112,075 research outputs found
3D scaffolds prepared from platelet rich plasma versus conventional plastic surface for culturing adipose-derived canine mesenchymal stem cells and stromal vascular fraction cells
No full text3D scaffolds prepared from Platelet Rich Plasma versus conventional plastic surface for culturing adipose-derived canine Mesenchymal Stromal Cells and Stromal Vascular Fraction cells.
Suelzu C., Conti V., Basini G., Ramoni R. and Grolli S.
Dipartimento di Scienze Mediche Veterinarie, Università di Parma
OBJECTIVE: Mesenchymal Stem Cells (MSCs) are considered a valuable tool for regenerative medicine applications. Stromal vascular fraction (SVF), a heterogeneous population of cells derived from adipose tissue, represents a valid source of MSCs. We evaluated the ability of canine MSCs and adipose-derived SVF cells to be amplified in vitro by exploiting, in addition to the canonical plastic adhesion, a three-dimensional matrix obtained by gelling Platelet Rich Plasma (PRP) or Platelet Poor Plasma (PPP). The aim was to evaluate the possibility of obtaining autologous preparations capable of supporting MSCs and SVF cells growth for the "point of care" application in the veterinary clinic.
MATERIALS AND METHODS: 1. MSCs growth on plastic surface vs 3D fibrin matrix. Total cell number, cell doubling time and cell doubling number of MSCs grown on plastic surface were compared to MSCs cells grown inside a 3D-fibrin matrix prepared by mixing PPP or PPP (50%v/v), DMEM, thrombin (10% v/v) and calcium gluconate (100mg/ml, 10%v/v). 2. SVF growth on plastic surface vs 3D fibrin matrix. SVF cells prepared by collagenase type-I digestion were cultured either on conventional culture dishes or within a 3D fibrin matrix. Total cell number, cell doubling time and cell doubling number were determined for both culture conditions until P3. 3.Phenotypic characterization by RT-PCR of MSCs and SVF cells. Gene expression was compared between MSCs and SVF grown in different culture conditions, i.e. plastic surface versus 3D fibrin matrix. A set of typical MSCs markers and gene involved in their biological properties were evaluated.
RESULTS: Canine MSCs grow within 3D fibrin-based matrices, demonstrating a shorter doubling time and a higher duplication rate when compared to cells grown on the plastic surface: the number of cells obtained is about seven-fold higher in the 3D environment after 144 hours of culture (p<0.01). Moreover, the cells included herein can be frozen and sub-cultured. In our experimental setup, the use of PRP instead of PPP in the preparation of the matrix does not modify cell replication rate. RT-PCR characterisation of cells cultured within the 3D matrix confirms the expression framework of MSCs markers. Furthermore, 3D environment improves SVF cells replication rate, producing at the first culture passage, a mean 1.5-fold increase in cell number.
CONCLUSION: 3D matrices prepared with autologous PRP or PPP are suitable for canine MSCs and SVF cells cultures. Cells grow faster than in standard 2D culture on the plastic surface, while they maintain their panel of gene expression. Furthermore, they can be sub-cultured and frozen for future applications. These results could contribute to set-up more effective MSCs-based therapies, with advantages regarding time shortening for the production of adequate amounts of cells to be applied in the clinical practice
Crystal structures of bovine odorant-binding protein in complex with odorant molecules
No full textThe structure of bovine odorant-binding protein (bOBP) revealed a striking feature of a dimer formed by domain swapping [Tegoni, M., Ramoni, R., Bignetti, E., Spinelli, S. & Cambillau, C. (1996) Nat. Struct. Biol.3, 863-867; Bianchet, M.A., Bains, G., Pelosi, P., Pevsner, J., Snyder, S.H., Monaco, H.L. & Amzel, L.M. (1996) Nat. Struct. Biol.3, 934-939] and the presence of a naturally occuring ligand [Ramoni, R., Vincent, F., Grolli, S., Conti, V., Malosse, C., Boyer, F.D., Nagnan-Le Meillour, P., Spinelli, S., Cambillau, C. & Tegoni, M. (2001) J. Biol. Chem.276, 7150-7155]. These features led us to investigate the binding of odorant molecules with bOBP in solution and in the crystal. The behavior of odorant molecules in bOBP resembles that observed with porcine OBP (pOBP), although the latter is monomeric and devoid of ligand when purified. The odorant molecules presented K(d) values with bOBP in the micromolar range. Most of the X-ray structures revealed that odorant molecules interact with a common set of residues forming the cavity wall and do not exhibit specific interactions. Depending on the ligand and on the monomer (A or B), a single residue--Phe89--presents alternate conformations and might control cross-talking between the subunits. Crystal data on both pOBP and bOBP, in contrast with binding and spectroscopic studies on rat OBP in solution, reveal an absence of significant conformational changes involving protein loops or backbone. Thus, the role of OBP in signal triggering remains unresolved
SAT0247 Efficacy and safety of modified-release prednisone in managing moderate activity systemic lupus erythematous during pregnancy: an implemented case-control study
No full textCossus ramoni – a new species from northeastern Iran (Lepidoptera, Cossidae, Cossinae) /
No full textNew species Cossus ramoni sp. n. is described from northeastern Iran. It is compared with closely distributed Iranian subspecies of Cossus cossus (Linnaeus, 1758) and Cossus afghanistana Daniel, 1953
Congenital heart block not associated with anti-Ro/La antibodies: comparison with anti-Ro/La-positive cases.
