Transplantation of mononuclear cells from human umbilical cord blood promotes functional recovery after traumatic spinal cord injury in Wistar rats

Cell transplantation is a promising experimental treatment for spinal cord injury. The aim of the present study was to evaluate the efficacy of mononuclear cells from human umbilical cord blood in promoting functional recovery when transplanted after a contusion spinal cord injury. Female Wistar rats (12 weeks old) were submitted to spinal injury with a MASCIS impactor and divided into 4 groups: control, surgical control, spinal cord injury, and one cell-treated lesion group. Mononuclear cells from umbilical cord blood of human male neonates were transplanted in two experiments: a) 1 h after surgery, into the injury site at a concentration of 5 x 10⁶ cells diluted in 10 µL 0.9% NaCl (N = 8-10 per group); b) into the cisterna magna, 9 days after lesion at a concentration of 5 x 10⁶ cells diluted in 150 µL 0.9% NaCl (N = 12-14 per group). The transplanted animals were immunosuppressed with cyclosporin-A (10 mg/kg per day). The BBB scale was used to evaluate motor behavior and the injury site was analyzed with immunofluorescent markers to label human transplanted cells, oligodendrocytes, neurons, and astrocytes. Spinal cord injury rats had 25% loss of cord tissue and cell treatment did not affect lesion extension. Transplanted cells survived in the injured area for 6 weeks after the procedure and both transplanted groups showed better motor recovery than the untreated ones (P < 0.05). The transplantation of mononuclear cells from human umbilical cord blood promoted functional recovery with no evidence of cell differentiation.     

High-intensity intermittent exercise increases adenosine hydrolysis in platelets and lymphocytes and promotes platelet aggregation in futsal athletes

Acute bouts of high-intensity intermittent exercise (HIIE) or sports are associated with changes in lymphocytes and platelet functions and we hypothesized that the purinergic system is involved with these alterations. We investigated the activity of ectonucleotidases in platelets and lymphocytes as well as the platelet aggregation of futsal players in response to an acute protocol of HIIE. Thus, 19 male semi-professional futsal players were submitted to 40 min of HIIE on a treadmill. Blood samples were collected three-time points: before exercise, immediately after, and 30 min after the end of the session. Platelet-rich plasma (PRP) and lymphocytes were isolated. ATP, ADP, AMP, and adenosine hydrolysis, NTPDase1 (CD39) expression as well as platelet aggregation were measured. Our results showed HIIE induced a decrease in ATP and ADP hydrolysis in platelets, an increase in adenosine hydrolysis and an increase in platelet aggregation immediately after exercise. After 30 min of recovery, enzymatic activity and platelet aggregation returned to baseline levels. In lymphocytes, adenosine hydrolysis was augmented immediately after exercise and remained increased even after 30 min of recovery. In conclusion, acute HIIE triggers a transient proaggregant status that is reverted after a 30 min of recovery. The effects of HIIE in lymphocytes remained after 30 min of recovery, indicating a pro-inflammatory response. This work elucidated some of the mechanisms by which purinergic system regulates lymphocytes and platelets activities related to HIIE, suggesting that the type of exercise may influence an increase in platelet aggregation even in trained individuals.     

Multilocular Cystic Leiomyoma of the Kidney

A large, multilocular cystic mass from the kidney of a 26-year-old woman was examined by light microscopy and transmission electron microscopy. Ultrastructurally the cysts were shown to be lined by smooth muscle cells, giving evidence that the cysts originated in smooth muscle. Thus the tumor is considered to be a multilocular cystic leiomyoma, an apparently unique renal tumor.     

Hematologic and immunophenotypic characterization of human umbilical cord blood.

