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The electron-nuclear Overhauser effect on spin ½ nuclei in solutions containing free radicals has been calculated from a method based on treatment used by Solomon. The magnitude and sign of the enhancement of the nuclear signal, which may be represented by a factor ρ depends on the nature and time dependence of the the electron-nuclear signal, which is assumed to be either dipolar or scalar. The time dependence for the dipole-dipole interactions is assumed to arise from the translational diffusion of the free radicals and the molecule containing the nuclei. three models are employed for the scalar interactions. In the first of these, termed the Sticking Model, the scalar interaction is assumed to be finite only while a molecule and free radical are "stuck" together. Thus sticking time is assumed to be short in comparison with the diffusional correlation time. However, an alternative model in which this assumption is not made has been postulated to explain certain experimental results. This is termed the Modified Sticking Model. In the third model, called the Diffusion Model, the hypothesis is made that the scalar interaction is a very short range interaction with magnitude ℏ B(d/R) exp. [-λ(r-d)]. where λ d ≫ 1, B and λ are constants, R is the distance between a nucleus and an electron, and d is the distances of the closest approach, inside of which the interaction is zero. In this model the time dependence of the scalar interaction, as in the case of the dipolar interactions, arises from the relative diffusion of the molecules and free radicals in the solution. For all the models of the scalar interaction, ρ may be plotted as a function of wsτc where ws is the electron Larmor frequency and τc the correlation time and it is found that the sign of ρ may change. The product wsτc may be altered by working at different magnetic fields or changing ρc. From measurements, at two different magnetic fields, of the nuclear spin-lattice relaxation times, the concentration of free radical in the solution and ρ, the theories allow the calculation of the mean diffusion coefficient of the radicals and molecules in solution, the correlation times and the measure of the relative importance of scalar and dipolar interactions. Experiments on fluorinated and protonic solvents containing the radical tri-tert-butyl phenoxyl (T.T.B.P) have been carried out at two different magnetic fields. The proton-free radical interactions may be explained in terms of dipole-dipole interactions. On the other hand with fluorine nuclei scalar coupling does seem to be important. The Sticking Model does not successfully describe the effect of this coupling. The other models give reasonable values for the calculable parameters, but it is difficult to decide unambiguously between them. If scalar coupling does occur, a scalar shift of the nuclear resonance is expected. this has been observed for hexaflourobenzene solutions containing T.T.B.P. The theoretical and experimental results are compared, and while either theory might explain the observed shift, less assumptions have to be made in using the Diffusion Model. The correlation time dependence of the Overhauser effect has also been studied for two systems, hexafluorobenzene and 1-1-1-trichloro-2-2-2-trifluoro-ethane, at two magnetic fields. The correlation times were altered by addition of carbon disulphide or a chlorinated hydrocarbon or by changing the temperature of the sample. Qualitatively the Diffusion Model was in best agreement with the experimental results, but quantitatively, neither model was satisfactory. In the Diffusion Model, the assumption λd≫1 is made and in an attempt to obtain better agreement with the experimental results ρ has been calculated using different values of λd in thescalar interaction function. The best agreement between experiment and theory was for values of λd≫ 1000. Measurements of the proton electron Overhauser effect can usually be interpreted in terms of dipolar interactions (positive ρ) between the electrons and the protons. However for the resonances of the tert-butyl groups of tri-tert-butyl phenol in solutions of T.T.B.P. ρ is negative. This has been shown to be associated with the proton exchange between the radical and phenol molecules. It is postulated that in the radical molecule the scalar coupling between the electron and the tert-buyl protons, occurs at least in part by a hyperconjugative mechanism. the random rotation of these tert-butyl groups could modulate this scalar coupling. This would lead to a positive polarization of the tert-butyl protons, which persists when the radical accepts a proton and becomes a tri-tert-butyl phenol molecule.

