Rapamycin does not adversely affect intrahepatic islet engraftment in mice and improves early islet engraftment in humans.

Objective: In this study we examined the effect of rapamycin (RAPA), a key component of the immunosuppressive regimen in clinical islet transplantation, on islet engraftment and function in vivo. Methods and results: Diabetic C57BL/6 or BALB/C recipient mice were transplanted with 350 syngeneic islets through the portal vein (PV-Tx; C57BL/6 n = 60; BALB/C n = 22) and treated with once-daily oral RAPA (1 mg/kg) or vehicle. No differences in post-transplant blood glucose concentrations and glucose tolerance were observed between RAPA-and vehicle-treated mice. The impact of RAPA on human islet engraftment was assessed in 10 patients with type 1 diabetes treated with 0.1 mg/kg/day rapamycin before islet transplantation. Compared to non pre-treated islet transplant recipents (n = 12), RAPA pre-treated patients had increased blood RAPA concentrations (p = 0.006) and fasting C-peptide concentrations (p = 0.005) in the two weeks post-transplant. RAPA pre-treatment was associated with a reduction in chemokines CCL2 and CCL3 concentrations pre-transplant (p < 0.01), and a dampened chemokine response (p = 0.005) post-transplant. Concordantly, in vitro RAPA inhibited the secretion of CCL2 and CCL3 by monocytes. Conclusion: Rapamycin does not adversely affect intrahepatic islet engraftment in the mouse, and potentially improves islet engraftment in humans by an anti-inflammatory mechanism

SERS spectroscopy as a tool for the study of thiopurine drug pharmacokinetics in a model of human B leukemia cells

: The thiopurine drugs are immunomodulatory antimetabolites that are characterized by dose-dependent adverse effects such as myelosuppression and hepatotoxicity, often related to inter-individual differences, involving the activity of important enzymes at the basis of their biotransformation, such as thiopurine S-methyltransferase (TPMT). Surface Enhanced Raman Scattering (SERS) spectroscopy is emerging as a bioanalytical tool and represents a valid alternative in terms of affordable costs, shorter analysis time and easier sample preparation in comparison to the most employed methods for pharmacokinetic analysis of drugs. The aim of this study is to investigate mercaptopurine and thioguanine pharmacokinetics by SERS in cell lysates of a B-lymphoblastoid cell line (NALM-6), that did (TPMT*1) or did not (MOCK) overexpress the wild-type form of TPMT as an in vitro cellular lymphocyte model to discriminate between cells with different levels of TPMT activity on the base of the amount of thioguanosine nucleotides (TGN) metabolites formed. SERS analysis of the cell lysates was carried out using SERS substrates constituted by Ag nanoparticles deposited on paper and parallel samples were used for quantification of thiopurine nucleotides with liquid chromatography-tandem mass-spectrometry (LC-MS/MS). A direct SERS detection method has been set up that could be a tool to study thiopurine drug pharmacokinetics in in vitro cellular models to qualitatively discriminate between cells that do and do not overexpress the TPMT enzyme, as an alternative to other more laborious techniques. Results underlined decreased levels of TGN and increased levels of methylated metabolites when TPMT was over expressed, both after mercaptopurine and thioguanine treatments. A strong positive correlation (Spearman's rank correlation coefficient rho = 0.96) exists between absolute quantification of TGMP (pmol/1x106 cells), obtained by LC-MS/MS, and SERS signal (intensity of TGN at 915 cm-1). In future studies, we aim to apply this method to investigate TPMT activity in patients' leukocytes

Co-Graft of Allogeneic Immune Regulatory Neural Stem Cells (NPC) and Pancreatic Islets Mediates Tolerance, while Inducing NPC-Derived Tumors in Mice

Data available on the immunomodulatory properties of neural stem/precursor cells (NPC) support their possible use as modulators for immune-mediated process. The aim of this study was to define whether NPC administered in combination with pancreatic islets prevents rejection in a fully mismatched allograft model.Diabetic Balb/c mice were co-transplanted under the kidney capsule with pancreatic islets and GFP(+) NPC from fully mismatched C57BL/6 mice. The following 4 groups of recipients were used: mice receiving islets alone; mice receiving islets alone and treated with standard immunosuppression (IL-2Ralpha chain mAbs + FK506 + Rapamycin); mice receiving a mixed islet/NPC graft under the same kidney capsule (Co-NPC-Tx); mice receiving the islet graft under the left kidney capsule and the NPC graft under the right kidney capsule (NPC-Tx). Our results demonstrate that only the co-transplantation and co-localization of NPC and islets (Co-NPC-Tx) induce stable long-term graft function in the absence of immunosuppression. This condition is associated with an expansion of CD4(+)CD25(+)FoxP3(+) T regulatory cells in the spleen. Unfortunately, stable graft function was accompanied by constant and reproducible development of NPC-derived cancer mainly sustained by insulin secretion.These data demonstrate that the use of NPC in combination with islets prevents graft rejection in a fully mismatched model. However, the development of NPC-derived cancer raises serious doubts about the safety of using adult stem cells in combination with insulin-producing cells outside the original microenvironment

Heating response patterns of Alpine soils: from a plot-scale to lab experiments.