No full textJ Rheumatol. 2009 Aug;36(8):1744-8. Epub 2009 Jun 30.
Congenital heart block not associated with anti-Ro/La antibodies: comparison with anti-Ro/La-positive cases.
Brucato A, Grava C, Bortolati M, Ikeda K, Milanesi O, Cimaz R, Ramoni V, Vignati G, Martinelli S, Sadou Y, Borghi A, Tincani A, Chan EK, Ruffatti A.
Source
Medicina Interna, Ospedali Riuniti, Bergamo, Italy. [email protected]
Abstract
OBJECTIVE:
To study anti-Ro/La-negative congenital heart block (CHB).
METHODS:
Forty-five fetuses with CHB were evaluated by analysis of anti-Ro/La antibodies using sensitive laboratory methods.
RESULTS:
There were 9 cases of anti-Ro/La-negative CHB; 3 died (33.3%). Only 3 (33.3%) were complete in utero and 5 (55.5%) were unstable. No specific etiology was diagnosed. Six infants (66.6%) were given pacemakers. There were 36 cases of anti-Ro/La-positive CHB. All except 2 infants (94.4%) had complete atrioventricular block in utero. Ten died (27.8%), one (2.7%) developed severe dilated cardiomyopathy, and 26 (72.2%) were given pacemakers.
CONCLUSION:
Nine of the 45 consecutive CHB cases (20%) were anti-Ro/La-negative with no known cause. They were less stable and complete than the anti-Ro/La positive cases.
PMID:
19567621
[PubMed - indexed for MEDLINE]
PMCID: PMC279858
Application studies of in-vitro bioactivity tests for the development of cell-based biosensors
No full textSingle turn-over mechanism of a Trypsin-reactor with high enzyme concentration
No full textJ. BIOTECHNOL
Pregnancy outcomes in patients with autoimmune diseases and anti-Ro/SSA antibodies
No full textAnti-Ro/SSA antibodies are associated with neonatal lupus (congenital heart block (CHB), neonatal transient skin rash, hematological and hepatic abnormalities), but do not negatively affects other gestational outcomes, and the general outcome of these pregnancies is now good, when followed by experienced multidisciplinary teams. The prevalence of CHB, defined as an atrioventricular block diagnosed in utero, at birth, or within the neonatal period (0-27 days after birth), in the offspring of an anti-Ro/SSA-positive women is 1-2%, of neonatal lupus rash around 10-20%, while laboratory abnormalities in asymptomatic babies can be detected in up to 27% of cases. The risk of recurrence of CHB is ten times higher. Most of the mothers are asymptomatic at delivery and are identified only by the birth of an affected child. Half of these asymptomatic women develop symptoms of a rheumatic disease, most commonly arthralgias and xerophtalmia, but few develop lupus nephritis. A standard therapy for CHB is still matter of investigation, although fluorinated corticosteroids have been reported to be effective for associated cardiomyopathy. Serial echocardiograms and obstetric sonograms, performed at least every 1-2 weeks starting from the 16th week of gestational age, are recommended in anti-Ro/SSA-positive pregnant women to detect early fetal abnormalities that might be a target of preventive therapy
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