In this work, cord blood cells from 30 healthy term newborns were analyzed for complete blood counts with an automated cytometer and, in part of the sample, for surface molecules in cord blood monocytes, lymphocytes and CD34+ cells by two-color flow cytometry. Hematological parameters were as follows: WBC = 12.85 (5.24-15.10) x10(9)/l; platelets = 304.33 (156.00-469.00) x 10(9)/l; Hb = 14.45 (11.90-17.82) g/dl; RBC = 3.99 (3.14-5.12) x 10(12)/l; MCV 107.25 (99.60-115.00) fl; reticulocytes = 157.80 (101.00-124.00) x 10(9)/l or 3.99 (2.45-6.01)%; erythroblasts = 0.88 (0.15-2.58) x 10(9)/l or 6.63 (2.86-16.80) per 100 WBC [corrected]. The mononuclear population, as evaluated by flow cytometry, was composed of 22.9 +/- 7.2% monocytes and 77.05 +/- 7.24% lymphocytes, among which 46.59 +/- 15.62% were T lymphocytes (43.94 +/- 16.94% CD3+/CD4+ and 13.45 +/- 7.46% CD3+/CD8+). CD34+ cells were on average 0.54 +/- 0.24% of the mononuclear fraction. CD11c, CD49e and HLA-DR were found mainly on monocytes, and CD31 and CD62L occurred in similar levels on monocytes and lymphocytes. CD117+ cells were less than 5% of these populations. Among CD34+ cells, CD31 and HLA-DR were the molecules with higher frequencies (79.7 +/- 19.9 and 65.7 +/- 23.0%, respectively), followed by CD62L (41.8 +/- 31.9%) and CD117 (20.1 +/- 15.8%). The presence of CD11c and CD49e on CD34+ cells was low (below 10%). The results stress the phenotypic heterogeneity of cord blood CD34+ cells, and the different behavior of the cells when manipulated in vitro in different degrees of isolation.     

Effects of cryopreservation on the characteristics of dental pulp stem cells of intact deciduous teeth

Objectives

The aim of this study was to isolate and cultivate cells from the pulp of 7-day-cryopreserved intact deciduous human teeth and evaluate the effect of cryopreservation on dental pulp stem cell (DPSC) characteristics.

Design

Twenty-six deciduous teeth were collected and allocated in two groups: immediate cell isolation (non-cryopreserved group) and intact cryopreserved (cryopreserved group). The teeth were cryopreserved in dimethylsulfoxide solution and recovered after 7 days. The success rate of isolation, proliferation, surface markers (CD14, CD29, CD34, CD45, CD73, CD90, and HLA-DR), differentiation capacity, and morphology were evaluated.

Results

Isolation success rate was 61% and 30% for the non-cryopreserved and cryopreserved groups, respectively. There were no statistical differences between the groups for the tested surface markers. The cells in both groups were capable of differentiating into three mesenchymal lineages. No statistical differences between the groups were observed through the time course proliferation assay (0, 1, 3, 5, and 7 days); however, the mean time between isolation and the fifth passage was shorter for the non-cryopreserved group (p = 0.035). The morphology of the cells was considered altered in the cryopreserved group.

Conclusion

DPSCs were obtained from cryopreserved intact deciduous teeth without changes in the immunophenotypical characteristics and differentiation ability; however, lower culture rates, proliferation potential, and morphological alterations were observed in relation to the control group.     

Nephroblastoma in adults

Nephroblastoma, or Wilms tumor, is the most common renal neoplasm in children and accounts for approximately a fifth of all malignant growths in this age group. However, the incidence of Wilms tumor in adults is much less common, with less than 200 cases having been reported in the literature. In addition, since there are at least 53 synonyms for this tumor an adequate computer search for articles relating to this subject is difficult. Because of the scarcity of this disease in adults definite treatment modalities have not been accepted thoroughly until recently. The rationale for the present treatment modalities is based largely on the National Wilms Tumor Study groups 1 to 3. We present radiologic and histologic findings of this tumor in 2 additional cases. The current treatment modalities, consisting of chemotherapy with or without adjunctive radiotherapy, are discussed.     

Cis variants identified in F508del complex alleles modulate CFTR channel rescue by small molecules

Molecules correcting the trafficking (correctors) and gating defects (potentiators) of the cystic fibrosis causing mutation c.1521_1523delCTT (p.Phe508del) begin to be a useful treatment for CF patients bearing p.Phe508del. This mutation has been identified in different genetic contexts, alone or in combination with variants in cis. Until now, 21 exonic variants in cis of p.Phe508del have been identified, albeit at a low frequency. The aim of this study was to evaluate their impact on the efficacy of CFTR‐directed corrector/potentiator therapy (Orkambi). The analysis by minigene showed that two out of 15 cis variants tested increased exon skipping (c.609C > T and c.2770G > A). Four cis variants were studied functionally in the absence of p.Phe508del, one of which was found to be deleterious for protein maturation c.1399C > T (p.Leu467Phe). In the presence of p.Phe508del, this variant was the only to prevent the response to Orkambi treatment. This study showed that some patients carrying p.Phe508del complex alleles are predicted to poorly respond to corrector/potentiator treatments. Our results underline the importance to validate treatment efficacy in the context of complex alleles.