ABSTRACT
Toxoplasma gondii can cause severe and even fatal disease in human beings and animals. Effective vaccines may contribute to control toxoplasmosis. GRA14, a novel secreted dense granule protein of T. gondii, has been proposed as a vaccine candidate due to its intervacuolar transport and unique topology in the parasitophorous vacuole membrane. In this study, we constructed a DNA vaccine encoding GRA14 of T. gondii. BALB/c mice were immunized intramuscularly three times at 2 week intervals and challenged with T. gondii RH strain 5 weeks later. The immune responses were evaluated using lymphocyte proliferation assay, cytokine and antibody measurements. In addition, the survival times and parasite load of mice challenged with the virulent T. gondii RH strain were evaluated. The results showed that the mice immunized with pcGRA14 induced both enhanced specific humoral and Th1 cellular immune responses, and also mice immunized with the pcGRA14 showed an increased survival time and decreased parasite load compared with control groups (P<.05). The results indicated, for the first time, that the GRA14 is a potential DNA vaccine against toxoplasmosis.
Subject(s)
Protozoan Vaccines/immunology , Toxoplasma/immunology , Toxoplasmosis/immunology , Animals , Antibodies, Protozoan/blood , Antigens, Protozoan/immunology , CHO Cells , Cricetulus , Cytokines/immunology , Humans , Immunity, Cellular , Immunity, Humoral , Injections, Intramuscular , Lymphocytes/cytology , Male , Mice , Mice, Inbred BALB C , Parasite Load , Protozoan Proteins/immunology , Protozoan Vaccines/administration & dosage , Toxoplasmosis/prevention & control , Vaccines, DNA/immunologyABSTRACT
UNLABELLED: Opportunistic infections caused by Non-Candida albicans. have been increasing. Traditional methods that are used to identify clinical isolates of Candida species are time-consuming and not appropriate for rapid, accurate and reliable identification. PURPOSE: To identify Candida spp isolated from cancer patients using PCR-restriction enzyme. MATERIALS AND METHODS: Using universal primers, ITS1 and ITS4, in this study, we could amplify ITS1-5.8S-ITS2 rDNA regions at both 80 clinical isolates and 3 standard strains. The PCR products were digested with two restriction enzymes MspI and BlnI separately. RESULT: We successfully identified all isolated species using two restriction enzymes (MspI, BlnI). Candida albicans was the most common species (77.5%), followed by C. glabrata (15%), C. tropicalis (5%), C. krusei (2.5%). Although the primers and enzyme had the ability to identify C. parapsilosis, C. guilliermondii, C. dubliniensis, present isolates did not include these among identified ones. CONCLUSION: RFLP-PCR using ITSI and ITS4 primers and restriction enzyme is a rapid, easy, reliable and also applicable method in clinical laboratory for identification of medically important Candida spp.
Subject(s)
Candida/classification , Candida/isolation & purification , Candidiasis/microbiology , Neoplasms/complications , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Candida/genetics , DNA Primers/genetics , DNA, Fungal/genetics , DNA, Ribosomal/genetics , DNA, Ribosomal Spacer/genetics , Deoxyribonuclease HpaII , Deoxyribonucleases, Type II Site-Specific , Humans , IranABSTRACT
With a view to examine the effects of defined doses of retinyl palmitate (Vit. A) on limb morphogenesis and their effects at the critical time in mouse embryos, pregnant Swiss Webster albino mice were administered retinyl palmitate (10000 or 15000 IU/kg, i.p.) on different days of pregnancy. Vitamin A in 15000 IU/kg, i.p. dose was most effective as produced malformations in the forelimbs by day 10 in 28.6% mice and in the hindlimbs by day 11 in 20.6% mice. Further, two injections in a day with the lower dose (10000 IU/kg, i.p.) had more teratogenic effects than single 15000 IU/kg, i.p. injection. Two injections of either dose on day 10 resulted in higher embryo absorption.
