5,753 research outputs found

    A description of the life stages of Echinoparyphium elegans (Trematoda: Echinostomatidae)

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    The life cycle of Echinoparyphium elegans Looss 1899 is described from the Free State, South Africa.The freshwater snail Bulinus tropicus (Krauss 1848), the intermediate host of Calicophorort microbothrium (Paramphistomum microbothrium Fischoeder, 1901) in this area, serves as first intermediate host. The same snail species also harbours the metacercaria stage. Adults of this parasite were obtained by feeding infected snails to laboratory-reared rats. All stages of the life cycle were described by means of light and scanning electron microscopy. A natural infection was found in the cattle egret, Bubulcus ibis (Linnaeus 1758)

    Elucidating the Role of Topological Constraint on the Structure of Overstretched DNA Using Fluorescence Polarization Microscopy

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    The combination of DNA force spectroscopy and polarization microscopy of fluorescent DNA intercalator dyes can provide valuable insights into the structure of DNA under tension. These techniques have previously been used to characterize S-DNA—an elongated DNA conformation that forms when DNA overstretches at forces ≥ 65 pN. In this way, it was deduced that the base pairs of S-DNA are highly inclined, relative to those in relaxed (B-form) DNA. However, it is unclear whether and how topological constraints on the DNA may influence the base-pair inclinations under tension. Here, we apply polarization microscopy to investigate the impact of DNA pulling geometry, torsional constraint, and negative supercoiling on the orientations of intercalated dyes during overstretching. In contrast to earlier predictions, the pulling geometry (namely, whether the DNA molecule is stretched via opposite strands or the same strand) is found to have little influence. However, torsional constraint leads to a substantial reduction in intercalator tilting in overstretched DNA, particularly in AT-rich sequences. Surprisingly, the extent of intercalator tilting is similarly reduced when the DNA molecule is negatively supercoiled up to a critical supercoiling density (corresponding to ∼70% reduction in the linking number). We attribute these observations to the presence of P-DNA (an overwound DNA conformation). Our results suggest that intercalated DNA preferentially flanks regions of P-DNA rather than those of S-DNA and also substantiate previous suggestions that P-DNA forms predominantly in AT-rich sequences

    A mechanistic modelling approach for the determination of the mechanisms of inhibition by cyclosporine on the uptake and metabolism of atorvastatin in rat hepatocytes using a high throughput uptake method

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    (1) Determine the inhibition mechanism through which cyclosporine inhibits the uptake and metabolism of atorvastatin in fresh rat hepatocytes using mechanistic models applied to data generated using a high throughput oil spin method. (2) Atorvastatin was incubated in fresh rat hepatocytes (0.05–150 nmol/ml) with or without 20 min pre-incubation with 10 nmol/ml cyclosporine and sampled over 0.25–60 min using a high throughput oil spin method. Micro-rate constant and macro-rate constant mechanistic models were ranked based on goodness of fit values. (3) The best fitting model to the data was a micro-rate constant mechanistic model including non-competitive inhibition of uptake and competitive inhibition of metabolism by cyclosporine (Model 2). The association rate constant for atorvastatin was 150-fold greater than the dissociation rate constant and 10-fold greater than the translocation into the cell. The association and dissociation rate constants for cyclosporine were 7-fold smaller and 10-fold greater, respectively, than atorvastatin. The simulated atorvastatin-transporter-cyclosporine complex derived using the micro-rate constant parameter estimates increased in line with the incubation concentration of atorvastatin. (4) The increased amount of data generated with the high throughput oil spin method, combined with a micro-rate constant mechanistic model helps to explain the inhibition of uptake by cyclosporine following pre-incubation

    Unravelling the mechanisms of Type 1A topoisomerases using single-molecule approaches

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    Topoisomerases are essential enzymes that regulate DNA topology. Type 1A family topoisomerases are found in nearly all living organisms and are unique in that they require single-stranded (ss)DNA for activity. These enzymes are vital for maintaining supercoiling homeostasis and resolving DNA entanglements generated during DNA replication and repair. While the catalytic cycle of Type 1A topoisomerases has been long-known to involve an enzyme-bridged ssDNA gate that allows strand passage, a deeper mechanistic understanding of these enzymes has only recently begun to emerge. This knowledge has been greatly enhanced through the combination of biochemical studies and increasingly sophisticated single-molecule assays based on magnetic tweezers, optical tweezers, atomic force microscopy and Förster resonance energy transfer. In this review, we discuss how single-molecule assays have advanced our understanding of the gate opening dynamics and strand-passage mechanisms of Type 1A topoisomerases, as well as the interplay of Type 1A topoisomerases with partner proteins, such as RecQ-family helicases. We also highlight how these assays have shed new light on the likely functional roles of Type 1A topoisomerases in vivo and discuss recent developments in single-molecule technologies that could be applied to further enhance our understanding of these essential enzymes

    The effect of combined supplementation of carbohydrates and creatine on anaerobic performance.

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    The purpose of the study was to examine the effect of creatine (Cr) supplementation on anaerobic performance when ingesting creatine and carbohydrates (CHO) together. Twenty male physical education students comprised the two experimental (CR and CRCHO) and one control (CON) groups of the study. All groups performed three 30 s anaerobic Wingate tests (AWTs) interspersed with 6 minutes of recovery. The CR group (n = 7) ingested 5 g of Cr 5 times per day for 4 days. Subjects in the CRCHO group (n = 6) ingested the same quantity but additionally after each 5 g dose of Cr consumed 500 ml of a commercially available energy drink containing 100 g of simple sugars. Over all three AWTs average mean power improved significantly compared to baseline for the CR group (5.51%) but not for the CRCHO group (3.06%). Mean power for the second AWT was improved following the acute loading for the CR group only (4.54%) and for the third AWT for both CR (8.49%) and CRCHO (5.75%) groups. Over all three AWTs a significant change was recorded in average peak power following the acute loading for the CR group (8.26%) but not for the CRCHO group (4.11%). Peak power was significantly improved following the loading only for the CR group during the third AWT (19.79%). No changes in AWT performance were recorded for the CON group after intervention. The findings of the present study suggest that ingesting creatine together with carbohydrates will not further improve performance compared to the ingestion of creatine only

    Universality of electron accumulation at wurtzite c- and a-plane and zinc-blende InN surfaces

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    Electron accumulation is found to occur at the surface of wurtzite (112¯0), (0001), and (0001¯) and zinc-blende (001) InN using x-ray photoemission spectroscopy. The accumulation is shown to be a universal feature of InN surfaces. This is due to the low Г-point conduction band minimum lying significantly below the charge neutrality level

    P2X receptors: epithelial ion channels and regulators of salt and water transport.

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    When the results from electrophysiological studies of renal epithelial cells are combined with data from in vivo tubule microperfusion experiments and immunohistochemical surveys of the nephron, the accumulated evidence suggests that ATP-gated ion channels, P2X receptors, play a specialized role in the regulation of ion and water movement across the renal tubule and are integral to electrolyte and fluid homeostasis. In this short review, we discuss the concept of P2X receptors as regulators of salt and water salvage pathways, as well as acknowledging their accepted role as ATP-gated ion channels
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