Kinetics of injected fluorescently labeled IgA in a fluorescence molecular tomography system.

<p>A fluorescence molecular tomography system (FMT) is capable of resolving size and concentration of fluorochromes in deep tissue <i>in vivo</i>. Fluorescein-labeled IgA samples from gddY and Balb/c mice were injected into nude mice and monitored from 10 min to 24 h postinjection by FMT. After 2 h, IgA signals in the liver and bladder were found in a similar manner in both the groups of nude mice. However, IgA signals in the kidneys clearly differed between them. Mice injected with gddY IgA showed strong signals in the kidneys, with a peak at 4 h.</p

The Kinetics of Glomerular Deposition of Nephritogenic IgA

<div><p>Whether IgA nephropathy is attributable to mesangial IgA is unclear as there is no correlation between intensity of deposits and extent of glomerular injury and no clear mechanism explaining how these mesangial deposits induce hematuria and subsequent proteinuria. This hinders the development of a specific therapy. Thus, precise events during deposition still remain clinical challenge to clarify. Since no study assessed induction of IgA nephropathy by nephritogenic IgA, we analyzed sequential events involving nephritogenic IgA from IgA nephropathy-prone mice by real-time imaging systems. Immunofluorescence and electron microscopy showed that serum IgA from susceptible mice had strong affinity to mesangial, subepithelial, and subendothelial lesions, with effacement/actin aggregation in podocytes and arcade formation in endothelial cells. The deposits disappeared 24-h after single IgA injection. The data were supported by a fluorescence molecular tomography system and real-time and 3D in vivo imaging. In vivo imaging showed that IgA from the susceptible mice began depositing along the glomerular capillary from 1 min and accumulated until 2-h on the first stick in a focal and segmental manner. The findings indicate that glomerular IgA depositions in IgAN may be expressed under the balance between deposition and clearance. Since nephritogenic IgA showed mesangial as well as focal and segmental deposition along the capillary with acute cellular activation, all glomerular cellular elements are a plausible target for injury such as hematuria.</p></div

IgA from gddY mice is deposited along the glomerular capillary wall in a focal and segmental manner.

<p>Detailed kinetics of IgA deposition analyzed from 1 min to 2 h postinjection using confocal laser microscopy. Alexa Fluor 633-labeled IgA from gddY and Balb/c mice (red) and 500-kDa fluorescein-labeled dextran (green) were injected for analyzing kinetics of IgA deposition and visualizing blood vessel wall integrity, respectively. (a) IgA signals were detectable even after 1 min and accumulated up to 2 h in a focal and segmental manner in mice with IgA from gddY mice. In contrast, mice who received Balb/c IgA did not show a signal even after 2 h. (b)(c) Serial images of a glomerulus in mice with IgA from gddY mice showed that these IgA molecules accumulated on top of the initial aggregates along the glomerular capillaries. These aggregates were found in a focal and segmental manner but not in a diffuse and global manner.</p

A single injection of serum from gddY mice induced glomerular IgA deposition with activation of glomerular podocytes and endothelial cells.

<p>(a) Glomerular IgA deposits were found at 2 h in mice injected with serum from gddY mice but not from Balb/c mice. These fluorescent signals disappeared after 24 h in this single-injection model. (b) These deposits and clearance were confirmed using electron microscopy. Electron-dense deposits were mainly detected in paramesangial lesions. (c) Some glomeruli showed subendothelial and subepithelial deposits (*) with arcade formation in glomerular endothelial cells (**) and effacement and actin aggregation in podocytes (***) 2 h after the injection.</p

Scavenging of reactive oxygen species by astaxanthin inhibits epithelial–mesenchymal transition in high glucose-stimulated mesothelial cells

<div><p>Background</p><p>High glucose concentrations influence the functional and structural development of the peritoneal membrane. We previously reported that the oral administration of astaxanthin (AST) suppressed peritoneal fibrosis (PF) as well as inhibited oxidative stress, inflammation, and epithelial–mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) in a chlorhexidine-induced PF rat model. This suggests that oxidative stress induction of EMT is a key event during peritoneal damage. The present study evaluated the therapeutic effect of AST in suppressing EMT, in response to glucose-induced oxidative stress.</p><p>Methods</p><p>Temperature-sensitive mesothelial cells (TSMCs) were cultured in the presence or absence of AST and then treated with 140 mM glucose for 3 or 12 hours. Expression levels of TNF-α, TGF-β, and VEGF were determined at the mRNA and protein levels, and nuclear factor kappa B (NF-κB) activity was evaluated. We measured NO₂⁻/NO₃⁻ concentrations in cellular supernatants and determined 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels in mitochondrial and nuclear DNA. The expressions of E-cadherin and alpha-smooth muscle actin (α-SMA) were evaluated by double immunofluorescence and protein levels.</p><p>Results</p><p>High glucose concentrations induced overproduction of reactive oxidative species (ROS), increasing 8-OHdG mitochondrial DNA and cytokine levels. The NF-κB pathway was activated in response to high glucose concentrations, whereas <i>de novo</i> α-SMA expression was observed with decreased E-cadherin expression. AST treatment attenuated ROS production, inflammatory cytokine production, NF-κB activation, and EMT.</p><p>Conclusion</p><p>The findings of the present study indicate that AST may have an anti-EMT effect due to anti-oxidative and anti-inflammatory activities by scavenging glucose-induced ROS from mitochondria in PMCs. AST may be an efficacious treatment for PF.</p></div

Effect of high glucose stimulation, AST, and AA to TSMCs as for ROS.

