bluuebunny/crossref_metadata_2025_split huggingface dataset:
P0019MICROVESICLES FROM MESENCHYMAL STEM CELLS OVEREXPRESSING MIR-34A INHIBIT TGF-BETA-1-INDUCED EPITHELIAL-MESENVHYMAL TRANSITION IN RENAL TUBULAR EPITHELIAL CELLS IN VITRO
Authors: He Juan , Jiang Yali , Wang Yan , Tian Xiujuan , Sun Shiren 
Published: June 2020

Abstract

Background and Aims
The application of microRNAs (miRNAs) in the therapy of kidney diseases is hampered by difficulties in delivering miRNAs effectively. Nano-sized microvesicles (MVs) are known as natural carriers of small RNAs. Our prior work has demonstrated that MVs isolated from mesenchymal stem cells (MSCs) attenuated kidney injuries induced by unilateral ureteral obstruction and 5/6 subtotal nephrectomy in mice. The present work aimed at evaluating the effects of miR-34a-5p (miR-34a)-modified MSC-MVs on transforming growth factor (TGF)-β1 induced fibrosis and apoptosis in vitro.

Method
Bone marrow MSCs were further modified by lentiviruses overexpressing miR-34a, and MVs were collected from these MSCs to treat HK-2 renal tubular cells exposed to TGF-β1. Alterations in epithelial-mesenchymal transition (EMT) and cell survival were further determined.

Results
We first demonstrated that MVs generated by miR-34a-modified MSCs contained more miR-34a. By analyzing the expression levels of epithelial markers (E-cadherin and Tight Junction Protein 1 (TJP1)) and mesenchymal markers (α-SMA and fibronectin), we found that the pro-fibrotic TGF-β1 induced EMT was remarkably suppressed by miR-34a-enriched MSC-MVs. Notch-1 receptor and Jagged-1 ligand, two major molecules of Notch signaling pathway, are predicted targets of miR-34a. We further found that the elevation in Notch-1 and Jagged-1 induced by TGF-β1 was inhibited by miR-34a-enriched MSC-MVs. The inhibitory effects of miR-34a-enriched MSC-MVs on EMT and Notch signaling pathway were stronger than the control MSC-MVs. In addition, TGF-β1 exposure also induced apoptosis in HK-2 cells. Although miR-34amofidied MSC-MVs could inhibit TGF-β1-triggered apoptosis in HK-2 cells, the effects were less significant than the control MSC-MVs. This phenomenon may be resulted from the pro-apoptotic effects of miR-34a.

Conclusion
Our study demonstrates that miR-34a-overexpressing MSC-MVs inhibit EMT induced by pro-fibrotic TGF-β1 in renal tubular epithelial cells, possibly through inhibiting Jagged-1/Notch-1 pathway. Genetic modification of MSC-MVs with anti-fibrotic molecule may present a novel strategy for treatment of renal injuries.
P0035URINARY CITRATE AS A MARKER OF RENAL FUNCTION IN PATIENTS WITH AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE
Authors: Borrego-Utiel Francisco-Jose , Herrera Isidoro , Merino Garcia Enoc , Moriana Dominguez Clara , Camacho Reina Victoria , Ocaña Perez Esther 
Published: June 2020

Abstract

Background and Aims
In autosomal dominant polycystic kidney disease (ADPKD) is frequent to find low urinary citrate levels. Recently it has been suggested that urinary citrate could be a marker of covert metabolic acidosis. Our aim was to analyze relationship between urinary citrate levels and renal functionality in ADPKD patients.

Method
We determined citrate, calcium and uric acid in 24-hour collected urine from 91 ADPKD patients

Results
Urinary citrate/creatinine ratio was 214±158 (range 5.3-678) mg/g Cr with levels significantly higher in females. When considering chronic kidney disease (CKD) stages we observed a progressive decrease in urinary osmolality and in urinary citrate, calcium and uric acid elimination. Low levels of citrate (&lt;300 mg/g Cr) were present in 40% in CKD-1 stage, in 69.7% in CKD-2 stage, 92% in CKD-3 stage and 100% in CKD-4 + 5 stages. Urinary citrate was correlated with serum creatinine (r= -0.66, p&lt;0.001) and eGFR (r= 0.56, p&lt;0.001). Urinary citrate significantly correlated with urinary calcium but correlation with urinary uric acid was weaker. We did not find any correlation with serum bicarbonate. Using multiple lineal regression analysis we found as predictors of urinary citrate to glomerular filtration rate, female gender and urinary calcium levels. In a subgroup of patients we measured total kidney volume and we found an inverse correlation with urinary citrate levels that dissappeared when it was corrected with glomerular filtration rate. We did not also find a relationship between urinary elimination of calcium or uric acid and TKV after adjusting with eGFR.

