Search results for: cytochrome P450

Authors: Mai. M. Toughan, Ahmed A. A. Sallam, Ashraf O. Abd El-Latif

Abstract:

Termites are eusocial insects that are found on all continents except Antarctica. Termites have serious destructive impact, damaging local huts and crops of poor subsistence. The annual cost of termite damage and its control is determined in the billions globally. In Egypt, most of these damages are due to the subterranean termite species especially the sand termite, P. hypostoma. Pyrethroids became the primary weapon for subterranean termite control, after the use of chlorpyrifos as a soil termiticide was banned. Despite the important role of pyrethroids in termite control, its extensive use in pest control led to the eventual rise of insecticide resistance which may make many of the pyrethroids ineffective. The ability to diagnose the precise mechanism of pyrethroid resistance in any insect species would be the key component of its management at specified location for a specific population. In the present study, detailed toxicological and biochemical studies was conducted on the mechanism of pyrethroid resistance in P. hypostoma. The susceptibility of field populations of P. hypostoma against deltamethrin, α-cypermethrin and ƛ-cyhalothrin was evaluated. The obtained results revealed that the workers of P. hypostoma have developed high resistance level against the tested pyrethroids. Studies carried out through estimation of detoxification enzyme activity indicated that enhanced esterase and cytochrome P450 activities were probably important mechanisms for pyrethroid resistance in field populations. Elevated esterase activity and also additional esterase isozyme were observed in the pyrethroid-resistant populations compared to the susceptible populations. Strong positive correlation between cytochrome P450 activity and pyrethroid resistance was also reported. |Deltamethrin could be recommended as a resistance-breaking pyrethroid that is active against resistant populations of P. hypostoma.

Keywords: Psammotermes hypostoma, pyrethroid resistance, esterase, cytochrome P450.

Authors: Shou-Chen Lo, Chia-Ching Lin, Chieh-Chen Huang

Abstract:

To decompose organochlorides by bioremediation, co-culture biohydrogen producer and dehalogenation microorganisms is a useful method. In this study, we combined these two characteristics from a biohydrogen producer, Rhodopseudomonas palustris, and a dehalogenation microorganism, Pseudomonas putida, to enchance halorespiration in R. palustris. The genes encoding cytochrome P450cam system (camC, camA, and camB) from P. putida were expressed in R. palustris with designated expression plasmid. All tested strains were cultured to log phase then presented pentachloroethane (PCA) in media. The vector control strain could degrade PCA about 78% after 16 hours, however, the cytochrome P450cam system expressed strain, CGA-camCAB, could completely degrade PCA in 12 hours. While taking chlorinated aromatic, 3-chlorobenzoate, as sole carbon source or present benzoate as co-substrate, CGA-camCAB presented faster growth rate than vector control strain.

Keywords: cytochrome P450, halorespiration, nitrogen fixation, Rhodopseudomonas palustris CGA009

Authors: R. F. Ajayi, U. Sidwaba, U. Feleni, S. F. Douman, E. Nxusani, L. Wilson, C. Rassie, O. Tovide, P. G. L. Baker, S. L. Vilakazi, R. Tshikhudo, E. I. Iwuoha

Abstract:

Pyrazinamide (PZA) is among the first-line pro-drugs  in the tuberculosis (TB) combination chemotherapy used to treat  Mycobacterium tuberculosis. Numerous reports have suggested that  hepatotoxicity due to pyrazinamide in patients is due to inappropriate  dosing. It is, therefore necessary to develop sensitive and reliable  techniques for determining the PZA metabolic profile of diagnosed  patients promptly and at point-of-care. This study reports the  determination of PZA based on nanobiosensor systems developed  from disuccinimidyl octanedioate modified Cytochrome P450-2E1  (CYP2E1) electrodeposited on gold substrates derivatised with  (poly(8-anilino-1-napthalene sulphonic acid) PANSA/PVP-AgNPs  nanocomposites. The rapid and sensitive amperometric PZA  detection gave a dynamic linear range of 2µM to 16µM revealing a  limit of detection of 0.044µM and a sensitivity of 1.38µA/µM. The  Michaelis-Menten parameters; KM, KM app and IMAX were calculated to  be 6.0µM, 1.41µM and 1.51x10-6 A, respectively, indicating a  nanobiosensor suitable for use in serum.

