Human AB serum for generation of mesenchymal stem cells from human chorionic villi: comparison with other source and other media including platelet lysate.

OBJECTIVES: We have investigated foetal mesenchymal stem cells (MSCs) obtained from first-trimester chorionic villi (CV) and second-trimester amniotic fluid (AF), comparing them to adult bone marrow-derived MSCs. MATERIALS AND METHODS: We report on cell population growth in human allogeneic serum (HS) and platelet lysate (PL), immunophenotype, cytokine expression profile and immunoregulatory activity, of these foetal MSCs on stimulated peripheral blood mononuclear and lymphocyte subpopulations. RESULTS: Chorionic villi cells grow rapidly in HS, with 20 populations doublings (PDs) after 59 days (six passages), and also in animal serum, with 27 PDs after 65 days (seven passages). PL allowed for expansion in 60% of the samples tested, although it was lower than in HS. HS supported an average of 40 PDs of expansion in 20% of AF cells after 90 days, whereas animal serum supported 28.5 PDs in 66 days. CV and AF cells inhibited proliferation of stimulated T lymphocytes, suppressing population growth of both CD4+ and CD8+ T subpopulations and sometimes also, CD19+ cells. CONCLUSIONS: Our results indicate that CV would be an optimal source of MSCs with high expansion potential in a HS propagation system and immunoregulatory capacity of T and B lymphocytes. More than 90% of CV samples achieved large-scale expansion in HS, which is encouraging for potential clinical applications of these cells.

Coenzyme Q10, antioxidant status and ApoE isoforms.

The aim of this study was to inquire the antioxidant status in plasma and lipoproteins isolated from normal subjects possessing different ApoE genotypes. For this purpose we investigated blood samples from 106 healthy blood donors: the distribution of ApoE alleles (E2/E2 = 0.9%, E2/E3 = 10.4%, E2/E4 = 2.8%, E3/E3 = 71.7%, E3/E4 = 12.3% and E4/E4 1.9% with 1, 11, 3, 76, 13, and 2 subjects respectively for each genotype) was in agreement with previous data. Almost no differences were found in the concentrations of both coenzyme Q10 (CoQ10) and vitamin E for the different genotypes. Concentration of CoQ10 in isolated lipoproteins was also similar, in the different genotypes, when referred to cholesterol; CoQ10 in LDL was higher for the E3/E3 subjects when referred to protein. Neither CoQ10 nor vitamin E correlated with paraoxonase (PON) activity or cholesteryl-ester hydroperoxides (CHP). Furthermore, there was no correlation between the same lipophilic antioxidants and CHP levels. The only E2 homozygous subject found had high levels of PON and low levels of CHP; the two E4/E4 subjects had low PON activity together with low levels of CHP.

Lower serum activity of prolyl endopeptidase in anorexia and bulimia nervosa.

The aim of this study was to examine whether anorexia and bulimia nervosa are accompanied by lower serum activity of prolyl endopeptidase (PEP;EC 3.4.21.26; post-proline cleaving enzyme), a cytosolic endopeptidase which cleaves peptide bonds on the carboxyl side of proline in proteins of relatively small molecular mass. Substrates of PEP are, amongst others, neuroactive peptides, such as arginine vasopressin, luteinizing hormone-releasing hormone, thyrotropin releasing hormone,alpha-melanocyte secreting hormone, substance P, oxytocin, bradykinin, neurotensin and angiotensin (Ag) I and II. Serum PEP activity was measured in the serum of 18 normal women, 21 anorexia nervosa and 21 bulimia nervosa women by means of a fluoremetric method. The Bulimic Investigatory Test, Edinburgh (BITE), the Eating Disorder Inventory (EDI) and the Hamilton Depression Rating Scale (HDRS) were scored. Serum PEP activity was significantly lower in patients with bulimia nervosa and anorexia nervosa, irrespective of the restricted or binging subtype, than in normal controls. There were significant and inverse correlations between serum PEP activity and the HDRS and BITE. In anorectic patients, but not in normal or bulimic patients, there was a significant correlation between serum PEP and body mass index. In bulimic patients, but not in normal or anorectic patients, there was a significant correlation between serum PEP and duration of illness. It is concluded that lowered serum PEP activity takes part in the pathophysiology of anorexia and bulimia nervosa. It is hypothesized that a combined dysregulation of PEP and neuroactive peptides, which are substrates of PEP, could be an integral component of eating disorders.

