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1.
Heliyon ; 9(7): e17918, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37539165

RESUMO

Thermoplastic composite pipes (TCP) in comparison to other pipes have proven beneficial features due to its flexibility which includes being fit for purpose, lightweight and no corrosion. However, during the manufacturing of TCP which involves the consolidation process, certain defects may be induced in it because of certain parameters, and this can affect the performance of the pipe in the long run as the induced defects might lead to in-service defects. Current techniques used in the industry are facing challenges with on-the-spot detection in a continuous manufacturing system. In TCP manufacturing process, the pipe is regularly monitored. When a defect is noticed, the whole process stops, and the appropriate action is taken. However, shutting down the process is costly; hence it is vital to decrease the downtime during manufacturing to the barest minimum. The solutions include optimizing the process for reduction in the manufacturing defects amount and thoroughly understanding the effect of parameters which causes certain defect types in the pipe. This review covers the current state-of-the-art and challenges associated with characterizing the identified manufacturing induced defects in TCP. It discusses and describes all effective consolidation monitoring strategies for early detection of these defects during manufacturing through the application of suitable sensing technology that is compatible with the TCP. It can be deduced that there is a correlation between manufacturing process to the performance of the final part and selection of characterization technique as well as optimizing process parameters.

2.
Polymers (Basel) ; 14(7)2022 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-35406328

RESUMO

Traditionally, the drilling waste generated in oil and gas exploration operations, including spent drilling fluid, is disposed of or treated by several methods, including burial pits, landfill sites and various thermal treatments. This study investigates drilling waste valorisation and its use as filler in polymer composites. The effect of the poor particle/polymer interfacial adhesion bonding of the suspended clay in oil-based mud (OBM) slurry and the LDPE matrix is believed to be the main reason behind the poor thermo-mechanical and mechanical properties of low-density polyethylene (LDPE)/OBM slurry nanocomposites. The thermo-mechanical and mechanical performances of LDPE)/OBM slurry nanocomposites without the clay surface treatment and without using compatibilizer are evaluated and discussed. In our previous studies, it has been observed that adding thermally treated reclaimed clay from OBM waste in powder form improves both the thermal and mechanical properties of LDPE nanocomposites. However, incorporating OBM clay in slurry form in the LDPE matrix can decrease the thermal stability remarkably, which was reported recently, and thereby has increased the interest to identify the mechanical response of the composite material after adding this filler. The results show the severe deterioration of the tensile and flexural properties of the LDPE/OBM slurry composites compared to those properties of the LDPE/MMT nanocomposites in this study. It is hypothesised, based on the observation of the different test results in this study, that this deterioration in the mechanical properties of the materials was associated with the poor Van der Waals force between the polymer molecules/clay platelets and the applied force. The decohesion between the matrix and OBM slurry nanoparticles under stress conditions generated stress concentration through the void area between the matrix and nanoparticles, resulting in sample failure. Interfacial adhesion bonding appears to be a key factor influencing the mechanical properties of the manufactured nanocomposite materials.

3.
Waste Manag ; 139: 362-380, 2022 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-35026697

RESUMO

Operational discharges from oil and gas exploration industry, accidental spillage, or improperly disposed drilling wastes has serious detrimental effects on human and the environment. The water- and oil-based fluids wastes are generated every year all over the world and remain a serious challenge in compliance with the requirements of zero discharge for the oil and gas industry. To meet environmental regulations, sustainable and effective waste management is critical yet mostly missing in the oil and gas industry. This work aims to provide the current state of art in drilling waste (drill cuttings and drilling fluids). An overview of the drilling fluid waste is first provided followed by its characteristics, environmental concerned constituents in this waste stream are then explored while considering the current waste management efforts. Environmental and regulatory issues regarding drilling waste and the shortcomings of regulations are also discussed. The work sums up with a perspective future trends on drilling waste management, opportunities and challenges ahead including the potential for recycling and re-use of waste drilling fluids and cuttings for commercial products development. There are opportunities for waste valorisation especially in raw materials recovery for valuable products utilisation rather than incurring burden to the environment.