Conclusion
Urinary citrate is very frequently reduced in ADPKD patients being present from very early CKD stages. Their levels are inversely correlated with glomerular filtration rate and directly with urinary calcium excretion. We did not found a relathionship with serum bicarbonate. We think that it would be interesting to study urinary citrate in other nephropathies and verify if it could be a marker of covert metabolic acidosis.
P0043SYMPTOMS IN PATIENTS WITH AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE: FINDINGS FROM QUALITATIVE PATIENT INTERVIEWS
Authors: DasMahapatra Pronabesh , Cline Jennifer , Pulikottil-Jacob Ruth , Maski Manish , Cantor Sarah , Hariri Ali , Krupnick Robert , Modur Vijay 
Published: June 2020

Abstract

Background and Aims
Patients with Autosomal Dominant Polycystic Kidney Disease (ADPKD) experience various symptoms due to increasing kidney volume and its associated complications. To understand symptom burden, qualitative interviews were conducted to characterize the ADPKD patient experience pertaining to the most common and disturbing symptoms.

Method
Combined concept elicitation and cognitive debriefing interviews were performed by trained interviewers following a preliminary conceptual model based on a targeted literature review. We currently report on the concept elicitation component, which explored relevant and bothersome symptoms in ADPKD patients, due to either the disease or associated treatments. Nineteen telephone interviews were conducted in four waves of 5, 4, 6, and 4 patients, respectively.

Results
The mean age of patients was 47 years (SD: 9 years); 68% (n=13) were female, and the mean estimated glomerular filtration rate (eGFR) was 41 ml/min/1.73m2 (SD: 25 ml/min/1.73m2), with the majority 47% (n=9) in stage 3 chronic kidney disease (CKD). The classes of treatments included antihypertensives (74%, n=14), vasopressin receptor antagonists (21%, n=4); diuretics (16%, n=3), analgesics (16%, n=3) and erythropoietin (5%, n=1). The most frequently mentioned symptoms were fatigue (89%, n=17), pain (84%, n=16), and nocturia (74%, n=14); all were considered at least moderately disturbing to patients’ lives. Most patients experienced constant fatigue (74%, n=14) that worsened over time. Patients’ descriptions of pain were variable, including back and side pain, severe cystic pain, and muscle pain; a few patients also mentioned shoulder pain, or pain due to kidney stones. Among patients reporting nocturia (n=14), about half (57%, n=8) rated nocturia on the upper end of the disturbance scale, regardless of number of voids per night, often accompanied by sleep disruption. While less frequently occurring, itchy skin, muscle cramping, abdominal swelling, and inability to eat were spontaneously mentioned among the most disturbing symptoms.

Conclusion
The novel component of this study is the significant disruption of nocturia in patients with ADPKD even in the absence of vasopressin receptor antagonist administration. In addition, it confirms the findings of the literature review and highlights the patient experience in ADPKD, most notably fatigue, nocturia, and pain as the most frequent and bothersome symptoms experienced by patients. Subjective evaluation of the ADPKD patient experience is critical to understand symptom severity and the burden of disease, to potentially quantify disease progression, and to assess the impact of emerging therapies.
P0086FOCAL SEGMENTAL GLOMERULOSCLEROSIS AND TROMBOCYTOPENIA WITHOUT BLEEDING DIATHESIS: WHEN GENETICS MATTER
Authors: Olivo Elisa , Leonardi Sabina , Di Maso Vittorio , Artero Mary Louise , Bianco Francesco 
Published: June 2020

Abstract

Background and Aims
A 45-year old patient presented to our renal unit because of increased serum creatinine (2 mg/dL), proteinuria (2g/24h collection) and haematuria; platelets were 12000/mcL. His family history was negative. He had already undergone cataract surgery and was wearing a hearing device. Alport syndrome was suspected and kidney biopsy performed. Light microscopy showed focal segmental glomerulosclerosis (FSGS), while immunofluorescence was negative. Electron microscopy was not available. No specific treatment was undertaken and he progressed to end-stage renal disease (ESRD) in 10 years. The patient chose peritoneal dialysis. He was considered at high risk of bleeding but the Tenchkoff catheter was inserted without problems. After 3 months he received kidney transplant and again no pathological bleeding occurred.

Method
A genetic test was requested using next generation sequencing (NGS).

