IDENTIFICATION OF CIS-ACTING ELEMENTS CONTROLLING GENE EXPRESSION IN S. neurona

Sarcocystis neurona is an apicomplexan parasite that is a major cause of equine protozoal myeloencephalitis (EPM). During intracellular development of S. neurona, many genes are temporally regulated. To better understand gene regulation, it is important to identify and characterize regulatory elements controlling gene expression in S. neurona. To perform this study, it was essential to establish transfection system for this parasite. Hence, the 5 flanking region of the SnSAG1 gene was isolated from a genomic library and used to construct expression plasmids. In transient assays, the reporter molecules -galactosidase (-gal) and yellow fluorescent protein (YFP) were expressed by electroporated S. neurona, thereby confirming the feasibility of performing molecular genetic experiments in this organism. Stable transformation of S. neurona was achieved using a mutant dihydrofolate reductase thymidylate synthase (DHFR-TS) gene of T. gondii that confers resistance to pyrimethamine. This selection system was used to create transgenic S. neurona that stably express -gal and YFP. These transgenic clones were shown to be useful for analyzing growth rate of parasites in-vitro and for assessing drug sensitivities. To uncover possible sequence elements involved in promoter activity, the 5 flanking regions of five S. neurona genes were subjected to comparative analysis. This revealed the presence of a 7-base conserved motif GCGTCTC. Using a dual luciferase assay system, the SnSAG1 promoter was subjected to functional analysis. The motif GAGACGC located between -136 and -129 upstream of the transcription start site was found to be essential for SnSAG1 expression. This motif functions in an orientation dependent manner and was shown to play a role in binding nuclear proteins of S. neurona

FEA BASED ANALYSIS OF SHACKLE FOR OFFSHORE APPLICATION

Nowadays Oil exploration in offshore area is responsible for increasingly greater percentage of production in the world. So mooring Systems play key role. Failure of a single component in a mooring line can produce incalculable damages. Due to environmental loading the mooring changes its position. Each component in the mooring are subjected to angular tensions from top and bottom. It produces strong non linearity in the components. Shackle is the critical part of this mooring system. Analysis of shackle is carried out by consideration of variation in angle from 1˚ to 5˚ in Plane and Out of Plane with ANSYS 12. An attempt has been made to analyze the effect of force angle on proof load. As the variation in angle increases from 1˚ to 5˚, proof load decreses. Validation of results are carried out by using horizontal UTM

Psychological Effects of Boko Haram Insurgencies in Some Liberated Communities across Yobe State, Nigeria

Millions of people have been exposed to Boko Haram insurgencies in the last decade, systematic studies on the mental health consequences in affected communities are still rare. This study investigates the psychological effects of the insurgency across several affected communities within Yobe state, Nigeria. Participants from five (5) war-affected local governments in Yobe state were recruited through a random-walk technique. In each of these regions, 3 affected localities were randomly selected, resulting in a selection of 15 localities across all 5 local governments. In each locality, 3 streets were randomly identified. Every fourth household was selected until a maximum of 5 interviews for one street were completed. The history of potentially traumatic experiences were assessed using a specifically amended version of the Life Stressor Checklist. The study shows a strong positive correlation between exposures to traumatic events as a result of the insurgency with the development of psychological stress. Over one third (36.01%) of respondents met symptom criteria for post-traumatic stress disorder (PTSD) and close to half (47.90%) of the respondents met symptom criteria for depression. Men were more exposed to trauma events, however women developed higher symptoms of psychological stress compared to men. Conclusively, the study provide evidence of mental health distress among populations exposed to Boko Haram insurgencies in Yobe state, Nigeria. Keywords: Boko Haram; Communities; Psychological Stress and Yobe State. DOI: 10.7176/DCS/11-7-04 Publication date:July 31st 202

Enterprise Systems Education: Where are We? Where are We Going?

Enterprise systems are used by companies worldwide. As the importance of enterprise systems has increased in the corporate world, so have their importance increased in IS education. As a result, enterprise systems education impacts the IS curriculum of many universities. The maturity of enterprise systems education has developed over the years, however, when compared to other maturity models, enterprise systems education has a long way to go. As part of its mission to support Information Systems (IS) education, JISE is pleased to publish this special issue devoted to enterprise systems education. A Teaching Tip, a Teaching Case, and eight papers on varying enterprise systems education topics are presented

Expression of the essential Kinase PfCDPK1 from Plasmodium falciparum in Toxoplasma gondii facilitates the discovery of novel antimalarial drugs

