165 research outputs found
Microfluidic tools for enhanced characterization of therapeutic stem cells and prediction of their potential antimicrobial secretome
Antibiotic resistance is creating enormous attention on the development of new antibiotic-free therapy strategies for bacterial diseases. Mesenchymal stromal stem cells (MSCs) are the most promising candidates in current clinical trials and included in several cell-therapy protocols. Together with the well-known immunomodulatory and regenerative potential of the MSC secretome, these cells have shown direct and indirect anti-bacterial effects. However, the low reproducibility and standardization of MSCs from different sources are the current limitations prior to the purification of cell-free secreted antimicrobial peptides and exosomes. In order to improve MSC characterization, novel label-free functional tests, evaluating the biophysical properties of the cells, will be advan-tageous for their cell profiling, population sorting, and quality control. We discuss the potential of emerging microfluidic technologies providing new insights into density, shape, and size of live cells, starting from heterogeneous or 3D cultured samples. The prospective application of these technologies to studying MSC populations may contribute to developing new biopharmaceutical strategies with a view to naturally overcoming bacterial defense mechanisms
Effect of Gabapentin in a Neuropathic Pain Model in Mice Overexpressing Human Wild-Type or Human Mutated Torsin A
Background: DYT1 dystonia is the most common form of early-onset inherited dystonia, which is caused by mutation of torsin A (TA) belonging to the "ATPases associated with a variety of cellular activities" (AAA + ATPase). Dystonia is often accompanied by pain, and neuropathic pain can be associated to peripherally induced movement disorder and dystonia. However, no evidence exists on the effect of gabapentin in mice subjected to neuropathic pain model overexpressing human normal or mutated TA. Methods: Mice subjected to L5 spinal nerve ligation (SNL) develop mechanical allodynia and upregulation of the alpha 2 delta-1 L-type calcium channel subunit, forming a validated experimental model of neuropathic pain. Under these experimental conditions, TA is expressed in dorsal horn neurons and astrocytes and colocalizes with alpha 2 delta-1. Similar to this subunit, TA is overexpressed in dorsal horn 7 days after SNL. This model has been used to investigate (1) basal mechanical sensitivity; (2) neuropathic pain phases; and (3) the effect of gabapentin, an alpha 2 delta-1 ligand used against neuropathic pain, in non-transgenic (NT) C57BL/6 mice and in mice overexpressing human wild-type (hWT) or mutant (hMT) TA. Results: In comparison to non-transgenic mice, the threshold for mechanical sensitivity in hWT or hMT does not differ (Kruskal-Wallis test = 1.478; p = 0.4777, although, in the latter animals, neuropathic pain recovery phase is delayed. Interestingly, gabapentin (100 mg/Kg) reduces allodynia at its peak (occurring between post-operative day 7 and day 10) but not in the phase of recovery. Conclusions: These data lend support to the investigation on the role of TA in the molecular machinery engaged during neuropathic pain
A new predictive technology for perinatal stem cell isolation suited for cell therapy approaches
The use of stem cells for regenerative applications and immunomodulatory effect is in-creasing. Amniotic epithelial cells (AECs) possess embryonic‐like proliferation ability and multipo-tent differentiation potential. Despite the simple isolation procedure, inter‐individual variability and different isolation steps can cause differences in isolation yield and cell proliferation ability, compromising reproducibility observations among centers and further applications. We investi-gated the use of a new technology as a diagnostic tool for quality control on stem cell isolation. The instrument label‐free separates cells based on their physical characteristics and, thanks to a micro-camera, generates a live fractogram, the fingerprint of the sample. Eight amniotic membranes were processed by trypsin enzymatic treatment and immediately analysed. Two types of profile were generated: a monomodal and a bimodal curve. The first one represented the unsuccessful isolation with all recovered cell not attaching to the plate; while for the second type, the isolation process was successful, but we discovered that only cells in the second peak were alive and resulted adherent. We optimized a Quality Control (QC) method to define the success of AEC isolation using the frac-togram generated. This predictive outcome is an interesting tool for laboratories and cell banks that isolate and cryopreserve fetal annex stem cells for research and future clinical applications
Learning intrinsic excitability in medium spiny neurons
We present an unsupervised, local activation-dependent learning rule for
intrinsic plasticity (IP) which affects the composition of ion channel
conductances for single neurons in a use-dependent way. We use a
single-compartment conductance-based model for medium spiny striatal neurons in
order to show the effects of parametrization of individual ion channels on the
neuronal activation function. We show that parameter changes within the
physiological ranges are sufficient to create an ensemble of neurons with
significantly different activation functions. We emphasize that the effects of
intrinsic neuronal variability on spiking behavior require a distributed mode
of synaptic input and can be eliminated by strongly correlated input. We show
how variability and adaptivity in ion channel conductances can be utilized to
store patterns without an additional contribution by synaptic plasticity (SP).
The adaptation of the spike response may result in either "positive" or
"negative" pattern learning. However, read-out of stored information depends on
a distributed pattern of synaptic activity to let intrinsic variability
determine spike response. We briefly discuss the implications of this
conditional memory on learning and addiction.Comment: 20 pages, 8 figure
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Angiogenic potential of human dental pulp stromal (stem) cells.
