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Capra E.,Biotrack | Beretta R.,Biotrack | Baldi A.,University of Milan
Proteome Science | Year: 2012

Background: Human mesenchymal stem cells (hMSC) have recently raised the attention because of their therapeutic potential in the novel context of regenerative medicine. However, the safety of these new and promising cellular products should be carefully defined before they can be used in the clinical setting, as. The protein expression profile of these cells might reveal potential hazards associated with senescence and tumoral transformation which may occur during culture. Proteomic is a valuable tool for hMSC characterization and identification of possible changes during expansion.Results: We used Surface Enhanced Laser Desorption/Ionization-Time Of Flight-Mass Spectrometry (SELDI-ToF-MS) to evaluate the presence of stable molecular markers in adipose tissue-derived mesenchymal stem cells (AD-MSC) produced under conditions of good manufacturing practices (GMP). Proteomic patterns of cells prepared were consistent, with 4 up-regulated peaks (mass-to-charge ratio (m/z) 8950, 10087, 10345, and 13058) through subculture steps (P0-P7) with similar trend in three donors. Among the differentially expressed proteins found in the cytoplasmic and nuclear fractions, a cytoplasmic 10.1 kDa protein was upregulated during culture passages and was identified as S100A6 (Calcyclin).Conclusions: This study suggests for the first time that common variation could occur in AD-MSC from different donors, with the identification of S100A6, a protein prevalently related to cell proliferation and cell culture condition. These results support the hypothesis of common proteomic changes during MSCs expansion and could give important insight in the knowledge of molecular mechanisms intervening during MSC expansion. © 2012 Capra et al.; licensee BioMed Central Ltd.


Tamminga G.G.,Center of Excellence for Sustainable Water Technology | Tamminga G.G.,Biotrack | Paulitsch-Fuchs A.H.,Center of Excellence for Sustainable Water Technology | Paulitsch-Fuchs A.H.,Medical University of Graz | And 2 more authors.
Journal of Microbiological Methods | Year: 2016

State of the art software methods (such as fixed value approaches or statistical approaches) to create a binary image of fluorescent bacterial cells are not as accurate and precise as they should be for counting bacteria and measuring their area. To overcome these bottlenecks, we introduce biological significance to obtain a binary image from a greyscale microscopic image. Using our biological significance approach we are able to automatically count about the same number of cells as an individual researcher would do by manual/visual counting. Using the fixed value or statistical approach to obtain a binary image leads to about 20% less cells in automatic counting. In our procedure we included the area measurements of the bacterial cells to determine the right parameters for background subtraction and threshold values. In an iterative process the threshold and background subtraction values were incremented until the number of particles smaller than a typical bacterial cell is less than the number of bacterial cells with a certain area. This research also shows that every image has a specific threshold with respect to the optical system, magnification and staining procedure as well as the exposure time. The biological significance approach shows that automatic counting can be performed with the same accuracy, precision and reproducibility as manual counting. The same approach can be used to count bacterial cells using different optical systems (Leica, Olympus and Navitar), magnification factors (200 × and 400 ×), staining procedures (DNA (Propidium Iodide) and RNA (FISH)) and substrates (polycarbonate filter or glass). © 2016 Elsevier B.V.


Grant
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: HEALTH-2007-1.4-7 | Award Amount: 5.13M | Year: 2009

Rapid progress in stem cell research pave the way for the development and use of new cell therapy products in regenerative medicine. The objective of CASCADE is to provide the European Community with unique expertise which focuses on the GMP production of mesenchymal stem cells (MSC) and their use in treating skin and corneal wounds, so that quality and safety criteria are paramount and match therapeutic expectations. Commencing with known procedures, CASCADE will develop innovative technologies based on the expertise of its partners. Among our research themes, the specific issue of multiple recipients and universal donors will be investigated. To this end, human MSC will be derived from 4 tissues, bone marrow, fat tissue, cord blood and amniotic membrane, according to good manufacturing practice (GMP). Phenotypic identity of GMP produced MSC, their genetic status and their potential for transformation will constitute the basis for establishing standards for controlling cell production. The efficacy of GMP produced MSC will be tested in in vitro and in vivo models, and the immunological consequences of their use will be studied. Moreover, during the whole CASCADE initiative, ethical and legal issues raised as a result of MSC therapies will be considered. Direct collaboration with two clinical networks will allow CASCADE to finely tune production specifications, and define optimal clinical protocols for these MSC-based cellular therapies. CASCADE will therefore gather together leading European cell therapy laboratories with unrivalled expertise in the production of therapeutic cells and in MSC biology, four SMEs with specific know-how in GMP production of cells and two clinical networks in dermatology and ophthalmology. The outcome will be a consortium that encompasses all the technologies and skills required to efficiently develop innovative technologies for the production of clinical grade MSC and to translate this production into a cell drug to repair wounds.


Bernardo C.S.S.,São Paulo State University | Cresswell B.,Biotrack | Lloyd H.,Northumbria University | Azeredo R.,Crax Brazil | Simpson J.,Crax Brazil
European Journal of Wildlife Research | Year: 2011

Long-term monitoring of reintroduced individuals is a central component of many endangered species reintroduction programs. Radio-telemetry techniques are rarely used to monitor reintroduced captive-bred Cracids and few data exist regarding possible adverse effects of radio-tagging Cracids. In this study, we identify an appropriate radio transmitter design and develop a suitable attachment method that minimizes anthropogenic influence and enables long-term, post-release monitoring (2-3 years) of reintroduced captive-bred Red-billed Curassows in the Brazilian Atlantic Rainforest. We also review studies about the effects of different VHF radio transmitter models on survival, reproduction, behavior, and physiology of Galliformes. © 2011 Springer-Verlag.


