Glutathione Ethyl Ester Supplementation during Pancreatic Islet Isolation Improves Viability and Transplant Outcomes in a Murine Marginal Islet Mass Model

Background

The success of pancreatic islet transplantation still faces many challenges, mainly related to cell damage during islet isolation and early post-transplant. The increased generation of reactive oxygen species (ROS) during islet isolation and the consumption of antioxidant defenses appear to be an important pathway related to islet damage.

Methodology/Principal Findings

In the present study we evaluated whether supplementation of glutathione-ethyl-ester (GEE) during islet isolation could improve islet viability and transplant outcomes in a murine marginal islet mass model. We also cultured human islets for 24 hours in standard CMRL media with or without GEE supplementation. Supplementation of GEE decreased the content of ROS in isolated islets, leading to a decrease in apoptosis and maintenance of islet viability. A higher percentage of mice transplanted with a marginal mass of GEE treated islets became euglycemic after transplant. The supplementation of 20 mM GEE in cultured human islets significantly reduced the apoptosis rate in comparison to untreated islets.

Conclusions/Significance

GEE supplementation was able to decrease the apoptosis rate and intracellular content of ROS in isolated islets and might be considered a potential intervention to improve islet viability during the isolation process and maintenance in culture before islet transplantation.     

Impairment of IFN-Gamma Response to Synthetic Peptides of Mycobacterium tuberculosis in a 7-Day Whole Blood Assay

Studies on Mycobacterium tuberculosis (MTB) antigens are of interest in order to improve vaccine efficacy and to define biomarkers for diagnosis and treatment monitoring. The methodologies used for these investigations differ greatly between laboratories and discordant results are common. The IFN-gamma response to two well characterized MTB antigens ESAT-6 and CFP-10, in the form of recombinant proteins and synthetic peptides, was evaluated in HIV-1 uninfected persons in both long-term (7 day) and 24 hour, commercially available QuantiFERON TB Gold in Tube (QFT-GIT), whole blood assays. Our findings showed differences in the IFN-gamma response between 24 hour and 7 day cultures, with recombinant proteins inducing a significantly higher response than the peptide pools in 7 day whole blood assays. The activity of peptides and recombinant proteins did not differ in 24 hour whole blood or peripheral blood mononuclear cell (PBMC) based assays, nor in the ELISpot assay. Further analysis by SELDI-TOF mass spectrometry showed that the peptides are degraded over the course of 7 days of incubation in whole blood whilst the recombinant proteins remain intact. This study therefore demonstrates that screening antigenic candidates as synthetic peptides in long-term whole blood assays may underestimate immunogenicity.     

Hemangiomas - current therapeutic strategies

Hemangiomas are benign neoplasms of the vasculature frequently encountered in children. Several studies have shown that these tumors are characterized by excessive angiogenesis. Although benign, the lesions can present with complications, and may thus require treatment. There are multiple therapeutic options available for patients with problematic or life threatening hemangiomas, some of which have serious side effects. Randomized clinical trials and evidence-based studies on the efficacy of these treatments is still lacking. The recognition that excessive angiogenesis underlies hemangiogenesis offers an opportunity for the development of safer therapeutic strategies that are based on the inhibition of angiogenesis. We review medical therapies currently employed in the management of hemangiomas and the role of angiogenesis inhibition in hemangioma therapy.     

Mechanical properties of mitral allografts are not reasonably influenced by cryopreservation in sheep model

A mitral allograft is used exceptionally in the mitral, as well as in the tricuspid position, mostly as an experimental surgical procedure. The authors decided to evaluate the possibility of inserting a cryopreserved mitral allograft into the tricuspid position in a sheep experimental model. Within the framework of this experimental project the mechanical properties of the cryopreserved mitral allograft were tested. A novel methodology studying the functional unit composed of mitral annulus, leaflet, chordae tendinaea, and papillary muscle is presented. A five-parameter Maxwell model was applied to characterize the viscoelastic behavior of sheep mitral valves. A control group of 39 fresh mitral specimens and a test group of 13 cryopreserved mitral allografts from tissue bank were tested. The testing protocol consisted of six loading cycles with 1 mm elongation every 5 min. There was no significant difference in the mean values of the determined parameters (p>0.05) which confirms the main hypothesis that cryopreservation does not influence significantly material parameters characterizing the tissue mechanics. Slight discrepancy is observed in variances of viscous parameters suggesting that the values of the test group may be spread over larger interval due to the treatment.     