Dental pulp is a heterogeneous microenviroment where unipotent progenitor and pluripotent mesenchymal stem cells cohabit. In this study we investigated whether human Dental Pulp Stromal (Stem) Cells (DP-SCs) committed to the angiogenic fate. DP-SCs showed the specific mesenchymal immunophenotypical profile positive for CD29, CD44, CD73, CD105, CD166 and negative for CD14, CD34, CD45, in accordance with that reported for bone marrow-derived SCs. The Oct-4 expression in DP-SCs, evaluated through RT-PCR analysis, increased in relation with the number of the passages in cell culture and decreased after angiogenic induction. In agreement with their multipotency, DP-SCs differentiated toward osteogenic and adipogenic commitments. In angiogenic experiments, differentiation of DP-SCs, through Vascular Endothelial Growth Factor (VEGF) induction, was evaluated by in vitro matrigel assay and by cytometric analysis. Accordingly, endothelial-specific markers like Flt-1 and KDR were basally expressed and they increased after exposure to VEGF together with the occurrence of ICAM-1 and von Willebrand Factor positive cells. In addition, VEGF-induced DP-SCs maintained endothelial cell-like features when cultured in a 3-D fibrin mesh, displaying focal organization into capillary-like structures. The DP-SC angiogenic potential may prove a remarkable tool for novel approaches to developing tissue-engineered vascular grafts which are useful when vascularization of ischemic tissues is required
Profiling of ubiquitination pathway genes in peripheral cells from patients with frontotemporal dementia due to C9ORF72 and GRN mutations
We analysed the expression levels of 84 key genes involved in the regulated degradation of cellular protein by the ubiquitin-proteasome system in peripheral cells from patients with frontotemporal dementia (FTD) due to C9ORF72 and GRN mutations, as compared with sporadic FTD and age-matched controls. A SABiosciences PCR array was used to investigate the transcription profile in a discovery population consisting of six patients each in C9ORF72, GRN, sporadic FTD and age-matched control groups. A generalized down-regulation of gene expression compared with controls was observed in C9ORF72 expansion carriers and sporadic FTD patients. In particular, in both groups, four genes, UBE2I, UBE2Q1, UBE2E1 and UBE2N, were down-regulated at a statistically significant (p < 0.05) level. All of them encode for members of the E2 ubiquitin-conjugating enzyme family. In GRN mutation carriers, no statistically significant deregulation of ubiquitination pathway genes was observed, except for the UBE2Z gene, which displays E2 ubiquitin conjugating enzyme activity, and was found to be statistically significant up-regulated (p = 0.006). These preliminary results suggest that the proteasomal degradation pathway plays a role in the pathogenesis of FTD associated with TDP-43 pathology, although different proteins are altered in carriers of GRN mutations as compared with carriers of the C9ORF72 expansion
Effect of Millettia ferruginea (Birbra) foliage supplementation on feed intake, digestibility, body weight change and carcass characterstics of Washera sheep fed natural pasture grass hay basal diet
New functional ingredients from agri-food by-product for fortified food
A wheat bran hydrolisate was obtained using commercial enzyme preparations selected in order to get all the enzymatic activities needed (Cellulases, Hemicellulases, Arylesterases, etc.) to get a liquid suspension containing 168,39 mg/L of ferulic acid and 703 ORAC/L as measure of the antioxidant activity.
Fortified breads were prepared by substituting different percentages of water (33,3%, 50% e 100% w/w ) with the hydrolised wheat bran during dough preparation. After bread making samples were extracted to recover the phenolic fraction in which the amount of ferulic acid was found proportional to the added quantity of wheat bran hydrolysate.
Surprisingly the antioxidant activity measured on the same extracts shown higher values than expected considering the antioxidant activities of each individual ingredient, up to 400% than the theoretical values. The ORAC/L values vs. wheat bran hydrolysate added had an exponential trend likely due to an interaction between some component of hydrolysed bran and the yeast during the bread making process
On the implementation of quadraphonic data recording
A recent patent in the acoustics field introduced a new approach to quadraphonic data recording and playback. Based on fixed objective criteria, this method seems a promising technique to be used in high-fidelity systems where a characterization of the recording environment is needed. This may be useful for instance when documenting "sound events" such as music performances in real acoustic spaces or when composing virtual acoustic environments as for cinema sound tracks. A major bottleneck for real-time application of this system is represented by the physical implementation, which has to cope both with feasible implementation and no detriment to the high-end performances offered by the quadraphonic technique. This paper presents the architectural design issues for implementation of the quadraphonic recording algorithm: after the algorithm profiling, the hardware/software architecture is derived as a trade-off between complexity, performance and flexibility among different target technologies ranging from Digital Signal Processor (DSP) throughApplication Specific Instruction set Processor (ASIP) up to Field ProgrammableGate Arrays (FPGA) and Application Specific Integrated Circuit (ASIC). As a conclusion, preliminary implementation results for the most promising technology will be provided
Constitutive and Stimulated Production of Vegf by Human Megakaryoblastic Cell Lines: Effect on Proliferation and Signaling Pathway
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