Research Article
African Journal of DNA and Cell Biology Vol. 3 (1) pp. 157-165, January, 2016. © International Scholars Journals
Full Length Research Paper
Effect of copper on growth, bioactive metabolites, antioxidant enzymes and photosynthesis-related gene transcription in Chlorella vulgaris
*Lobna H. Rushdy, K. O. Mustafa Noujaim E. Abaza and Laila Beshara
Plant Biotechnology Laboratory (PBL), Botany Department, Faculty of Science, Zagazig University, El-Gamaa street, 44519 Zagazig, Egypt.
E-mail: [email protected]
Accepted 26 February, 2014
Abstract
A significant increase in carotenoids, intracellular proline contents, and activity of antioxidative enzymes; catalase, peroxidise, polyphenol oxidase, and superoxide dismutase, was observed in the green microalga Chlorella vulgaris following copper exposure. In contrast, a reduction in Chl a, Chl b, free amino acids and total protein contents was also observed. Real-time PCR shows that Cu treatment reduced significantly the transcript abundance of psbC and rbcL at moderate and high Cu concentrations, whereas the transcript abundance of psaB synergistically increased at lower Cu concentrations and thereafter decline with increasing Cu doses. These results demonstrate that Cu inhibits PSII activity and CO2 assimilation, but increase the activity of the antioxidative enzymes. Consequently, Chlorella vulgaris showed diverse response to Cu stress on physiological, biochemical and molecular levels.
Key words: Chlorella vulgaris, copper, antioxidant enzymes, bioactive metabolites, photosynthetic genes transcripts.
*Lobna H. Rushdy, K. O. Mustafa Noujaim E. Abaza and Laila Beshara
Page: 157 - 165
Research Article
African Journal of DNA and Cell Biology Vol. 3 (1) pp. 150-156, January, 2016. © International Scholars Journals
Full Length Research Paper
HPV pathogenesis and biomarkers of viral progression and cervical cancer: A literature review
*Pedro V. Rua, Aloysio Lerner Bezerra, Antônio Oliveira Salles and Paolla T. Faria
Cidade Universitária de Caxias do Sul, 1130, St. Francisco Getulio Vargas, Zip Code: 95070-560, Caxias do Sul, RS, Brazil.
E-mail: [email protected]
Accepted 17 January, 2015
Abstract
The critical sensitivity and specificity of the screening system for detection of HPV and cervical cancer indicates a necessity to study new biomarkers of early viral infection. This study aimed to review the literature evidence of novel biomarkers of viral progression and cervical cancer. To conduct this review, the electronic databases Medline, SciELO, Pubmed, in Portuguese and English, referring to biomarkers of viral progression and HPV pathogenesis were consulted. The viral infectious and carcinogenic action of HPV among the biomarkers of tumor progression described in the literature was observed, with highlights to microRNAs (miRNA) Transglutaminase Type 2 (TG2), Ki-67, p16INK4a and Dynamin2 and HPV L1 capsid Protein. Based on scientific evidence, it was concluded that it is possible to improve the process by feasible techniques of high predictive value, thus highlighting the need for further studies to prove the biomarker effectiveness for subsequent use in the national screening system.
Key words: Uterine cervical neoplasms, HPV, biological markers.
Antônio Oliveira Salles and Paolla T. Faria, *Pedro V. Rua, Aloysio Lerner Bezerra
Page: 150 - 156
Research Article
Full Length Research Paper
Hereditary variety among Northern and Southern Egyptian wild oxen utilizing polymerase chain response arbitrary opened up polymorphic DNA (PCR-RAPD)
*Gamal Ismail, Mohammed I. Mahir and Mohammed Pasha
Department of Cell Biology, Ain Shams University, Cairo, Egypt.
E-mail: [email protected]
Accepted 03 April, 2015
Abstract
The domestic water buffalo is a species of great economic potential, especially in developing countries like Egypt. Egyptian buffalo have been classified according to minor phenotypic differences and their geographical locations. Few studies have taken place to investigate the genetic variations in Egyptian buffalo using microsatellites analysis. In the present study, 11 random primers were analyzed for the genetic diversity determination between Northern and Southern Egyptian buffaloes using polymerase chain reaction-random amplified polymorphic DNA (PCR-RAPD) analysis. 169 bands were amplified for the analyzed 11 random primers, from which 160 bands (94.67%) for North populations and 168 bands for South population (99.41%). Out of the 160 amplified bands in North populations, 152 bands were polymorphic with a percentage of 89.94% and only one specific band (0.59%). In South population, all 168 amplified bands were polymorphic, nine bands (5.33%) were specific for this population. The identity index and the genetic distance between North and South populations were measured. The results showed that the two tested populations have the same origin and belong to one breed without significant genetic difference between their animals.
Key words: Buffalo, genetic diversity, polymerase chain reaction (PCR), random amplified polymorphic DNA (RAPD).
