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Fernando Alonso

Publications and source records attributed to Fernando Alonso.

12 recordsLinked to original sources

Molecular cloning characterization and expression of porcine immunoreceptor SIRPalpha.

SWC3 is a porcine CD that has been the reference marker of myeloid lineage. It is expressed in every myelomonocytic cell from early bone marrow precursors. We have identified the molecule recognized by anti-SWC3 antibodies as a member of the signal-regulatory proteins (SIRPs)alpha family. Here, we describe the cloning of a cDNA coding for a porcine SIRPalpha protein. The sequence is 2470 nucleotides long and contains an open reading frame encoding a 507 amino acid sequence. The predicted polypeptide was composed of a 30 amino acids putative signal peptide, a 342 amino acid extracellular region, a 23 amino acid transmembrane segment and a 112 amino acid cytoplasmic domain. Analysis of the sequence reveals a high degree of homology with known SIRPs in other species, being easily identified the three extracellular Ig type domains and two cytoplasmic ITIM motifs characteristic of this molecule. The gene coding for porcine SIRPalpha has been mapped to porcine chromosome 17, in a region syntenic to the human chromosome 20 where SIRP genes have been mapped. During the analysis of SIRP gene expression in tissues by RT-PCR, we noticed the existence of a shorter mRNA, and cloned the corresponding cDNA. This coded for a splicing variant of SIRPalpha that lacked the two membrane proximal Ig domains. In transfection experiments, we have been able to show that anti-SWC3 antibodies recognize both forms of the molecule, mapping the SWC3 epitopes to the N-terminal IgV type domain.

Animals↗

Impact of fatigue in Parkinson's disease: the Fatigue Impact Scale for Daily Use (D-FIS).

The Fatigue Impact Scale for Daily Use (D-FIS) was used in a cross-sectional study including 142 consecutive Parkinson's disease (PD) patients. Usual clinical measures for PD, the Montgomery-Asberg Depression Rating Scale and the Parkinson's Disease Questionnaire-8 items were applied. In addition to the D-FIS, patients with fatigue (67.6%, PWF) completed the Multidimensional Fatigue Inventory (MFI), a visual analogue scale for fatigue (VAS-F) and a Global Perception of Fatigue scale (GPF). Relevant psychometric D-FIS results were: floor effect = 4.2%; ceiling effect = 1.1%; skewness = 0.44; item homogeneity = 0.63; Cronbach's alpha = 0.93; item-total correlation = 0.68 (item 1)-0.82 (item 8); standard error of measurement = 2.15; convergent validity with other fatigue measures = 0.54 [GPF]-0.62 [VAS-F] (p<0.001). In a multiple linear regression model, fatigue, depression, and disability independently influenced HRQoL, as measured by the PDQ-8. Patients on amantadine had lower prevalence of fatigue. In PD, D-FIS is a consistent and valid measure for fatigue, a frequent symptom previously found to impair patients' HRQoL. Fatigue was also linked to depression and disability in this study.

Aged↗

Molecular cloning, characterization and tissue expression of porcine Toll-like receptor 4.

A cDNA containing the porcine Toll-like receptor 4 (TLR4) coding sequence has been cloned by RT-PCR from alveolar macrophages mRNA, and its complete sequence has been determined. The predicted amino acid sequence comprises an extracellular domain with 21 leucine-rich repeats (LRR) and a LRR-C-terminal (LRR-CT) motif, followed by a 30 amino acid transmembrane segment, and a 179 amino acid intracytoplasmic region containing the Toll/IL-1R domain. Pig TLR4 shows 63-80% amino acid sequence identity with those of cow, horse, cat, human, rabbit and mouse. The degree of sequence identity rises to over 90% in the TIR domain. The whole TLR4 sequence and its ectodomain were expressed as GFP fusion proteins in CHO cells. Using RT-PCR analysis, porcine TLR4 transcripts were detected in DCs, monocytes and macrophages, and in tissue samples of bone marrow, thymus, lymph node, spleen, brain, liver, kidney and ovary. The expressed protein will be used for the development of reagents. Knowledge of TLR4 expression will help to address mechanisms of immune induction by antigens and vaccines.

