Mesenchymal stem cells from umbilical cord blood: parameters for isolation, characterization and adipogenic differentiation.

Isolation of mesenchymal stem cells (MSCs) from umbilical cord blood (UCB) from full-term deliveries is a laborious, time-consuming process that results in a low yield of cells. In this study we identified parameters that can be helpful for a successful isolation of UCB-MSCs. According to our findings, chances for a well succeeded isolation of these cells are higher when MSCs were isolated from UCB collected from normal full-term pregnancies that did not last over 37 weeks. Besides the duration of pregnancy, blood volume and storage period of the UCB should also be considered for a successful isolation of these cells. Here, we found that the ideal blood volume collected should be above 80 mL and the period of storage should not exceed 6 h. We characterized UCB-MSCs by morphologic, immunophenotypic, protein/gene expression and by adipogenic differentiation potential. Isolated UCB-MSCs showed fibroblast-like morphology and the capacity of differentiating into adipocyte-like cells. Looking for markers of the undifferentiated status of UCB-MSCs, we analyzed the UCB-MSCs' protein expression profile along different time periods of the differentiation process into adipocyte-like cells. Our results showed that there is a decrease in the expression of the markers CD73, CD90, and CD105 that correlates to the degree of differentiation of UCB-MSCs We suggest that CD90 can be used as a mark to follow the differentiation commitment degree of MSCs. Microarray results showed an up-regulation of genes related to the adipogenesis process and to redox metabolism in the adipocyte-like differentiated MSCs. Our study provides information on a group of parameters that may help with successful isolation and consequently with characterization of the differentiated/undifferentiated status of UCB-MSCs, which will be useful to monitor the differentiation commitment of UCB-MSC and further facilitate the application of those cells in stem-cell therapy.

Migraine and motion sickness independently contribute to visual discomfort.

The objective of this study was to evaluate, in patients with migraine and healthy volunteers, with and without a history of motion sickness, the degree of discomfort elicited by drifting striped patterns. Eighteen healthy volunteers (HV) and 30 migraine patients participated in the study. Discomfort was greater in migraine patients than in HV, and in individuals with a history of motion sickness than in those without, but the effect of history of migraine was independent of history of motion sickness. Generalized Estimating Equations models for binary correlated data revealed that these differences did not depend on levels of duty cycle, spatial and temporal frequencies. Visual discomfort in migraine patients was associated with worse performance. There was a significant correlation between median degree of discomfort across conditions and number of migraine attacks in the past month. Discomfort to drifting striped patterns may be related to central sensitization in migraine patients.

Brazilian experience with two conditioning regimens in patients with multiple sclerosis: BEAM/horse ATG and CY/rabbit ATG.

Studies have shown that autologous hematopoietic SCT (HSCT) can be used as an intensive immunosuppressive therapy to treat refractory patients and to prevent the progression of multiple sclerosis (MS). This is a prospective multicentric Brazilian MS trial comparing two conditioning regimens: BEAM/horse ATG and CY/rabbit ATG. Most (80.4%) of the 41 subjects in the study had the secondary progressive MS subtype and the mean age was 42 years. The baseline EDSS score in 58.5% of the subjects was 6.5 and 78% had a score of 6.0 or higher, respectively. The complication rate during the intra-transplantation period was 56% for all patients: 71.4% of the patients in the BEAM/hATG group and 40% in the CY/rATG group (P=0.04). Three subjects (7.5%) died of cardiac toxicity, sepsis and alveolar hemorrhage, all of them in the BEAM/ATG group. EFS was 58.54% for all patients: 47% in the BEAM/hATG group and 70% in the CY/rATG group (P=0.288). In conclusion, the CY/rATG regimen seems to be associated with similar outcome results, but presented less toxicity when compared with the BEAM/hATG regimen. Long-term follow-up would be required to fully assess the differences in therapeutic effectiveness between the two regimens.

Analysis of liver transplantation outcome in patients with MELD Score > or = 30.

Since July 2006, the liver graft allocation has been changed from the waiting time to the Model for End-stage Liver Disease (MELD), prioritizing the sickest patients, who have a higher risk of dying on the waiting list, and sometimes in such poor clinical condition that it compromises transplantation outcomes. The aim of this study was to analyze the impact of a MELD score > or = 30 on 30-day survival after liver transplantation (OLT). We prospectively collected the data on 178 liver transplants on 163 patients performed from March 2003 to August 2007. The subjects were divided in two groups according to their MELD scores: group 1, MELD > or = 30 (n = 15) and group 2, MELD < 30 (n = 96). The groups were compared with regard to hospital and intensive care unit (ICU) length of stay, intraoperative blood products transfusion, early survival (30 days), and need for retransplantation. We excluded, patients with prioritization criteria, those receiving extra points for any special situation, and six other patients without significant data for MELD calculation (of whom only one has died after transplantation). Patients under a "special situation" were those with hepatocelular carcinoma, hepatopulmonary syndrome, and metabolic diseases, who initially received a MELD/PELD score 20, and 24, and 29. The mean MELD score at group I was 34 (range, 30 to 42), and for group II it was 16 (range, 6 to 29). Group I displayed a mean hospital length of stay of 24 days (4 to 155), with 12.60 days (ranges, 1 to 103) in the ICU versus 15.55 (range, 1 to 48) and 5.13 (range, 1 to 45) days, respectively, for group II. The need for blood component transfusions were greater in group I; 25.28% of patients in group II did not receive any transfusion during the entire inpatient period. There were nine retransplants in group II, and none in group I. The 30-day survivals were 93.3% for group I and 84.37% for group II. Besides the increased complexity of these sickest patients, there was no negative impact on early survival rates.

