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Objective: To evaluate new compounds synthesized by integrating quinoline, quinazoline, and acridine rings with the active moiety of (5-nitroheteroaryl) methylene hydrazine. Methods: A new series of compounds (1a, 1b, 2a, 2b, 3a, and 3b) were synthesized and evaluated for cytotoxicity against COS-7 cells using the MTT assay. In vitro anti-plasmodial activity of the compounds was measured against CQ-sensitive (3D7) and CQ-resistant (K1) Plasmodium (P.) falciparum strains. β-hematin assay was performed to assess the inhibitory effects of β-hematin formation for new compounds. Results: The synthetic compounds had anti-plasmodial activity against blood-stage of 3D7 [IC
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Background: With considering the importance of natural products for their remedial and therapeutic value, this research was aimed to analyze the chemical compositions and antimicrobial activity of four propolis samples from different areas of Iran [Chenaran, Taleghan, Morad Beyg, and Kalaleh] with various climates and flora
Methods: Ethanolic [70% EtOH] and dichlromethane [DCM] extracts of Iranian propolis were analyzed by gas chromatography-mass spectrometry [GC-MS] methods, and antimicrobial activity was evaluated against Candida albicans, Escherichia coli, and Staphylococcus aureus using disk diffusion antimicrobial method
Results: The results of GC-MS analysis showed the presence of fatty acids, flavonoids, terpenes, aromatic-aliphatic acids, and their related esters. The total flavonoids in DCM extract of Chenaran, Taleghan, Morad Beyg, and Kalaleh propolis were pinocembrin and pinostrobin chalcone. The common phenolic and terpene compounds detected in all four tested EtOH extracts were P-cumaric acid and dimethyl -1,3,5,6-tetramethyl-[1,3-[13C2]] bicycloce [5.5.0] dodeca- 1,3,5,6,8,10-hexaene-9,10-dicarboxylate, respectively. The highest inhibitory diameter zone of the Iranian propolis against C. albicans, E. coli, and S. aureus was for DCM extract of Kalaleh propolis [13.33 mm], Morad Beyg propolis [12 mm], and Kalaleh [11.67 mm], respectively
Conclusion: Iranian propolis showed antimicrobial activities against C. albicans, E. coli, and S. aurous, perhaps due to the presence of flavonoids, phenolic acids, and terpenes as active components that can be used alone or in combination with the selected antibiotics to synergize antibiotic effect, as well as to prevent microbial resistance to available antimicrobial drugs
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Non-invasive therapeutic methods have recently been used in medical sciences. Enzymes have shown high activity at very low concentrations in laboratories and pharmaceutical, enabling them to play crucial roles in different biological phenomena related to living organism, especially human medicine. Recently, using the therapeutic methods based on non-invasive approaches has been emphasized in medical society. Researchers have focused on producing medicines and tools reducing invasive procedures in medical. Collagenases are proteins which catalyze chemical processes and break the peptide bonds in collagen. Collagen may be generated more than the required amount or produced in unsuitable sites or may not degrade after a certain time. In such cases, using an injectable collagenase or its ointment can be helpful in collagen degradation. In both in vitro and in vivo tests, it has been revealed that collagenases have several therapeutic properties in wound healing, burns, nipple pain and some diseases including intervertebral disc herniation, keloid, cellulite, lipoma among others. This review describes the therapeutic application of collagenase in medical sciences and the process for its production using novel methods, paving the way for more effective and safe applications of collagenases.
