Characterization of seed nuclei in glucagon aggregation using light scattering methods and field-flow fractionation

<p>Abstract</p> <p>Background</p> <p>Glucagon is a peptide hormone with many uses as a therapeutic agent, including the emergency treatment of hypoglycemia. Physical instability of glucagon in solution leads to problems with the manufacture, formulation, and delivery of this pharmaceutical product. Glucagon has been shown to aggregate and form fibrils and gels <it>in vitro</it>. Small oligomeric precursors serve to initiate and nucleate the aggregation process. In this study, these initial aggregates, or seed nuclei, are characterized in bulk solution using light scattering methods and field-flow fractionation.</p> <p>Results</p> <p>High molecular weight aggregates of glucagon were detected in otherwise monomeric solutions using light scattering techniques. These aggregates were detected upon initial mixing of glucagon powder in dilute HCl and NaOH. In the pharmaceutically relevant case of acidic glucagon, the removal of aggregates by filtration significantly slowed the aggregation process. Field-flow fractionation was used to separate aggregates from monomeric glucagon and determine relative mass. The molar mass of the large aggregates was shown to grow appreciably over time as the glucagon solutions gelled.</p> <p>Conclusion</p> <p>The results of this study indicate that initial glucagon solutions are predominantly monomeric, but contain small quantities of large aggregates. These results suggest that the initial aggregates are seed nuclei, or intermediates which catalyze the aggregation process, even at low concentrations.</p

Population pharmacokinetics of artesunate and dihydroartemisinin following single- and multiple-dosing of oral artesunate in healthy subjects

<p>Abstract</p> <p>Background</p> <p>The population pharmacokinetics of artesunate (AS) and its active metabolite dihydroartemisinin (DHA) were studied in healthy subjects receiving single- or multiple-dosing of AS orally either in combination with pyronaridine (PYR) or as a monotherapy with or without food.</p> <p>Methods</p> <p>Data from 118 concentration-time profiles arising from 91 healthy Korean subjects were pooled from four Phase I clinical studies. Subjects received 2-5 mg/kg of single- and multiple-dosing of oral AS either in combination with PYR or as a monotherapy with or without food. Plasma AS and DHA were measured simultaneously using a validated liquid chromatography- mass spectrometric method with a lower limit of quantification of 1 ng/mL for both AS and DHA. Nonlinear mixed-effect modelling was used to obtain the pharmacokinetic and variability (inter-individual and residual variability) parameter estimates.</p> <p>Results</p> <p>A novel parent-metabolite pharmacokinetic model consisting of a dosing compartment, a central compartment for AS, a central compartment and a peripheral compartment for DHA was developed. AS and DHA data were modelled simultaneously assuming stoichiometric conversion to DHA. AS was rapidly absorbed with a population estimate of absorption rate constant (Ka) of 3.85 h^-1. The population estimates of apparent clearance (CL/F) and volume of distribution (V2/F) for AS were 1190 L/h with 36.2% inter-individual variability (IIV) and 1210 L with 57.4% IIV, respectively. For DHA, the population estimates of apparent clearance (CLM/F) and central volume of distribution (V3/F) were 93.7 L/h with 28% IIV and 97.1 L with 30% IIV, respectively. The population estimates of apparent inter-compartmental clearance (Q/F) and peripheral volume of distribution (V4/F) for DHA were 5.74 L/h and 18.5 L, respectively. Intake of high-fat and high-caloric meal prior to the drug administration resulted in 84% reduction in Ka. Body weight impacted CLM/F, such that a unit change in weight resulted in 1.9-unit change in CLM/F in the same direction.</p> <p>Conclusions</p> <p>A novel simultaneous parent-metabolite pharmacokinetic model with good predictive power was developed to study the population pharmacokinetics of AS and DHA in healthy subjects following single- and multiple-dosing of AS with or without the presence of food. Food intake and weight were significant covariates for Ka and CLM/F, respectively.</p

Toward the Restoration of Hand Use to a Paralyzed Monkey: Brain-Controlled Functional Electrical Stimulation of Forearm Muscles