A questionnaire assessment of unmet needs for rehabilitation services and resources for people with multiple sclerosis: results of a pilot survey in five European countries. Needs Task group of MARCH (Multiple Sclerosis and Rehabilitation, Care and Health Services Research in Europe).

OBJECTIVE: To develop an international services and needs assessment instrument (SUN) for people with multiple sclerosis and their carers and to pilot this in different countries of the European Community. DESIGN: Interview study of people with multiple sclerosis, their carers and nominated key professionals examining the unmet needs of patients and carers. SETTING: Belgium, Estonia, Greece, Italy and the United Kingdom. MAIN OUTCOME MEASURES: Needs assessment questionnaire. RESULTS: The study comprised 137 people with multiple sclerosis, 125 carers and 111 professionals. Patients reported on average 2.9 unmet needs for themselves; their carers and professionals reported on average 2.4. Needs were categorized into seven broad categories. Due to difficulties experienced by the local researchers in distinguishing between needs and objectives a large proportion of needs had to be assigned to the 'other' category. CONCLUSIONS: The SUN is a valuable and practicable tool for the identification of unmet needs for people with multiple sclerosis and their carers. Formal validation and reliability testing of the different language versions is recommended.

Low number of complement C3b/C4b receptors (CR1) on erythrocytes from patients with essential mixed cryoglobulinemia, systemic lupus erythematosus and rheumatoid arthritis: relationship with disease activity, anticardiolipin antibodies, complement activation and therapy.

Our aim was to assess whether the amount of complement C3b/C4b receptors (CR1) on erythrocytes shows a correlation to disease activity in various connective tissue diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and essential mixed cryoglobulinemia (EMC). Using an anti-CR1 monoclonal antibody, 26 patients with SLE, 34 with RA and 22 patients with EMC were investigated for erythrocyte CR1 expression. The control group consisted of 30 healthy individuals. The mean number of CR1/erythrocyte in the control group was 568 +/- 197 (range 174-1060), significantly higher than studied (EMC:379 +/- 248; p = 0.0005;SLE 147 +/- 56, p less than 0.0001; RA 298 +/- 177, p less than 0.0001). In patients with RA and in SLE, but not in patients with EMC, the number of CR1 numbers and anticardiolipin antibody (aCl) titers (r2 = 0.493; p = 0.034). A statistically significant correlation between CR1 numbers and CH50 values was found in patients with SLE, while in 3 patients with RA 4 months of therapy with cyclosporine A led to a further 30% reduction in CR1 number. Our conclusions are that (a) the decreased expression of erythrocyte CR1 is apparently a common feature of patients with various connective tissue diseases; (b) several acquired factors such as disease activity, complement activation, aCl and drugs may contribute to the loss of CR1 from erythrocytes; (c) in patients with RA and SLE, but not in patients with EMC, CR1 enumeration on erythrocytes may serve as a variable for clinical monitoring.

The effect of electrode position on flash visual evoked potentials in the newborn.

We examined how changes in electrode position affected the visual evoked potential in 74 high-risk newborns using a multiple electrode technique. The variation in the visual evoked potential across the occiput was documented. Visual evoked potentials that were visible at one occipital electrode position were absent at a different electrode position in 21 of 87 recordings (24%). Changes of greater than 20 msec between electrodes in latency of waves P1 or N2 occurred in 34% and 29% of recordings, respectively. The amplitude of response varied by a factor of 2.36 between mid occipital and lateral occipital electrodes. The results demonstrated that large changes in morphologic characteristics, latency, and amplitude in the visual evoked potential of the newborn result from small changes in the position of recording electrodes. These findings underscored the importance of electrode position and accurate electrode placement. Our observations also indicated that recordings from a single electrode are inadequate in providing an accurate representation of the neonatal visual evoked potential.

