Search results for: sequencing

Authors: Ahmed F. Azmy , Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

Abstract:

Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosa (PS1), and Pseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazinon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 10^12CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, Malathion, organophosphate pesticides.

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Authors: Prasad Senadheera, Younousse Saidi, Frans JM Maathuis

Abstract:

Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes vector. Sequencing of the cDNA inserts revealed that they encode for the putative proteins with the sequence homologous to rice putative protein PROLM24 (Os06g31070), a prolamin precursor. Expression of this cDNA did not affect yeast growth in absence of salt. Axt3k and G19 strains expressing the PROLM24 were able to grow upto 400 mM and 600 mM of NaCl respectively. Similarly, Axt3k mutant with PROLM24 expression showed comparatively higher growth rate in the medium with excess LiCl (50 mM). The observation that expression of PROLM24 rescued the salt sensitive phenotypes of G19 and Axt3k indicates the existence of a regulatory system that ameliorates the effect of salt stress in the transformed yeast mutants. However, the exact function of the cDNA sequence, which shows partial sequence homology to yeast UTR1 is not clear. Although UTR1 involved in ferrous uptake and iron homeostasis in yeast cells, there is no evidence to prove its role in Na+ homeostasis in yeast cells. Absence of transmembrane regions in Os06g31070 protein indicates that salt tolerance is achieved not through the direct functional complementation of the mutant genes but through an alternative mechanism.

Keywords: Rice seed expression, salt stress, prolamin, salinitytolerance, Oryza sativa

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Authors: Miguel A. Gómez, Enrique Hilario, Francisco J. Alvarez, Elena Gastiasoro, Antonia Alvarez, Jose A. Casla, Jorge Arguinchona, Juan L. Larrabe

Abstract:

Total liquid ventilation can support gas exchange in animal models of lung injury. Clinical application awaits further technical improvements and performance verification. Our aim was to develop a liquid ventilator, able to deliver accurate tidal volumes, and a computerized system for measuring lung mechanics. The computer-assisted, piston-driven respirator controlled ventilatory parameters that were displayed and modified on a real-time basis. Pressure and temperature transducers along with a lineal displacement controller provided the necessary signals to calculate lung mechanics. Ten newborn lambs (<6 days old) with respiratory failure induced by lung lavage, were monitored using the system. Electromechanical, hydraulic and data acquisition/analysis components of the ventilator were developed and tested in animals with respiratory failure. All pulmonary signals were collected synchronized in time, displayed in real-time, and archived on digital media. The total mean error (due to transducers, A/D conversion, amplifiers, etc.) was less than 5% compared to calibrated signals. Improvements in gas exchange and lung mechanics were observed during liquid ventilation, without impairment of cardiovascular profiles. The total liquid ventilator maintained accurate control of tidal volumes and the sequencing of inspiration/expiration. The computerized system demonstrated its ability to monitor in vivo lung mechanics, providing valuable data for early decision-making.

Keywords: Immature lamb, perfluorocarbon, pressure-limited, total liquid ventilation, ventilator, volume-controlled.

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Authors: Amr A. El Hanafy, Muhammad Qureshi, Jamal Sabir, Mohamed Mutawakil, Mohamed M. Ahmed, Hassan El Ashmaoui, Hassan Ramadan, Mohamed Abou-Alsoud, Mahmoud Abdel Sadek

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Domestic goats (Capra hircus) are extremely diverse species and principal animal genetic resource of the developing world. These facilitate a persistent supply of meat, milk, fibre, and skin and are considered as important revenue generators in small pastoral environments. This study aimed to fingerprint β-LG gene at PCR-RFLP level in native Saudi goat breeds (Ardi, Habsi and Harri) in an attempt to have a preliminary image of β-LG genotypic patterns in Saudi breeds as compared to other foreign breeds such as Indian and Egyptian. Also, the Phylogenetic analysis was done to investigate evolutionary trends and similarities among the caprine β-LG gene with that of the other domestic specie, viz. cow, buffalo and sheep. Blood samples were collected from 300 animals (100 for each breed) and genomic DNA was extracted. A fragment of the β-LG gene (427bp) was amplified using specific primers. Subsequent digestion with Sac II restriction endonuclease revealed two alleles (A and B) and three different banding patterns or genotypes i.e. AA, AB and BB. The statistical analysis showed a general trend that β-LG AA genotype had higher milk yield than β-LG AB and β-LG BB genotypes. Nucleotide sequencing of the selected β-LG fragments was done and submitted to GenBank NCBI (Accession No. KJ544248, KJ588275, KJ588276, KJ783455, KJ783456 and KJ874959). Phylogenetic analysis on the basis of nucleotide sequences of native Saudi goats indicated evolutional similarity with the GenBank reference sequences of goat, Bubalus bubalis and Bos taurus. However, the origin of sheep which is the most closely related from the evolutionary point of view, was located some distance away.

Keywords: β-Lactoglobulin, Saudi goats, PCR-RFLP, Phylogenetic analysis.