Assuntos
Indústria de Petróleo e Gás , Gerenciamento de Resíduos , Reciclagem
4.
Polymers (Basel) ; 12(11)2020 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-33202958

RESUMO

There is a rising demand for replacement, regeneration of tissues and organ repairs for patients who suffer from diseased/damaged bones or tissues such as hip pains. The hip replacement treatment relies on the implant, which may not always meet the requirements due to mechanical and biocompatibility issues which in turn may aggravate the pain. To surpass these limitations, researchers are investigating the use of scaffolds as another approach for implants. Three-dimensional (3D) printing offers significant potential as an efficient fabrication technique on personalized organs as it is capable of biomimicking the intricate designs found in nature. In this review, the determining factors for hip replacement and the different fabrication techniques such as direct 3D printing, Fused Deposition Modelling (FDM), Selective Laser Sintering (SLS) and stereolithography (SLA) for hip replacement. The study also covers surface modifications of 3D printed implants and provides an overview on 3D tissue regeneration. To appreciate the current conventional hip replacement practices, the conventional metallic and ceramic materials are covered, highlighting their rationale as the material of choice. Next, the challenges, ethics and trends in the implants' 3D printing are covered and conclusions drawn. The outlook and challenges are also presented here. The knowledge from this review indicates that 3D printing has enormous potential for providing a pathway for a sustainable hip replacement.

5.
Waste Manag Res ; 38(12): 1331-1344, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32720590

RESUMO

Oil-based mud (OBM) waste from the oil and gas exploration industry can be valorised to tailor-made reclaimed clay-reinforced low-density polyethylene (LDPE) nanocomposites. This study aims to fill the information gap in the literature and to provide opportunities to explore the effective recovery and recycling techniques of the resources present in the OBM waste stream. Elemental analysis using inductively coupled plasma-optical emission spectrometry (ICP-OES) and X-ray fluorescence analysis, chemical structural analysis by Fourier transform infrared (FTIR) spectroscopy, and morphological analysis of LDPE/organo-modified montmorillonite (LDPE/MMT) and LDPE/OBM slurry nanocomposites by scanning electron microscopy (SEM) have been conducted. Further analysis including calorimetry, thermogravimetry, spectroscopy, microscopy, energy dispersive X-ray analysis and X-ray diffraction (XRD) was carried out to evaluate the thermo-chemical characteristics of OBM waste and OBM clay-reinforced LDPE nanocomposites, confirming the presence of different clay minerals including inorganic salts in OBM slurry powder. The microscopic analysis revealed that the distance between polymer matrix and OBM slurry filler is less than that of MMT, which suggests better interfacial adhesion of OBM slurry compared with the adhesion between MMT and LDPE matrix. This was also confirmed by XRD analysis, which showed the superior delamination structure OBM slurry compared with the structure of MMT. There is a trend noticeable for both of these fillers that the nanocomposites with higher percentage filler contents (7.5 and 10.0 wt% in this case) were indicated to act as a thermal conductive material. The heat capacity values of nanocomposites decreased about 33% in LDPE with 7.5 wt% MMT and about 17% in LDPE with 10.0 wt% OBM slurry. It was also noted, for both nanocomposites, that the residue remaining after 1000°C increases with the incremental wt% of fillers in the nanocomposites. There is a big difference in residue amount (in %) left after thermogravimetric analysis in the two nanocomposites, indicating that OBM slurry may have significant influence in decomposing LDPE matrix; this might be an interesting area to explore in the future. The results provide insight and opportunity to manufacture waste-derived renewable nanocomposites with enhanced structural and thermal properties.


Assuntos
Nanocompostos , Polietileno , Bentonita , Cromatografia Gasosa-Espectrometria de Massas , Difração de Raios X
6.
Amino Acids ; 52(5): 693-710, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32367435

RESUMO

In this study, a determination of Troponin I and creatine kinase activity in whole-blood samples in a cohort of 100 small infants in the age of 2-5 years from Uganda with complicated Plasmodium falciparum malaria suggests the prevalence of cardiac symptoms in comparison to non-infected, healthy patients. Troponin I and creatine kinase activity increased during infection. Different reports showed that complicated malaria coincides with hypoxia in children. The obtained clinical data prompted us to further elucidate the underlying regulatory mechanisms of cardiac involvement in human cardiac ventricular myocytes. Complicated malaria is the most common clinical presentation and might induce cardiac impairment by hypoxia. Eukaryotic initiation factor 5A (eIF-5A) is involved in hypoxia induced factor (HIF-1α) expression. EIF-5A is a protein posttranslationally modified by hypusination involving catalysis of the two enzymes deoxyhypusine synthase (DHS) and deoxyhypusine hydroxylase. Treatment of human cardiomyocytes with GC7, an inhibitor of DHS, catalyzing the first step in hypusine biosynthesis led to a decrease in proinflammatory and proapoptotic myocardial caspase-1 activity in comparison to untreated cardiomyocytes. This effect was even more pronounced after co-administration of GC7 and GPI from P. falciparum simulating the pathology of severe malaria. Moreover, in comparison to untreated and GC7-treated cardiomyocytes, co-administration of GC7 and GPI significantly decreased the release of cytochrome C and lactate from damaged mitochondria. In sum, coadministration of GC7 prevented cardiac damage driven by hypoxia in vitro. Our approach demonstrates the potential of the pharmacological inhibitor GC7 to ameliorate apoptosis in cardiomyocytes in an in vitro model simulating severe malaria. This regulatory mechanism is based on blocking EIF-5A hypusination.