Proteomics can help to discover and quantify novel biomarkers and we show how this is achieved using liver fibrosis as an example. Liver biopsy is the reference standard for assessing liver fibrosis and serum biomarkers are less invasive. The severity of liver fibrosis can be determined using immunoassays. However biomarkers may be degraded due to sample storage conditions and therefore may not be detected using these antibody-based assays. Detection of biomarkers by mass spectrometry overcomes this disadvantage and the approach can be applied to all diseases. Two dimensional gel electrophoresis (2DE) was used to find differences in proteins in plasma samples from patients with varying liver fibrosis stages. The identified proteins were potential liver fibrosis biomarkers. Mass spectrometry can assay for biomarkers by detecting their tryptic peptides and fragments and was used to develop an antibody-free assay. We are the first and only lab in the UK to use a novel absolute quantitation method which is the only biomarker assay using a universal calibration mix. Using 2DE, we identified several potential biomarkers for liver fibrosis which were analysed by Western blotting using plasma samples from patients with varying stages of liver fibrosis. Our novel biomarkers were promising when compared to the markers used in current liver fibrosis tests. A mass spectrometry assay was developed for the best novel liver fibrosis biomarker. We are working towards the first ever antibody-free biomarker assay for liver fibrosis. Our assay is nine times faster than conventional quantitation by mass spectrometry making our approach for absolute biomarker quantitation applicable for clinical use. This is also the only assay which can analyse all points of the calibration curve and determine the absolute concentration of the biomarker in a single acquisition. Our assay may help reduce the need for invasive liver biopsies and the approach could be used for any other disease.

Immunologically self carbohydrates protect the human immunodeficiency virus type -1 (HIV-1) surface glycoprotein, gp120 from antibody recognition. However, one broadly neutralising antibody, 2G12, can protect against primary viral challenge by direct recognition of these "self" glycans on gp120.

The morphogenesis of HCV belongs to steps of the viral cycle that have not yet been targeted by antiviral strategies. With the lack of an efficient and reliable culture system able to produce and secrete HCV virions, we use two complementary approaches to study the effect of c~-gluensidase inhibitors on HCV morphogenesis, secretion and entry. These two approaches have allowed us to demonstrate tile mechanism of tile antiviral effect of O,-glucosidase inhibitors on HCV glyenprotein folding and assembly, morphogenesis, viral secretion, and viral infectivity.

The antiviral properties of iminosugars have been reported previously in vitro and in small animal models against Ebola virus (EBOV); however, their effects have not been tested in larger animal models such as guinea pigs. We tested the iminosugars N-butyl-deoxynojirimycin (NB-DNJ) and N-(9- methoxynonyl)-1deoxynojirimycin (MON-DNJ) for safety in uninfected animals, and for antiviral efficacy in animals infected with a lethal dose of guinea pig adapted EBOV. 1850 mg/kg/day NB-DNJ and 120 mg/kg/day MON-DNJ administered intravenously, three times daily, caused no adverse effects and were well tolerated. A pilot study treating infected animals three times within an 8 hour period was promising with 1 of 4 infected NB-DNJ treated animals surviving and the remaining three showing improved clinical signs. MON-DNJ showed no protective effects when EBOV-infected guinea pigs were treated. On histopathological examination, animals treated with NB-DNJ had reduced lesion severity in liver and spleen. However, a second study, in which NB-DNJ was administered at equally- spaced 8 hour intervals, could not confirm drug-associated benefits. Neither was any antiviral effect of iminosugars detected in an EBOV glycoprotein pseudotyped virus assay. Overall, this study provides evidence that NB-DNJ and MON-DNJ do not protect guinea pigs from a lethal EBOV-infection at the dose levels and regimens tested. However, the one surviving animal and signs of improvements in three animals of the NB-DNJ treated cohort could indicate that NB-DNJ at these levels may have a marginal beneficial effect. Future work could be focused on the development of more potent iminosugars

Provided are novel iminosugars and methods of treating and/or preventing a disease or condition caused by or associated with a virus belonging to the Orthomyxoviridae family using iminosugars, such as DNJ derivatives.

The inventors have proposed a novel panel of human serum protein biomarkers for diagnosing hepatic fibrosis and cirrhosis. Presently there is no reliable non-invasive way of assessing liver fibrosis. A 2D-PAGE based proteomics study was used to identify potential fibrosis biomarkers. Serum from patients with varying degrees of hepatic scarring induced by infection with the hepatitis C virus (HCV) was analysed. Several proteins associated with liver scarring and/or viral infection were identified. These proteins include the inter-alpha-trypsin inhibitor heavy chain H4 fragments, complement factor H-related protein 1, CD5L, Apo L1, and beta2GPI. Increased and decreased thiolester cleavage of a2M and Complement C3, respectively, was also detected.; The concentrations of these novel biomarkers can be determined using an immunoassay where the concentrations would reflect the extent of fibrosis. A fibrosis scoring scale for each of the novel biomarkers is proposed. The additive result from the scores of all the novel biomarkers would give a more reliable indication of the degree of fibrosis rather than examining individual biomarkers.

Provided are methods of treating or preventing viral infections caused by or associated with a Dengue virus using iminosugars.