Results
A mutation of myosin heavy chain 9 (MYH9) gene which encodes the non-muscle myosin heavy chain-IIA (NMMH-IIA) was found. This mutation is associated with thrombocytopenia and proteinuria; platelets are giant and display normal aggregation, even in those patients who have a very low count. Proteinuria can be mild or in the nephrotic range, with or without microhaematuria. Kidney involvement is highly heterogeneous and leads to ESRD in half of the cases. Light microscopy shows FSGS as the prevalent pattern whereas electron microscopy is characterized by podocyte foot process effacement. In fact, NMMH-IIA is expressed in podocytes and controls cytoskeleton and cell migration. The differential diagnosis with Alport disease is challenging because additional features of MYH9 mutation are sensorineural deafness and pre-senile cataract. For all these reasons, accurate kidney biopsy study, family history and genetics are decisive.

Conclusion
Genetic testing has a pivotal role in understanding kidney diseases and should be more and more used.
P0089KEAP1/NRF2 PATHWAY REGULATES GFR BY INCREASING THE GLOMERULAR EFFECTIVE AREA WITHOUT AFFECTING THE AFFERENT/EFFERENT ARTERIOLE RATIO
Authors: KIDOKORO KENGO , Wada Yoshihisa , Kondo Megumi , Tokuyama Atsuyuki , Itano Seiji , Sasaki Tamaki , Wigley Chris , Proksch Joel , Meyer Colin , Kashihara Naoki 
Published: June 2020

Abstract

Background and Aims
The Keap1/Nrf2 pathway regulates the expression of a series of cytoprotective, anti-inflammatory and antioxidant genes. The Nrf2 activator, bardoxolone methyl (BARD), has consistently increased estimated GFR (eGFR) in clinical studies in patients with chronic kidney disease. BARD demonstrated improvement of renal function assessed by inulin clearance, the clinical gold standard for measuring GFR, in diabetic kidney disease patients. These findings suggest the Keap1/Nrf2 system is deeply involved in the regulatory mechanisms of GFR. However, the precise mechanisms are not fully elucidated. We pharmacologically and genetically investigated the mechanisms of GFR regulation by Keap1/Nrf2 system using in vivo multiphoton microscope (MPM) imaging techniques.

Method
C57BL/6 (Cont), Nrf2 knockout (Nrf2-KO), and Nrf2-activated Keap1-knockdown mice (Keap1-KD) were used. The mice were treated the synthetic triterpenoid RTA dh404 (10 mg/kg/day by gavage) which is a Nrf2 activator for rodents, for a week. We successfully developed the technique to evaluate single-nephron GFR (SNGFR) using MPM (Circulation 2019). The glomerular hemodynamics, diameter of the afferent/efferent arterioles and glomerular permeability were also evaluated. The calcium influx into cells in response to ATP and angiotensin II stimulation and the effect on [Ca2+]i by RTAdh404 were evaluated using Fluo 4 and Fura red in cultured mesangial cells and podocytes. Production of reactive oxygen species and nitric oxide (NO) availability were assessed by fluorescent method using CellROX® Deep Red and diaminofluorescein-FM diacetate (DAF-FM DA) upon the exposure to these stimuli.

Results
SNGFR in Keap1-KD mice was significantly higher than in the control (9.13±0.55 vs 4.40±0.39 nl/min, Figure 1). RTA dh404 increased SNGFR in the control but not in the Nrf2-KO mice (6.00±0.40 vs 4.66±0.35 nl/min, Figure 1). There was no significant change in the ratio of the glomerular afferent/efferent arteriole diameter in all groups. RTA dh404 treatment increased glomerular volume but did not affect the glomerular permeability of albumin and 40kd-dextran. RTA dh404-treatment inhibited calcium influx into cultured podocytes and mesangial cells induced by angiotensin II or ATP, thereby affecting contractile responses. Oxidative stress and NO-bioavailablity were also ameliorated with RTA dh404.

Conclusion
The Keap1/Nrf2 pathway plays a pivotal role in controlling GFR and presumably underlies the effect of BARD on GFR in patients.
P0095MOLECULAR MECHANISMS OF THE CARDIOVASCULAR EFFECTS OF LANTHIONINE, A NEW UREMIC TOXIN, AND ITS INTERACTIONS WITH THE REDOX MICROENVIRONMENT
Authors: Vigorito Carmela , Anishchenko Eugenia , Coppola Annapaola , Lombari Patrizia , Ingrosso Diego , Perna Alessandra 
Published: June 2020

Abstract

Background and Aims
The non-proteinogenic amino acid lanthionine is a byproduct of the biosynthesis of hydrogen sulfide (H2S), an endogenously produced gas with cardiovascular properties). Lanthionine concentration is increased in uremia and it has been proposed as a new uremic toxin. In the zebrafish model, lanthionine induces effects on cardiac embriogenesis, arrhythmia, and locomotor alterations. Some effects are counteracted by glutathione, the well known antioxidant. In a human endothelial cell model, lanthionine significantly reduces H2S release, both protein content and glutathionylation of cystathione beta-synthase (CBS), one of the main H2S-producing enzymes, miR-200c and miR-423 levels, as well as vascular endothelial growth factor expression, while it also increases intracellular calcium levels. We investigated in the present abstract the actions of glutathione on some of lanthionine effects on zebrafish and endothelial cells. We utilized what is considered to be a more stable form of glutathione, acetylglutathione (AcGSH).

