AS101 prevents diabetic nephropathy progression and mesangial cell dysfunction: regulation of the AKT downstream pathway.

Diabetic nephropathy (DN) is characterized by proliferation of mesangial cells, mesangial expansion, hypertrophy and extracellular matrix accumulation. Previous data have cross-linked PKB (AKT) to TGFβ induced matrix modulation. The non-toxic compound AS101 has been previously shown to favorably affect renal pathology in various animal models and inhibits AKT activity in leukemic cells. Here, we studied the pharmacological properties of AS101 against the progression of rat DN and high glucose-induced mesangial dysfunction. In-vivo administration of AS101 to Streptozotocin injected rats didn't decreased blood glucose levels but ameliorated kidney hypotrophy, proteinuria and albuminuria and downregulated cortical kidney phosphorylation of AKT, GSK3β and SMAD3. AS101 treatment of primary rat glomerular mesangial cells treated with high glucose significantly reduced their elevated proliferative ability, as assessed by XTT assay and cell cycle analysis. This reduction was associated with decreased levels of p-AKT, increased levels of PTEN and decreased p-GSK3β and p-FoxO3a expression. Pharmacological inhibition of PI3K, mTORC1 and SMAD3 decreased HG-induced collagen accumulation, while inhibition of GSK3β did not affect its elevated levels. AS101 also prevented HG-induced cell growth correlated to mTOR and (rp)S6 de-phosphorylation. Thus, pharmacological inhibition of the AKT downstream pathway by AS101 has clinical potential in alleviating the progression of diabetic nephropathy

Immunosuppression reduction when administering a booster dose of the BNT162b2 mRNA SARS-CoV-2 vaccine in kidney transplant recipients without adequate humoral response following two vaccine doses: protocol for a randomised controlled trial (BECAME study)

INTRODUCTION: Inadequate antibody response to mRNA SARS-CoV-2 vaccination has been described among kidney transplant recipients. Immunosuppression level and specifically, use of antimetabolite in the maintenance immunosuppressive regimen, are associated with inadequate response. In light of the severe consequences of COVID-19 in solid organ transplant recipients, we believe it is justified to examine new vaccination strategies in these patients. METHODS AND ANALYSIS: BECAME is a single-centre, open-label, investigator-initiated randomised controlled, superiority trial, aiming to compare immunosuppression reduction combined with a third BNT162b2 vaccine dose versus third dose alone. The primary outcome will be seropositivity rate against SARS-CoV-2. A sample size of 154 patients was calculated for the seropositivity endpoint assuming 25% seropositivity in the control group and 50% in the intervention group. A sample of participants per arm will be also tested for T-cell response. We also plan to perform a prospective observational study, evaluating seropositivity among ~350 kidney transplant recipients consenting to receive a third vaccine dose, who are not eligible for the randomised controlled trial. ETHICS AND DISSEMINATION: The trial is approved by local ethics committee of Rabin Medical Center (RMC-0192-21). All participants will be required to provide written informed consent. Results of this trial will be published; trial data will be available. Protocol amendments will be submitted to the local ethics committee

Changes in Serum Creatinine May Cause Hypoglycemia among Non-Critically Ill Patients Admitted to Internal Medicine Units

Background: The association between changes in serum creatinine levels and hypoglycemia during hospitalization was investigated. Methods: This was a retrospective analysis of medical charts. Patients were categorized as having significant change in creatinine (SCIC) when serum creatinine levels rose or dropped ≥ 0.3 mg/dL from admission values at any time during their hospitalization. Patients were considered hypoglycemic if they had at least one documented glucose level ≤ 70 mg/dL during the hospitalization. Multiple logistic, linear and Cox regression analyses were used to ascertain the association between incident SCIC, severity and timing with incident hypoglycemia. Results: Included were 25,400 (mean age 69.9 ± 18.0, 49.3% were males). The rate of SCIC was 22.2%, and 62.2% of them were diagnosed upon admission. Patients with SCIC had a higher incidence of hypoglycemia compared to patients without (13.1% vs. 4.1%, respectively, p < 0.001). Patients with SCIC had an increased risk of hypoglycemia (OR 1.853, 95% CI 1.586–2.166, p < 0.001). The magnitude of SCIC was associated with the incidence (OR 1.316, 95% CI 1.197–1.447, p < 0.001) and the number of events (HR 0.054, 95% CI 0.021–0.087, p = 0.001). More than 60% of patients with hypoglycemia had their first event documented during days 0–6 after SCIC occurrence. Of those, the majority of events occurred on day 0–1, and the rate showed a gradual decrease throughout the first 5 days from SCIC occurrence. The results were similar for patients with and without DM. Conclusions: Changes in creatinine during hospitalization may cause hypoglycemia among patients admitted to internal medicine departments, regardless of DM status

Novel Protein to Phosphorous Ratio Score Predicts Mortality in Hemodialysis Patients.

ObjectiveLowering serum phosphorus in people on hemodialysis may improve their survival. However, prior studies have shown that restricting dietary protein intake, a major source of phosphorus, is associated with higher mortality. We hypothesized that a novel metric that incorporates both these values commensurately can improve survival prediction.MethodsWe used serum phosphorous and normalized protein catabolic rate (nPCR), a surrogate of dietary protein intake, to form a new metric R that was used to examine the associations with mortality in 63,016 people on hemodialysis (HD) of one year after treatment initiation. Survival models were adjusted for case-mix, malnutrition-inflammation cachexia syndrome (MICS), and residual kidney function (RKF).ResultsIndividuals treated with hemodialysis were divided into five groups in accordance with R value. Group 1 included sick individuals with high phosphorous and low nPCR. Group 5 included individuals with low phosphorous and high nPCR. After 1-year follow-up, survival difference between the groups reflected R value, where an increase in R was associated with improved survival. The association of R with mortality was strengthened by adjustment in demographic variables and attenuated after adjustment to MICS. Mortality associations in accordance with R were not influenced by residual kidney function (RKF).ConclusionThe novel protein to phosphorus ratio score R predicts mortality in people on dialysis, probably reflecting both nutrition and inflammation state independent of RKF. The metric enables better phosphorus monitoring, although adequate dietary protein intake is ensured and may improve the prediction of outcomes in the clinical setting

AS101 attenuates high glucose induced mesangial cell proliferation.