Loss of hand use is considered by many spinal cord injury survivors to be the most devastating consequence of their injury. Functional electrical stimulation (FES) of forearm and hand muscles has been used to provide basic, voluntary hand grasp to hundreds of human patients. Current approaches typically grade pre-programmed patterns of muscle activation using simple control signals, such as those derived from residual movement or muscle activity. However, the use of such fixed stimulation patterns limits hand function to the few tasks programmed into the controller. In contrast, we are developing a system that uses neural signals recorded from a multi-electrode array implanted in the motor cortex; this system has the potential to provide independent control of multiple muscles over a broad range of functional tasks. Two monkeys were able to use this cortically controlled FES system to control the contraction of four forearm muscles despite temporary limb paralysis. The amount of wrist force the monkeys were able to produce in a one-dimensional force tracking task was significantly increased. Furthermore, the monkeys were able to control the magnitude and time course of the force with sufficient accuracy to track visually displayed force targets at speeds reduced by only one-third to one-half of normal. Although these results were achieved by controlling only four muscles, there is no fundamental reason why the same methods could not be scaled up to control a larger number of muscles. We believe these results provide an important proof of concept that brain-controlled FES prostheses could ultimately be of great benefit to paralyzed patients with injuries in the mid-cervical spinal cord

Imaging Primary Mouse Sarcomas After Radiation Therapy Using Cathepsin-Activatable Fluorescent Imaging Agents

Purpose: Cathepsin-activated fluorescent probes can detect tumors in mice and in canine patients. We previously showed that these probes can detect microscopic residual sarcoma in the tumor bed of mice during gross total resection. Many patients with soft tissue sarcoma (STS) and other tumors undergo radiation therapy (RT) before surgery. This study assesses the effect of RT on the ability of cathepsin-activated probes to differentiate between normal and cancerous tissue. Methods and Materials: A genetically engineered mouse model of STS was used to generate primary hind limb sarcomas that were treated with hypofractionated RT. Mice were injected intravenously with cathepsin-activated fluorescent probes, and various tissues, including the tumor, were imaged using a hand-held imaging device. Resected tumor and normal muscle samples were harvested to assess cathepsin expression by Western blot. Uptake of activated probe was analyzed by flow cytometry and confocal microscopy. Parallel in vitro studies using mouse sarcoma cells were performed. Results: RT of primary STS in mice and mouse sarcoma cell lines caused no change in probe activation or cathepsin protease expression. Increasing radiation dose resulted in an upward trend in probe activation. Flow cytometry and immunofluorescence showed that a substantial proportion of probe-labeled cells were CD11b-positive tumor-associated immune cells. Conclusions: In this primary murine model of STS, RT did not affect the ability of cathepsin-activated probes to differentiate between tumor and normal muscle. Cathepsin-activated probes labeled tumor cells and tumor-associated macrophages. Our results suggest that it would be feasible to include patients who have received preoperative RT in clinical studies evaluating cathepsin-activated imaging probes.Damon Runyon Cancer Research Foundation (Damon Runyon-Rachleff Innovation Award

Physical Response Functions of Strongly Coupled Massive Quantum Liquids

We study physical properties of strongly coupled massive quantum liquids from their spectral functions using the AdS/CFT correspondence. The generic model that we consider is dense, heavy fundamental matter coupled to SU(N_c) super Yang-Mills theory at finite temperature above the deconfinement phase transition but below the scale set by the baryon number density. In this setup, we study the current-current correlators of the baryon number density using new techniques that employ a scaling behavior in the dual geometry. Our results, the AC conductivity, the quasi-particle spectrum and the Drude-limit parameters like the relaxation time are simple temperature-independent expressions that depend only on the mass-squared to density ratio and display a crossover between a baryon- and meson-dominated regime. We concentrated on the (2+1)-dimensional defect case, but in principle our results can also be generalized straightforwardly to other cases.Comment: 21 pages, 10 figures, extra paragraph and figure are added in response to referee's comment

