Molecular characterization of matrix metalloproteinase-1 (MMP-1) in Lucilia sericata larvae for potential therapeutic applications

Background: The salivary glands of Lucilia sericata are the first organs to express specific endopeptidase enzymes. These enzymes play a central role in wound healing, and they have potential to be used therapeutically. Methods: Rapid amplification of cDNA ends and rapid amplification of genomic ends were used to identify the coding sequence of MMP-1 from L. sericata. Different segments of MMP1 gene, namely the middle part, 3' end, and 5' end, were cloned, sequenced, and analyzed using bioinformatics tools to determine the distinct features of MMP-1 protein. Results: Assembling the different segments revealed that the complete mRNA sequence of MMP-1 is 1932 bp long. CDS is 1212 bp long and is responsible for the production of MMP-1 of 404 amino acid residues with a predicted molecular weight of 45.1 kDa. The middle part, 3' end, and 5' end sequences were 933, 503, and 496 bp. In addition, it was revealed that the MMP-1 genomic sequence includes three exons and two introns. Furthermore, the three-dimensional structure of L. sericata MMP-1 protein was evaluated, and its alignment defined that it has high similarity to chain A of human MMP-2 with 100% confidence, 72% coverage, and 38% identity according to the SWISS-MODEL modeling analysis. Conclusions: MMP-1 of L. sericata has a close relationship with its homologs in invertebrates and other insects. The present study significantly contributes to understanding the function, classification, and evolution of the characterized MMP-1 from L. sericata and provides basic required information for the development of an effective medical bioproduct.

Low Prevalence of Plasmodium vivax - Plasmodium falciparum Mixed - Infection in Patients from Central and Eastern Part of Sudan: Implication for Case Management in Sudan.

Accurate diagnosis of malaria parasite species is crucial for rational treatment that is a key success for a malaria control and elimination programmes. The main objective of this investigation was to correct species identification and re-assessment of diagnosis method in central and eastern part of Sudan. The blood samples were collected from 71 febrile cases infected with P. vivax in Eastern and Central Sudan, diagnosed by light microscopy and also by nested-PCR assay, using 18S small sub-unit ribosomal RNA (ssrRNA) gene. The nested-PCR were detect 92.9% (66/71) and 2.8% (2/71) P. vivax and P. falciparum mono-infection, respectively. Based on microscopy method, the level of mixed - Infection was zero; however, nested-PCR assay detected 4.2% (3/71) mixed infections in collected samples. In detecting P. vivax infection, microscopy had high sensitivity (97%) and specificity (50%). In conclusion, the present data point to the need of improving microscopy diagnosis method in malaria endemic region and also suggest that although molecular techniques are not practical for diagnosis of P. vivax and P. falciparum mixed infections in any areas; these could be used to collect epidemiological facts for control and elimination of the disease in Sudan.

Detection of mixed Plasmodium falciparum & P. vivax infections by nested-PCR in Pakistan, Iran & Afghanistan.

Background & objectives: Species identification and information on transmission pattern of malaria parasite in any malaria endemic area is key to success for a malaria control programme. In this investigation, malaria diagnosis using molecular method was used to assess the transmission pattern of malaria parasite in three malaria endemic regions: Afghanistan, Iran and Pakistan. Methods: Blood samples were collected from the patients presenting with vivax malaria from Afghanistan (n = 108), Iran (n = 200) and Pakistan (n = 199). Malaria parasite detection was made by the gold standard (microscopy) and also nested-PCR assay, using 18S small sub-unit ribosomal RNA (ssrRNA) gene. Results: Based on microscopy method, the level of mixed infection was zero to 2.5 per cent; however, nested-PCR assay detected 6.5, 22 and 23.5 per cent mixed infections in samples collected from Afghanistan, Iran and Pakistan, respectively. The present results showed that the co-infection of P. vivax with P. falciparum was frequent in malaria endemic regions of Iran and Pakistan. Interpretation & conclusion: The present data suggest the need for improving microscopy diagnosis method and the clinician should also have careful clinical observation, along with the reports on Giemsa-stained thick blood films, particularly in summer time when P. vivax is predominant. Also sharing information on transmission pattern of mixed infection among these countries may help in designing better control strategies for malaria.