Wildfires play the role of ecosystem shapers in the majority of terrestrial biomes. Nowadays, their regimes are changing as a consequence of land abandonment and climate change. After-fire dynamics are widely studied in North America and Mediterranean environments. However, soils developed in different biomes might not unequivocally respond to fire-induced heating, and forests of the Western Italian Alps are not unfamiliar to fire occurrence. For these reasons, we conducted several experiments (at plot and lab scale) at environmentally realistic conditions to systematically assess the impacts of fire on the physico-chemical properties of soils belonging to the Italian Alpine ecological region. A homogenous pine forest (Pinus sylvestris L.) located in a mountain region near Torino experienced the passage of a severe and large wildfire in fall 2017. The field survey carried out in 2020 revealed that lower organic carbon (OC) contents and higher bulk density (BD) values were associated to a greater fire severity. Abundance of pyrogenic carbon was related to the steepness degree, as a consequence of erosion. In the superficial horizons, the naturally high WR expected from soils developed under a conifer stand was not present. To elucidate mechanisms regulating WR occurrence and evolution, the thermal transformations borne by Alpine soils were investigated at controlled laboratory conditions. Topsoil samples displayed extremely different wettable behaviors upon increasing temperatures (Ts), with or without WR build-up. This occurred mainly in relation to content and composition of organic matter (OM), particle size distribution and abundance of iron (Fe) oxides. Notwithstanding the initial sample hydrophobicity, WR was dramatically lost above 200 °C due to increasing pH values, inducing OM de-sorption from the negatively charged mineral surfaces. In the same T range, the thermal transformation of soil Fe oxides were found to be primarily directed towards oxidative processes (hematite formation). Ts up to 300 °C could have potentially promoted the stabilization of the remaining (non-combusted) OM, with the synthesis of defect-rich Fe oxides and enrichment in condensed and aromatic compounds, and yet OM was highly dispersible at the high pH values resulting from the thermal treatment, such that OC might be weakly retained on mineral phases in an after-fire scenario

Apollo. A. 9(1993)

A. 9(1993): Ardovino, A. M., Un’olpe di bronzo con graffiti da Padula, P. 3 ; Bonifacio, R., I rilievi su una base marmorea del Museo Archeologico Provinciale di Salerno: un’ipotesi di lettura, P. 14 ; Johannowsky, W., Appunti sulla tipologia e lo sviluppo architettonico della ‘villa urbana’, P. 30 ; Romito, M., Elementi per una documentazione di Cava dei Tirreni in età romana: una ‘villa’ e una statua funeraria ritrovata. Con un’appendice di Françoise Brien-Poitevin, P. 35 ; Di Muro, A., Organizzazione territoriale e modi della produzione nell’alto Medioevo. Il caso del ‘locus’ Tusciano, P. 60 ; Peduto, P., Rapporti tra Salerno e la Sicilia alla luce dei recenti rinvenimenti ceramici del secolo XII, P. 108 ; Pastore, I., La ceramica a ‘bande rosse’ del castello di Salerno, P. 123 ; Capriolo, G., Pergamene e diplomi conservati nell’archivio dei Musei Provinciali del Salernitano, P. 123 ; De Dominicis, A., La posa della prima pietra del nuovo edificio della Università degli Studi di Napoli al Rettifilo, P. 128 ; Guarino, D. P., Intervento conservativo su un elemento lapideo del Museo Archeologico Provinciale di Salerno, P. 136.Dono Alfonso Andria, nov. 201

Mesenchymal cells appearing in pancreatic tissue culture are bone marrow-derived stem cells with the capacity to improve transplanted islet function

Adherent fibroblast-like cells have been reported to appear in cultures of human endocrine or exocrine pancreatic tissue during attempts to differentiate human β cells from pancreatic precursors. A thorough characterization of these mesenchymal cells has not yet been completed, and there are no conclusive data about their origin. We demonstrated that the human mesenchymal cells outgrowing from cultured human pancreatic endocrine or exocrine tissue are pancreatic mesenchymal stem cells (pMSC) that propagate from contaminating pMSC. The origin of pMSC is partly extrapancreatic both in humans and mice, and by using green fluorescent protein (GFP+) bone marrow transplantation in the mouse model, we were able to demonstrate that these cells derive from the CD45+ component of bone marrow. The pMSC express negligible levels of islet-specific genes both in basal conditions and after serum deprivation or exogenous growth factor exposure, and might not represent optimal candidates for generation of physiologically competent β-cells. On the other hand, when cotransplanted with a minimal pancreatic islet mass, pMSC facilitate the restoration of normoglycemia and the neovascularization of the graft. These results suggest that pMSCs could exert an indirect role of "helper" cells in tissue repair processes