(De)constructing Intersecting M5-branes

We describe intersecting M5-branes, as well as M5-branes wrapping the holomorphic curve xy=c, in terms of a limit of a defect conformal field theory with two-dimensional (4,0) supersymmetry. This dCFT describes the low-energy theory of intersecting D3-branes at a C^2/Z_k orbifold. In an appropriate k -> infinity limit, two compact spatial directions are generated. By identifying moduli of the M5-M5 intersection in terms of those of the dCFT, we argue that the SU(2)_L R-symmetry of the (4,0) defect CFT matches the SU(2) R-symmetry of the N =2, d=4 theory of the M5-M5 intersection. We find a 't Hooft anomaly in the SU(2)_L R-symmetry, suggesting that tensionless strings give rise to an anomaly in the SU(2) R-symmetry of intersecting M5-branes.Comment: latex, 25 pages, 4 figure

Intersecting D3-branes and Holography

We study a defect conformal field theory describing D3-branes intersecting over two space-time dimensions. This theory admits an exact Lagrangian description which includes both two- and four-dimensional degrees of freedom, has (4,4) supersymmetry and is invariant under global conformal transformations. Both two- and four-dimensional contributions to the action are conveniently obtained in a two-dimensional (2,2) superspace. In a suitable limit, the theory has a dual description in terms of a probe D3-brane wrapping an AdS_3 x S^1 slice of AdS_5 x S^5. We consider the AdS/CFT dictionary for this set-up. In particular we find classical probe fluctuations corresponding to the holomorphic curve wy=c\alpha^{\prime}. These fluctuations are dual to defect fields containing massless two-dimensional scalars which parameterize the classical Higgs branch, but do not correspond to states in the Hilbert space of the CFT. We also identify probe fluctuations which are dual to BPS superconformal primary operators and to their descendants. A non-renormalization theorem is conjectured for the correlators of these operators, and verified to order g^2.Comment: 46 pages, 5 figures, Latex, minor corrections to section 4.2, version published in Phys. Rev.

Thermodynamics of Holographic Defects

Using the AdS/CFT correspondence, we study the thermodynamic properties and the phase diagram of matter fields on (2+1)-dimensional defects coupled to a (3+1)-dimensional N=4 SYM "heat bath". Considering a background magnetic field, (net) quark density, defect "magnitude" $\delta N_c$ and the mass of the matter, we study the defect contribution to the thermodynamic potentials and their first and second derivatives to map the phases and study their physical properties. We find some features that are qualitatively similar to other systems e.g. in (3+1) dimensions and a number of features that are particular to the defect nature, such as its magnetic properties, unexpected properties at T->0 and finite density; and the finite $\delta N_c$ effects, e.g. a diverging susceptibility and vanishing density of states at small temperatures, a physically consistent negative heat capacity and new types of consistent phases.Comment: 33 pages, 16 figures (jpg and pdf), typos fixed and references added, final version published in JHE

The Similarity of Broad Iron Lines in X-ray Binaries and Active Galactic Nuclei

We have compared the 2001 XMM-Newton spectra of the stellar mass black hole binary XTE J1650-500 and the active galaxy MGC-6-30-15, focusing on the broad, excess emission features at ~4--7 keV displayed by both sources. Such features are frequently observed in both low mass X-ray binaries and active galactic nuclei. For the former case it is generally accepted that the excess arises due to iron emission, but there is some controversy over whether their width is partially enhanced by instrumental processes, and hence also over the intrinsic broadening mechanism. Meanwhile, in the latter case, the origin of this feature is still subject to debate; physically motivated reflection and absorption interpretations are both able to reproduce the observed spectra. In this work we make use of the contemporaneous BeppoSAX data to demonstrate that the breadth of the excess observed in XTE J1650-500 is astrophysical rather than instrumental, and proceed to highlight the similarity of the excesses present in this source and MGC-6-30-15. Both optically thick accretion discs and optically thin coronae, which in combination naturally give rise to relativistically-broadened iron lines when the disc extends close to the black hole, are commonly observed in both class of object. The simplest solution is that the broad emission features present arise from a common process, which we argue must be reflection from the inner regions of an accretion disc around a rapidly rotating black hole; for XTE J1650-500 we find spin constraints of 0.84 < a* < 0.98 at the 90 per cent confidence level. Other interpretations proposed for AGN add potentially unnecessary complexities to the theoretical framework of accretion in strong gravity.Comment: Accepted for publication in MNRAS; 22 pages, 17 figure