Paracrine delivery of therapeutic biologics for cancer

A fundamental goal of cancer drug delivery is to achieve sufficient levels within the tumour without leading to high systemic concentrations that might cause off-target toxicities. In situ production of protein-based therapeutics by tumour cells provides an attractive alternative to treatment with repeated high bolus injections, as secretion by the tumour itself could provide high local concentrations that act in a paracrine fashion over an extended duration. For this purpose, we have developed a non-oncolytic adenoviral delivery system that allows for targeting of Ad5 to discrete cell types by redirecting viral tropism to cell surface biomarkers through the use of interchangeable adapters. Furthermore, we recently described the engineering of a protein-based ‘shield’ that is coated on the Ad5 capsid, which, together with the retargeting adapters, allows for improved tumour specificity and prevention of viral clearance. To test this delivery strategy in vivo, SCID-beige mice bearing orthotopic BT474 xenografts were treated with three doses of either a cancerspecific, non-replicative Ad5 that encodes a secreted anti-HER2 antibody, trastuzumab, in its genome, or with the protein therapeutic itself (Herceptin®). We have employed state-of-the-art whole tumour clearing and imaging with confocal microscopy at high spatial resolution in 3D to assess biodistribution, and large volumetric imaging has revealed that the secreted therapeutic diffuses significantly throughout the tumour leading to a therapeutic effect and delayed tumour outgrowth. Moreover, the systemic concentration of antibody is significantly reduced with viral delivery, suggesting that paracrine delivery may be a promising strategy for delivery of biologics with narrow therapeutic indices

Application of Discrete Differential Forms to Spherically Symmetric Systems in General Relativity

In this article we describe applications of Discrete Differential Forms in computational GR. In particular we consider the initial value problem in vacuum space-times that are spherically symmetric. The motivation to investigate this method is mainly its manifest coordinate independence. Three numerical schemes are introduced, the results of which are compared with the corresponding analytic solutions. The error of two schemes converges quadratically to zero. For one scheme the errors depend strongly on the initial data.Comment: 22 pages, 6 figures, accepted by Class. Quant. Gra

Intersection local times of independent fractional Brownian motions as generalized white noise functionals

In this work we present expansions of intersection local times of fractional Brownian motions in $\R^d$, for any dimension $d\geq 1$, with arbitrary Hurst coefficients in $(0,1)^d$. The expansions are in terms of Wick powers of white noises (corresponding to multiple Wiener integrals), being well-defined in the sense of generalized white noise functionals. As an application of our approach, a sufficient condition on $d$ for the existence of intersection local times in $L^2$ is derived, extending the results of D. Nualart and S. Ortiz-Latorre in "Intersection Local Time for Two Independent Fractional Brownian Motions" (J. Theoret. Probab.,20(4)(2007), 759-767) to different and more general Hurst coefficients.Comment: 28 page

Correlation between erythropoietin serum levels and erythrocyte susceptibility to lipid peroxidation in elderly with type 2 diabetes

Erythropoietin (EPO), a key hormone involved in red blood cell formation has been recently acknowledged for its pleiotropic actions and protective role in ageing and various pathological conditions concurrent with oxidative stress, vascular diseases and metabolic abnormalities such as diabetes mellitus. The aim of the study was to evaluate the relationship between circulating erythropoietin levels and oxidative stress biomarkers, in elderly with type 2 diabetes (T2DM). The study was carried out in 67 subjects with T2DM (69 ± 5 years; n = 37) without anemia, and aged-matched controls (70 ± 6 years; n = 30). EPO serum levels, erythrocyte susceptibility to lipid peroxidation (ESP) and total antioxidant capacity (TAC) were evaluated. Lower EPO levels (p < 0.01) and higher ESP values (p < 0.001) were found in T2DM group, compared to healthy subjects. EPO levels showed significant negative associations with ESP, both in T2DM subjects (r = −0.565; p < 0.001) and in all study population (r = –0,600; p < 0,001; n = 67). In conclusion, we provide new data regarding the cytoprotective effect of EPO exerted at systemic level on erythrocyte membrane, in the particular state of impaired glucose metabolism associated with oxidative stress, in the elderly

