Virally and physically transgenized equine adipose-derived stromal cells as a cargo for paracrine secreted factors

<p>Abstract</p> <p>Background</p> <p>Adipose-Derived Stromal Cells have been shown to have multiple lineage differentiation properties and to be suitable for tissues regeneration in many degenerative processes. Their use has been proposed for the therapy of joint diseases and tendon injuries in the horse. In the present report the genetic manipulation of Equine Adipose-Derived Stromal Cells has been investigated.</p> <p>Results</p> <p>Equine Adipose-Derived Stromal Cells were successfully virally transduced as well as transiently and stably transfected with appropriate parameters, without detrimental effect on their differentiation properties. Moreover, green fluorescent protein alone, fused to <it>neo </it>gene, or co-expressed as bi-cistronic reporter constructs, driven by viral and house-keeping gene promoters, were tested. The better expressed cassette was employed to stably transfect Adipose-Derived Stromal Cells for cell therapy purposes. Stably transfected Equine Adipose-Derived Stromal Cells with a heterologous secreted viral antigen were able to immunize horses upon injection into the lateral wall of the neck.</p> <p>Conclusion</p> <p>This study provides the methods to successfully transgenize Adipose-Derived Stromal Cells both by lentiviral vector and by transfection using optimized constructs with suitable promoters and reporter genes. In conclusion these findings provide a working platform for the delivery of potentially therapeutic proteins to the site of cells injection via transgenized Equine Adipose-Derived Stromal Cells.</p

Humoral and cellular response to BoHV-1 in buffalo and cattle treated with an inactivated marker vaccine

The study is aimed at assessing and comparing the immune response to BoHV-1 elicited by an inactivated marker vaccine in buffaloes and cattle. Vaccination did not produced any local or general reactions in buffaloes. Seroneutralizing antibodies and cellular response by IFN-γ- test have been detected in buffaloes and cattle after a prime/ booster vaccination strategy. Humoral and cellular responses were significantly higher in cattle than in buffaloes. Data pointed out the possibility to use the marker vaccine in buffaloes. However, further studies must be planned to assess the immune pressure of marker vaccines in terms of IBR eradicative attitude in infected buffalo herds

Lunar lander conceptual design

This paper is a first look at the problems of building a lunar lander to support a small lunar surface base. A series of trade studies was performed to define the lander. The initial trades concerned choosing number of stages, payload mass, parking orbit altitude, and propellant type. Other important trades and issues included plane change capability, propellant loading and maintenance location, and reusability considerations. Given a rough baseline, the systems were then reviewed. A conceptual design was then produced. The process was carried through only one iteration. Many more iterations are needed. A transportation system using reusable, aerobraked orbital transfer vehicles (OTV's) is assumed. These OTV's are assumed to be based and maintained at a low Earth orbit (LEO) space station, optimized for transportation functions. Single- and two-stage OTV stacks are considered. The OTV's make the translunar injection (TLI), lunar orbit insertion (LOI), and trans-Earth injection (TEI) burns, as well as midcourse and perigee raise maneuvers

A Comparison of the Energy Demands of Quadrupedal Movement Training to Walking

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Evidence for a Novel Reaction Mechanism of a Prompt Shock-Induced Fission Following the Fusion of 78Kr and 40Ca Nuclei at E/A =10 MeV

An analysis of experimental data from the inverse-kinematics ISODEC experiment on 78Kr+40Ca reaction at a bombarding energy of 10 AMeV has revealed signatures of a hitherto unknown reaction mechanism, intermediate between the classical damped binary collisions and fusion-fission, but also substantially different from what is being termed in the literature as fast fission or quasi fission. These signatures point to a scenario where the system fuses transiently while virtually equilibrating mass asymmetry and energy and, yet, keeping part of the energy stored in a collective shock-imparted and, possibly, angular momentum bearing form of excitation. Subsequently the system fissions dynamically along the collision or shock axis with the emerging fragments featuring a broad mass spectrum centered around symmetric fission, relative velocities somewhat higher along the fission axis than in transverse direction, and virtually no intrinsic spin. The class of massasymmetric fission events shows a distinct preference for the more massive fragments to proceed along the beam direction, a characteristic reminiscent of that reported earlier for dynamic fragmentation of projectile-like fragments alone and pointing to the memory of the initial mass and velocity distribution.Comment: 5 PAGES, 6 FIGURE

