Electrospun EVOH/AST-120 hybrid nanofiber membranes for removal of indoxyl sulfate from blood.

Nanofibers containing activated carbon using poly(ethylene-co-vinyl alcohol) (EVOH) were prepared to remove indoxyl sulfate (IS) from the blood. IS is a urinary toxin that is highly toxic and triggers the progression of chronic kidney disease (CKD). Here, nanofibers containing activated carbon (AST-120), which has been used practically as an adsorbent for indole (a precursor of IS), were fabricated via electrospinning for the adsorption and removal of IS from the blood. EVOH containing different ethylene ratios was used as the nanofiber material; moreover, the effect of the ethylene ratio on various properties of the nanofibers, such as surface wettability and the IS adsorption rate, was investigated. As a result, EVOH/AST-120 nanofibers comprising EVOH with a low ethylene ratio exhibited faster IS adsorption behavior. This adsorption behavior agreed well with the pseudo-second-order model, suggesting that the diffusion of IS into the nanofibers is the rate-limiting step of the process of adsorption. Furthermore, the nanofibers successfully reduced the IS concentration in the blood under circulating conditions. Therefore, these EVOH/AST-120 nanofibers are expected to greatly improve the prognosis of patients with CKD when used in combination with the current hemodialysis therapy as an IS-adsorbing filter.

Pruritus and protein-bound uremic toxins in patients undergoing hemodialysis: a cross-sectional study.

Background: Patients undergoing hemodialysis frequently experience pruritus; its severity is associated with poor quality of life and mortality. Recent progress in hemodialysis treatment has improved the removal of small- and middle-molecular-weight molecules; however, the removal of protein-bound uremic toxins (PBUTs) remains difficult. It is possible that pruritus is associated with serum PBUTs in patients undergoing hemodialysis. Methods: We conducted a multicenter cross-sectional study in patients undergoing hemodialysis (n = 135). The severity of pruritus was assessed using the 5D-itch scale and medication use. Serum PBUTs, including indoxyl sulfate, p-cresyl sulfate, indole acetic acid, phenyl sulfate, and hippuric acid, were measured using mass spectrometry; the PBUT score was calculated from these toxins using principal component analysis. Univariate and multiple regression analyses were performed to examine independent predictors of pruritus. Results: Pruritus was reported by 62.2%, 21.5%, and 13.3%, 1.5% and 0.7% as 5 (not at all), 6-10, 11-15, 16-20, and 21-25 points, respectively. The PBUT score was higher in patients undergoing dialysis having pruritus than those without pruritus (0.201 [-0.021 to 0.424] vs -0.120 [-0.326 to 0.087]; P = 0.046). The PBUT score was shown to have an association with the presence of pruritus (coefficient 0.498[Formula: see text]0.225, odds ratio: 1.65 [1.06-2.56]; P = 0.027). Conclusion: Uremic pruritus was frequently found and associated with the PBUT score in patients undergoing hemodialysis. Further studies are required to clarify the impact of PBUTs on uremic pruritus and to explore therapeutic strategies in patients undergoing hemodialysis.

Correlation Between NKG2DL Expression and Antitumor Effect of Protein-bound Polysaccharide-K in Tumor-bearing Mouse Models.

BACKGROUND/AIM: We investigated the relationship between the expression of natural killer group 2, member D ligands (NKG2DLs) and the antitumor effects of protein-bound polysaccharide-K (PSK). MATERIALS AND METHODS: PSK was administered to evaluate its effectiveness against tumor growth. The expression of Rae-1 and H60 were analyzed in multiple cell lines. RESULTS: PSK showed the highest antitumor effects in mice implanted with cells expressing neither Rae-1 nor H60. PSK had little antitumor effect in mice implanted with cells expressing both Rae-1 and H60. A correlation between the expression of NKG2DLs and the antitumor effect of PSK was observed. After PSK administration, INF-γ production in CD8+ T cells increased in mice with cells expressing neither Rae-1 nor H60, but did not change in mice implanted with cells expressing both Rae-1 and H60. CONCLUSION: We demonstrated that the expression of NKG2DLs affects tumor immunity and the efficacy of immuno therapy in tumor-bearing mouse model.

PolyQ proteins interfere with nuclear degradation of cytosolic proteins by sequestering the Sis1p chaperone.

Dysfunction of protein quality control contributes to the cellular pathology of polyglutamine (polyQ) expansion diseases and other neurodegenerative disorders associated with aggregate deposition. Here we analyzed how polyQ aggregation interferes with the clearance of misfolded proteins by the ubiquitin-proteasome system (UPS). We show in a yeast model that polyQ-expanded proteins inhibit the UPS-mediated degradation of misfolded cytosolic carboxypeptidase Y(∗) fused to green fluorescent protein (GFP) (CG(∗)) without blocking ubiquitylation or proteasome function. Quantitative proteomic analysis reveals that the polyQ aggregates sequester the low-abundant and essential Hsp40 chaperone Sis1p. Overexpression of Sis1p restores CG(∗) degradation. Surprisingly, we find that Sis1p, and its homolog DnaJB1 in mammalian cells, mediates the delivery of misfolded proteins into the nucleus for proteasomal degradation. Sis1p shuttles between cytosol and nucleus, and its cellular level limits the capacity of this quality control pathway. Upon depletion of Sis1p by polyQ aggregation, misfolded proteins are barred from entering the nucleus and form cytoplasmic inclusions.

Direct inhibition of the transforming growth factor-ß pathway by protein-bound polysaccharide through inactivation of Smad2 signaling.

Transforming growth factor-ß (TGF-ß) is involved in the regulation of cell proliferation, differentiation, and apoptosis and is associated with epithelial-mesenchymal transition (EMT). Inhibition of the TGF-ß pathway is an attractive strategy for the treatment of cancer. We recently screened for novel TGF-ß inhibitors among commercially available drugs and identified protein-bound polysaccharide (PSK) as a strong inhibitor of the TGF-ß-induced reporter activity of 3TP-lux, a TGF-ß1-responsive luciferase reporter. Protein-bound polysaccharide is used as a non-specific immunostimulant for the treatment of gastric and colorectal cancers in Japan. The anticancer activity of this agent may involve direct regulation of growth factor production and enzyme activity in tumors in addition to its immunomodulatory effect. Although several clinical studies have shown the beneficial therapeutic effects of PSK on various types of tumors, its mechanism of action is not clear. In the present study, Western blot analysis showed that PSK suppressed the phosphorylation and nuclear localization of the Smad2 protein, thereby suggesting that PSK inhibits the Smad and MAPK pathways. Quantitative PCR analysis showed that PSK decreased the expression of several TGF-ß pathway target genes. E-cadherin and vimentin immunohistochemistry showed that PSK suppressed TGF-ß1-induced EMT, and FACS analysis showed that PSK inhibited the EMT-mediated generation of CD44(+) /CD24(-) cells. These data provide new insights into the mechanisms mediating the TGF-ß-inhibiting activity of PSK and suggest that PSK can effectively treat diseases associated with TGF-ß signaling.