Assuntos
Apoptose , Malária/patologia , Miócitos Cardíacos/patologia , Parasitemia/patologia , Fatores de Iniciação de Peptídeos/metabolismo , Plasmodium berghei/isolamento & purificação , Proteínas de Ligação a RNA/metabolismo , Animais , Pré-Escolar , Feminino , Humanos , Lactente , Malária/metabolismo , Malária/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/parasitologia , Parasitemia/metabolismo , Parasitemia/parasitologia , Fatores de Iniciação de Peptídeos/genética , Proteínas de Ligação a RNA/genética , Fator de Iniciação de Tradução Eucariótico 5A
7.
Sensors (Basel) ; 18(1)2018 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-29351221

RESUMO

This work focuses on the proof-mass mechanical structural design improvement of a tri-axial piezoresistive accelerometer specifically designed for head injuries monitoring where medium-G impacts are common; for example, in sports such as racing cars or American Football. The device requires the highest sensitivity achievable with a single proof-mass approach, and a very low error (<1%) as the accuracy for these types of applications is paramount. The optimization method differs from previous work as it is based on the progressive increment of the sensor proof-mass mass moment of inertia (MMI) in all three axes. Three different designs are presented in this study, where at each step of design evolution, the MMI of the sensor proof-mass gradually increases in all axes. The work numerically demonstrates that an increment of MMI determines an increment of device sensitivity with a simultaneous reduction of cross-axis sensitivity in the particular axis under study. This is due to the linkage between the external applied stress and the distribution of mass (of the proof-mass), and therefore of its mass moment of inertia. Progressively concentrating the mass on the axes where the piezoresistors are located (i.e., x- and y-axis) by increasing the MMI in the x- and y-axis, will undoubtedly increase the longitudinal stresses applied in that areas for a given external acceleration, therefore increasing the piezoresistors fractional resistance change and eventually positively affecting the sensor sensitivity. The final device shows a sensitivity increase of about 80% in the z-axis and a reduction of cross-axis sensitivity of 18% respect to state-of-art sensors available in the literature from a previous work of the authors. Sensor design, modelling, and optimization are presented, concluding the work with results, discussion, and conclusion.


Assuntos
Sistemas Microeletromecânicos , Aceleração , Acelerometria , Traumatismos Craniocerebrais , Desenho de Equipamento , Humanos
8.
J Hazard Mater ; 340: 57-66, 2017 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-28711833

RESUMO

The risk assessment, exposure and understanding of the release of embedded carbon nanotubes (CNTs) and carbon nanofibers (CNFs) from commercial high performance composites during machining processes are yet to be fully evaluated and quantified. In this study, CNTs and CNFs were dispersed in epoxy matrix through calendaring process to form nanocomposites. The automated drilling was carried out in a specially designed drilling chamber that allowed elimination of background noise from the measurements. Emission measurements were taken using condensed particle counter (CPC), scanning mobility particle sizer (SMPS) and DMS50 Fast Particulate Size Spectrometer. In comparison to the neat epoxy, the study results revealed that the nano-filled samples produced an increase of 102% and 227% for the EP/CNF and EP/CNT sample respectively in average particle number concentration emission. The particle mass concentration indicated that the EP/CNT and EP/CNF samples released demands a vital new perspective on CNTs and CNFs embedded within nanocomposite materials to be considered and evaluated for occupational exposure assessment. Importantly, the increased concentration observed at 10nm aerosol particle sizes measurements strongly suggest that there are independent CNTs being released at this range.

