Background and Objectives
The aim of our study was to test a platelet‐rich plasma releasate (PRP‐R/SRGF) from CaCl2‐activated platelets as a source of growth factors for the expansion of mesenchymal ...stromal cells (MSCs). PRP‐R/SRGF, obtained with a low‐cost procedure, is characterized by a reduced variability of growth factor release.
Materials and Methods
PRP‐R/SRGF is a clinical‐grade quality solution obtained from CaCl2‐activated platelets. Its activity was evaluated by measuring the proliferation, the phenotype, the differentiation potential and the immunosuppressive properties of MSCs derived from bone marrow (BM) and adipose tissue (AT).
Results
PRP‐R/SRGF was more active than FBS to expand BM‐ and AT‐derived MSCs. PRP‐R/SRGF treatment did not affect the expression of typical MSCs surface markers, neither MSCs differentiation potential nor their capability to inhibit activated T‐cell proliferation.
Conclusions
The clinical‐grade PRP‐R/SRGF may be used in the clinical setting for the expansion of MSCs.
Aim of the work was to characterize the peels of novel tomato lines (with red, orange, pink, black-red, and dark-red fruit) for the content of bioactive substances and for some parameters linked to ...no-food use. Chlorophyll, polyphenols, and carotenoid content, energy value, theoretical biochemical methane potential (TBMP) through stochiometric method were evaluated. The peel color mirrored a different content of the bioactive substances: the dark-red fruits showed a significant increase of the carotenoid content, while the peel of the black-red fruits had the highest values of polyphenols. The preferential accumulation of functional compounds influenced the values of the other parameters. The colored lines compared to the red-fruited control (C) contained less carbon and hydrogen mass fractions and, subsequently, a lower LHV. This, in turn, led to a substantial difference in the estimation of the TBMP. Interestingly, this production was higher in the highly pigmented line, which contained a gene present in some high-lycopene commercial varieties. A circular production model for exhaustive use of peel is proposed.
•Wastes of tomato processing (peels mainly) may be re-used for additional process.•Peels of new tomato lines with red, yellow or black color were analyzed.•Carotenoids and polyphenols content varied greatly depending by the peel color.•The content of the antioxidant compound affected in turn the potential BMP.•Once extracted the biocompounds peels may enter in the anaerobic digestion route.
Negative magnetic shear is found to suppress electron turbulence and improve electron thermal transport for plasmas in the National Spherical Torus Experiment (NSTX). Sufficiently negative magnetic ...shear results in a transition out of a stiff profile regime. Density fluctuation measurements from high-k microwave scattering are verified to be the electron temperature gradient (ETG) mode by matching measured rest frequency and linear growth rate to gyrokinetic calculations. Fluctuation suppression under negligible E×B shear conditions confirm that negative magnetic shear alone is sufficient for ETG suppression. Measured electron temperature gradients can significantly exceed ETG critical gradients with ETG mode activity reduced to intermittent bursts, while electron thermal diffusivity improves to below 0.1 electron gyro-Bohms.
Increased β-cell senescence contributes to the development of type 2 diabetes (T2D). Exercise is critical in the treatment of T2D and can attenuate aging-associated cellular changes, but exercise ...effects on β-cell senescence are unknown. We tested the effects of treadmill running (1h/day for 10 days) on two insulin resistance models: high-fat diet and insulin receptor antagonist (S961). Exercise improved glucose responsiveness and decreased senescence markers (SA-βGal activity, p21Cip1 and p16Ink4a mRNA and protein levels). RNAseq from pancreatic islets of exercised mice revealed decreased expression of senescence effector genes and senescence associated secretory phenotype factors. Exercise increased AMPK activity in islets as assessed by alpha catalytic subunit phosphorylation. Islets from sedentary mice cultured for 2 days with serum from exercised mice showed decreased senescence, suggesting a circulatory factor was responsible for these effects. Pathway analysis of RNASeq revealed that exercise upregulated glucagon type ligand receptors. Glucagon treatment of mouse and human islets activated AMPK and decreased senescence in β-cells, an affect attenuated by an AMPK inhibitor (compound C). Exercised mice treated with a glucagon antagonist (adomeglivant) blocked AMPK phosphorylation and increased p21Cip1 and p16Ink4. Exercise increased nuclear translocation of the AMPK substrate NRF2 in mouse islets while downregulation of Nrf2 with siRNA in MIN6 cells increased transcription of p16Ink4a, suggesting a potential mechanism linking exercise, AMPK activation and downregulation of β-cell senescence. Treatment of human donor islets with serum obtained pre- and post-exercise training for 10 weeks in people with T2D showed that training activated AMPK and decreased senescence markers. These findings demonstrate that exercise training decreases β-cell senescence and suggest that this important response to exercise occurs through circulating factors mediating an AMPK-dependent pathway.