Impact of cumulative risk factors for expanded criteria donors on early survival after liver transplantation.

There are various options to help overcome the organ shortage, including performing transplants using grafts from marginal donors with characteristics previously described as unacceptable because of the high risk of graft failure. Nowadays, expanded criteria donors for liver transplantation (OLT) is a strategy used routinely by many teams. Some donor features have been suggested to jeopardize initial function or survival; when these features are aggregated, they may impact prognosis. The aim of this study was to evaluate the impact of donor risk factors on early patient survival and retransplantation. Donor risk factors were considered to be older than 60 years, body mass index > 30, serum sodium level > 155 mEq/L, cold ischemia time > 12 hours, and intensive care unit stay > 4 days. We prospectively recorded data from 139 patients who underwent 152 OLT from March 2003 to May 2007. Patients were classified into four groups: I, no risk factors; II, one risk factor; III, two risk factors; IV, three or more risk factors. Retransplantation or OLT due to acute liver failure was considered to be an exclusion criterion. Early overall survival rate was 83.76%; 12 retransplantations were required (10.25%). Comparing the four groups, patient survivals (P = .41) and retransplantation rates (P = .518) were similar. In conclusion, cumulative risk factors showed no impact on early (30-day) recipient survival and or on the necessity of retransplantation after OLT.

Inspiratory fraction and exercise impairment in COPD patients GOLD stages II-III.

The inspiratory-to-total lung capacity ratio or "inspiratory fraction" (inspiratory capacity(IC)/total lung capacity (TLC)) may be functionally more representative than traditional indices of resting airflow limitation and lung hyperinflation in patients with chronic obstructive pulmonary disease (COPD). In the present retrospective study, a comparison was made of the individual performance of post-bronchodilator IC, IC/TLC and forced expiratory volume in one second (FEV(1)) in predicting a severely reduced peak oxygen uptake (V'(O(2)); <60% predicted) in 44 COPD patients Global Initiative for Chronic Obstructive Lung Disease stages II-III (post-bronchodilator FEV(1) ranging from 31-79% pred). Patients with lower IC/TLC values (

The impact of syncope during clinical presentation of sustained ventricular tachycardia on total and cardiac mortality in patients with chronic Chagasic heart disease.

OBJECTIVE: To assess the impact of syncope during sustained ventricular tachycardia on total and cardiac mortality in patients with chronic chagasic heart disease. METHODS: We assessed 78 patients with sustained ventricular tachycardia and chronic Chagas' heart disease. The mean age was 53+/-10 years, 45 were males, and the mean ejection fraction was 49.6+/-13%. The patients were divided into 2 groups according to the presence (GI=45) or absence (GII=33) of syncope during sustained ventricular tachycardia. RESULTS: After a mean follow-up of 49 months, total mortality was 35% (28 deaths), 22 deaths having a cardiac cause (78.6%). No difference was observed in total (33.3% x 39.4%) and cardiac (26.7% x 30.3%) mortality, or in nonfatal sustained ventricular tachycardia between GI and GII patients (57.6% x 54.4%, respectively). However, the presence of syncope during recurrences was significantly greater in those patients who had had the symptom from the beginning (65.4% x 18.1%, p<0.01). CONCLUSION: Syncope during the presentation of sustained ventricular tachycardia is not associated with an increase in total or cardiac mortality in patients with chronic Chagas' heart disease. However, syncope during the recurrence ventricular tachycardia is greater in patients experiencing syncope in the first episode, of sustained ventricular tachycardia.

Risk of linear growth retardation during the first two years of life: a new approach.

OBJECTIVE: Estimate the risk of linear growth retardation during the first two years of life as a result of household social vulnerability. SETTING: Families who participated in the National Supplementary Feeding Program in the Health Units of the metropolitan area of the city of São Paulo, Brazil. SUBJECTS: Four hundred and thirty-one index-babies, weighing more than 2500 grams and who had at least one young sibling under the age of five who participated in the Program for a minimum of two years. DESIGN: The index-babies were divided into two cohorts: 74.9% coming from 'non-stunted families' (those with normal height siblings) and 25.1% from 'stunted families' (those with stunted siblings). The study design allowed the observation of growth patterns over a period of time and over a childhood growth range. It also allowed the estimation of the stunting and the recovery probabilities at each moment, not only within a given age range. The transition probabilities between 'stunted' and 'non-stunted' index-babies were estimated. The relative risk ratio (RR) was also calculated. RESULTS: The prevalence of stunting in the index-babies at 12 and 24 months of age was significantly greater in 'stunted families' (P < 0.001). Probabilities of becoming stunted began to differ from the fourth month on (confidence intervals non-superposed), and were higher for index-babies from 'stunted families'. The recovery probability of a stunted child was smaller in the 'stunted families' cohort after the 12th month of age. From the third month on, the (RR) was always above 1.5. CONCLUSION: The family context exposes children to failure in growth in the first two years of life when there are already stunted children in the household.