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Objective: The apical membrane antigen-1 [AMA-1] is considered as a promising candidate for development of a malaria vaccine against Plasmodium parasites. The correct conformation of this protein appears to be necessary for the stimulation of parasite-inhibitory responses, and these responses, in turn, seem to be antibody-mediated. Therefore, in the present investigation, we expressed the Plasmodium vivax AMA-1 [PvAMA-1] ectodomain in Escherichia coli [E. coli], purified it using standard procedures and characterized it to determine its biological activities for it to be used as a potential target for developing a protective and safe vivax malaria vaccine
Materials and Methods: In this experimental investigation, the ectodomain of PvAMA-1 antigen [GenBank accession no. JX624741] was expressed in the E. coli M15- pQE30 expression system and purified with immobilized-metal affinity chromatography. The correct conformation of the recombinant protein was evaluated by Western blotting and indirect immunofluorescence antibody [IFA] test. In addition, the immunogenic properties of PvAMA-1 were evaluated in BALB/c mice with the purified protein emulsified in Freund's adjuvant
Results: In the present study, the PvAMA-1 ectodomain was expressed at a high-level [65 mg/L] using a bacterial system. Reduced and non-reduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE] as well as Western blot analysis confirmed the appropriate conformation and folding of PvAMA-1. The evaluation of immunogenic properties of PvAMA-1 showed that both T helper-1 and 2 cells [Th1 and Th2] responses were present in mice after three immunizations and persisted up to one year after the first immunization. Moreover, the antibodies raised against the recombinant PvAMA-1 in injected mice could recognize the native protein localized on P. vivax parasites
Conclusion: We demonstrate that our recombinant protein had proper conformation and folding. Also, there were common epitopes in the recombinant forms corresponding to native proteins. These results; therefore, indicate that the expressed PvAMA-1 has the potential to be used as a vivax malaria vaccine
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Objective: Apical membrane antigen-1 [AMA-1] is one of the most promising bloodstage candidate antigens for production of a malaria vaccine against the Plasmodium parasite. Genetic diversity in protective antigens, which is a common phenomenon in a complex pathogen such as the Plasmodium parasite, is responsible for problems with the development of an effective malaria vaccine. This phenomenon will increase the parasite's ability to evade immune responses. Therefore, malaria vaccine development requires the evaluation of immune responses to different allelic forms of the vaccine candidate antigens
Methods: In this investigation, the two variant forms of PvAMA-1 [PvAMA-1A and B] were expressed in an Escherichia coli M15-pQE30 system using genomic DNA from Iranian individuals with patent Plasmodium vivax infection. The IgG responses of two antigens were evaluated in BALB/c mice with the purified protein emulsified in Freund's adjuvant. In addition, the correct conformation of the recombinant proteins was evaluated by the indirect immunofluorescence antibody test [IFA]
Results: The evaluation of immunogenic responses of two variant forms of PvAMA-1 showed the presence of IgG responses in mice after three immunizations. Cross-reactions were observed. Monitoring of IgG responses showed the persistence up to one year after the last immunization. The antibodies raised against recombinant PvAMA-1s in injected mice recognized the native protein [PvAMA-1] localized on Plasmodium vivax merozoites
Conclusion: The present outcomes confirmed the presence of common epitopes in recombinant forms of the protein that corresponded to native proteins. These emulsified proteins in Freund's adjuvant were immunogenic in BALB/c mice and IgG responses persisted for up to one year. The IgG responses to two PvAMA-1 variants did not differ significantly. The presence of cross-reactive antibodies has implied that one of these two forms of protein could be used in a universal blood-stage vaccine based on the PvAMA-1 antigen
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Different studies have shown an association of TLR4 polymorphisms with susceptibility/resistance to malaria disease. In the current immunogenetic study, we assessed the TLR4 genotypes formed by the two common single nucleotide polymorphisms [SNPs] [Asp299Gly and Thr399Ile] in the co-segregate state in Baluchi Plasmodium falciparum infected and healthy populations from malaria hypoendemic areas of Iran. The study was performed to evaluate the distribution and correlation of TLR4 co-segregating genotypes in patients with mild malaria. Moreover, the frequency of these genotypes was compared with reported results from other populations in similar or contrasting malaria settings around the world. In this case control study, the presence of 2 SNPs in the TLR4 gene [Asp299Gly and Thr399Ile] were analyzed in 350 Baluchi patients with mild malaria and 350 unrelated healthy controls by using polymerase chain reaction/restriction fragment length polymorphism [PCR/RFLP] techniques followed by sequencing analysis. Differences in the TLR4 co-segregate genotype frequencies among the studied group were determined by Fisher's exact test. Although the distribution of the two commonly co-segregating TLR4 genotypes presented a diverse and distinct pattern in the Baluchi population, no significant difference was detected between the cases and controls [p>0.05]. A lower frequency of TLR4 Asp299Gly/Thr399Thr was observed in Baluchis with mild malaria compared to African populations [p<0.05]. Differences in the co-segregation patterns of TLR4 Asp299Gly/Thr399Ile genotypes in the Baluchi population compared to other malaria endemic populations may suggest different local evolutionary pressure on TLR4 polymorphisms by malaria in this region. The higher frequency of Asp299Gly/Thr399Ile genotypes among the Baluchi population compared with the African population [p<0.05] which suffers from a larger number of severe cases might suggest that this genotype has a role in protecting against severe malaria. These findings are useful for further understanding the pathogenesis of severe malaria
Subject(s)
Humans , Toll-Like Receptor 4 , Genotype , Case-Control Studies , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNAABSTRACT
Because of the lack of an effective and economical control strategy against malaria [the most devastating infectious disease in developing countries] Transmission-Blocking Vaccines [TBVs] concept has been raised in recent years, promising a more efficient way to malaria control. TBVs aim at interfering and/or blocking pathogen development within the vector, halting transmission to non-infected vertebrate host. Aminopeptidase N [APN] is one of the most potent proteins in parasite development in Anopheles malaria vectors, which is strongly co-localized with human malaria parasites in the mosquito midgut epithelium. Therefore, Aminopeptidase N is one of the best choices for a new TBV. In this study for the first time we used 3'-RACE to amplify APN gene in Anopheles stephensi [An.stephensi], a major malaria vector in Iran, Indian subcontinent up to China by using different sets of primers including exon junction, conserved and specific region primers. Full length of APN was sequenced stepwise, which could be applied in designing a new regional TBV and act as an essential component of malaria elimination program in An. stephensi distribution areas. Primers design and method modification should be set up exactly in approach based amplifications. From results we came to this conclusion that that 3'-RACE could be applied to amplified key regions which are be-yond reach
Subject(s)
Malaria , Insect Proteins , Anopheles , Communicable Disease Control , Insect VectorsABSTRACT
Calpastatin is an endogenous inhibitor of calpain [calcium-dependent cysteine protease]. Calpastatin activity is highly related to the rate of protein turnover and rate of meat tenderization. In order to characterize the structure of calpastatin in Iranian Afshari breed of sheep, intron 6 and partial exon 7 of the L domain were amplified and sequenced. A fragment of approximately 1.5 kb was identified. In this study, an Afshari calpastatin gene fragment that encoded L Domain amino acids was detected. Hence by detection of such conserved mutations, it is possible to use these polymorphisms in Marker-Assisted Selection [MAS]
Subject(s)
Animals , Sheep/genetics , Sequence AnalysisABSTRACT
Anopheles culicifacies is a main malaria vector in southeastern part of Iran, bordering Afghanistan and Pakistan. So far, resistance to DDT, dieldrin, malathion and partial tolerance to pyrethroids has been reported in An. stephensi, but nothing confirmed on resistance status of An. culicifacies in Iran. In current study, along with WHO routine susceptibility test with DDT [4%], dieldrin [0.4%], malathion [5%], permethrin [0.25%], lambadacyhalothrin [0.1%], and deltamethrin 0.025, we cloned and sequenced segment VI of domain II [SII6] in voltage-gated sodium channel [vgsc] gene of An. Culicifacies specimens collected in Sistan and Baluchistan province [Iran]. A 221-bp amplified fragment showed 91% and 93% similarity with exon I and exon II of An. gambiae. The size of intron II in An. culicifacies is 62 bp, while in An. gambiae is 57 bp. The major difference within An. culicifacies specimens and also with An. gambiae is in position 29 of exon I, which led to substitution of Leu to His amino acid. This data will act as first report on partial sequence of vgsc gene and its polymorphism in An. culicifacies. A Leu to His amino acid substitution detected upstream the formerly known knockdown resistance [kdr] mutation site could be an indication for other possible mutations related to insecticide resistance. However, the result of WHO susceptibility test carried out in Baluchistan of Iran revealed a level of tolerance to DDT and dieldrin, but almost complete susceptibility to pyrethroids in An. culicifacies. We postulate that the molecular diagnostic tool developed for detection and identification of kdr-related mutations in An. culicifacies, could be useful in monitoring insecticide resistance in Iran and neighbouring countries such as Pakistan and Afghanistan. A phylogenetic tree also constructed based on the sequence of exon I and II, which readily separated An. culicifacies populations from An. stephensi, An. fluviatilis and An. gambiae
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Background The knowledge on population structure of the parasite isolates has contributed greatly to understanding the dynamics of the disease transmission for designing and evaluating malaria vaccines as well as for drug applications. msp-1 and msp-3? genes have been used as a genetic marker in population studies of Plasmodium vivax isolates. In this study; msp-3? was compared and assessed with msp-1marker in order to find whether msp-3? is a reliable genetic marker for P. vivax population studies. Methods This comparative study was designed and carried out as the first assessment of diversity in Pvmsp-3? gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in the 50 northern and 94 southern P. vivax isolates from Iran; which had been analysed before for msp-1 gene. Results Three allele size as; Type A (1.8 kb); Type B (1.5 kb) and Type C (1.2 kb) have been detected among both northern and southern isolates based on PCR results. Type C (70
Subject(s)
Disease Transmission, Infectious , Malaria , Plasmodium vivaxABSTRACT
Mixed malaria infections, Plasmodium falciparum and P. vivax, are suspected to occur at a greater frequency than is detected by conventional light microscopy. In order to determine the year round pattern of transmission and the frequency of mixed infections in malaria endemic area, we carried out a prospective comparison of diagnosis by conventional light microscopy and nested PCR in Chahbahar district, south-eastern part of Iran. Out of 280 Giemsa-stained slides, 158 [56.42%] were identified as having only P. vivax and 89 [31.78%] were P. falciparum infection by microscopy. Only eight slides [2.8%] were interpreted as having mixed P. vivax-P. falciparum infections and 25 [8.9%] were negative. Comparing to the microscopy results, the PCR detected 33 more mixed infections. These results showed that the number, of mixed infections was increased during April to September and reduced after September, although malaria cases with only P. falciparum were increased. The possibility that malaria patients in Chahbahar district may have undetected mixed infections during first peak of transmission should be kept in mind because of the specific therapy required both for P. falciparum and for radical cure of P. vivax