Self-avoiding fractional Brownian motion - The Edwards model

In this work we extend Varadhan's construction of the Edwards polymer model to the case of fractional Brownian motions in $\R^d$, for any dimension $d\geq 2$, with arbitrary Hurst parameters $H\leq 1/d$.Comment: 14 page

Genetically Encoded Spy Peptide Fusion System to Detect Plasma Membrane-Localized Proteins In Vivo

Membrane proteins are the main gatekeepers of cellular state, especially in neurons, serving either to maintain homeostasis or instruct response to synaptic input or other external signals. Visualization of membrane protein localization and trafficking in live cells facilitates understanding the molecular basis of cellular dynamics. We describe here a method for specifically labeling the plasma membrane-localized fraction of heterologous membrane protein expression using channelrhodopsins as a case study. We show that the genetically encoded, covalent binding SpyTag and SpyCatcher pair from the Streptococcus pyogenes fibronectin-binding protein FbaB can selectively label membrane-localized proteins in living cells in culture and in vivo in Caenorhabditis elegans. The SpyTag/SpyCatcher covalent labeling method is highly specific, modular, and stable in living cells. We have used the binding pair to develop a channelrhodopsin membrane localization assay that is amenable to high-throughput screening for opsin discovery and engineering

Optical dopamine monitoring with dLight1 reveals mesolimbic phenotypes in a mouse model of neurofibromatosis type 1

Neurofibromatosis type 1 (NF1) is an autosomal dominant disorder whose neurodevelopmental symptoms include impaired executive function, attention, and spatial learning that could be due to perturbed mesolimbic dopaminergic circuitry. However, these circuits have never been directly assayed in vivo. We employed the genetically encoded optical dopamine sensor dLight1 to monitor dopaminergic neurotransmission in the ventral striatum of NF1 mice during motivated behavior. Additionally, we developed novel systemic AAV vectors to facilitate morphological reconstruction of dopaminergic populations in cleared tissue. We found that NF1 mice exhibit reduced spontaneous dopaminergic neurotransmission that was associated with excitation/inhibition imbalance in the ventral tegmental area and abnormal neuronal morphology. NF1 mice also had more robust dopaminergic and behavioral responses to salient visual stimuli, which were stimulus-dependent, independent of learning, and rescued by optogenetic inhibition of non-dopaminergic neurons in the VTA. Overall, these studies provide a first in vivo characterization of dopaminergic circuit function in the context of NF1 and reveal novel pathophysiological mechanisms

From constructive field theory to fractional stochastic calculus. (II) Constructive proof of convergence for the L\'evy area of fractional Brownian motion with Hurst index α∈(1/8,1/4)\alpha\in(1/8,1/4)

{Let $B=(B_1(t),...,B_d(t))$ be a $d$-dimensional fractional Brownian motion with Hurst index $\alpha<1/4$, or more generally a Gaussian process whose paths have the same local regularity. Defining properly iterated integrals of $B$ is a difficult task because of the low H\"older regularity index of its paths. Yet rough path theory shows it is the key to the construction of a stochastic calculus with respect to $B$, or to solving differential equations driven by $B$. We intend to show in a series of papers how to desingularize iterated integrals by a weak, singular non-Gaussian perturbation of the Gaussian measure defined by a limit in law procedure. Convergence is proved by using "standard" tools of constructive field theory, in particular cluster expansions and renormalization. These powerful tools allow optimal estimates, and call for an extension of Gaussian tools such as for instance the Malliavin calculus. After a first introductory paper \cite{MagUnt1}, this one concentrates on the details of the constructive proof of convergence for second-order iterated integrals, also known as L\'evy area