9.
Bioorg Med Chem ; 22(15): 4338-46, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24909679

RESUMO

East Coast fever (ECF) is a tick-borne disease caused by the parasite Theileria parva which infects cattle. In Sub-Saharan Africa it leads to enormous economic costs. After a bite of a tick, sporozoites invade the host lymphocytes and develop into schizonts. At this stage the parasite transforms host lymphocytes resulting in the clonal expansion of infected lymphocytes. Animals develop a lymphoma like disorder after infection which is rapidly fatal. Hitherto, a few drugs of the quinone type can cure the disease. However, therapy can only be successful after early diagnosis. The genera Theileria and Plasmodium, which includes the causative agent of human malaria, are closely related apicomplexan parasites. Enzymes of the hypusine pathway, a posttranslational modification in eukaryotic initiation factor EIF-5A, have shown to be druggable targets in Plasmodium. We identified the first enzyme of the hypusine pathway from T. parva, the deoxyhypusine synthase (DHS), which is located on chromosome 2 of the Muguga strain. Transcription is significantly increased in schizonts. The expressed T. parva DHS reveals an open reading frame (ORF) of 370 amino acids after expression in Escherichia coli Rosetta cells with a molecular size of 41.26 kDa and a theoretical pI of 5.26. Screening of the Malaria Box which consists of 400 active compounds resulted in a novel heterocyclic compound with a guanyl spacer which reduced the activity of T. parva DHS to 45%. In sum, the guanyl residue seems to be an important lead structure for inhibition of Theileria DHS. Currently, more different guanyl analogues from the Malaria Box are tested in inhibitor experiments to determine their efficacy.


Assuntos
Oxirredutases atuantes sobre Doadores de Grupo CH-NH/antagonistas & inibidores , Plasmodium/enzimologia , Theileria parva/enzimologia , Sequência de Aminoácidos , Animais , Bovinos , Clonagem Molecular , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Escherichia coli/metabolismo , Guanina/química , Compostos Heterocíclicos/química , Compostos Heterocíclicos/metabolismo , Humanos , Cinética , Linfócitos/parasitologia , Dados de Sequência Molecular , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Plasmodium/genética , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Alinhamento de Sequência , Theileria parva/genética
10.
Toxicol Sci ; 138(2): 354-64, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24449423

RESUMO

As nanoparticles could form aggregates in biological systems, the dynamics of their dispersity drives the temporal effect of nanoparticles in vitro. To test this hypothesis, the fumed silica nanoparticles (SiNPs) that have primary sizes of 7-14 nm and form aggregates in culture medium were selected for toxicity study in human lung A549 cells. The dispersity of SiNPs was analyzed by dynamic light scattering and transmission of electron microscopy. Cytotoxicity assays including mitochondrial activity, intracellular level of reactive oxygen species (ROS), and membrane damage together with the ¹H-NMR-based extracellular metabonomic assay were conducted to determine the temporal dose-effect relationship of SiNPs. In cell culture medium, SiNPs dispersed well initially at 25-100 µg/ml; however, they sedimented rapidly in a concentration-dependent manner. SiNPs caused a dose-dependent increase of intracellular ROS and cell membrane damage at 4 h and a loss of cell viability after 48 h. SiNPs also induced an elevation of extracellular glucose, lactate, phenylalanine, histidine, and tyrosine levels in a time- and concentration-dependent manner. The dose-effect patterns at 4 h were different from that at 12 and 24 h as assessed by both cytotoxicity and metabonomic assays. Both fitted better with polynomial regression than linear regression, implying multimode action of SiNPs at different concentrations. The early NP-cell interaction and the late sedimentation could be attributable to the temporal effects of SiNPs. The extracellular ¹H-NMR-based metabonomics demonstrated a potential as a robust nondestructive tool for monitoring the temporal effect of NPs that tend to aggregate in nature.


Assuntos
Poluentes Atmosféricos/toxicidade , Líquido Extracelular/metabolismo , Pulmão/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Nanopartículas/química , Dióxido de Silício/toxicidade , Poluentes Atmosféricos/química , Técnicas de Cultura de Células , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Meios de Cultura , Relação Dose-Resposta a Droga , Humanos , Pulmão/metabolismo , Pulmão/patologia , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Transmissão , Análise Multivariada , Tamanho da Partícula , Espécies Reativas de Oxigênio/metabolismo , Espalhamento de Radiação , Dióxido de Silício/química , Propriedades de Superfície
11.
Materials (Basel) ; 6(11): 5171-5198, 2013 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-28788383

RESUMO

This work describes flax fibre reinforced polymeric composites with recent developments. The properties of flax fibres, as well as advanced fibre treatments such as mercerization, silane treatment, acylation, peroxide treatment and coatings for the enhancement of flax/matrix incompatibility are presented. The characteristic properties and characterizations of flax composites on various polymers including polypropylene (PP) and polylactic acid, epoxy, bio-epoxy and bio-phenolic resin are discussed. A brief overview is also given on the recent nanotechnology applied in flax composites.