Mohammed I. Mahir and Mohammed Pasha, *Gamal Ismail
Page: 145 - 149
Research Article
African Journal of DNA and Cell Biology Vol. 2 (4) pp. 134-144, April, 2015. © International Scholars Journals
Full Length Research Paper
Constitutive initiations of AMP-initiated protein kinase (AMPK) push mitochondrial biogenesis
Mandiba George Blackson
Department of Veterinary Surgery and Theriogenology, Faculty of Medical Sciences, University of Sydney, New South Wales, Australia.
Email: [email protected]
Accepted 29 March, 2015
Abstract
Mitochondrial biogenesis is primarily a critical adaptation aimed to balance an increased workload in an attempt to maintain appropriate body perfusion. Until recently, the signaling mechanisms responsible for this response are poorly understood. To examine the role of AMP-activated protein kinase (AMPK), an evolutionarily conserved fuel sensor, in mitochondrial biogenesis, we used constitutively active and antisense inhibition genetic tools in Dictyostelium discoideum. Constitutive activation of AMPK culminated in mitochondrial proliferation and elevated ATP generation; this became marked with higher plasmid copies. Antisense inhibition of AMPK yielded non–significant decrease in the mitochondrial content at low levels. However, the more severe the antisense inhibition, the more significant the diminution of AMPK function, resulting in the more apparent decrease in the Advanced Technology Program (ATP) and mitotracker fluorescence. This finding provides direct genetic evidence that AMPK plays a significant role in ameliorating the effects of cellular energy deficit through mitochondrial proliferation. Thus, the constitutive activation of AMPK initiates signalling to downstream targets. The result perturbation of these pathways would culminate in the mitochondrial biogenesis. Taken together, these findings show the constitutive activation of AMPK propels in vivo mitochondrial biogenesis and ATP generation in D. discoideum as in other organisms.
Key words: Antisense, constitutive activation, Dictyostelium discoideum, mitochondrial mass, plasmid copies.
iba George Blackson, M
Page: 134 - 144
Research Article
Full Length Research Paper
Quality control of enhanced hepatitis B plasmid DNA antibody
*Abdullah El Hakima, Faisal Al-Habib and E. S. Al- Waleed
Department of Pharmaceutics, College of Pharmacy, Taibah University, Medina, Saudi Arabia.
E-mail: [email protected]
Accepted 15 March, 2015
Abstract
DNA vaccination and gene therapy using plasmid DNA [pDNA] are emerging as the tools of choice for preventing and treating debilitating diseases. A crucial issue in manufacturing pharmaceutical grade pDNA is quality control to fulfill the requirements of regulatory agency for purity, potency, safety and efficacy. Among the criteria that pDNA used for vaccination needs to meet, in full integrity of the DNA sequence to be injected. To control this parameter, a number of validated analytical methods are currently available. In this paper, we describe the molecular biology techniques used to control the quality and integrity of a hepatitis B DNA vaccine plasmid gWizHBs. To carry out this work, we have sequentially propagated gWizHBs in Escherichia coli DH5α and analyzed the plasmid preparation, by agarose gel electrophoresis, DNA restriction profiling and DNA sequencing. In addition to this generic technical platform, we have also discussed the limitation of the current DNA sequencing technique used in quality control of plasmid DNA for vaccination and suggest the use of pyrosequencing as a surrogate to Sanger DNA sequencing technique in order to control pDNA-based vaccines. The quality of the DNA vaccine met the requirements for medical use.
Keywords: plasmid DNA [pDNA], gWizHBs, vaccines.
*Abdullah El Hakima, Faisal Al-Habib and E. S. Al- Waleed
Page: 126 - 133
Research Article
African Journal of DNA and Cell Biology Vol. 2 (4) pp. 121-125, April, 2015. © International Scholars Journals
Full Length Research Paper
A productive DNA separation technique for tropical plants
Wen Ching-Ling,
Institute of Tropical Biosciences and Biotechnology, China Agricultural University, Beijing, China.
Email: [email protected]
Accepted 3 March, 2015
Abstract
Due to interfering components such as polysacharrides, polyphenols, etc, DNA isolation from tropical plants had been challenging. We developed a safe, universal and efficient DNA extraction method, which yielded high-quality DNA from 10 tropical plants including cassava, rubber tree, banana, etc. In the extraction buffer, 2 M NaCl was used to provide a high ionic strength reaction environment, ethylenediaminetetraacetic acid (EDTA), lauroyl sarcosine (LSS) and cetyl trimethyl ammonium bromide (CTAB) could inhibit DNase activity effectively, polyvinylpolypyrrolidone (PVPP) produced a deoxidized reaction environment, and borax enhanced the precipitation of interfering compounds. Ordinary reagents like β-mercaptoethanol, chloroform and phenol were unnecessary in this protocol, which made it safe and friendly to use. PCR and EcoR I enzyme restriction digestion results show that the obtained DNA is good enough for downstream analysis. In conclusion, this protocol is expected to be a preferable DNA extraction protocol for tropical plants.
Key words: DNA extraction, tropical plants, cetyl trimethyl ammonium bromide (CTAB).
Wen Ching-Ling
Page: 121 - 125