235 research outputs found
Optimization of Naked DNA Delivery for Interferon Subtype Immunotherapy in Cytomegalovirus Infection
Type I interferon (IFN) gene therapy modulates the immune response leading to inflammatory heart disease following cytomegalovirus (CMV) infection in a murine model of post-viral myocarditis. Efficacy of different immunisation protocols for the IFN constructs was influenced by the dose of DNA, subtype choice, combination use, pre-medication, and timing of DNA administration. Optimal efficacy was found with bupivacaine treatment prior to DNA inoculation of 200mg IFN DNA 14 days prior to virus challenge. Maximal antiviral and antimyocarditic effects were achieved with this vaccination schedule. Furthermore, inoculation of synergistic IFN subtypes demonstrated enhanced efficacy when delivered either alone or with CMV gB DNA vaccination in the CMV model. Thus naked DNA delivery of IFN provides an avenue of immunotherapy for regulating herpesvirus-induced diseases
Expression of Distal-less, dachshund, and optomotor blind in Neanthes arenaceodentata (Annelida, Nereididae) does not support homology of appendage-forming mechanisms across the Bilateria
The similarity in the genetic regulation of
arthropod and vertebrate appendage formation has been
interpreted as the product of a plesiomorphic gene
network that was primitively involved in bilaterian
appendage development and co-opted to build appendages
(in modern phyla) that are not historically related
as structures. Data from lophotrochozoans are needed to
clarify the pervasiveness of plesiomorphic appendage forming
mechanisms. We assayed the expression of three
arthropod and vertebrate limb gene orthologs, Distal-less
(Dll), dachshund (dac), and optomotor blind (omb), in
direct-developing juveniles of the polychaete Neanthes
arenaceodentata. Parapodial Dll expression marks premorphogenetic
notopodia and neuropodia, becoming restricted
to the bases of notopodial cirri and to ventral
portions of neuropodia. In outgrowing cephalic appendages,
Dll activity is primarily restricted to proximal
domains. Dll expression is also prominent in the brain. dac
expression occurs in the brain, nerve cord ganglia, a pair
of pharyngeal ganglia, presumed interneurons linking a
pair of segmental nerves, and in newly differentiating
mesoderm. Domains of omb expression include the brain,
nerve cord ganglia, one pair of anterior cirri, presumed
precursors of dorsal musculature, and the same pharyngeal
ganglia and presumed interneurons that express dac.
Contrary to their roles in outgrowing arthropod and
vertebrate appendages, Dll, dac, and omb lack comparable
expression in Neanthes appendages, implying independent
evolution of annelid appendage development. We infer
that parapodia and arthropodia are not structurally or
mechanistically homologous (but their primordia might
be), that Dll’s ancestral bilaterian function was in sensory
and central nervous system differentiation, and that
locomotory appendages possibly evolved from sensory
outgrowths
Drosophila errantiviruses
Retroelements with long-terminal repeats (LTRs) inhabit nearly all eukaryotic genomes. During the time of their rich evolutionary history they have developed highly diverse forms, ranging from ordinary retrotransposons to complex pathogenic retroviruses such as HIV-I. Errantiviruses are a group of insect endogenous LTR elements that share structural and functional features with vertebrate endogenous retroviruses. The errantiviruses illustrate one of the evolutionary strategies of retrotransposons to become infective, which together with their similarities to vertebrate retroviruses make them an attractive object of research promising to shed more light on the evolution of retroviruses
The influence of skeletal muscle anisotropy on electroporation: in vivo study and numerical modeling
The aim of this study was to theoretically and experimentally investigate electroporation of mouse tibialis cranialis and to determine the reversible electroporation threshold values needed for parallel and perpendicular orientation of the applied electric field with respect to the muscle fibers. Our study was based on local electric field calculated with three-dimensional realistic numerical models, that we built, and in vivo visualization of electroporated muscle tissue. We established that electroporation of muscle cells in tissue depends on the orientation of the applied electric field; the local electric field threshold values were determined (pulse parameters: 8 × 100 μs, 1 Hz) to be 80 V/cm and 200 V/cm for parallel and perpendicular orientation, respectively. Our results could be useful electric field parameters in the control of skeletal muscle electroporation, which can be used in treatment planning of electroporation based therapies such as gene therapy, genetic vaccination, and electrochemotherapy
Reduced basis approximation and a posteriori error estimation for the time-dependent viscous Burgers’ equation
In this paper we present rigorous a posteriori L 2 error bounds for reduced basis approximations of the unsteady viscous Burgers’ equation in one space dimension. The a posteriori error estimator, derived from standard analysis of the error-residual equation, comprises two key ingredients—both of which admit efficient Offline-Online treatment: the first is a sum over timesteps of the square of the dual norm of the residual; the second is an accurate upper bound (computed by the Successive Constraint Method) for the exponential-in-time stability factor. These error bounds serve both Offline for construction of the reduced basis space by a new POD-Greedy procedure and Online for verification of fidelity. The a posteriori error bounds are practicable for final times (measured in convective units) T≈O(1) and Reynolds numbers ν[superscript −1]≫1; we present numerical results for a (stationary) steepening front for T=2 and 1≤ν[superscript −1]≤200.United States. Air Force Office of Scientific Research (AFOSR Grant FA9550-05-1-0114)United States. Air Force Office of Scientific Research (AFOSR Grant FA-9550-07-1-0425)Singapore-MIT Alliance for Research and Technolog
Amorphous formulations of indomethacin and griseofulvin prepared by electrospinning
Following an array of optimization
experiments, two series of electrospun
polyvinylpyrrolidone (PVP) fibers were prepared. One set of fibers
contained various loadings of indomethacin, known to form stable glasses,
and the other griseofulvin (a poor glass former). Drug loadings of
up to 33% w/w were achieved. Electron microscopy data showed the fibers
largely to comprise smooth and uniform cylinders, with evidence for
solvent droplets in some samples. In all cases, the drug was found
to exist in the amorphous physical state in the fibers on the basis
of X-ray diffraction and differential scanning calorimetry (DSC) measurements.