Animals↗

IMRT planning on adaptive volume structures--a decisive reduction in computational complexity.

The objective of radiotherapy planning is to find a compromise between the contradictive goals of delivering a sufficiently high dose to the target volume while widely sparing critical structures. The search for such a compromise requires the computation of several plans, which mathematically means solving several optimization problems. In the case of intensity modulated radiotherapy (IMRT) these problems are large-scale, hence the accumulated computational expense is very high. The adaptive clustering method presented in this paper overcomes this difficulty. The main idea is to use a preprocessed hierarchy of aggregated dose-volume information as a basis for individually adapted approximations of the original optimization problems. This leads to a decisively reduced computational expense: numerical experiments on several sets of real clinical data typically show computation times decreased by a factor of about 10. In contrast to earlier work in this field, this reduction in computational complexity will not lead to a loss in accuracy: the adaptive clustering method produces the optimum of the original optimization problem.

Algorithms↗

Quality of life of caregivers in Parkinson's disease.

OBJECTIVE: To assess the impact of PD on informal caregivers of patients and identify the main factors related to caregiver strain. PATIENTS AND METHODS: Pairs of PD patients and their caregivers. Evaluation by neurologists included the Hoehn and Yahr, Schwab and England, UPDRS (parts 1-3), ISAPD, and Pfeiffer's SPMSQ rating scales. Patients completed the Euro-QoL 5D, PDQ-8, and Hospital Anxiety and Depression Scale. The SQLC was used to assess caregivers' quality of life (QoL), with caregivers, in turn, applying the Euro-QoL and PDQ-8 to assess patients' health-related quality of life (HRQoL). Multiple linear regression models were fitted to ascertain factors linked to the SQLC. RESULTS: Significant correlations were in evidence between the following scores: SQLC and clinical rating scales and SQLC and patients' HRQoL. Based on multiple regression analysis, patients' functional state (ADL) proved to be the main predictor of caregivers' QoL. Self- and caregiver-assessed patients' HRQoL also proved to be a relevant factor. CONCLUSIONS: (1) Patients' functional state was significantly related to caregivers' psychosocial burden; (2) patients' HRQoL proved to be an additional factor linked to caregiver QoL; (3) improvement of patient disability and HRQoL might alleviate caregiver strain.

Aged↗

Phenotypic and functional heterogeneity of porcine blood monocytes and its relation with maturation.

Swine monocytes constitute a heterogeneous population of cells which can be divided into four subsets based on the expression of SWC3, CD14, CD163 and swine leucocyte antigen (SLA) DR markers. These subsets appear to represent different maturation stages in a pathway along which these cells up-regulate the expression of SLA DR and CD163 antigens and reduce that of CD14. Differences in the expression of adhesion and costimulatory molecules are also patent, with a progressive increase in the expression of CD11a, wCD11R1, CD29, CD49d, CD61, CD1a and CD80/86, and a concomitant decrease in that of wCD11R2. Besides, these subsets differ in their capacity for tumour necrosis factor-alpha (TNF-alpha) production in response to lipopolysaccharide + interferon-gamma. The CD163(+) CD14(-) SLA DR(+) subset produces higher amounts of TNF-alpha than the CD163(-) CD14(+) SLA DR(-) subset, whereas CD163(+) CD14(+) SLA DR(+) and CD163(-) CD14(+) SLA DR(+) subsets show intermediate values. CD163(+) monocytes also display a higher ability to present soluble antigens to T cells than CD163(-) monocytes.

Animals↗

Health-related quality of life evaluation by proxy in Parkinson's disease: approach using PDQ-8 and EuroQoL-5D.

Patient- and caregiver-based scores were compared and agreement levels ascertained to determine the reliability of proxy evaluation of Parkinson's Disease (PD) patients' health-related quality of life (HRQoL) using the EuroQoL and PD questionnaire (PDQ)-8. Of 72 patient-caregiver pairs, 64 (88.88%) returned the questionnaires. The degree of agreement varied for individual dimensions. Proxy evaluation of PD patients' HRQoL showed limitations mainly with assessments using the EuroQoL and especially in patients with severe disease and depression.

Adolescent↗

In vitro differentiation of porcine blood CD163- and CD163+ monocytes into functional dendritic cells.