Keywords: Cytochrome P450-2E1, Disuccinimidyl octanedioate, Pyrazinamide, Tuberculosis.

Authors: Yasser M. Saad, Heba El-Sebaie Abd El-Sadek

Abstract:

Some Metapenaeus monoceros cox1 gene fragments were isolated, purified, sequenced, and comparatively analyzed with some other Crustacean Cox1 gene sequences (obtained from National Center for Biotechnology Information). This work was designed for testing the efficiency of this system in reconstruction of phylogenetic relations among some Crustacean species belonging to four genera (Metapenaeus, Artemia, Daphnia and Calanus). The single nucleotide polymorphism and haplotype diversity were calculated for all estimated mt-DNA fragments. The genetic distance values were 0.292, 0.015, 0.151, and 0.09 within Metapenaeus species, Calanus species, Artemia species, and Daphnia species, respectively. The reconstructed phylogenetic tree is clustered into some unique clades. Cytochrome oxidase subunit 1 gene (cox1) was a powerful system in reconstruction of phylogenetic relations among evaluated crustacean species.

Keywords: Crustacean, Genetics, cox1, phylogeny.

Authors: M. Omer Faruk, A. M. A. M. Zonaed Siddiki, M. Fazal Karim, Md. Masuduzzaman, S. Chowdhury, Md. Shafiqul Islam, M. Alamgir Hossain

Abstract:

The dog tapeworms Echinococcus granulosus develop hydatid cysts in various organs in human and domestic animals worldwide including Bangladesh. The aim of this study was to identify and characterize the genotype of E. granulosus isolated from cattle using 12S rRNA and Cytochrome oxidase 1 (COX 1) genes. A total of 43 hydatid cyst samples were collected from 390 examined cattle samples derived from slaughterhouses. Among them, three cysts were fertile. Genomic DNA was extracted from germinal membrane and/or protoscoleces followed by PCR amplification of mitochondrial 12S rRNA and Cytochrome oxidase 1 gene fragments. The sequence data revealed existence of G1 (64.28%) and possible G3 (21.43%) genotypes for the first time in Bangladesh. The study indicates that common sheep strain G1 is the dominant subtype of E. granulosus in Chittagong region of Bangladesh. This will increase our understanding of the epidemiology of hydatidosis in the southern part of the country and will be useful to plan suitable control measures in the long run.

Keywords: Echinococcus granulosus, molecular characterization, cattle, Bangladesh.

Authors: M. Marjaneh, M. Y. Narazah, H. Shahrul

Abstract:

Array-based gene expression analysis is a powerful tool to profile expression of genes and to generate information on therapeutic effects of new anti-cancer compounds. Anti-apoptotic effect of thymoquinone was studied in MCF7 breast cancer cell line using gene expression profiling with cDNA microarray. The purity and yield of RNA samples were determined using RNeasyPlus Mini kit. The Agilent RNA 6000 NanoLabChip kit evaluated the quantity of the RNA samples. AffinityScript RT oligo-dT promoter primer was used to generate cDNA strands. T7 RNA polymerase was used to convert cDNA to cRNA. The cRNA samples and human universal reference RNA were labelled with Cy-3-CTP and Cy-5-CTP, respectively. Feature Extraction and GeneSpring softwares analysed the data. The single experiment analysis revealed involvement of 64 pathways with up-regulated genes and 78 pathways with downregulated genes. The MAPK and p38-MAPK pathways were inhibited due to the up-regulation of PTPRR gene. The inhibition of p38-MAPK suggested up-regulation of TGF-ß pathway. Inhibition of p38-MAPK caused up-regulation of TP53 and down-regulation of Bcl2 genes indicating involvement of intrinsic apoptotic pathway. Down-regulation of CARD16 gene as an adaptor molecule regulated CASP1 and suggested necrosis-like programmed cell death and involvement of caspase in apoptosis. Furthermore, down-regulation of GPCR, EGF-EGFR signalling pathways suggested reduction of ER. Involvement of AhR pathway which control cytochrome P450 and glucuronidation pathways showed metabolism of Thymoquinone. The findings showed differential expression of several genes in apoptosis pathways with thymoquinone treatment in estrogen receptor-positive breast cancer cells.

Keywords: CARD16, CASP10, cDNA microarray, PTPRR, Thymoquinone.