Method
Zebrafish behavior and heart size were analyzed by the Danio Vision system and confocal microscopy, respectively. Gene and protein expression were analyzed usimg qPCR and Western Blot, respectively. CBS glutathionylation has been assessed by immunoprecipitation with a magnetic beads-coated anti-GSH. Interleukins levels were evaluated by ELISA. Lanthionine acetylation was evaluated by LC/MS.

Results
In zebrafish, we demonstrated that AcGSH significantly increases heart size and the expression of atrium specific proteins. Increased heart rate and heart rhythm plasticity determined by AcGSH were partially counteracted by lanthionine. Monitoring of larval movements showed that this behavior was affected by AcGSH supplementation. In endothelial cells, AcGSH increased H2S release, and, utilized in combination with lanthionine, it was able to partially counteract its effects on H2S release. AcGSH was able to increase CBS glutathionylation and to offset lanthionine effects on glutathionylation. In addition, it increased tubulin, histone, and NFKB acetylation and IL-8 and IL-13 levels. We also demonstrated with mass spectrometry that lanthionine was non-enzymatically acetylated by AcGSH, suggesting a possible mechanism.

Conclusion
In conclusion, our findings support the notion that lanthionine is a uremic toxin, which is strongly modified in its effects by the redox microenvironment.
P0096RENAL EXPRESSION AND CIRCULATING LEVELS OF CANONICAL WNT PATHWAY INHIBITORS IN THE EXPERIMENTAL MODEL OF EARLY CHRONIC KIDNEY DISEASE
Authors: Bogdanova Evdokia , Beresneva Olga , Galkina Olga , Zubina Irina , Ivanova Galina , Parastaeva Marina , Dobronravov Vladimir 
Published: June 2020

Abstract

Background and Aims
Molecular mechanisms implicated in the initial stages of inorganic phosphate (Pi) imbalance in chronic kidney disease (CKD) remain poorly understood.
The aim of the study was to evaluate whether canonical Wnt pathway inhibitors (iWnt) involved in early response to Pi retention in CKD.

Method
Mild CKD was induced by 3/4 nephrectomy (NE, n=10) in spontaneously hypertensive rats (SHR) fed rat chow diet containing 0.8 % phosphate. Controls were sham operated SHR (SO, n=9). Duration of experimental exposure (NE or SO) was 4 months. Creatinine (Cr), albumin/ creatinine ratio (Alb/Cr), inorganic phosphate (Pi), fractional (FEPi) and daily Pi excretion (uPi24), intact parathyroid hormone (PTH), fibroblast growth factor 23 (FGF23), Klotho protein, sclerostin (SOST) and Dickkopf-1 (DKK1), SOST and DKK1 mRNA expression in kidney, renal interstitial fibrosis (RF) were analyzed.

Results
Levels of Alb/Cr, serum Cr, RF and indices of Pi exchange in the experimental model corresponded to mild CKD with Pi retention (Table). No differences were observed in serum levels of Klotho, FGF23 and PTH in experimental group vs controls (Table). Serum SOST and DKK1 concentrations were significantly higher in NE group while the kidney expression of SOST and DKK1 mRNAs decreased compared to SO (Table).

Conclusion
Early stages of experimental CKD are characterized by increased serum concentrations of sclerostin and Dickkopf-1 in the absence of Klotho, FGF23 or PTH levels alterations. The increase of circulating iWnt levels seems not to be associated with their renal biosynthesis and likely have a bone origin.
P0099CALCINEURIN INHIBITION, BUT NOT DEHYDRATION, IN RATS MIMICS HUMAN RENAL HISTOPATHOLOGY OF PATIENTS WITH CHRONIC INTERSTITIAL NEPHRITIS IN AGRICULTURAL COMMUNITIES (CINAC)
Authors: Schreurs Gerd , Nast Cynthia , Maudsley Stuart , D'Haese Patrick , De Broe Marc , Vervaet Benjamin 
Published: June 2020

Abstract

Background and Aims
CINAC patients present a newly discovered constellation of proximal tubular lysosomal lesions which is also observed in patients experiencing calcineurin inhibitor (CNI) nephrotoxicity, suggesting that CINAC is a toxin-induced nephropathy involving calcineurin inhibition. An alternate hypothesis advocates chronic heat stress/dehydration as the major etiological factor for CINAC. Here, we evaluated in rats to what extent heat stress/dehydration versus CNI exposure reflects proximal tubular CINAC histopathology.