Introduction: Liver biopsy is the reference standard for assessing liver fibrosis, and no reliable non-invasive diagnostic approach is available to discriminate between the intermediate stages of fibrosis. Therefore suitable serological biomarkers of liver fibrosis are urgently needed.Aim: The objective of this study was to use proteomics to identify novel fibrosis biomarkers in hepatitis C patients with different degrees of liver fibrosis.Method: Proteins in plasma samples from healthy control individuals and patients with hepatitis C virus (HCV) induced cirrhosis were analysed using a proteomics technique—two dimensional gel electrophoresis (2-DE). This technique separated the proteins in plasma samples of control and cirrhotic patients and by visualising the separated proteins we were able to identify proteins which were increasing or decreasing in hepatic cirrhosis. Identified markers were validated across all Ishak stages and compared to the markers used in FibroTest, Enhanced Liver Fibrosis (ELF) test, Hepascore and FIBROSpect by western blotting.Results: 44 candidate biomarkers for hepatic fibrosis were identified of which 20 were novel. western blot validation of all candidate markers using plasma samples from patients across all Ishak fibrosis scores showed that the markers which changed with increasing fibrosis most consistently included lipid transfer inhibitor protein, complement C3d, corticosteroid-binding globulin, apolipoprotein J and apolipoprotein L1. These five novel markers which are secreted in blood showed a promising consistent change with increasing fibrosis stage when compared to the markers used for the FibroTest, ELF test, Hepascore and FIBROSpect.Conclusion: This study identifies 20 novel fibrosis biomarker candidates. The proteins identified by these improved approaches may help to assess hepatic fibrosis and eliminate the need for invasive liver biopsies.

The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Poxviridae family.

The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Filoviridae family.

The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Bunyaviral or Togaviral family.

The present application relates to iminosugars and methods of treating viral infections with iminosugars and, in particular, to the use of iminosugars for treatment and/or prevention of viral infections caused by or associated with a virus belonging to the Arenaviridae family.

The inventors have proposed a novel panel of human plasma protein biomarkers for diagnosing hepatic fibrosis and cirrhosis. Presently there is no reliable non-invasive way of assessing liver fibrosis. A 2D-PAGE based proteomics study was used to identify potential fibrosis biomarkers. Plasma from patients with hepatic cirrhosis induced by infection with the hepatitis C virus (HCV) were analysed. Several proteins associated with liver scarring and potentially also related to viral infection were identified. These proteins include 14-3-3 protein zeta/delta, adiponectin, afamin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, apolipoprotein C-III, apolipoprotein E, C4b-binding protein beta chain, intact/cleaved complement C3dg, corticosteroid-binding globulin, fibrinogen gamma chain, beta haptoglobin at pH 5.46-5.49, haptoglobin-related protein, hemopexin, immunoglobulin J chain, leucine-rich alpha-2-glycoprotein, lipid transfer inhibitor protein, retinol-binding protein 4, serum paraoxonase/arylesterase 1, sex hormone-binding globulin and zinc-alpha-2-glycoprotein. These biomarkers can be used in conjunction with polypeptides in WO/2008/031051. The concentrations of these novel biomarkers can be determined using an immunoassay where the concentrations would reflect the extent of fibrosis. A fibrosis scoring scale for each of the novel biomarkers is proposed. The additive result from the scores of all the novel biomarkers would give a more reliable indication of the degree of fibrosis rather than examining individual biomarkers.

Disclosed are compositions and methods useful for inducing an immunogenic response in a subject or host. In particular, the compositions and methods may be directed to carbohydrate HIV vaccines and to methods of producing a carbohydrate HIV vaccine by introducing antigenic sugars into mimics of the glycans of the HIV envelope glycoproteins gp120 and gp41.

The Hepatitis C virus (HCV) genome encodes a polyprotein that is processed co- and posttranslationally. Incomplete processing at the E2/p7 junction generates the E2p7 product. Using a recombinant system, we analysed the processing, localization and topology of E2p7. By immunoprecipitation of proteins expressed by metabolically labelled cells, we confirm that E2p7 is a precursor of E2. E2p7 forms a native-like heterodimer with E1, and it is localized entirely to the endoplasmic reticulum, in contrast to fully processed E2 and p7 that leak to the plasma membrane. No change in the topology of p7 was observed upon processing of E2p7, indicating that incomplete cleavage at the E2/p7 site is not regulated by changes in p7 membrane topology.

The present application relates to treatment of tumors and, more particularly, to treatment of tumors with iminosugars.

Novel anti-HCV molecules are still needed to better combat HCV infection. Molecules targeting specifically viral activities are the most attractive in terms of drug development and are therefore the most studied. The antiviral properties of glucosidase inhibitors were established using the most relevant models currently available.