Swine monocytes constitute a heterogeneous cell population containing subsets with distinct functional capacities or representing different maturational stages. Based on the expression of CD163, we have recently identified two monocyte subpopulations. In this study, we investigate the ability of both CD163- and CD163+ monocytes to differentiate into dendritic cells (DCs) in the presence of GM-CSF and IL-4. Monocyte differentiation into DC is accompanied by an up-regulation of the expression of swine leukocyte antigen (SLA) I, SLA II and CD80/86 molecules, and a decrease in the expression of CD14, CD16 and CD163. These DC express the pan-myeloid marker SWC3 and display typical dendritic cytoplasmic projections. When monocytes are split into CD163+ and CD163- cells, both subsets give rise to DC. However, compared to CD163- monocyte-derived DC (MoDC), CD163+ MoDC appear to have reached a more advanced stage of maturation, expressing higher levels of SLA II and CD80/86 and inducing more efficiently proliferation of T cells to recall antigens and alloantigens.

Animals↗

Patients', doctors', and caregivers' assessment of disability using the UPDRS-ADL section: are these ratings interchangeable?

This multicenter study sought to analyze the validity and reliability of the Unified Parkinson's Disease Rating Scale (UPDRS)-section 2 (Activities of Daily Living, ADL) as applied by patients and caregivers. Sixty pairs of PD patients-caregivers were enrolled for study purposes. Neurologists used a set of scales to determine disease severity and patients' functional state. Patients and caregivers used adapted versions of the UPDRS-section 2 in tandem with other measures. Wilcoxon and Mann-Whitney tests, weighted kappa, intraclass and Spearman's correlation coefficients, as well as multivariate linear regression models were applied. On the whole, PD patient self-assessment and caregiver evaluation of patients' disability showed close concordance with neurologists' ratings. Correlation between caregiver ratings and clinical evaluation tended to be slightly lower than that for patient-based self-assessment. Depression showed a positive correlation with disability and had a nonsystematic influence on UPDRS-section 2 (ADL) scores. As expected, there was a significant correlation between perceived disability and health-related quality of life measures. Caregiver burden did not reduce the level of agreement with neurologists as to the overall rating of any given patient's disability. In PD, UPDRS-section 2-based assessment of disability by patients themselves and caregivers is a valid and reliable outcome.

Aged↗

Characterization of a novel activation antigen on porcine lymphocytes recognized by monoclonal antibody 5A6/8.

In this report, we describe the characterization of a novel activation antigen on porcine lymphocytes recognized by mAb 5A6/8. This antigen was detected on B and T cells 24 h after treatment with various stimuli. It was also found on alveolar macrophages, and at low levels on untreated monocytes. MAb 5A6/8 precipitated two bands of 45 and 50 kDa under non-reducing conditions, and of 22 and 28 kDa under reducing conditions. The cellular distribution, expression kinetics and/or molecular size of the 5A6/8 antigen differ from those of other known lymphocyte activation antigens. MAb 5A6/8 was able to inhibit lymphocyte proliferative responses driven by different stimuli, suggesting a role for this molecule in the events that lead to lymphocyte activation.

Animals↗

In vitro effect of classical swine fever virus on a porcine aortic endothelial cell line.

The effect of classical swine fever (CSF) virus on some phenotypic and functional features of an established porcine aortic endothelial cell (AOC) line was investigated. AOC cells show most of the characteristics of primary endothelial cells, avoiding the alterations and senescence that these cells undergo after a few passages in culture. AOC cells were susceptible to CSF virus infection to a high degree, reaching 90% of CSF virus positive cells after 24 h of infection; however as with other porcine susceptible cells, no cytopathic effect could be observed. In these conditions none of the surface molecules studied, including SLA-II MHC antigens, adhesion or co-stimulatory molecules, were altered by virus infection after 24 or 48 h. Functionally CSF virus infection induced a decrease in the pro-coagulant activity of the AOC cells, determined by the increase in the clot formation time shown by the lysates of these cells. This contrasts with the increase observed in the expression of mRNA corresponding to IL-1 alpha and IL-6, two proinflammatory and pro-coagulant cytokines, in CSF virus-infected AOC cells.

Animals↗