<p>Mesangial cells were cultured in DMEM without serum for 24 h. Cells were then transferred to fresh DMEM without serum in the presence or absence of HG treatment (30 mM). For osmotic control, cells were treated with Mannitol (24.5 mM). Medium for the serum deprived control (Starved) or the osmotic control (Mannitol) contained normal levels of glucose (5.5 mM). (a) Cells treated with HG were supplemented with different concentrations of AS101 (0.1, 0.5, 1, 2, 5 µg/ml). After 24 h, XTT assay was performed. OD levels were compared to control which was normalized to 100%. Results represent mean±SEM from three experiments. #p<0.05 decrease vs. HG. *p<0.05 increase vs. starved. &p = 0.057 increase vs. mannitol. (b) Cells treated with Normal glucose levels were supplemented with AS101 at the concentrations indicated in panel a. After 24 h, XTT assay was performed. OD levels were compared to control which was normalized to 100%. Results represent three experiments. (c) Cells treated with HG were treated with different concentrations of AS101 (0.1, 1, 2, 5 µg/ml). After 48 h of treatment, cells were stained with PI buffer; cell cycle analysis was performed by FACS to determine the level of cell accumulation in each cell cycle phase: sub-G1, G1, S, G2. Results represent two experiments. (d) Cells treated with HG were treated with different concentrations of AS101 (0.1, 0.5, 1, 2 µg/ml) or LY294002 (50 µM). After 24 h, mesangial cell expansion was examined by light microscopy (X40). Results shown are from a single experiment representative of seven. (e) Cells treated with HG were treated with different concentrations of AS101 (0.1, 0.5, 1, 2 µg/ml) or LY294002 (50 µM). After 24 h, cell lysates were extracted for detection of PTEN, p-AKT, p-GSK3β, p-FoxO3a by western blot analysis, with actin as loading control. Results represent three experiments.</p

Post renal transplant anemia: severity, causes and their association with graft and patient survival

Abstract Background Post transplantation anemia (PTA) is common among kidney transplant patients. PTA is associated with increased graft loss and in most studies with increased mortality. However, the effect of the severity of anemia on this associations was not thoroughly evaluated. Methods Patients who underwent kidney transplantation in Rabin Medical Center (RMC) were included in the study. Data were collected during the years 2002–2016. Anemia was defined as hemoglobin (Hb) level less than 12 g/dL in women and less than 13 g/dL in men, in accordance with World Health Organization (WHO) criteria. Severe anemia was defined as hemoglobin lower than 11 g/dL. Primary outcome was a composite of patient and graft survival. We used univariate and multivariate models to evaluate association between severity and specific causes of anemia with the outcomes. As the risk associated with anemia changed over time we analyzed the risk separately for the early and the late period (before and after 1251 days). Results Our cohort included 1139 patients, 412 (36.2%) of which had PTA and 134 (11.7%) had severe anemia. On multivariable analysis, severe anemia was highly associated with the primary outcome at the early period (HR 6.26, 95% CI 3.74–10.5, p < 0.001). Anemia due to either AKI & acute rejection (11.9% of patients) or infection (16.7%), were associated with primary outcome at the early period (HR 9.32, 95% CI 5.3–26.41, p < 0.001 and HR 3.99, 95% CI 2.01–7.95, p < 0.001, respectively). There was non-significant trend for association between anemia due to Nutritional deficiencies (29.1%) and this outcome (HR 3.07, 95% CI 0.93–10.17, p = 0.067). Conclusion PTA is associated with graft loss and mortality especially during the first three years. Anemia severity affects this association. An anemia workup is recommended for PTA

AS101 downregulates AKT, GSK3β and SMAD3 phosphorylation on cortical kidney cells in-vivo.

<p>Diabetes was induced by single intraperitoneal injection of STZ (65 mg/kg). Mice were treated with AS101 at 0.5 mg/kg (LD) or 1 mg/kg (HD) by intraperitoneal injection every other day. Control animals were treated with PBS alone. Western blot analysis of cortical tissue was performed 4 weeks after diabetes induction. Lysates were extracted for detection of p-AKT, p-GSK3β, and p-SMAD3 by western blot analysis. Total AKT and SMAD antibodies were used as a loading control. Results shown are from one experiment representative of three.</p

Effect of PI3K, mTOR, GSK3β and SMAD3 inhibitors on total collagen production in-vitro.

<p>Mesangial cells were pre-treated as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114287#pone-0114287-g004" target="_blank">figure 4</a>. (a, b) Cells treated with HG were separately treated with AS101 (2 µg/ml), LY294002 (50 µM), Rapamycin (0.1 ng/ml), PD98059 (100 µM), SB216763 (30 µM) and SIS3 (20 µM). After 48 h, cells were stained with Sirius red to detect collagen. Results are representative of two experiments. After washing the dye (a) Light microscope examination (X40) and (b) photograph of culture plates was performed.</p