Disclosure
P.Carapeto: None. J.Kahng: None. A.T.Alves wagner: None. R.J.W.Middelbeek: Research Support; Novo Nordisk. M.F.Hirshman: Stock/Shareholder; Abbott, AbbVie Inc., Amgen Inc., Colgate-Palmolive, Eli Lilly and Company, Medtronic. G.A.Rutter: Consultant; Sun Pharmaceutical Industries Ltd. L.Goodyear: None. C.Aguayo-mazzucato: None.
Funding
Joslin Diabetes Center (to C.A-M.); Richard and Susan Smith Family Foundation; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-Brasil (001)
We investigated the role of gibberellins (GAs) in the phenotype of parthenocarpic fruit (pat), a recessive mutation conferring parthenocarpy in tomato (Solanum lycopersicum L.). Novel phenotypes that ...parallel those reported in plants repeatedly treated with gibberellic acid or having a GA-constitutive response indicate that the pat mutant probably expresses high levels of GA. The retained sensitivity to the GA-biosynthesis inhibitor paclobutrazol reveals that this condition is dependent on GA biosynthesis. Expression analysis of genes encoding key enzymes involved in GA biosynthesis shows that in normal tomato ovaries, the GA20ox1 transcript is in low copy number before anthesis and only pollination and fertilization increase its transcription levels and, thus, GA biosynthesis. In the unpollinated ovaries of the pat mutant, this mechanism is de-regulated and GA20ox1 is constitutively expressed, indicating that a high GA concentration could play a part in the parthenocarpic phenotype. The levels of endogenous GAs measured in the floral organs of the pat mutant support such a hypothesis. Collectively, the data indicate that transcriptional regulation of GA20ox1 mediates pollination-induced fruit set in tomato and that parthenocarpy in pat results from the mis-regulation of this mechanism. As genes involved in the control of GA synthesis (LeT6, LeT12 and LeCUC2) and response (SPY) are also altered in the pat ovary, it is suggested that the pat mutation affects a regulatory gene located upstream of the control of fruit set exerted by GAs.
The steep increase in human West Nile virus (WNV) infections in 2011–2012 in north-eastern Italy prompted a refinement of the surveillance plan. Data from the 2010–2012 surveillance activities on ...mosquitoes, equines, and humans were analysed through Bernoulli space–time scan statistics, to detect the presence of recurrent WNV infection hotspots. Linear models were fit to detect the possible relationships between WNV occurrence in humans and its activity in mosquitoes. Clusters were detected for all of the hosts, defining a limited area on which to focus surveillance and promptly identify WNV reactivation. Positive relationships were identified between WNV in humans and in mosquitoes; although it was not possible to define precise spatial and temporal scales at which entomological surveillance could predict the increasing risk of human infections. This stresses the necessity to improve entomological surveillance by increasing both the density of trapping sites and the frequency of captures.
Peripheral blood stem cell cryopreservation is associated with cell damage and decreased viability. We evaluated the impact of up to 10 years of cryopreservation (5% DMSO) on viability of CD34+ cells ...utilizing graft samples of consecutive patients (2002–2012) with different malignancies who underwent stem cell collection and transplantation. Viability of CD34+ cells from oncohaematological patients measured after 5 weeks (97·2 ± 0·6%) or after 9–10 years of cryopreservation (95·9 ± 0·5%) was unaffected. Haemoglobin, granulocyte and platelet recovery after transplantation of long‐term cryopreserved grafts occurred within 8–13 days. CD34+ stem cells can be safely stored up to 9–10 years, without affecting cell viability and clinical effectiveness.
