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Purpose@#Toxoplasmosis, transmitted by Toxoplasma gondii, is a worldwide parasitic disease that affects approximately one-third of the world’s inhabitants. Today, there are no appropriate drugs to deter tissue cysts from developing in infected hosts. So, developing an effective vaccine would be valuable to avoid from toxoplasmosis. Considering the role of microneme antigens such as microneme protein 4 (MIC4) in T. gondii pathogenesis, it can be used as potential candidates for vaccine against T. gondii. @*Materials and Methods@#In this study several bioinformatics methods were used to assess the different aspects of MIC4 protein such as secondary and tertiary structure, physicochemical characteristics, the transmembrane domains, subcellular localization, B-cell, helper-T lymphocyte, cytotoxic-T lymphocyte epitopes, and other notable characteristic of this protein design a suitable vaccine against T. gondii. @*Results@#The studies revealed that MIC4 protein includes 59 potential post-translational modification sites without any transmembrane domains. Moreover, several probable epitopes of Band T-cells were detected for MIC4. The secondary structure comprised 55.69% random coil, 5.86% beta-turn, 19.31% extended strand, and 19.14% alpha helix. According to the Ramachandran plot results, 87.42% of the amino acid residues were located in the favored, 9.44% in allowed, and 3.14% in outlier regions. The protein allergenicity and antigenicity revealed that it was non-allergenic and antigenic. @*Conclusion@#This study gives vital basic on MIC4 protein for further research and also established an effective vaccine with different techniques against acute and chronic toxoplasmosis.
ABSTRACT
Purpose@#Toxoplasma gondii is an opportunistic parasite infecting all warm-blooded animals including humans. The dense granule antigens (GRAs) play an important role in parasite survival and virulence and in forming the parasitophorous vacuole. Identification of protein characteristics increases our knowledge about them and leads to develop the vaccine and diagnostic studies. @*Materials and Methods@#This paper gave a comprehensive definition of the important aspects of GRA12 protein, including physico-chemical features, a transmembrane domain, subcellular position, secondary and tertiary structure, potential epitopes of B-cells and T-cells, and other important features of this protein using different and reliable bioinformatics methods to determine potential epitopes for designing of a high-efficient vaccine. @*Results@#The findings showed that GRA12 protein had 53 potential post-translational modification sites. Also, only one transmembrane domain was recognized for this protein. The secondary structure of GRA12 protein comprises 35.55% alpha-helix, 19.50% extended strand, and 44.95% random coil. Moreover, several potential B- and T-cell epitopes were identified for GRA12. Based on the results of the Ramachandran plot, 79.26% of amino acid residues were located in favored, 11.85% in allowed and 8.89% in outlier regions. Furthermore, the results of the antigenicity and allergenicity assessment noted that GRA12 is immunogenic and nonallergenic. @*Conclusion@#This research provided important basic and conceptual data on GRA12 to develop an effective vaccine against acute and chronic toxoplasmosis for further in vivo investigations. More studies are required on vaccine development using the GRA12 alone or combined with other antigens in the future.
ABSTRACT
Purpose@#Toxoplasma gondii is an opportunistic parasite infecting all warm-blooded animals including humans. The dense granule antigens (GRAs) play an important role in parasite survival and virulence and in forming the parasitophorous vacuole. Identification of protein characteristics increases our knowledge about them and leads to develop the vaccine and diagnostic studies. @*Materials and Methods@#This paper gave a comprehensive definition of the important aspects of GRA12 protein, including physico-chemical features, a transmembrane domain, subcellular position, secondary and tertiary structure, potential epitopes of B-cells and T-cells, and other important features of this protein using different and reliable bioinformatics methods to determine potential epitopes for designing of a high-efficient vaccine. @*Results@#The findings showed that GRA12 protein had 53 potential post-translational modification sites. Also, only one transmembrane domain was recognized for this protein. The secondary structure of GRA12 protein comprises 35.55% alpha-helix, 19.50% extended strand, and 44.95% random coil. Moreover, several potential B- and T-cell epitopes were identified for GRA12. Based on the results of the Ramachandran plot, 79.26% of amino acid residues were located in favored, 11.85% in allowed and 8.89% in outlier regions. Furthermore, the results of the antigenicity and allergenicity assessment noted that GRA12 is immunogenic and nonallergenic. @*Conclusion@#This research provided important basic and conceptual data on GRA12 to develop an effective vaccine against acute and chronic toxoplasmosis for further in vivo investigations. More studies are required on vaccine development using the GRA12 alone or combined with other antigens in the future.
