Transparency of the cornea is essential for vision and is maintained by the corneal endothelium. Consequently, corneal endothelial decompensation arising from irreversible damage to the corneal ...endothelium causes severe vision impairment. Until recently, transplantation of donor corneas was the only therapeutic choice for treatment of endothelial decompensation. In 2013, we initiated clinical research into cell-based therapy involving injection of a suspension of cultured human corneal endothelial cells (HCECs), in combination with Rho kinase inhibitor, into the anterior chamber. The aim of the present study was to establish a protocol for cryopreservation of HCECs to allow large-scale commercial manufacturing of these cells. This study focused on the effects of various cryopreservation reagents on HCEC viability. Screening of several commercially available cryopreservation reagents identified Bambanker hRM as an effective agent that maintained a cell viability of 89.4% after 14 days of cryopreservation, equivalent to the cell viability of 89.2% for non-cryopreserved control cells. The use of Bambanker hRM and HCECs at a similar grade to that used clinically for cell based therapy (passage 3-5 and a cell density higher than 2000 cells/mm2) gave a similar cell density for cryopreserved HCECs to that of non-preserved control HCECs after 28 days of cultivation (2099 cells/mm2 and 2111 cells/mm2, respectively). HCECs preserved using Bambanker hRM grew in a similar fashion to non-preserved control HCECs and formed a monolayer sheet-like structure. Cryopreservation of HCECs has multiple advantages including the ability to accumulate stocks of master cells, to transport HCEC stocks, and to manufacture HCECs on demand for use in cell-based treatment of endothelial decompensation.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Abstract
We performed statistical analysis on the flare emission data to examine parameters related to the flare extreme-ultraviolet (EUV) spectra. This study used the data from the Geostationary ...Operational Environmental Satellite X-ray Sensors to determine the fundamental flare parameters. The relationship between soft X-ray data and EUV emission data observed by the Extreme Ultraviolet Variability Experiment on board the Solar Dynamics Observatory (SDO) MEGS-A was investigated for 50 events. The results showed the hotter Fe line emissions have strong correlation with soft X-ray data in many cases. However, our statistical study revealed that EUV flare peak flux of Fe
xv
, Fe
xvi
and He
ii
lines have weak correlation with soft X-ray peak flux. In EUV line light curves, there was time difference in peak time, however the tendency to reach the peak in order from the hotter line to cooler line was not so clear. These results indicate that the temporal evolution of EUV emission can be roughly explained by soft X-ray data. However, the time changes of temperature and density distributions in the flare loop must be needed for accurate reproduction. Moreover, we compared the geometrical features of solar flares observed by the Atmospheric Imaging Assembly on board the SDO with the fundamental flare parameters for 32 events. The ribbon distance strongly correlated with both soft X-ray flare rise and decay times. This results indicate that the geometrical feature is essential parameter for predicting flare emission duration.
Abstract
Understanding the dissolution kinetics of resist materials is essential for their efficient development. In this study, we investigated the dissolution kinetics of poly(4-hydroxystyrene) ...(PHS) with a weight-average molecular weight (
M
w
) of 8000–30 000 and a polydispersity index (
M
w
/M
n
) of 1.07–1.20. The dissolution kinetics of PHS films was observed in tetramethylammonium hydroxide (TMAH) aqueous developers by a quartz crystal microbalance (QCM) method. The TMAH concentration was changed from 0 to 2.38 wt%. The formation of a thick transient swelling layer at these
M
w
/M
n
values was suppressed compared with that at
M
w
/M
n
> 2. QCM data were analyzed using the polynomial regression to clarify the effects of
M
w
and
M
w
/M
n
on the dissolution kinetics in a narrow polydispersity region. Both dissolving and swelling kinetics largely depended on
M
w
/
M
n
.
M
w
had little effect in 2.38 wt% TMAH developer; however, it had a large effect on the swelling when 2.38 wt% TMAH developer was diluted.
