bluuebunny/crossref_metadata_2025 huggingface dataset:
Altered Dynamic Information Flow through the Cortico-Basal Ganglia Pathways Mediates Parkinson’s Disease Symptoms
Authors: Chiken Satomi , Takada Masahiko , Nambu Atsushi
Published: May 2021

Parkinson’s disease (PD) is a progressive neurodegenerative disorder caused by dopamine deficiency. To elucidate network-level changes through the cortico-basal ganglia pathways in PD, we recorded neuronal activity in PD monkeys treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. We applied electrical stimulation to the motor cortices and examined responses in the internal (GPi) and external (GPe) segments of the globus pallidus, the output and relay nuclei of the basal ganglia, respectively. In the normal state, cortical stimulation induced a triphasic response composed of early excitation, inhibition, and late excitation in the GPi and GPe. In the PD state, cortically evoked inhibition in the GPi mediated by the cortico-striato-GPi “direct” pathway was largely diminished, whereas late excitation in the GPe mediated by the cortico-striato-GPe-subthalamo (STN)-GPe pathway was elongated. l-DOPA treatment ameliorated PD signs, particularly akinesia/bradykinesia, and normalized cortically evoked responses in both the GPi and GPe. STN blockade by muscimol injection ameliorated the motor deficit and unmasked cortically evoked inhibition in the GPi. These results suggest that information flow through the direct pathway responsible for the initiation of movements is largely reduced in PD and fails to release movements, resulting in akinesia/bradykinesia. Restoration of the information flow through the direct pathway recovers execution of voluntary movements.
Joining Formal Concept Analysis to Feature Extraction for Data Pruning in Cloud of Things
Authors: Moulahi Tarek
Published: May 2021

The enormous increase in the number of Internet of Things data sources (sensors, personal devices and embedded systems) has given rise to a huge amount of unnecessary and redundant data being sent to the cloud. This makes the task of processing and storing this volume of information a very hard one. Therefore, data pre-processing and filtering closer to data sources, such as in fog computing, is necessary. In particular, data reduction in the fog nodes may play a significant role in preventing the dramatic decrease in the Cloud of Things performance, especially in energy consumption, storage space, bandwidth and throughput. However, existing solutions are still lacking, considering they do not achieve the optimal data reduction performance especially in terms of delay and accuracy. In this article, we introduce an approach aiming to eliminate useless and redundant data captured by things basing on some intelligent information extraction techniques. We also evaluate the performance of the proposed solution on a real data set sample to demonstrate that it achieves a good features reduction.
CARAF: Crypto Agility Risk Assessment Framework
Authors: Ma Chujiao , Colon Luis , Dera Joe , Rashidi Bahman , Garg Vaibhav
Published: May 2021

Crypto agility refers to the ability of an entity to replace existing crypto primitives, algorithms, or protocols with a new alternative quickly, inexpensively, with no or acceptable risk exposure. These changes may be driven by regulatory action, advances in computing, or newly discovered vulnerabilities. Yet everyday operational needs may put crypto agility considerations on the back burner when deploying technology, designing processes, or developing products/services. Consequently, changes are often performed in an ad hoc manner. Transition from one crypto solution to another can then take a long time and expose organizations to unnecessary security risk. This paper presents a framework to analyze and evaluate the risk that results from the lack of crypto agility. The proposed framework can be used by organizations to determine an appropriate mitigation strategy commensurate with their risk tolerance. We demonstrate the application of this framework with a case study of quantum computing and related threats to cryptography in the context of TLS for Internet of Things.
Surface modification by superimposing piecewise shape function satisfying hull variation given by arbitrary characteristic curve on surface
Authors: Luong Quang-Phap , Nam Jong-Ho
Published: May 2021