ABSTRACT
Trichomonas species usually reside in the mouth and occasionally in the respiratory tract. These species can be found in the lungs of humans. Although the pathogenicity of the parasite in the respiratory system has not been proven, it is more prevalent in people who lack good oral health or suffer from asthma or chronic pulmonary diseases. In the present descriptive study, we have identified Trichomonas by direct microscopic observation of stained smears and by PCR on the lung sputum of patients with asthma and chronic lung diseases that include lung cancer, bronchiectasis, COPD, and malignant pulmonary disease who were admitted to Masih Daneshvari Hospital, Tehran, Iran. For direct examination of 133 sputum samples, we stained the smears with giemsa. In addition a total of 60 samples were used for DNA extraction by an extraction kit [Cinnagen]. Nested-PCR was used for amplifying the ITS1 target gene of the parasite. Finally the DNA sequence of the gene was determined. According to the results, in direct examination of the sputum smears there were only 4 positive cases identified, whereas 22 [36.66%] of the samples were identified as Trichomonasby nested PCR. According to gender, 33.33% of the female samples and 38.46% of the male samples were found to be positive. Considering the high prevalence of this parasite in the study group, chronic pulmonary disease and asthmatic patients may be more susceptible to Trichomonas
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Background and purpose:Microsporidia infections occurvirtually in all in vertebrate and vertebrate hosts, in cluding humans. The this study is aimed atcomparison of distribution of microsporidiosis in various samples of Iranian immunocompromised patients and healthy individuals by molecular methods.
Materials and Methods: Inthiscase - control study, 258stool, blood, bronchoalveolar lavage [BAL] and urine sampleswere obtained fromimmunocompromisedpatients [bone marrowtransplant, kidney transplant and respiratory complications] referred to Masih Daneshvari and Shariati Hospitalduring 2010-2011.
After recording clinical data, DNA extraction was performed on all samples. To identifyhuman related microsporidiosis [Encephalitozoon spp. And Enterocytozoon spp.], polymerase chain reactionwas performedusingspecific primers andmultiplex-PCR on allsamples.
Results: Overall, the prevalence of microsporidiosis in immunocompromised patients were 4.3 and 3.9 percent in cases and control group, respectively.Incidence of microsporidiosis in patients undergoing bone marrow transplantation [5 isolates from 70 samples] was 7.1%, in patients with respiratory complications [4 isolates from 150 samples] 2.7 percent and patients undergoing renal transplantation [2 isolates from 38 samples] 5.3 percent. In the case group, most cases of infection occurred among men at the age of40-60 years. In bronchoalveolar lavage samples 3 isolates of Encephalitozoon [2%] and one isolate of Entrocytozoon [0.7%], were identified, respectively. In cancer patients undergoing bone marrow transplantation 4 isolates of Encephalitozoon [5.7%], 1 isolate of Enterocytozoon [1.4%] and in patients with renal transplants 2 isolates of Enterocytozoon [5.3%]were detected. 4 isolates of Enterocytozoon [2.2%] and 3 isolates of Encephalitozoon [1.7%] were identified in the control group that most infections occurred among menat the age of30-45years.
Conclusion:The most cases of human microsporidiosis are associated with human immunodeficiency virus infection or other states of immunosuppression, particularly in organ transplant recipients; The obtained results confirm this claim.