Fuchs endothelial corneal dystrophy (FECD) is a slowly progressive bilateral disease of corneal endothelium in which accumulation of extracellular matrix (ECM) and loss of corneal endothelial cells ...(CECs) are phenotypic features. The corneal endothelium maintains corneal transparency by regulating water hydration; consequently, corneal endothelial dysfunction causes serious vision loss. The only therapy for corneal haziness due to corneal endothelial diseases, including FECD, is corneal transplantation using donor corneas, and no pharmaceutical treatment is available. We provide evidence that the expression levels of transforming growth factor-β (TGF-β) isoforms and TGF-β receptors are high in the corneal endothelium of patients with FECD. A cell model based on patients with FECD shows that TGF-β signaling induced a chronic overload of ECM proteins to the endoplasmic reticulum (ER), thereby enhancing the formation of unfolded protein and triggering the intrinsic apoptotic pathway through the unfolded protein response (UPR). We propose that inhibition of TGF-β signaling may represent a novel therapeutic target that suppresses cell loss as well as the accumulation of ECM in FECD.
Background/Aim: Glioblastoma multiforme (GBM) is an intractable tumor that has a very poor prognosis despite intensive treatment with temozolomide plus radiotherapy. Patients and Methods: Sixteen ...newly diagnosed patients with high-grade gliomas were enrolled in a phase II study of the α-type-1 DC vaccine. Briefly, DCs obtained from the culture of enriched monocytes in the presence of a cytokine cocktail, were pulsed with a cocktail of 5 synthetic peptides and cryopreserved until injection into patients. Results: The amount of IL-12 produced by activated DCs was higher than that previously reported. Among 15 evaluable patients, 10 showed positive CTL responses to any peptides in an ELISPOT assay. After 6 years of observation, five patients were still alive, and two of these patients were relapse-free. Moreover, a significant survival-prolonging effect was verified in DC-treated glioma patients. Conclusion: Peptide-cocktail-pulsed α-type-1 DC vaccines have a potential therapeutic effect on survival when used in combination with the standard regimen, which is partly based on IL-12-IFN-γ-mediated T-cell activation.
X-rays and extreme ultraviolet (EUV) emissions from solar flares rapidly change the physical composition of the Earth’s thermosphere and ionosphere, thereby causing space weather phenomena such as ...communication failures. Numerous empirical and physical models have been developed to estimate the effects of flare emissions on the Earth’s upper atmosphere. We verified the reproduction of the flare emission spectra using a one-dimensional hydrodynamic calculation and the CHIANTI atomic database. To validate the proposed model, we used the observed EUV spectra obtained by the Extreme ultraviolet variability experiment (EVE) on board the Solar Dynamics Observatory (SDO). We examined the “EUV flare time-integrated irradiance” and “EUV flare line rise time” of the EUV emissions for 21 events by comparing the calculation results of the proposed model and observed EUV spectral data. The proposed model successfully reproduced the EUV flare time-integrated irradiance of the Fe VIII 131 Å, Fe XVIII 94 Å, and Fe XX133 Å, as well as the 55–355 Å and 55–135 Å bands. For the EUV flare line rise time, there was an acceptable correlation between the proposed model estimations and observations for all Fe flare emission lines. These results demonstrate that the proposed model can reproduce the EUV flare emission spectra from the emitting plasma with a relatively high formation temperature. This indicates that the physics-based model is effective for the accurate reproduction of the EUV spectral irradiance.
The X-rays and extreme ultraviolet (EUV) emitted during solar flares can rapidly change the physical composition of Earth’s ionosphere, causing space weather phenomena. It is important to develop an ...accurate understanding of solar flare emission spectra to understand how it affects the ionosphere. We reproduced the entire solar flare emission spectrum using an empirical model and physics-based model, and input it into the Earth’s atmospheric model, GAIA to calculate the total electron content (TEC) enhancement due to solar flare emission. We compared the statistics of nine solar flare events and calculated the TEC enhancements with the corresponding observed data. The model used in this study was able to estimate the TEC enhancement due to solar flare emission with a correlation coefficient greater than 0.9. The results of this study indicate that the TEC enhancement due to solar flare emission is determined by soft X-ray and EUV emission with wavelengths shorter than 35 nm. The TEC enhancement is found to be largely due to the change in the soft X-ray emission and EUV line emissions with wavelengths, such as Fe XVII 10.08 nm, Fe XIX 10.85 nm and He II 30.38 nm.