Modulated temperature DSC showed that the relationship between a formulation’s
glass transition temperature (<i>T</i><sub>g</sub>) and
the drug loading follows the Gordon–Taylor equation, but not
the Fox equation. The results of Gordon–Taylor analysis indicated
that the drug/polymer interactions were stronger with indomethacin.
The interactions between drug and polymer were explored in more detail
using molecular modeling simulations and again found to be stronger
with indomethacin; the presence of significant intermolecular forces
was further confirmed using IR spectroscopy. The amorphous form of
both drugs was found to be stable after storage of the fibers for
8 months in a desiccator (relative humidity <25%). Finally, the
functional performance of the fibers was studied; in all cases, the
drug-loaded fibers released their drug cargo very rapidly, offering
accelerated dissolution over the pure drug
A new vesicle trafficking regulator CTL1 plays a crucial role in ion homeostasis
Ion homeostasis is essential for plant growth and environmental adaptation, and maintaining ion homeostasis requires the precise regulation of various ion transporters, as well as correct root patterning. However, the mechanisms underlying these processes remain largely elusive. Here, we reported that a choline transporter gene, CTL1, controls ionome homeostasis by regulating the secretory trafficking of proteins required for plasmodesmata (PD) development, as well as the transport of some ion transporters. Map-based cloning studies revealed that CTL1 mutations alter the ion profile of Arabidopsis thaliana. We found that the phenotypes associated with these mutations are caused by a combination of PD defects and ion transporter misregulation. We also established that CTL1 is involved in regulating vesicle trafficking and is thus required for the trafficking of proteins essential for ion transport and PD development. Characterizing choline transporter-like 1 (CTL1) as a new regulator of protein sorting may enable researchers to understand not only ion homeostasis in plants but also vesicle trafficking in general
Summary Report of PQRI Workshop on Nanomaterial in Drug Products: Current Experience and Management of Potential Risks
At the Product Quality Research Institute (PQRI) Workshop held last January 14-15, 2014, participants from academia, industry, and governmental agencies involved in the development and regulation of nanomedicines discussed the current state of characterization, formulation development, manufacturing, and nonclinical safety evaluation of nanomaterial-containing drug products for human use. The workshop discussions identified areas where additional understanding of material attributes, absorption, biodistribution, cellular and tissue uptake, and disposition of nanosized particles would continue to inform their safe use in drug products. Analytical techniques and methods used for in vitro characterization and stability testing of formulations containing nanomaterials were discussed, along with their advantages and limitations. Areas where additional regulatory guidance and material characterization standards would help in the development and approval of nanomedicines were explored. Representatives from the US Food and Drug Administration (USFDA), Health Canada, and European Medicines Agency (EMA) presented information about the diversity of nanomaterials in approved and newly developed drug products. USFDA, Health Canada, and EMA regulators discussed the applicability of current regulatory policies in presentations and open discussion. Information contained in several of the recent EMA reflection papers was discussed in detail, along with their scope and intent to enhance scientific understanding about disposition, efficacy, and safety of nanomaterials introduced in vivo and regulatory requirements for testing and market authorization. Opportunities for interaction with regulatory agencies during the lifecycle of nanomedicines were also addressed at the meeting. This is a summary of the workshop presentations and discussions, including considerations for future regulatory guidance on drug products containing nanomaterials
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