ABSTRACT
Echinococcosis is a zoonotic parasitic disease of humans and various herbivorous domestic animals transmitted by the contact with domestic and wild carnivores, mainly dogs and foxes. The aim of this study is the production, purification and evaluation immunogenicity of new construction of EG95 protein. The recombinant plasmid pET32-a+ used for Eg95 expression was constructed with the EG95 gene of Echinococcus granulosus fused with the thioredoxin tag. This recombinant clone was over expressed in Escherichia coli BL-21 [DE-3]. The expressed fusion protein was found almost entirely in the insoluble form [inclusion bodies] in cell lysate. The purification was performed under denaturing conditions in the presence of 8M urea by Ni-NTA column and dialysis. The purified recombinant proteins were confirmed with western blot analysis using polyclonal antiserum. To find out the immunogenicity of the purified protein, the BALB/c mice [10 mice/group] were immunized by injecting 20 micro g rEG95 protein formulated in Freund's and alum adjuvant. Immunization of mice with rEG95 using CFA/IFA and alum adjuvant generated high level of total antibody. In proliferation assay, the lymphocytes were able to mount a strong proliferative response with related production of IFN-gamma, IL-12 and TNF-alpha but with low secretion of either IL-4 or IL-10. The humoral and cellular immune responses against rEG95 suggested a mixed Th1/Th2 response with high intensity toward Th1. Our findings suggest that new construct of rEG95 formulated with CFA/IFA and alum adjuvant elicited strong cellular and humoral responses supporting further development of this vaccine candidate
Subject(s)
Animals, Laboratory , Antigens, Helminth , Helminth Proteins , Mice, Inbred BALB CABSTRACT
Curcumin as a yellow natural compound extracted from turmeric root is known it as an antibacterial agent. One of the nanoparticles ability is to decrease the defects of usual drug delivery systems. Chitosan is a low toxic, biodegradable, biocompatible and safe polymer which is used in production of nanoparticles. Nanoparticles like chitosan-tripolyphosphate [TPP] are able to increase antibacterial properties of curcumin. Curcumin-loaded chitosan-TPP nanoparticles containing chitosan, curcumin and TPP salt were synthesized by ionotropic gelation methods. First, the skin of anesthetized mice was inoculated with staphylococcus aureus and pseudomonas aeruginosa suspension. Then the infected mice were treated with curcumin-loaded chitosan-TPP nanoparticles for 3 days. Following that, antibacterial characteristics of the mice treated with curcumin-loaded chitosan- TPP nanoparticles were evaluated by bacterial culture of these mice. Our results showed the size of 160 +/- 10 nm and the charge of +7 +/- 2 mV in curcumin-loaded chitosan-TPP nanoparticles. These nanoparticles were also spiral shape. The encapsulation efficiency of curcumin in chitosan-TPP nanoparticles was 75 +/- 2%. Bacterial culture showed that curcumin-loaded chitosan-TPP nanoparticles inhibited staphylococcus aureus and pseudomonas aeruginosa growth. Our study demonstrated that curcumin-loaded chitosan-TPP nanoparticles can be utilized as a potent agent in treatment of Staphylococcus aureus and Pseudomonas aeruginosa infections
Subject(s)
Animals, Laboratory , Curcumin/pharmacology , Chitosan/pharmacology , Mice, Inbred BALB C , Nanoparticles , Pseudomonas aeruginosa , Infections , Staphylococcus aureus , Anti-Bacterial AgentsABSTRACT
Microsporidia are ubiquitous opportunistic pathogens infecting all animal phyla. The purpose of this study was to characterization human-associated microsporidia in pigeons of Tehran by staining and molecular methods. In the year 2010 a total of 147 pigeon's fecal samples were randomly collected from bird stores and public parks of Tehran and screened for the existence of human pathogenic microsporidia by staining method and multiplex/Nested-PCR and RFLP techniques. Nineteen [12.92%] of the studied samples were positive by microscopic examination, and 31 [21.08%] isolates were detected with specific primers. Genotyping based on the ITS regions of the rRNA gene were done for the Entrocytozoon bieneusi, Encephalitozoon intestinalis, Enc. hellem and Enc. cuniculi, respectively. The genotypes of Ent. bieneusi were identical to the D, M and J; genotypes of Enc. hellem were similar to the genotype 1 and 3 and genotypes of Enc. cuniculi were equal to I and II genotypes which previously characterized in human and animal origins. These results revealed that there is no limits to microsporidia transmission between pigeons of Tehran and humans for human infective species. This study points to the hygienic importance of this bird, because feces of pigeons are one of the sources of infection with microsporidia in human and easily pollutes our environment; on the other hand, children and elderly people comprise the principal visitors of public parks and they are the populations at risk for microsporidiosis