The ship hull form is directly related to each design phase of a ship. This form is thus frequently designed based on a variation of a well-made parent ship to avoid new design risks. However, in hull form variation, satisfactory geometric preservation and good performance continuity of the parent ship are considered competing requirements. To effectively facilitate the variation, an appropriate compromise between these requirements should be achieved. Instead of altering hull form parameters, we propose a flexible and intuitive approach to interactively vary the hull surfaces with arbitrary curve constraints and to focus on preserving the properties of the original hull surface. The advantage of the superposition of the curve or surface is leveraged to modify the hull shape. An intermediate surface that satisfies an intermediate curve constraint and has a smooth shape is constructed and superimposed on the top of the original surface to yield the desired surface. A shape control mechanism using a shape function simplifies the control of the surface modification with a curve constraint. The developed method was applied to modify the characteristic curves and section curves situated on the stern part of a container vessel. The changes in the geometric shape and hydrodynamic performance due to the curve-constraint variation were assessed to demonstrate the practicality and efficiency of the proposed method. The results show that the proposed method produces a smooth shape and almost unchanging hydrodynamic patterns. The proposed variation provides an effective tool for varying the hull in a linear fashion.
Survival of chemo-naïve patients with EGFR mutation-positive advanced non-small cell lung cancer after treatment with afatinib and bevacizumab: updates from the Okayama Lung Cancer Study Group Trial 1404
Authors: Ninomiya Takashi , Nogami Naoyuki , Kozuki Toshiyuki , Harada Daijiro , Kubo Toshio , Ohashi Kadoaki , Ichihara Eiki , Kuyama Shoichi , Kudo Kenichiro , Bessho Akihiro , Sakugawa Makoto , Fujimoto Nobukazu , Aoe Keisuke , Minami Daisuke , Sugimoto Keisuke , Ochi Nobuaki , Takigawa Nagio , Hotta Katsuyuki , Maeda Yoshinobu , Kiura Katsuyuki
Published: May 2021

Background

In a phase I study, afatinib (30 mg/body daily) plus bevacizumab (15 mg/kg every 3 weeks) was well tolerated and showed favourable outcomes in patients with epidermal growth factor receptor (EGFR)-mutant advanced non-small-cell lung cancer. Herein, we report the 2-year progression-free survival, overall survival and safety profile of these patients.

Methods

Chemo-naïve patients with EGFR-mutant advanced non-small-cell lung cancer were enrolled. One group of patients received 40 mg afatinib daily and 15 mg/kg bevacizumab every 3 weeks (level 0) until disease progression or severe toxicity. Another group of patients received 30 mg afatinib daily and the same dose of bevacizumab (level 1). Dose-limiting toxicity was the primary endpoint, whereas long-term progression-free survival, overall survival and tolerability were secondary endpoints. Survival rates were estimated using the Kaplan–Meier method.

Results

The study included 19 patients (level 0: 5; level − 1: 14). Until the data cut-off date, seven patients continued the treatment, whereas 12 discontinued due to disease progression (n = 5) or toxicity (n = 7). The median PFS was 24.2 months, while the median overall survival was not reached. All patients developed adverse effects. Diarrhoea and skin rash were frequently observed as severe adverse events (grade 3). A secondary EGFR mutation (T790M) was detected in two patients after progression.

Conclusions

Prolonged follow-up revealed that combination therapy with afatinib and bevacizumab might improve survival outcomes in EGFR-mutant advanced non-small-cell lung cancer patients and seems to be promising.

Trial registration

UMIN000015944.
Digital Communication at the Time of COVID-19: Relieve the Refugees’ Psychosocial Burden and Protect Their Wellbeing
Authors: Mancini Tiziana , Imperato Chiara , Vesco Silvia , Rossi Michele
Published: May 2021

The study aims to evaluate the usefulness and effectiveness of digital communication tools implemented by an Italian NGO during the coronavirus disease 2019 (COVID-19) emergency period, considering their relations with perception of health-related risk, sense of belonging to host community, psychological distress, and wellbeing of the refugees taken in charge by the association. The study was conducted through an anonymous questionnaire proposed to a group of 102 refugees. The results confirmed that by increasing the sense of belonging to the Italian community, the time refugees spent on digital communication negatively associated with their psychological distress and positively associated with their wellbeing. Refugees’ access to digital communication also increased their health-related risk perception that in turn raised their psychological distress. It specifically occurred when digital communication has been perceived as less affordable, i.e. as less clear, easy and reassuring. The results will be used to improve digital communication for health, educational, and proximity services dedicated to refugees.
Adenosine 5′ phosphosulfate reductase and sulfite oxidase regulate sulfite-induced water loss in Arabidopsis
Authors: Bekturova Aizat , Oshanova Dinara , Tiwari Poonam , Nurbekova Zhadyrassyn , Kurmanbayeva Assylay , Soltabayeva Aigerim , Yarmolinsky Dmitry , Srivastava Sudhakar , Turecková Veronika , Strnad Miroslav , Sagi Moshe
Published: May 2021

