Metabolomics-Based Study of Logarithmic and Stationary Phases of Promastigotes in Leishmania major by 1H NMR Spectroscopy     

Mohammad Arjmand  Azadeh Madrakian  Ghader Khalili  Ali Najafi  Zahra Zamani  Ziba Akbari 《Iranian Biomedical Journal》2016,20(2):77-83

Background:

Cutaneous leishmaniasis is one of the most important parasitic diseases in humans. In this disease, one of the responsible organisms is Leishmania major, which is transmitted by sandfly vector. There are specific differences in biochemical profiles and metabolite pathways in logarithmic and stationary phases of Leishmania parasites. In the present study, ¹H NMR spectroscopy was used to examine the metabolites outliers in the logarithmic and stationary phases of promastigotes in L. major to enlighten more about the transmission mechanism in metacyclogenesis of L. major.

Methods:

Promastigote was cultured, logarithmic and stationary phases were separated by the peanut agglutinin, and cell metabolites were extracted. ¹H NMR spectroscopy was applied, and outliers were analyzed using principal component analysis.

Results:

The most altered metabolites in stationary and logarithmic phases were limited to citraconic acid, isopropylmalic acid, L-leucine, ornithine, caprylic acid, capric acid, and acetic acid.

Conclusion:

¹H NMR spectroscopy could play an important role in the characterization of metabolites in biochemical pathways during a metacyclogenesis process. These metabolites and their pathways can help in exploiting a transmission mechanism in metacyclogenesis, and outcoming data might be used in the metabolic network reconstruction of L. major modeling. Key Words: Leishmania major, Metabolomics, Principal component analysis  相似文献   

Comparison of distribution of virulence determinants in clinical and environmental isolates of Vibrio cholera     

Bakhshi B  Pourshafie MR  Navabakbar F  Tavakoli A  Shahcheraghi F  Salehi M  Faradjzadegan Z  Zahraei SM 《Iranian Biomedical Journal》2008,12(3):159-165

Background: The virulence of a pathogenic Vibrio cholerae is dependent on a discrete set of genetic determinants. In this study, we determined the distribution of virulence determinants among the clinical and environmental isolates of V. cholerae. Methods: The antibiotic resistance profiles of the isolates were determined using standard disk diffusion assay. PCR assay was performed to analyze the presence of toxin genes of ctx, zot and ace. The composition of cholera toxin encoding element (CTX) region flanking of the V. cholerae isolates was also analyzed. Results: All of the clinical isolates (100%) showed a complete set of virulence genes and also the attachment site of the filamentous bacteriophage CTXphi. None of the environmental isolates contained the virulence genes and the attachment site of the CTXphi. Analysis of the flanking regions including the toxin-linked cryptic element and repeat in toxin genes revealed their integrity in the clinical isolates while in the environmental isolates they were absent or contained incomplete sequences. Comparison of the antibiotic resistance assay of the environmental and clinical isolates showed a significant difference in the resistance profiles of the isolates obtained from the two sites. High rates of resistance to co-trimoxosol, streptomycin and chloramphenicol were found with clinical isolates. Conclusion: The absence of all virulence determinants in the environmental strains may suggest that certain ecological features must be present for V. cholerae to acquire a complete set of virulence determinants and to turn them into pathogenic strains.  相似文献   

Metal Extractability in Binary and Multi-metals Spiked Calcareous Soils     

Mohsen Jalali  Ziba Hourseresht 《Communications in Soil Science and Plant Analysis》2017,48(9):1089-1104

In the present study, a laboratory experiment was designed to compare the 0.01 M calcium chloride (CaCl₂) and diethylenetriaminepentaacetic acid (DTPA) extraction methods for their ability to predict cadmium (Cd), copper (Cu), iron (Fe), Manganese (Mn), nickel (Ni), and zinc (Zn) availability and mobility in five calcareous soils. The soils were spiked with different amounts of metals (0, 50, 100, 200, and 400 mg kg^?1) both in binary (Cu and Zn; Ni and Cd; Fe and Mn) and in multi-systems (Cd, Cu, Fe, Mn, Ni, and Zn) and incubated for 1 months at field capacity. In metal-spiked soils, both extraction methods showed a linear relationship of extractable to total metals for all soils. The fraction of total metals extracted by DTPA was much higher than the fraction extracted by CaCl₂, which was attributed to the formation of soluble metal-complexes in the complexing extracts calculated by the Visual Minteq program. DTPA extraction method showed higher selectivity for Cu over other metals both in binary and in multi-systems. Different order of metals extractability was found in binary and multi-systems for both extraction methods. Solid/solution distribution coefficient (K_d) was calculated by the ratio of the solid phase to soil solution concentration of metals extracted by CaCl₂ or DTPA extraction methods. Both in binary and in multi-systems, the average K_d (l kg^?1) of metals by soils were in the order of Mn (5398) > Fe (4413) > Zn (3376) > Cu (2520) > Ni (969) > Cd (350) in the CaCl₂-extractable metals and Fe (35) ≥ Ni (34) > Zn (18) > Mn (11.2) > Cu (6.3) > Cd (4) in the DTPA-extractable metals. Results showed that among the six studied metals, Cd had the lowest K_d, implying a relative higher mobility in these calcareous soils. The Visual Minteq indicated that in the CaCl₂-extraction method and in both binary and multi-systems the dominant species for Cu, Mn, Ni, and Zn were Cu²⁺, Mn²⁺, Ni²⁺ and Zn²⁺, respectively, while for Cd and Fe, the dominant species were CdCl⁺ and Fe(OH)²⁺, respectively.  相似文献