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Authors: Rowland M. Brucken

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This paper argues nation-building theories that prioritize democratic governance best explain the successful postindependence development of Botswana. Three main competing schools of thought exist regarding the sequencing of policies that should occur to re-build weakened or failed states. The first posits that economic development should receive foremost attention, while democratization and a binding sense of nationalism can wait. A second group of experts identified constructing a sense of nationalism among a populace is necessary first, so that the state receives popular legitimacy and obedience that are prerequisites for development. Botswana, though, transitioned into a multi-party democracy and prosperous open economy due to the utilization of traditional democratic structures, enlightened and accountable leadership, and an educated technocratic civil service. With these political foundations already in place when the discovery of diamonds occurred, the resulting revenues were spent wisely on projects that grew the economy, improved basic living standards, and attracted foreign investment. Thus democratization preceded, and therefore provided an accountable basis for, economic development that might otherwise have been squandered by greedy and isolated elites to the detriment of the greater population. Botswana was one of the poorest nations in the world at the time of its independence in 1966, with little infrastructure, a dependence on apartheid South Africa for trade, and a largely subsistence economy. Over the next thirty years, though, its economy grew the fastest of any nation in the world. The transparent and judicious use of diamond returns is only a partial explanation, as the government also pursued economic diversification, mass education, and rural development in response to public needs. As nation-building has become a project undertaken by nations and multilateral agencies such as the United Nations and the North Atlantic Treaty Organization, Botswana may provide best practices that others should follow in attempting to reconstruct economically and politically unstable states.

Keywords: Botswana, democratization, economic development, nation-building.

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Authors: Vishal V. R. Nandigana, Mohammad Heiranian, Narayana R. Aluru

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Graphene material has found tremendous applications in water desalination, DNA sequencing and energy storage. Multiple nanopores are etched to create opening for water desalination and energy storage applications. The nanopores created are of the order of 3-5 nm allowing multiple ions to transport through the pore. In this paper, we present for the first time, molecular dynamics study of single ion transport, where only one ion passes through the graphene nanopore. The diameter of the graphene nanopore is of the same order as the hydration layers formed around each ion. Analogous to single electron transport resulting from ionic transport is observed for the first time. The current-voltage characteristics of such a device are similar to single electron transport in quantum dots. The current is blocked until a critical voltage, as the ions are trapped inside a hydration shell. The trapped ions have a high energy barrier compared to the applied input electrical voltage, preventing the ion to break free from the hydration shell. This region is called “Coulomb blockade region”. In this region, we observe zero transport of ions inside the nanopore. However, when the electrical voltage is beyond the critical voltage, the ion has sufficient energy to break free from the energy barrier created by the hydration shell to enter into the pore. Thus, the input voltage can control the transport of the ion inside the nanopore. The device therefore acts as a binary storage unit, storing 0 when no ion passes through the pore and storing 1 when a single ion passes through the pore. We therefore postulate that the device can be used for fluidic computing applications in chemistry and biology, mimicking a computer. Furthermore, the trapped ion stores a finite charge in the Coulomb blockade region; hence the device also acts a super capacitor.

Keywords: Graphene, single ion transport, Coulomb blockade, fluidic computer, super capacitor.

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Authors: Fiona Wahr, Sitalakshmi Venkatraman

Abstract:

Industry employers require new graduates to bring with them a range of knowledge, skills and abilities which mean these new employees can immediately make valuable work contributions. These will be a combination of discipline and professional knowledge, skills and abilities which give graduates the technical capabilities to solve practical problems whilst interacting with a range of stakeholders. Underpinning the development of these disciplines and professional knowledge, skills and abilities, are “enabling” knowledge, skills and abilities which assist students to engage in learning. These are academic and learning skills which are essential to common starting points for both the learning process of students entering the course as well as forming the foundation for the fully developed graduate knowledge, skills and abilities. This paper reports on a project created to introduce and strengthen these enabling skills into the first semester of a Bachelor of Information Technology degree in an Australian polytechnic. The project uses an action research approach in the context of ongoing continuous improvement for the course to enhance the overall learning experience, learning sequencing, graduate outcomes, and most importantly, in the first semester, student engagement and retention. The focus of this is implementing the new curriculum in first semester subjects of the course with the aim of developing the “enabling” learning skills, such as literacy, research and numeracy based knowledge, skills and abilities (KSAs). The approach used for the introduction and embedding of these KSAs, (as both enablers of learning and to underpin graduate attribute development), is presented. Building on previous publications which reported different aspects of this longitudinal study, this paper recaps on the rationale for the curriculum redevelopment and then presents the quantitative findings of entering students’ reading literacy and numeracy knowledge and skills degree as well as their perceived research ability. The paper presents the methodology and findings for this stage of the research. Overall, the cohort exhibits mixed KSA levels in these areas, with a relatively low aggregated score. In addition, the paper describes the considerations for adjusting the design and delivery of the new subjects with a targeted learning experience, in response to the feedback gained through continuous monitoring. Such a strategy is aimed at accommodating the changing learning needs of the students and serves to support them towards achieving the enabling learning goals starting from day one of their higher education studies.

Keywords: Enabling skills, student retention, embedded learning support, continuous improvement.

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