Graphical Abstract
The metabolomic profiles of rat primary hepatocytes following treatment with rotenone, FCCP, or (+)-usnic acid were determined using liquid chromatography-mass spectrometry/mass spectrometry and gas ...chromatography-mass spectrometry. Significant and similar changes in the levels of 283 biochemical metabolites were associated with the three treatments compared with solvent control samples. Overall, the three treatments generated similar global biochemical profiles, with some minor differences associated with rotenone treatment. All three treatments resulted in a shift in energy metabolism as demonstrated by decreased glycogen stores and glycolysis. A reduced antioxidant response was detected in cells following all treatments. In addition, bile acid biosynthesis decreased as a potential consequence of increased oxidative stress by all three treatments. Conversely, rotenone treatment induced a number of changes after 1 hr, which were not detected in FCCP- or (+)-usnic acid-treated samples; these changes were not sustained over time and included increased NAD+ salvage and lysine degradation. In conclusion, these biochemical profiles could provide new insights into the mechanism(s) of mitochondrial toxicity.
Sterol regulatory element-binding proteins (SREBPs) are transcription factors that regulate the expression of genes involved in fatty acid and cholesterol biosynthetic pathways. The present study ...showed that the flavonoid chrysin impairs the fatty acid synthase promoter. Chrysin reduces the expression of SREBP target genes, such as fatty acid synthase, in human hepatoma Huh-7 cells and impairs de novo synthesis of fatty acids and cholesterol. Moreover, it reduces the endogenous mature, transcriptionally active forms of SREBPs, which are generated by the proteolytic processing of precursor forms. In addition, chrysin reduces the enforced expressing mature forms of SREBPs and their transcriptional activity. The ubiquitin-proteasome system is not involved in the chrysin-mediated reduction of SREBPs mature forms. These results suggest that chrysin suppresses SREBP activity, at least partially, via the degradation of SREBPs mature forms.
Abbreviations: ACC1: acetyl-CoA carboxylase 1; DMEM: Dulbecco's modified Eagle's medium; FAS: fatty acid synthase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; 25-HC: 25-hydroxycholesterol; HMGCS: HMG-CoA synthase; LDH: lactate dehydrogenase; LPDS: lipoprotein-deficient serum; PI3K: phosphatidylinositol 3-kinase; SCD1: stearoyl-CoA desaturase; SREBPs: sterol regulatory element-binding proteins.
Chrysin decreases the protein level of SREBPs mature forms regardless of the presence of MG132, a proteasome inhibitor.
Abstract
Shibata and Yokoyama (1999, ApJ, 526, L49; 2002, ApJ, 577, 422) proposed a method of estimating the coronal magnetic field strength (B) and magnetic loop length (L) of solar and stellar ...flares, on the basis of magnetohydrodynamic simulations of the magnetic reconnection model. Using the scaling law provided by Shibata and Yokoyama (1999, ApJ, 526, L49; 2002, ApJ, 577, 422), we obtain B and L as functions of the emission measure (EM = n
2
L
3) and temperature (T) at the flare peak. Here, n is the coronal electron density of the flares. This scaling law enables the estimation of B and L for unresolved stellar flares from the observable physical quantities EM and T, which is helpful for studying stellar surface activities. To apply this scaling law to stellar flares, we discuss its validity for spatially resolved solar flares. Quantities EM and T are calculated from GOES (Geostationary Operational Environmental Satellite) soft X-ray flux data, and B and L are theoretically estimated using the scaling law. For the same flare events, B and L were also observationally estimated with images taken by the Solar Dynamics Observatory (SDO)/Helioseismic and Magnetic Imager (HMI) Magnetogram and Atmospheric Imaging Assembly (AIA) 94 Å pass band. As expected, a positive correlation was found between the theoretically and observationally estimated values. We interpret this result as indirect evidence that flares are caused by magnetic reconnection. Moreover, this analysis makes us confident about the validity of applying this scaling law to stellar flares as well as solar flares.
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