New role for tRNA and its fragment purified from human urinary bladder carcinoma conditioned medium: Inhibition of endothelial cell growth

The growth of endothelial cells is necessary for angiogenesis, which in turn is required for later steps of tumor progression. In an attempt to purify new modulators of endothelial cell growth from the conditioned medium of human urinary bladder carcinoma cells, we isolated a small and stable oligonucleotide containing 10 to 16 bases. This oligonucleotide inhibited the growth of endothelial cells in vitro and was identified as a fragment of transfer RNA (tRNA). When unfractionated bovine tRNA was added to the cell culture, it specifically inhibited growth of endothelial cells, but not smooth muscle cells, bovine kidney cells, 3T3 fibroblasts, and several cancer cell lines. In contrast, ribosomal RNA, total yeast RNA, and single nucleosides from tRNA hydrolysate had no effect. These results demonstrate a new role for tRNA and its fragment as a selective endothelial cell inhibitor in vitro. J. Cell. Biochem. 76:109–117, 1999. © 1999 Wiley‐Liss, Inc.     

Molecular phylogeny of the Barnadesioideae (Asteraceae)

The small but morphologically diverse subfamily Barnadesioideae of the sunflower family, Asteraceae, is of special interest as it constitutes the sister-group to the rest of the family. Therefore it is of critical importance for elucidating the origin and early evolution of Asteraceae. Cladistic analyses of DNA sequence variation in the trnL intron and nuclear ribosomal ITS regions strongly support five major clades in the subfamily: Schlechtendalia, Chuquiraga-Doniophyton, Barnadesia-Huarpea, Dasyphyllum subgenus Dasyphyllutn and a clade comprising Dasyphyllum subgenus Archidasyphyllum, Arnaldoa and Fulcaldea. Within Dasyphyllum subgenus Dasyphyllum, D. hystrix has a basal position, and sect. Macrocephala is supported as monophyletic, while sect. Microcephala lacks jackknife support. Within Barnadesia, B. parviflora has a very divergent ITS sequence and a basal position in the genus. The phylogenetic trees make some sense of the great morphological variation within the subfamily, although some clades identified here lack obvious defining morphological characteristics. Optimisation of geographical distributions onto the molecular phylogenies shows that the Barnadesioideae most likely originated in southern South America.     

Plant root distributions and nitrogen uptake predicted by a hypothesis of optimal root foraging

CO(2)-enrichment experiments consistently show that rooting depth increases when trees are grown at elevated CO(2) (eCO(2)), leading in some experiments to increased capture of available soil nitrogen (N) from deeper soil. However, the link between N uptake and root distributions remains poorly represented in forest ecosystem and global land-surface models. Here, this link is modeled and analyzed using a new optimization hypothesis (MaxNup) for root foraging in relation to the spatial variability of soil N, according to which a given total root mass is distributed vertically in order to maximize annual N uptake. MaxNup leads to analytical predictions for the optimal vertical profile of root biomass, maximum rooting depth, and N-uptake fraction (i.e., the proportion of plant-available soil N taken up annually by roots). We use these predictions to gain new insight into the behavior of the N-uptake fraction in trees growing at the Oak Ridge National Laboratory free-air CO(2)-enrichment experiment. We also compare MaxNup with empirical equations previously fitted to root-distribution data from all the world's plant biomes, and find that the empirical equations underestimate the capacity of root systems to take up N.