Electrophysiologic studies, computed tomography, and neurologic outcome in acute bacterial meningitis.

To determine the value of computed tomography and electrophysiologic studies in predicting neurologic outcome, we prospectively studied 41 children with acute bacterial meningitis, using clinical examination, computed tomography of the head, electroencephalography, brain-stem auditory evoked response, and visual evoked potential mapping during the acute illness. Two children died; 32 of the remaining 39 children were reviewed clinically, electrophysiologically, and with computed tomography between 5 and 38 months after the illness. The electrophysiologic data obtained during the illness were not found to alter the acute-stage management. Focal or generalized suppression, demonstrated on the electroencephalogram, was associated with a poor outcome. Cerebral infarction and edema, demonstrated by computed tomography of the head, were predictive of a poor outcome, but enlarged ventricular and subarachnoid spaces and increased subdural effusions were of no predictive value. Neither computed tomographic scans nor electrophysiologic data were better indicators of neurologic prognosis than the clinical examination.

[Evaluation of cellular immunity in a selected sample of aged subjects]. / Valutazione dell'immunita' cellulare in un campione "selezionato" di soggetti anziani.

The immune system in the aging man is known to show age-related defects; the study of the subpopulations in human blood has yielded conflicting results. This mostly seems to be due to the heterogeneity of the subjects admitted to immunogerontological studies. We investigated a group of aged persons fulfilling the admission criteria described in the "SENIEUR" protocol and strictness criteria dietetics. We found a decrease in the number of T cells, while the number of B cells and, above all, the report of the T4/T8 remained constant. There individuals should be considered as optimally healthy and immunologically uncompromised.

Quantitative VEP analysis in children with cortical visual impairment.

Children with cortical visual impairment (CVI) usually have a typical clinical presentation. However, in some cases, it may be useful to have confirmatory evidence based on objective electrophysiologic information. To achieve this, we examined some mathematically derived parameters constructed from 20 channel visual evoked potential (VEP). A group of 30 children diagnosed with CVI by clinical and CT findings was compared to a normal control group of 52 children. Each recorded VEP was mathematically transformed using Hjorth's source derivation, to reduce reference contamination and enhance local features. The area under the response curve, computed for each channel within a fixed time window, was used as a measure of the response activity at that channel. These areas were then used to construct several parameters ("R values") describing ratios of activities between different recording electrode areas. Some of these ratios provided good separation between patient and control groups, especially for children older than 5 years of age; in particular CVI patients were found to have a low occipital-to-parietal activity ratio. This finding, together with the observed age independence of the R values in the normal population, their ease of computation and possible physiological interpretability, suggest that R values could be used as confirmatory diagnostic measures.

Statistical classification of spikes in benign rolandic epilepsy.

The occurrence of spikes during an electroencephalogram is a basic feature of Benign Rolandic Epilepsy of Childhood (BREC). In this study we addressed the question of whether the interictal spike structure is different between "typical" and "atypical" BREC patients. Atypical BREC patients are characterized by having other neurological abnormalities in addition to the typical BREC symptoms. This question is of interest given the good prognosis associated with the typical form of BREC. We analyzed data from 12 typical and 12 atypical BREC patients using ten variables to describe spike morphology and topography. The non-parametric method of "classification and regression trees" (CART) was used to detect discriminating features and construct a classification rule. In this way the location and amplitude of the spike were found to provide satisfactory discrimination, suggesting that the two groups may be affected by different epileptic processes. The study showed the effectiveness of the CART methodology in dealing with the categorical and non-normal variables that can be obtained from EEG records. It illustrates a simple but logical approach to the analysis of potentially complex topographic data.