We have previously shown that genetic disruption of Toxoplasma gondii calcium-dependent protein kinase 3 (TgCDPK3) affects calcium ionophore-induced egress. We examined whether Plasmodium falciparum CDPK1 (PfCDPK1), the closest homolog of TgCDPK3 in the malaria parasite P. falciparum, could complement a TgCDPK3 mutant strain. PfCDPK1 is essential and plays critical roles in merozoite development, motility, and secretion. We show that expression of PfCDPK1 in the TgCDPK3 mutant strain rescues the egress defect. This phenotypic complementation requires the localization of PfCDPK1 to the plasma membrane and kinase activity. Interestingly, PfCDPK1-expressing Toxoplasma becomes more sensitive to egress inhibition by purfalcamine, a potent inhibitor of PfCDPK1 with low activity against TgCDPK3. Based on this result, we tested eight small molecules previously determined to inhibit the kinase activity of recombinant PfCDPK1 for their abilities to inhibit ionophore-induced egress in the PfCDPK1-expressing strain. While two of these chemicals did not inhibit egress, we found that six drugs affected this process selectively in PfCDPK1-expressing Toxoplasma. Using mutant versions of PfCDPK1 and TgCDPK3, we show that the selectivities of dasatinib and PLX-4720 are regulated by the gatekeeper residue in the ATP binding site. Importantly, we have confirmed that the three most potent inhibitors of egress in the PfCDPK1-expressing strain effectively kill P. falciparum. Thus, we have established and validated a recombinant strain of Toxoplasma that can be used as a surrogate for the discovery and analysis of PfCDPK1-specific inhibitors that can be developed as antimalarials

Identity and the absent mother in Atta's Everything Good will Come

Everything good will come presents the trope of the absent mother which scholars have identified as a significant feature of third generation Nigerian women prose fiction writings. Besides the trope of the absent mother, religion and identity also feature prominently in Atta's Everything good will come. This article harmonises these three dominant motifs in the narrative towards an examination of the complexity of identity formation in Everything good will come. The article focuses on Mike's sculptures as an artistic depiction of the dynamics that ultimately influence Enitan's identity formation. Due to the plurality of religious ideologies in the postcolonial Nigeria depicted in the narrative, the motifs of Christianity and traditional religion present in the narrative are explored towards illumination of key elements of the text. Christian motifs provide deeper comprehension of the dynamics that influence the relationship of Enitan and Sheri against the backdrop of the trope of the absent mother. Victoria and Enitan's characters and experiences find parallels in the being and characteristics of Ala, the Earth Goddess and Obatala

Phosphorylation of a Myosin Motor by TgCDPK3 Facilitates Rapid Initiation of Motility during Toxoplasma gondii egress

Members of the family of calcium dependent protein kinases (CDPK's) are abundant in certain pathogenic parasites and absent in mammalian cells making them strong drug target candidates. In the obligate intracellular parasite Toxoplasma gondii TgCDPK3 is important for calcium dependent egress from the host cell. Nonetheless, the specific substrate through which TgCDPK3 exerts its function during egress remains unknown. To close this knowledge gap we applied the proximity-based protein interaction trap BioID and identified 13 proteins that are either near neighbors or direct interactors of TgCDPK3. Among these was Myosin A (TgMyoA), the unconventional motor protein greatly responsible for driving the gliding motility of this parasite, and whose phosphorylation at serine 21 by an unknown kinase was previously shown to be important for motility and egress. Through a non-biased peptide array approach we determined that TgCDPK3 can specifically phosphorylate serines 21 and 743 of TgMyoA in vitro. Complementation of the TgmyoA null mutant, which exhibits a delay in egress, with TgMyoA in which either S21 or S743 is mutated to alanine failed to rescue the egress defect. Similarly, phosphomimetic mutations in the motor protein overcome the need for TgCDPK3. Moreover, extracellular Tgcdpk3 mutant parasites have motility defects that are complemented by expression of S21+S743 phosphomimetic of TgMyoA. Thus, our studies establish that phosphorylation of TgMyoA by TgCDPK3 is responsible for initiation of motility and parasite egress from the host-cell and provides mechanistic insight into how this unique kinase regulates the lytic cycle of Toxoplasma gondii

Cell cycle-dependent, intercellular transmission of Toxoplasma gondii is accompanied by marked changes in parasite gene expression

Intracellular microbes have evolved efficient strategies for transitioning from one cell to another in a process termed intercellular transmission. Here we show that host cell transmission of the obligate intracellular parasite Toxoplasma gondii is closely tied to specific cell cycle distributions, with egress and reinvasion occurring most proficiently by parasites in the G1 phase. We also reveal that Toxoplasma undergoes marked changes in mRNA expression when transitioning from the extracellular environment to its intracellular niche. These mRNA level changes reflect a modal switch from expression of proteins involved in invasion, motility and signal transduction in extracellular parasites to expression of metabolic and DNA replication proteins in intracellular parasites. Host cell binding and signalling associated with the discharge of parasite secretory proteins was not sufficient to induce this switch in gene expression, suggesting that the regulatory mechanisms responsible are tied to the establishment of the intracellular environment. The genes whose expression increased after parasite invasion belong to a progressive cascade known to underlie the parasite division cycle indicating that the unique relationship between the G1 phase and invasion effectively synchronizes short-term population growth. This work provides new insight into how this highly successful parasite competently transits from cell to cell.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/79311/1/MMI_7441_sm_FigS1-2_TableS6-8.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/79311/2/j.1365-2958.2010.07441.x.pd