12.
Artigo em Inglês | MEDLINE | ID: mdl-19574142

RESUMO

This paper presents, for the first time, a coupled piezoelectric-circuit finite element model (CPC-FEM) to analyze the power output of a vibration-based piezoelectric energy-harvesting device (EHD) when it is connected to a load resistor. Special focus is given to the effect of the load resistor value on the vibrational amplitude of the piezoelectric EHD, and thus on the current, voltage, and power generated by the device, which are normally assumed to be independent of the load resistor value to reduce the complexity of modeling and simulation. The presented CPC-FEM uses a cantilever with a sandwich structure and a seismic mass attached to the tip to study the following characteristics of the EHD as a result of changing the load resistor value: 1) the electric outputs: the current through and voltage across the load resistor; 2) the power dissipated by the load resistor; 3) the displacement amplitude of the tip of the cantilever; and 4) the shift in the resonant frequency of the device. It is found that these characteristics of the EHD have a significant dependence on the load resistor value, rather than being independent of it as is assumed in most literature. The CPC-FEM is capable of predicting the generated output power of the EHD with different load resistor values while simultaneously calculating the effect of the load resistor value on the displacement amplitude of the tip of the cantilever. This makes the CPC-FEM invaluable for validating the performance of a designed EHD before it is fabricated and tested, thereby reducing the recurring costs associated with repeat fabrication and trials. In addition, the proposed CPC-FEM can also be used for producing an optimized design for maximum power output.

13.
SAHARA J ; 6(4): 170-8, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20485856

RESUMO

This paper examines the viability and effectiveness of a pilot farming initiative in reversing impacts of HIV/AIDS on the most affected households in Homa Bay, Kenya. The paper argues that once patients are stable, they can effectively be engaged in farming with minimal financial and technical support, resulting in enhanced food security of the affected households. More importantly, it helps to reduce HIV/AIDS-related stigma and improve the individual's self-esteem. Some of the key challenges of the pilot initiative were the limited number of agricultural extension workers and absence of facilities to enable them to deliver services to the farmers, the high cost of farm inputs, the unavailability of farm inputs when they were needed, poorly developed agricultural markets, and the absence of irrigation facilities. The paper recommends the sensitive scaling-up of this approach. However, farming initiatives by HIV/AIDS service NGOs should be linked to at least three key aspects: (a) treatment, care and support to HIV/AIDS affected households; (b) micro grant schemes or subsidies to enable farmers to purchase farming tools and farm inputs; and (c) comprehensive on-farm training support. To ensure effectiveness and wider reach, government needs to view agriculture through an HIV lens and promote a multisectoral approach that recognises the relationship between HIV/AIDS and food security. A number of immediate actions are required to strengthen this relationship, such as increased public investment to augment extension services, subsidise farm inputs, and develop infrastructure including agricultural markets.


Assuntos
Características da Família , Abastecimento de Alimentos/economia , Infecções por HIV/economia , Síndrome da Imunodeficiência Adquirida/economia , Adulto , Criança , Produtos Agrícolas/economia , Produtos Agrícolas/provisão & distribuição , Países em Desenvolvimento/economia , Feminino , Organização do Financiamento , Abastecimento de Alimentos/normas , HIV , Infecções por HIV/epidemiologia , Humanos , Quênia/epidemiologia , Masculino , Cooperação do Paciente , Fatores de Risco , Grupos de Autoajuda , Apoio Social , Fatores Socioeconômicos
14.
SAHARA J ; 5(2): 94-102, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18709212

RESUMO

Consequences of HIV and AIDS are exponential in Kenya, touching not only the health of those infected, but also depleting socioeconomic resources of entire families. Access to financial services is one of the important ways to protect and build economic resources. Unfortunately, the norm of financial viability discourages microfinance institutions from targeting people severely impacted by HIV and AIDS. Thus, HIV and AIDS service NGOs have been increasingly getting involved in microcredit activity in recent years for economic empowerment of their clients. Despite limited human resources and funding in the area of microcredit activity, these NGOs have demonstrated that nearly 50% of their microcredit beneficiaries invested money in income-generating activities, resulting in enhancement to their livelihood security. In the short term these NGOs need to improve their current practices. However, this does not mean launching microfinance initiatives within their AIDS-focused programmes, as financial services are best provided by specialised institutions. Longer-term cooperation between microfinance institutions and other AIDS service organisations and donors is necessary to master appropriate and rapid responses in areas experiencing severe impacts of HIV and AIDS.