•Competitive photoreactions of n-styryloxazole (n=2, 4 and 5) were investigated.•Positional isomerism affects photocyclization more than photoisomerization.•Effects of conformers and intramolecular ...Hbonds evidenced by QM calculations.•Analog compound bearing two heteroaromatic rings (oxazole and furan) was also studied.
This paper describes the results obtained in the study of the photobehaviour of heteroanalogs of stilbene bearing an oxazole ring. The competitive relaxation processes (fluorescence, isomerization and cyclization) of the excited states of n-styryloxazoles (n=2 and 4) were investigated and compared with the behaviour previously reported for n=5. After preparation of the unknown compound with n=4 and of its positional isomer with n=2 (the latter with a new method of synthesis), their photobehaviour was firstly investigated in preparative conditions by NMR analysis to measure the chemical yields of their photoproducts. The study was then continued in mild irradiation conditions to measure the quantum yields of the competitive photoreactions in the primary irradiation steps.
The effects of the position of the styryl group at the oxazole ring, the relative abundance of the various conformers and the possible formation of intramolecular Hbonds on the deactivation pathways are described. Quantum-mechanical Hyperchem calculations proved to be very useful to describe the conformational equilibria and the role of conformers on photoreactivity while more refined DFT calculations on the Z isomers allowed to explain the structure dependent competition between their isomerization/cyclization processes. The effect of the replacement of the phenyl ring with a second heteroaromatic group of electron donor character was investigated for the 5-(2-(furan-2-yl) ethenyl) oxazole.
Aging is a major risk factor for type 2 diabetes (T2D) and cellular senescence plays a critical role in β-cell dysfunction leading to T2D. Exercise is a central component in the treatment of T2D and ...has been shown to attenuate many of the biochemical changes that occur with aging including senescence, but nothing is known about the effects of exercise on β-cells and their aging process. To test the hypothesis that exercise decreases β-cell senescence we used two mouse models of insulin resistance: high-fat diet (HFD, 60% kcal from fat, 6 wks) and insulin receptor antagonist S961 (40nM/wk for 2 wks) . Exercise (treadmill, 1 h/day, 5 times/wk for 2 wks at 17 cm/s and 5% incline) prevented entrance of β-cells into senescence and reversed established senescence in both males and females across the lifespan (started at 1.5-9m or 14m old mice) as seen by a decrease in one or several of the following markers: βGal, p21Cip1 and p16Ink4a. This decrease in senescence was accompanied by improved glucose stimulated insulin secretion characterized by recovery of glucose responsiveness in islets from HFD and S961 exercised animals. Islets cultured with serum from trained animals showed decreased p21Cip1 mRNA, indicating a circulatory factor (s) was responsible for these effects. Exercise increased serum glucagon expression, identified as such factor. Incubation of human and mouse islets with glucagon (1nM) decreased p21Cip1 mRNA, and using AMPK agonist (1uM C99) and antagonist (5uM Compound C) , we found that AMPK mediates the glucagon-stimulated decrease in p21Cip1. Human cadaveric donor islets treated with serum obtained before and after 10-weeks of exercise-training in people with T2D demonstrated that training decreased senescence markers through AMPK activation.
In conclusion, exercise has anti-aging effects on mouse and human pancreatic islets by decreasing senescence and improving function. This is a novel mechanism of the effects of exercise on β-cell aging with implications for the treatment of T2D.
Disclosure
P.Carapeto: None. J.Kahng: None. A.T.Alves wagner: None. R.Middelbeek: Research Support; Novo Nordisk. M.F.Hirshman: Stock/Shareholder; Abbott, AbbVie Inc., Amgen Inc., Colgate-Palmolive, Eli Lilly and Company, Medtronic. L.J.Goodyear: None. C.Aguayo-mazzucato: Consultant; eGenesis.
Funding
American Diabetes Association (1-17-PMF-009) ; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001; Short-Term Research Experience for Underrepresented Persons (1R25DK113652) ; NIDDK grant (K23-DK114550) ; NIH grant (R01DK099511) and the Joslin Diabetes Center DRC (P30 DK36836) .