Method
Wistar rats were divided in 3 groups. Group 1 (n=6) was given water ad libitum (control group). Group 2 (n=8) was water deprived for 10 hours per 24h, 5 days/week and were placed in an incubator (37°C) for 30 min/hr of water deprivation. Group 3 (n=8) underwent daily oral gavage with cyclosporine (50mg/kg body weight). Animals were weighed daily and urine was collected at day 3, 17 and 28. After 28 days, rats were sacrificed. Kidneys were collected for light (LM) and electron microscopic (EM) histopathological analysis as well as for cortical renal tissue proteomics.

Results
Cyclosporine rats developed focal cortical lesions mimicking those of CINAC patients: i.e. atrophic proximal tubuli with thickened basement membranes and associated tubulo-interstitial fibrosis, PASM staining demonstrating enlarged argyrophyllic granules in the affected proximal tubuli, LAMP1 immunofluorescent staining identifying a subset of these granules as lysosomes, and EM confirming the presence of enlarged lysosomes, some dysmorphic approaching CINAC lysosomes. In dehydrated rats, confirmed by urinary osmolality and fluctuating body weight associated with water deprivation, none of the cyclosporine features were observed. Proteomic analysis confirmed cellular toxicity by cyclosporine, whereas the dehydration group lacked any markers of such.

Conclusion
The histopathological analogy between CNI nephrotoxicity in rats and humans and CINAC suggests a toxicological etiology for CINAC. In rats, dehydration/heat stress alone does not lead to the constellation of proximal tubular lesions as observed in CINAC patients.
P0122THE MECHANISM OF DNA INJURY INDUCED COLLAGEN TYPE VI EXCRETION IN GLOMERULAR ENDOTHERIAL CELLS
Authors: Ai Fujii , Sunatani Yumi , Kengo Furuichi , Fujimoto Keiji , ADACHI HIROKI , Iwabushi Kuniyoshi , Yokoyama Hitoshi 
Published: June 2020

Abstract

Background and Aims
Collagen deposition is observed in various part of kidney tissue and is serious pathological damage in kidney. Nodular glomerulosclerosis is one of a phenotype of collagen deposition in glomerular lesion, which affects the prognosis of renal function, but there is no effective treatment at present. Recently, we identified that glomerular endothelial cells with DNA double-strand breaks (DSB) are involved in the long-term accumulation of ECM molecules, especially collagen type VI(COL6). However, the mechanism of glomerular COL6 accumulation is still unclear.

Method
We examined the phospho-histone H2AX (γ-H2AX) of the DSB marker and COL6 accumulation in biopsied human kidneys to investigate their association. In addition, we investigated in vitro the relationship between DSB and COL6 excretion and the intracellular signal pathways in human glomerular endothelial cells (HRGEc) using mitomycin C (MMc)-induced DNA damage. In this study, we focused on the expression of DSB response signal pathways, i.e. ATM, ATR and DNA-PK using their specific kinase inhibitors (KU55933, VE-821, Nu7441).

Results
COL6 deposits and γ-H2AX expression in nuclei were detected in human glomerular capillary loop and glomerular nodular lesions. The multiple regression analysis showed that γ-H2AX positive area is the most independent factor for COL6 accumulation in glomeruli (β: 0.539, t=2.668, p=0.014). Furthermore, COL6 was the independent factor of nodular lesion in various pathologic diseases such as diabetic nephropathy, hypertensive glomerulosclerosis and the long-term transplanted allografts (β: 0.373, t=2.268, p=0.032).
In vitro study, COL6 excretion detected by the decrease of COL6 positive cells was suppressed only in the ATR-inhibited group (control vs. kinase inhibitors; 2 h, 40.1 ± 23.5 vs. 59.0 ± 23.2%, p &lt;0.01; 24 h, 18.8 ± 23.6 vs. 56.3 ± 16.5%, p &lt;0.001).

Conclusion
This study suggested that nodular glomerulosclerosis is induced by DNA damage and COL6 deposition through ATR pathway in glomerular capillary endothelial cells. These findings will provide great insights into the development of new therapies that would suppress glomerularsclerosis in various kidney diseases.