<p>(A) NO₂⁻/NO₃⁻ concentration in medium supernatant of each group. (B) Intracellular ROS levels of each group. (C) 8-OHdG ratio in mitochondrial DNA of each group. (D) 8-OHdG ratio in nuclear DNA of each group. AST concentration: 5 μM, AA concentration: 100 μM. NS: no significant change. *: p < 0.05. **: p < 0.01. ***: p < 0.0005. ****: p < 0.0001. Error bars represent SD. (E) ROS fluorescence of each group (1) C3h group, (2) C12h group, (3) AST3h group, (4) AST12h group, (5) AA3h group, (6) AA12h group, (7) G3h group, (8) G12h group, (9) AST-G3h group, (10) AST-G12h group, (11) AA-G3h group, and (12) AA-G12h group. Fluorescence solution was 2′, 7′-dichlorodihydrofluorescin diacetate (DCFH-DA).</p

Effect of high glucose stimulation, AST and AA to TSMCs as for EMT.

<p>(A) E-cadherin mRNA and protein expression levels in each groups. (B) α-SMA mRNA and protein expression levels in each group. :p < 0.05. **: p < 0.01. ***: p < 0.0005. ****: p < 0.0001. Error bars represent SD. (C) Effect of high glucose stimulation, AST, and AA to TSMCs as for EMT in double immunofluorescence. (1) C3h group, (2) C12h group, (3) AST3h group, (4) AST12h group, (5) AA3h group, (6) AA12h group, (7) G3h group, (8) G12h group, (9) AST-G3h group, (10) AST-G12h group, (11) AA-G3h group, and (12) AA-G12h group. Alexa 488 green (E-cadherin) and Alexa 555 red (α-SMA) were used as secondary antibodies. DAPI was used for nuclear staining.</p

Microwave-Driven Asbestos Treatment and Its Scale-up for Use after Natural Disasters

Asbestos-containing debris generated by the tsunami after the Great East Japan Earthquake of March 11, 2011, was processed by microwave heating. The analysis of the treated samples employing thermo gravimetry, differential thermal analysis, X-ray diffractometry, scanning electron microscopy, and phase-contrast microscopy revealed the rapid detoxification of the waste by conversion of the asbestos fibers to a nonfibrous glassy material. The detoxification by the microwave method occurred at a significantly lower processing temperature than the thermal methods actually established for the treatment of asbestos-containing waste. The lower treatment temperature is considered to be a consequence of the microwave penetration depth into the waste material and the increased intensity of the microwave electric field in the gaps between the asbestos fibers resulting in a rapid heating of the fibers inside the debris. A continuous treatment plant having a capacity of 2000 kg day^–1 of asbestos-containing waste was built in the area affected by the earthquake disaster. This treatment plant consists of a rotary kiln to burn the combustible waste (wood) and a microwave rotary kiln to treat asbestos-containing inorganic materials. The hot flue gas produced by the combustion of wood is introduced into the connected microwave rotary kiln to increase the energy efficiency of the combined process. Successful operation of this combined device with regard to asbestos decomposition is demonstrated

Study groups and experimental design.

<p>Unstimulated control groups (Control group): Incubated in M199 medium alone for 3 h (C3h group) or 12 h (C12h group). G groups: Incubated in M199 medium with 140 mM glucose for 3 h (G3h group) or 12 h (G12h group). Ascorbic acid (AA; AA group.): Incubated in M199 medium with 140 mM glucose and 100 μM AA for 3 h (AA-G3h group) or 12 h (AA-G12h group). AST pre-treated groups (AST group): Incubated in M199 medium alone for 3 h (AST3h group) or 12 h (AST12h group) after incubated in M199 medium with 5μM AST for 24h. AST-G group: Incubated in M199 medium with 140 mM glucose for 3 h (AST-glucose [AST-G] 3h group) or for 12 h (AST-G12h group) after incubated in M199 medium with 5μM AST for 24h.</p

Effect of high glucose stimulation to TSMCs as for NF-κB pathway.

<p>(A) NF-κB p65 protein subunit expression in TSMCs in each group. *: p < 0.05. **: p < 0.01. ***: p < 0.0005. ****: p < 0.0001. Error bars represent SD. (B) NF-κB p65 subunit immunofluorescence. (1) C3h group, (2) C12h group, (3) AST3h group, (4) AST12h group, (5) AA3h group, (6) AA12h group, (7) G3h group, (8) G12h group, (9) AST-G3h group, (10) AST-G12h group, (11) AA-G3h group, and (12) AA-G12h group. Alexa 488 green were used as secondary antibodies.</p