residues and two fragments of 171 bp and 383 bps for the 5’and 3’ UTRs, respectively. Significant cross-reactivity was found between melon profilin and Cynodon dactylon, tomato, peach, and grape profilins in cross-inhibition assays. Although the highest degree of amino acid identity was revealed with watermelon profilin, there was no significant cross-reactivity between melon and watermelon profilins. Conclusion: Melon profilin is the major IgE-binding component in melon extract, and the recombinant and natural forms exhibited similar IgE-binding capacities. A part of the fruit-fruit and pollen-fruit cross-reactions could be explained by the presence of this conserved protein; however, sequence homology provides insufficient information to predict IgE cross-reactivity of profilins. http://rbmb.net/article-1-30-en.pdf 2013-04-30 49 63 Cross-reactivity Fruit allergy Melon Profilin Recombinant allergen Mojtaba Sankian 1 Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Mahmoud Mahmoudi 2 Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Abdol-Reza Varasteh varasteha@mums.ac.ir 3 Allergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Relation of Type 2 Diabetes Mellitus with Gender, Education, and Marital Status in an Iranian Urban Population Background: Type 2 diabetes mellitus is one of the most important cardiovascular risk factors. Objectives: This study was performed to assess the relationship of diabetes with gender, education, and marital status in an Iranian urban population. Methods: A total of 892 men and women aged 30-85 were recruited using a cluster-stratified sampling method from an urban population. Using a questionnaire, demographical data including gender, education, and marital status were collected. A blood sample after fasting for at least eight hours was collected from each subject. Associations of type 2 diabetes mellitus and studied variables were tested for significance. Results: The prevalence of diabetes mellitus was 11.6%; 11.1% in men and 12.1% in women with no significant difference between them. Diabetes mellitus was most prevalent in the oldest age (age more than 60 years, 22.9%) and low education groups (17.9%, P < 0.001). Marital status was not significantly related to diabetes mellitus (P= 0.37). Conclusion: The prevalence of diabetes mellitus is related to education within the Iranian population. Thus preventive strategies should be based on the affective factors. http://rbmb.net/article-1-31-en.pdf 2013-04-30 64 68 Diabetes mellitus Education Gender Marital status Karamatollah Rahmanian 1 Department of Community Medicine, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR Mohammad Shojaei Shojaei1300@yahoo.com 2 Department of Internal Medicine, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR Abdolreza Sotoodeh Jahromi 3 Department of Immunology, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR
ORIGINAL_ARTICLE Identification of Leishmania Species Causing Cutaneous Leishmaniasis Using Random Amplified Polymorphic DNA (RAPD-PCR) in Kharve, Iran Background: Leishmaniasis, especially cutaneous leishmaniasis, is considered an important health problem in many parts of Iran including Kharve, Khorasan Razavi province. Cutaneous leishmaniasis is caused by various species of Leishmania, each having a different secondary host. Thus, identifying the parasites’ specie is of paramount importance for containment strategy planning. The morphological differentiation of Leishmania species is not possible, rendering the molecular methods as the sole means to this purpose. Therefore, to identify the causative agent of cutaneous leishmaniasis in Kharve, Random Amplified Polymorphic DNA-PCR (RAPD-PCR) was used. Methods: The disease was first confirmed by direct smears. Samples were gathered from 22 patients with established cutaneous leishmaniasis. The samples were immediately cultured in NNN medium, followed by sub-culture in RPMI-1640. Afterwards, DNA was extracted and amplified using RAPD-PCR. Electrophoresis patterns from each isolate were compared with reference strains of Leishmania major (L. major) and Leishmania tropica (L. tropica). Results: The results of this study indicated that the parasite causing cutaneous leishmaniasis in Kharve is L. tropica. Conclusion: It seems that L. tropica is the only causative agent of cutaneous leishmaniasis in Kharve, and RAPD-PCR is a suitable tool for Leishmania characterization in epidemiological studies. http://rbmb.net/article-1-32-en.pdf 2013-04-30 69 73 Kharve Leishmania major Leishmania tropica RAPD-PCR Khorasan Fatemeh Saadabadi 1 Parasitology & Medical Mycology Department, School of Medicine, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR Masoud Mohajery 2 Parasitology & Medical Mycology Department, School of Medicine, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR Elham Poostchi 3 Parasitology & Medical Mycology Department, School of Medicine, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR Seyyed Ali Akbar Shamsian shamsianaa@mums.ac.ir 4 Parasitology & Medical Mycology Department, School of Medicine, Mashhad University of Medical Sciences Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE The Role of Anti-CCD Antibodies in Grape Allergy Diagnosis Background: Allergens are mostly composed of glycoprotein structures. It is believed that glycan-specific antibodies may lead to false-positive reactions in immunoassays. In this study we investigated the glycosylation state of grape allergens as well as the presence of antibodies to cross-reactive carbohydrate determinants (anti-CCDs) in sera from grape-sensitive individuals. Methods: Grape extract proteins were electrotransferred onto PVDF membranes and their glycosylation states were analyzed by blotting methods. To assess the presence of anti-CCDs, natural and mildly deglycosylated proteins were immunoblotted with grape-allergic subjects' sera. We also measured the IgE reactivity of each subject’s sera with other fruit extracts via an indirect ELISA. Results: Immunoblotting studies showed that mildly deglycosylated grape proteins had lower IgE-binding capacity than their intact natural counterparts, which could be due to the presence of anti-CCDs. Biotinylation studies confirmed that the glycosylation levels of the 24, 32, and 60 kDa IgE-reactive proteins were higher than those of the 38 and 45 kDa ones. Lectin blotting showed that the 24 and 60 kDa bands were highly mannosylated, with the highest level of mannosylation on the 24 kDa allergen. Conclusion: This study showed that some grape allergens are glycosylated and that anti-CCD antibodies may cause weakly false-positive results during assessment of IgE reactivity to grape allergens. http://rbmb.net/article-1-33-en.pdf 2013-04-30 74 81 Allergy Antibody Cross-reactive carbohydrate determinants Grape Reza Falak 1 Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran AUTHOR Mojtaba Sankian 2 Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Hanieh Ketabdar 3 Department of Biology, Faculty of Science, Islamic Azad University, Mashhad Branch, Mashhad, Iran AUTHOR Malihe Moghadam 4 Allergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR Abdol-Reza Varasteh varasteha@mums.ac.ir 5 Allergy Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran AUTHOR
ORIGINAL_ARTICLE Construction of an Expression Plasmid (Vector) Encoding Brucella melitensis Outer Membrane Protein, a Candidate for DNA Vaccine Background: DNA vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. Bacteria of the genus Brucella are facultative intracellular pathogens for which new and efficient vaccines are needed. Methods: To evaluate the use of a DNA immunization strategy for protection against brucellosis, a plasmid containing the DNA encoding the Brucella melitensis (B. melitensis) 31 kDa outer membrane protein, as a potent immunogenic target, was constructed. Results: The constructed plasmid, pcDNA3.1+omp31, was injected intramuscularly into mice and the expression of omp31 RNA was assessed by RT-PCR. The integrity of the pcDNA3.1+omp31 construct was confirmed with restriction analysis and sequencing. Omp31 mRNA expression was verified by RT-PCR. Conclusion: Our results indicate that the pcDNA3.1+omp31 eukaryotic expression vector expresses omp31 mRNA and could be useful as a vaccine candidate. http://rbmb.net/article-1-34-en.pdf 2013-04-30 82 86 Brucella melitensis DNA Vaccine Omp31 PcDNA3.1 Fatemeh Vahedi f.vahedi@rvsri.ac.ir 1 Razi Vaccine and Serum Research Institute, Mashhad, Iran AUTHOR Elnaz Ghorbani 2 Microbiology Department, Al-Zahra University, Tehran, Iran AUTHOR Tahereh Falsafi 3 Microbiology Department, Al-Zahra University, Tehran, Iran AUTHOR
ORIGINAL_ARTICLE Partial Distal 10q Trisomy Due to De Novo Amplification: A new Case Without Furrows or Ridges in Fingers and Palms Background: Here we describe a new case of partial distal 10q trisomy in a 6-year-old Iranian girl from healthy parents with mental, growth, and psychomotor retardations. Methods: Additional clinical features include dysmorphic craniofacial features, microcephaly, bilateral hydronephrosis without heart problems, small and rotated low-set ears, bow-shaped mouth, abnormal teeth, short neck, and as a first case reported, fingers with camptodactly (i.e., without any furrows or ridges in the palms and fingers). Results: Cytogenetic analysis (GTG-banding) revealed an unbalanced female karyotype with additional bands at the end of the long arm of chromosome 10, karyotype: 46,XX,dup(10)(q25q26). Conclusion: According to the banding pattern it is most likely that a duplication of the distal part of the long arm of chromosome 10 occurred. http://rbmb.net/article-1-35-en.pdf 2013-04-30 87 90 De novo Distal Trisomy 10q Aliakbar Rahbarimanesh 1 Bahrami hospital, Medical Sciences/University of Tehran, Iran, PO Box 14155-1595 AUTHOR Pupak Derakhshandeh-Peykar derakhshandeh@tums.ac.ir 2 Bahrami hospital, Medical Sciences/University of Tehran, Iran, PO Box 14155-1595-Department of Medical Genetics, Medical Sciences/University of Tehran, Iran, PO Box 14155-1595-Medizinisch Genetisches Zentrum (MGZ), Munich, Germany, P.O. Box 201412 AUTHOR Amirhassan Barkhordari 3 Bahrami hospital, Medical Sciences/University of Tehran, Iran, PO Box 14155-1595 AUTHOR Reza Ebrahimzadeh-Vesal 4 Department of Medical Genetics, Medical Sciences/University of Tehran, Iran, PO Box 14155-1595 AUTHOR Soja Shamizadeh Kalkhoran 5 Bahrami hospital, Medical Sciences/University of Tehran, Iran, PO Box 14155-1595 AUTHOR