Once-weekly hemodialysis combined with low-protein and low-salt dietary treatment as a favorable therapeutic modality for selected patients with end-stage renal failure: a prospective observational study in Japanese patients

Abstract Background For patients with end-stage renal failure (ESFR), thrice-weekly hemodialysis is a standard care. Once-weekly hemodialysis combined with low-protein and low-salt dietary treatment (OWHD-DT) have been rarely studied. Therefore, here, we describe our experience on OWHD-DT, and assess its long-term effectiveness. Methods We instituted OWHD-DT therapy in 112 highly motivated patients with creatinine clearance below 5.0 mL/min. They received once-weekly hemodialysis on a diet of 0.6 g/kg/day of protein adjusted for sufficient energy intake, and less than 6 g/day of salt intake. Serial changes in their clinical, biochemical and nutritional parameters were prospectively observed, and the weekly time spent for hospital visits as well as their monthly medical expenses were compared with 30 age, sex- and disease-matched thrice-weekly hemodialysis patients. Results The duration of successfully continued OWHD-DT therapy was more than 4 years in 11.6% of patients, 3 years in 16.1%, 2 years in 24.1% and 1 year in 51.8%. Time required per week for hospital attendance was 66.7% shorter and monthly medical expenses were 50.5% lower in the OWHD-DT group than in the thrice-weekly hemodialysis group (both p < 0.001). Patient survival rates in the OWHD-DT group were better than those in the Japan Registry (p < 0.001). Serum urea nitrogen significantly decreased; hemoglobin significantly increased; and albumin and body mass index were not significantly different from baseline values. In the OWHD-DT patients, serum albumin at 1 and 2 years after initiation of therapy was significantly higher compared with prevalent thrice-weekly hemodialysis patients. Furthermore, residual urine output was significantly higher in the OWHD-DT patients than in those receiving thrice-weekly hemodialysis (p < 0.05). Interdialytic weight gain over the course of the entire week between treatments in patients on OWHD-DT were 0.9 ± 1.0, 2.0 ± 1.3, 1.9 ± 1.2, 1.9 ± 1.5 and 1.8 ± 1.0 kg at 1, 6, 12, 18 and 24 months, respectively, though the weekly weight gain for thrice-weekly hemodialysis group (summed over all 3 treatments) was 8.6 ± 0.63 kg, p < 0.001. Conclusions OWHD-DT may be a favorable therapeutic modality for selected highly motivated patients with ESRF. However, this treatment cannot be seen as a general maintenance strategy. Trial registration UMIN000027555, May 30, 2017 (retrospectively registered)

Citrus limon L.-Derived Nanovesicles Show an Inhibitory Effect on Cell Growth in p53-Inactivated Colorectal Cancer Cells via the Macropinocytosis Pathway

Edible plant-derived nanovesicles have been explored as effective materials for preventing colorectal cancer (CRC) incidence, dependent on gene status, as a K-Ras-activating mutation via the macropinocytosis pathway. Approximately 70% of CRC harbors the p53 mutation, which is strongly associated with a poor prognosis for CRC. However, it has not been revealed whether p53 inactivation activates the macropinocytosis pathway or not. In this study, we investigated parental cells, wild-type or null for p53 treated with Citrus limon L.-derived nanovesicles, as potential materials for CRC prevention. Using ultracentrifugation, we obtained C. limon L.-derived nanovesicles, the diameters of which were approximately 100 nm, similar to that of the exosomes derived from mammalian cells. C. limon L.-derived nanovesicles showed inhibitory effects on cell growth in not p53-wild, but also in p53-inactivated CRC cells. Furthermore, we revealed that the macropinocytosis pathway is activated by p53 inactivation and C. limon L.-derived nanovesicles were up taken via the macropinocytosis pathway. Notably, although C. limon L.-derived nanovesicles contained citrate, the inhibitory effects of citrate were not dependent on the p53 status. We thus provide a novel mechanism for the growth inhibition of C. limon L.-derived nanovesicles via macropinocytosis and expect to develop a functional food product containing them for preventing p53-inactivation CRC incidence

Expression of SIRT1 in human fetal and adult liver and isolated hepatocytes.

<p>(A) Immunohistochemistry of SIRT1 in human fetal and adult liver, (B) Immunofluorescence staining of SIRT1 in fetal and adult hepatocytes after 2 days of culture. (C) Quantification of hepatic SIRT1 mRNA expression measured by qRT- PCR in human fetal and adult hepatocytes in vitro and in vivo (n≥3/group).</p

Lipids and glucose levels in human fetal hepatocytes after Sirt1 inhibition.

<p>(A) Fluorescent staining of lipids (red droplets, white arrow) inside human fetal hepatocytes (HFH) with or without 50uM Sirtinol for 3 days (20x). (B) Triglycerides quantification of human fetal hepatocytes with or without +50uM Sirtinol for 3 days analyzed by a colorimetric assay. (C) Glucose concentration between normal HFH and HFH +50uM Sirtinol for 3 days. (n≥7/group; *, P < .05).</p

Expression of SIRT1 in human fetal and adult liver and isolated hepatocytes.

<p>(A) Immunohistochemistry of SIRT1 in human fetal and adult liver, (B) Immunofluorescence staining of SIRT1 in fetal and adult hepatocytes after 2 days of culture. (C) Quantification of hepatic SIRT1 mRNA expression measured by qRT- PCR in human fetal and adult hepatocytes in vitro and in vivo (n≥3/group).</p

Increased activation of Glucogenesis pathway in human fetal hepatocytes after exposure to Sirtinol.

<p>(A) Expression of hepatic PEPCK and G6PC mRNA measured by qRT- PCR in human fetal hepatocytes exposed to +50uM Sirtinol compared to controls. (B) Immunofluorescence of S-473 AKT in human fetal hepatocytes with or without 50uM Sirtinol (10x). (C) Fluorescence intensity quantification for S-473 AKT signal (arbitrary units) using ImageJ (n≥7/group; *, P < .05). (D) Western blot analysis for S-473 AKT/AKT, S-256 FOXO1/FOXO1 in human fetal hepatocytes exposed to 50uM Sirtinol compared to controls. GAPDH was used as a loading control. (E) Immunofluorescence staining of S-256 FOXO1 in human fetal hepatocytes with or without 50uM Sirtinol (20x).</p

Donor Demographics.

<p>Donor Demographics.</p

Representation of SIRT1 regulation of lipid and glucose balances in human fetal hepatocytes.

<p>In normal conditions, SIRT1 inhibits De Novo Lipogenesis and Gluconeogenesis through the AKT/FOXO1 pathway inside human fetal hepatocytes, keeping a normal balance inside the cells. Upon SIRT1 inhibition, both the lipid and glucose balance will be disrupted, leading to an increase of lipid and carbohydrates levels in human fetal hepatocytes.</p