Disclosed are methods and kits to treat HCV infection by administering an iminosugar derivative compound that is effective to inhibit the activity of HCV p7 protein, and methods by which to screen for compounds that inhibit the activity of p7 protein or variants thereof. The disclosed N-substituted imino compounds, and pharmaceutical compositions thereof, inhibit the capability of HCV p7 to permeabilize membranes. Particularly efficacious compounds are imino sugars derived from N-alkylated piperidines. Also disclosed are methods for screening for potential HCV antiviral agents.

The proportion of oligosaccharide chains on the Fc fragment of IgG which terminate with N-acetylglucosamine and not galactose (%GO) has previously been shown to be raised in rheumatoid arthritis (RA), Crohn's disease (CD) and tuberculosis (Tb), but to be normal in sarcoidosis (SA), and in both lepromatous and tuberculoid leprosy. However we have now studied %GO in sequential serum samples collected from lepromatous leprosy patients undergoing episodes of erythema nodosum leprosum (ENL). During ENL %GO is transiently raised, and this rise parallels an increase in circulating interleukin 2 receptors (IL-2R). These findings confirm that changes in T cell function occur during ENL. Moreover it appears that %GO rises when there is, simultaneously, T-cell-mediated tissue damage and an acute phase response (RA, CD, Tb, ENL), but not when there is an acute phase response without major T cell involvement, or chronic T cell activity alone (SA, and tuberculoid leprosy). We suggest therefore that %GO is an indicator of a type of T cell activity with broad immunopathological implications.

Two-dimensional gel electrophoresis (2-DE) is often used to separate plasma or serum proteins in an attempt to identify novel disease biomarkers. A major problem with this approach is the presence of high abundant plasma/serum proteins which limit the detection of low abundance features. We used two proteomic approaches to identify new fibrosis biomarkers in patients with different stages of liver fibrosis. Plasma samples from healthy individuals and patients with hepatitis C virus (HCV) induced cirrhosis were analysed using 2-DE over a narrow pH 3-5.6 range, arrange outside the pH of highly abundant albumin, transferrin and immunoglobulins. Several novel markers were identified by this approach and were validated across all fibrosis stages by Western blotting. This is the first time the pH 3-5.6 range has been used to separate plasma by 2-DE and was found to be beneficial for biomarker discovery. In addition, we used in-solution isoelectric focusing followed by SDS- PAGE to find biomarkers in HCV-induced liver cirrhosis. This approach was found to be beneficial for identifying basic, high molecular weight protein biomarkers. These approaches have successfully helped to identify novel biomarkers for hepatic fibrosis and these approaches are also useful for discovering novel biomarkers in other diseases.

Long chain N-alkyl amino and imino compounds, oxa-substituted derivatives thereof, and pharmaceutical compositions including such compounds are described. The long chain N-alkyl group is a C¿8?-C¿16? alkyl group. The long chain N-alkyl compounds and oxa-substituted derivatives thereof can be used in the treatment of viral infections, in particular hepatitis B virus or hepatitis C virus, in a cell or an individual. For example, the long chain N-alkyl compounds or oxa-substituted derivatives thereof can be derived from piperidines, pyrrolidines, phenylamines, pyridines, pyrroles, or amino acids.

Methods for inhibiting morphogenesis of host cell membrane-budding viruses and infections caused thereby using compounds that inhibit host cell glucosidase or glucosyltransferase enzymes. Methods for treating lipid storage diseases using compounds that inhibit glucosyltransferase enzymes.

Two-dimensional gel electrophoresis (2-DE) is often used to separate plasma or serum proteins in an attempt to identify novel biomarkers. A major difficulty with this approach is due to high abundant plasma/serum proteins which limits the detection of low abundance features. To overcome this problem a novel proteomics approach was developed and used to identify new fibrosis biomarkers in patients with different stages of liver fibrosis. Plasma samples from healthy individuals and patients with hepatitis C virus (HCV) induced cirrhosis were analysed using 2-DE over a narrow pH 3-5.6 range, a range outside the pH of highly abundant albumin, transferrin and immunoglobulins. Novel markers identified by this approach were validated across all fibrosis stages by Western blotting. 44 candidate biomarkers were revealed of which 20 were novel. Western blot analysis with newly identified biomarkers showed a consistent change with increasing fibrosis stage and were promising when compared to the markers used in established fibrosis tests. This is the first time the pH 3−5.6 range has been used to separate plasma by 2-DE. This pH range is useful for discovering novel biomarkers in all diseases. The novel fibrosis markers identified by this new proteomics approach may help to assess hepatic fibrosis and eliminate the need for invasive liver biopsies.