Dragoni I.,University of Milan | Balzaretti C.,University of Milan | Rossini S.,Biotrack | Rossi L.,University of Milan | And 2 more authors.
Food Analytical Methods | Year: 2011

Lysozyme is used in cheese manufacture in order to prevent blowing in cheeses caused by Clostridium tyrobutyricum. Being an egg derivative, the presence of lysozyme must be included on the label for residual allergenic risk (2003/89/CE). The aim of this study was to evaluate the presence of lysozyme on proteic profiles of typical Italian cheeses such as Grana Padano through surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. The proteolytic activity of ripening (from 0 to 24 months), confirmed by a decrease in casein, did not influence the intensity of lysozyme peaks. Furthermore, ripened Grana Padano cheese could be differentiated on mass profiling from immature Grana Padano by the presence of particular signals that are probably related to casein proteolysis. © 2010 Springer Science+Business Media, LLC.


PubMed | Parco Tecnologico Padano, Italy; Agroalimentare Sud SpA, Biotrack, Catholic University of the Sacred Heart and 2 more.
Type: | Journal: Journal of plant physiology | Year: 2015

Plant responses to herbivore insects involve direct and indirect defense with the production of signal molecules including jasmonic acid (JA) and its derivatives (e.g. methyl jasmonate, MeJA). In maize (Zea mays), root feeding by Diabrotica virgifera larvae activates an indirect defense mechanism, through enthomopathogenic nematodes that are recruited after Terpene Synthase 23 (tps23) upregulation and (E)--caryophyllene root emission. In order to gain insight into the correlation between JA signaling and response to Diabrotica attack, we analyzed tps23 expression and protein profiles in maize roots in response to MeJA treatment and insect infestation. Similar to herbivore feeding, MeJA treatment was found to increase tps23 transcript accumulation, with consistent variations for both treatments in maize lines differing in (E)--caryophyllene production. Analysis of root protein profiles showed specific alterations leading to the identification of three proteins that were induced by MeJA treatment. We focused on a peroxidase-like protein (Px-like) showing that the corresponding transcripts accumulated in all tested lines. Results show that exogenous application of MeJA upregulates tps23 expression and specifically alters protein patterns in maize roots. Parallel effects on tps23 transcript accumulation were observed upon hormone exposure and insect infestation in different maize lines. In contrast, Px-like transcript profiling showed differences between treatments. These results support the possible involvement of MeJA in mediating the upregulation of tps23 in response to Diabrotica attack.


Uzal A.,Bournemouth University | Uzal A.,University of Saskatchewan | Walls S.,Biotrack | Stillman R.A.,Bournemouth University | Diaz A.,Bournemouth University
European Journal of Wildlife Research | Year: 2013

Large populations of sika deer occur in lowland heath, woodland, and grassland mosaics in southern England. Previous studies have focused on understanding single factors potentially affecting distribution and habitat selection of sika deer rather than considering simultaneously effects of landscape configuration and human disturbance on their distribution and habitat selection. This study measured effects of habitat availability, landscape structure, and human disturbance on where sika deer placed their home ranges and habitat selection within those ranges. Two main hypotheses were tested: (1) habitat selection differs according to landscape structure and habitat availability at both landscape and home range scales and (2) distribution of sources of human disturbance within the home range of deer affects their distribution. Results from radiotracking 31 females provided support for the first hypothesis and partial support for the second. Habitat selection at the landscape and home range scales differed between landscapes with different habitat structure and availability and was driven by distribution and availability of food and cover and a perceived risk linked to disturbance. Furthermore, deer selected open areas close to cover and this selection was stronger with presence of human disturbance, although results differed between study areas with different habitat distribution and level of disturbance. The study highlights the importance for managing deer of a balance between grazing and cover resources and the distribution of human disturbance. © 2013 Springer-Verlag Berlin Heidelberg.


Cresswell B.,Biotrack | Edwards D.,Biotrack
Bird Study | Year: 2013

Capsule European Nightjars Caprimulgus europaeus breeding in southern England were found to over-winter in the Democratic Republic of Congo. Aims To ascertain the wintering areas and migration routes of European Nightjars breeding in southern England. Methods The wintering areas of three Nightjars were mapped using light geolocation tags (two in 2008 and one in 2010). For one of these birds, details of the timing and route of migration were determined. The impact of the birds' behaviour on location accuracy was measured and data on the timing of emergence and roosting was collected. Results All three Nightjars were found to be wintering in the south and east of the Democratic Republic of Congo, in an area not previously considered to be part of the wintering range of this species. The route of migration differed in each period. Autumn migration was across central Sahara, whereas in spring the route was to the west of the Sahara. Aberrations in the light curve caused by the roosting and emergence of the birds were found to affect the estimated location of the wintering areas, shifting them approximately 1° south, and reducing the estimated accuracy of the locations. The timing of these aberrations showed that roosting and emergence roughly follow the timing of dawn and dusk. Conclusions Current distribution maps for the wintering areas of Nightjars in Africa probably under-represent the true distribution of the species in the continent. The wide dispersal of birds from the same breeding area in the UK may be an indication of mixing of breeding populations during the wintering period. Further study is needed to understand how these results fit into the larger picture of Nightjar migration both from the UK and the wider Eurasian breeding range, and to determine locations of stopovers. © 2013 British Trust for Ornithology.


Methods for detecting micro-organisms and/or biological substances in a fluid are provided.


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