Assuntos
Organização do Financiamento/organização & administração , Infecções por HIV/economia , Renda , Organizações/organização & administração , Atitude Frente a Saúde , Comportamento Cooperativo , Efeitos Psicossociais da Doença , Países em Desenvolvimento , Grupos Focais , Infecções por HIV/epidemiologia , Infecções por HIV/psicologia , Necessidades e Demandas de Serviços de Saúde , Humanos , Renda/estatística & dados numéricos , Relações Interinstitucionais , Investimentos em Saúde/organização & administração , Quênia/epidemiologia , Modelos Organizacionais , Pobreza , Poder Psicológico , Avaliação de Programas e Projetos de Saúde , Fatores de Risco , Grupos de Autoajuda/organização & administração , Fatores Socioeconômicos , Estereotipagem , Inquéritos e Questionários , Populações Vulneráveis
15.
BMC Microbiol ; 6: 91, 2006 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-17042947

RESUMO

BACKGROUND: Plasmodium vivax is the most widespread human malaria parasite. However, genetic information about its pathogenesis is limited at present, due to the lack of a reproducible in vitro cultivation method. Sequencing of the Plasmodium vivax genome suggested the presence of a homolog of deoxyhypusine synthase (DHS) from P. falciparum, the key regulatory enzyme in the first committed step of hypusine biosynthesis. DHS is involved in cell proliferation, and thus a valuable drug target for the human malaria parasite P. falciparum. A comparison of the enzymatic properties of the DHS enzymes between the benign and severe Plasmodium species should contribute to our understanding of the differences in pathogenicity and phylogeny of both malaria parasites. RESULTS: We describe the cloning of a 1368 bp putative deoxyhypusine synthase gene (dhs) sequence from genomic DNA of P. vivax PEST strain Salvador I (Accession number AJ549098) after touchdown PCR. The corresponding protein was expressed and functionally characterized as deoxyhypusine synthase by determination of its specific activity and cross-reactivity to human DHS on a Western blot. The putative DHS protein from P. vivax displays a FASTA score of 75 relative to DHS from rodent malaria parasite, P. yoelii, and 74 relative to that from the human parasite, P. falciparum strain 3D7. The ORF encoding 456 amino acids was expressed under control of IPTG-inducible T7 promoter, and expressed as a protein of approximately 50 kDa (theoretically 52.7 kDa) in E. coli BL21 DE3 cells. The N-terminal histidine-tagged protein was purified by Nickel-chelate affinity chromatography under denaturing conditions. DHS with a theoretical pI of 6.0 was present in both eluate fractions. The specific enzymatic activity of DHS was determined as 1268 U/mg protein. The inhibitor, N-guanyl-1, 7-diaminoheptane (GC7), suppressed specific activity by 36-fold. Western blot analysis performed with a polyclonal anti-human DHS antibody revealed cross-reactivity to DHS from P. vivax, despite an amino acid identity of 44% between the proteins. CONCLUSION: We identify a novel DHS protein in the more benign malaria parasite,P. vivax, on the basis of specific enzymatic activity, cross-reactivity with a polyclonal antibody against human DHS, and amino acid identity with DHS homologs from the rodent malaria parasite, P. yoelii, and human P. falciparum strains.


Assuntos
Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Plasmodium vivax/enzimologia , Proteínas de Protozoários/genética , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Western Blotting , Cromatografia de Afinidade/métodos , Clonagem Molecular , Reações Cruzadas/imunologia , Regulação Enzimológica da Expressão Gênica/genética , Humanos , Dados de Sequência Molecular , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/imunologia , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Fatores de Iniciação de Peptídeos/metabolismo , Plasmodium vivax/genética , Proteínas de Protozoários/imunologia , Proteínas de Protozoários/metabolismo , Proteínas de Ligação a RNA/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Fator de Iniciação de Tradução Eucariótico 5A
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