Bovine endometrial stromal cells display osteogenic properties

The endometrium is central to mammalian fertility. The endometrial stromal cells are very dynamic, growing and differentiating throughout the estrous cycle and pregnancy. In humans, stromal cells appear to have progenitor or stem cell capabilities and the cells can even differentiate into bone. It is not clear whether bovine endometrial stromal cells exhibit a similar phenotypic plasticity. So, the present study tested the hypothesis that bovine endometrial stromal cells could be differentiated along an osteogenic lineage. Pure populations of bovine stromal cells were isolated from the endometrium. The endometrial stromal cell phenotype was confirmed by morphology, prostaglandin secretion, and susceptibility to viral infection. However, cultivation of the cells in standard endometrial cell culture medium lead to a mesenchymal phenotype similar to that of bovine bone marrow cells. Furthermore, the endometrial stromal cells developed signs of osteogenesis, such as alizarin positive nodules. When the stromal cells were cultured in a specific osteogenic medium the cells rapidly developed the characteristics of mineralized bone. In conclusion, the present study has identified that stromal cells from the bovine endometrium show a capability for phenotype plasticity similar to mesenchymal progenitor cells. These observations pave the way for further investigation of the mechanisms of stroma cell differentiation in the bovine reproductive tract

Evaluation of the finger wrinkling test: a pilot study

Purpose: Tilt table testing mainly evaluates the systemic cardiovascular part of the autonomic nervous system, while it is assumed that the finger wrinkling test assesses the peripheral part of the autonomic nervous system. In this study we explored whether the finger wrinkling test could be a useful test for autonomic dysfunction and whether the clinical evaluation of wrinkling can be improved by digital analysis of photographs. Methods: As much as 20 healthy subjects and 15 patients underwent tilt table testing and finger wrinkling testing. During the finger wrinkling test the right hand was immersed in water at 40°C. The degree of wrinkling was assessed with a 5-point clinical scale at baseline, 5, 15 and 30 min of immersion. Photographs were taken at the same intervals. Several features were evaluated using digital analysis: length and gradient of automatically detected wrinkle and mean, maximum, minimum, variance and derivative of grey value of pixels. Results: Clinical scoring of wrinkling allowed differentiation between healthy subjects and patients with a normal and an abnormal response to tilt table testing. Relevant features obtained with digital analysis were mean grey value and the gradient of automatically detected wrinkle. McNemar’s test showed no difference in test results between the tilt table test and the finger wrinkling test with a kappa of 0.68. Conclusion: The finger wrinkling test can be used as a screening test before tilt table testing. Visual evaluation of wrinkling is still superior to digital analysis of photographs

SNPLims: a data management system for genome wide association studies

<p>Abstract</p> <p>Background</p> <p>Recent progresses in genotyping technologies allow the generation high-density genetic maps using hundreds of thousands of genetic markers for each DNA sample. The availability of this large amount of genotypic data facilitates the whole genome search for genetic basis of diseases.</p> <p>We need a suitable information management system to efficiently manage the data flow produced by whole genome genotyping and to make it available for further analyses.</p> <p>Results</p> <p>We have developed an information system mainly devoted to the storage and management of SNP genotype data produced by the Illumina platform from the raw outputs of genotyping into a relational database.</p> <p>The relational database can be accessed in order to import any existing data and export user-defined formats compatible with many different genetic analysis programs.</p> <p>After calculating family-based or case-control association study data, the results can be imported in SNPLims. One of the main features is to allow the user to rapidly identify and annotate statistically relevant polymorphisms from the large volume of data analyzed. Results can be easily visualized either graphically or creating ASCII comma separated format output files, which can be used as input to further analyses.</p> <p>Conclusions</p> <p>The proposed infrastructure allows to manage a relatively large amount of genotypes for each sample and an arbitrary number of samples and phenotypes. Moreover, it enables the users to control the quality of the data and to perform the most common screening analyses and identify genes that become “candidate” for the disease under consideration.</p

Angiocentric glioma-associated seizures: The possible role of EATT2, pyruvate carboxylase and glutamine synthetase

Purpose: Our purpose was to better understand the pathogenesis of seizures associated with angiocentric glioma. Angiocentric glioma is an indolent and rare low-grade glioma. Its typical clinical presentation is with epileptic seizures. The pathogenesis of tumor-associated seizures is poorly understood. Among the possible pathomechanisms, the increased neurotoxic concentrations of the glutamate has been proposed. Glutamate transporters, pyruvate carboxylase and glutamine synthetase are involved in maintaining the physiological concentration of glutamate in the inter synaptic spaces. Methods: We evaluated the immunohistochemical expression of EAAT2 (the most important glutamate transporter), pyruvate carboxylase and glutamine synthetase in 17 angiocentric gliomas. Results: EAAT2 was never expressed (0%) in the neoplastic cells in none of the cases studied. Pyruvate carboxylase was expressed in the cytoplasm of the neoplastic cells in 16/17 cases (94 %). Glutamine synthetase was expressed in the cytoplasm of the neoplastic cells in 15/17 cases (88 %). Conclusion: The net result of this enzymatic expression, in particular considering the loss of EAAT2, could be an increased glutamate concentration in the synaptic clef, which might increase local network excitability initially involving intratumoral neurons. The observation that the angiocentric glioma-associated epilepsy might be at least in part related to EAAT2 deficiency opens up interesting therapeutic perspectives