Influence of impeller blade rounding and surface roughness on the internal hydraulics and performance of pump as turbine

The Pump As Turbine (PAT) is an important technology for low-cost micro-hydropower and energy recovery, and hence the internal hydraulics of PAT needs to be clearly understood. Additionally, during its operation, the sediments in the water increase the roughness of the internal surfaces and may alter the internal hydraulics and PAT performance similar to a centrifugal pump or Francis turbine. The researchers tried hard to perform simple modifications such as impeller blade rounding to increase the efficiency of PAT. In this paper, the developed test rig is used to analyze the performance of the impeller blade rounding and is validated with a numerical model. This numerical model is further used to study the influence of impeller blade rounding and surface roughness on internal hydraulics and PAT performance. The impeller blade rounding at the most increased the PAT efficiency by 1-1.5 % at the Best efficiency point (Q=16.8 lps), mainly due to the wake reduction on the suction side and increased flow area. With increasing the surface roughness from 0-70 μm, the PAT efficiency is decreased maximum by 4 %. The efficiency was mainly reduced due to increased hydraulic losses at flow zone and disk friction losses at the non-flow zone

Beta-trace protein as a marker of renal dysfun -ction in patients with chronic kidney disease: comparison with other renal markers.

Kratak sadr`aj: Beta-trace protein (BTP), tako|e poznat kao prostaglandin D sintaza, je protein male molekulske mase koji pripada familiji lipokalinskih proteina. Prona|eno je da je pove}an u serumu pacijenata sa renalnim bolestima. Cilj ovog rada je bio da se uporedi klini~ka korisnost serumskih nivoa beta-trace proteina u detekciji renalne disfunkcije kod pacijenata sa hroni~nom bole{}u bubrega (CKD) sa nivoima drugih renalnih markera: krea tinina, cistatina C i b 2 -mikroglo bu lina (B2M). Studija je uklju~ila 134 pacijenta sa {irokim opsegom renalne disfunkcije koji obuhvata svih 5 CKD stadijuma. Dobijeni podaci su pokazali da beta-trace pro tein veoma dobro koreli{e (Spearmanov test) sa kreatininom (r = 0,890), cistatinom C (r = 0,904) i B2M (r = 0,933) i da se njegovi nivoi u serumu zna~ajno pove }avaju od (1. do 5.) CKD stadijuma. Osim to ga, vrednosti ja~ine glomerulske filtracije (GFR) procenjene iz formule ba zirane na BTP zna~ajno koreli{u sa GFR izra~unatom iz formula baziranih na kreatininu i cistatinu C. ROC ana liza je pokazala da BTP ima sli~nu dijagnosti~ku ta~nost za detekciju redukovane renalne funkcije u CKD stadiju mima kao drugi renalni markeri, pri procenjenim GFR od < 30, < 60 i < 90 mL/min/1,73 m 2 . Povr{ine ispod ROC krivih (AUC) za BTP, za ove nivoe GFR, bile su od 0,983 do 0,917 i one se nisu zna~ajno razlikovale od AUC za druge renalne markere. Rezultati ove studije su pokazali da BTP mo`e biti koristan i pouzdan serumski marker za identi fi kaciju obima renalne disfunkcije kod pacijenata sa CKD i mo`e se koristiti uz serumski cistatin C i kreatinin kao alternativni endogeni GFR marker. Klju~ne re~i: beta-trace protein, kreatinin, cistatin C, b 2 -mikroglobulin, hroni~na bolest bubrega Summary: Beta-trace protein (BTP), also known as prostaglandin D synthase, is a low-molecular-mass protein which belongs to the lipocalin protein family. It was found to be increased in the serum of patients with renal diseases. The aim of this study was to compare the clinical usefulness of serum levels of beta-trace protein for the detection of renal dysfunction in patients with chronic kidney disease (CKD) with levels of other renal markers: creatinine, cystatin C and b 2 -microglobulin (B2M). The study included 134 patients with a wide range of renal dysfunction that encom passed all five CKD stages. Obtained data showed that beta-trace protein highly correlated (Spearman test) with creatinine (r = 0.890), cystatin C (r = 0.904) and B2M (r = 0.933) and its levels in serum significantly increased from CKD stage 1 to 5. Furthermore, the values of glomerular filtra tion rate (GFR) estimated from a BTP-based formula signi fi cantly correlated with GFR calculated from creatinine-based and cystatin Cbased formulas. ROC analyses showed that BTP had similar diagnostic accuracy for detection of reduced renal function in CKD stages as other renal markers, for estimated GFRs of < 30, < 60 and < 90 mL/min/1.73 m 2 . The areas under the ROC curves (AUC) for BTP, for these GFR limits, were from 0.983 to 0.917 and they were not significantly different from AUCs for other renal markers. The results of this study showed that BTP may be a useful and reliable serum marker for identifying the magnitude of renal dysfunction in patients with CKD and may have its place be si de serum cystatin C and creatinine as an alternative endog e nous GFR marker