Chloroplast-localized adenosine-5’-phosphosulphate reductase (APR) generates sulfite and plays a pivotal role in reduction of sulfate to cysteine. The peroxisome-localized sulfite oxidase (SO) oxidizes excess sulfite to sulfate. Arabidopsis wild type, SO RNA-interference (SO Ri) and SO overexpression (SO OE) transgenic lines infiltrated with sulfite showed increased water loss in SO Ri plants, and smaller stomatal apertures in SO OE plants compared with wild-type plants. Sulfite application also limited sulfate and abscisic acid-induced stomatal closure in wild type and SO Ri. The increases in APR activity in response to sulfite infiltration into wild type and SO Ri leaves resulted in an increase in endogenous sulfite, indicating that APR has an important role in sulfite-induced increases in stomatal aperture. Sulfite-induced H2O2 generation by NADPH oxidase led to enhanced APR expression and sulfite production. Suppression of APR by inhibiting NADPH oxidase and glutathione reductase2 (GR2), or mutation in APR2 or GR2, resulted in a decrease in sulfite production and stomatal apertures. The importance of APR and SO and the significance of sulfite concentrations in water loss were further demonstrated during rapid, harsh drought stress in root-detached wild-type, gr2 and SO transgenic plants. Our results demonstrate the role of SO in sulfite homeostasis in relation to water consumption in well-watered plants.
Magnetohydrodynamic stability of magnetars in the ultrastrong field regime I: the core
Authors: Rau Peter B , Wasserman Ira
Published: May 2021

We study magnetohydrodynamic stability of neutron star core matter composed of neutrons, protons, and leptons threaded by a magnetar-strength magnetic field 1014–1017 G, where quantum electrodynamical effects and Landau quantization of fermions are important. Stability is determined using the Friedman–Schutz formalism for the canonical energy of fluid perturbations, which we calculate for a magnetizable fluid with H ≠ B. Using this and the Euler–Heisenberg–Fermi–Dirac Lagrangian for a strongly magnetized fluid of Landau-quantized charged fermions, we calculate the local stability criteria for a neutron star core with a spherical axisymmetric geometry threaded by a toroidal field, accounting for magnetic and composition gradient buoyancy. We find that, for sufficiently strong fields B ≳ 1015 G, the magnetized fluid is unstable to a magnetosonic-type instability with growth times of the order of 10−3 s. The instability is triggered by sharp changes in the second-order field derivative of the Euler–Heisenberg–Fermi–Dirac Lagrangian that occur where additional Landau levels start being populated. These sharp changes are divergent at zero temperature, but are finite for non-zero temperature, so realistic neutron star core temperatures 5 × 107 K < T < 5 × 108 K are used. We conjecture that this mechanism could promote the formation of magnetic domains as predicted by Blandford and Hernquist and Suh and Mathews.
MO001THE EUROPEAN DRTA REGISTRY: AN INITIAL DATA ANALYSIS*
Authors: Bockenhauer Detlef , Emma Francesco , Talhi Amina , Papizh Svetlana , Atmis Bahriye , Homan Nakisa , Aksu Bagdagul , Santos Fernando , Boyer Olivia , Fila Marc , Bayrakci Umut Selda , Ouda Eid Loai Akram , VanDyck Maria , Moguel Carlos , Yüksel Selçuk , Canpolat Nur , Schaefer Franz , Oh Jun , Liebau Max , Ariceta Iraola Maria Gema , Vargas-Poussou Rosa , Levtchenko Elena , Lopez-Garcia Sergio Camilo , Wlodkowski Tanja , Topologlu Rezan
Published: May 2021

Background and Aims

Distal renal tubular acidosis (dRTA) is a rare disorder characterised by an inability of the distal tubule to secrete acid, leading to metabolic acidosis. Clinical consequences typically include hypokalaemia, hypercalciuria with nephrocalcinosis and/or urolithiasis, as well as bone disease. Treatment with adequate alkali supplementation corrects the acidosis and hypercalciuria, but there are few data on long-term outcome. In 2018, a registry for dRTA was established by the European Society for Paediatric Nephrology, hosted by the European Rare Kidney Disease Reference Network. Here, we present an initial analysis of data in the registry.

Method

Analysis of data entered into the registry by the cut-off data of 18/11/2020.

Results

A total of 135 patients had been entered, of which 106 had additional data from an annual follow-up visit. Median age at last visit was 10 years (range 0-54), including 16 adults (>17y). Genetic testing had been performed in 91 subjects and causative variants were reported in 74 (81%). Pertinent clinical details according to genetic group are listed in table 1.

Treatment was provided with at least 15 different preparations, containing citrate or bicarbonate, given in 1-10 (median 3) daily doses.

Adequate treatment at last follow-up, as judged by a plasma bicarbonate level >21 mmol/l and a urine calcium-creatinine ratio in the age-specific normal range was present in 46% of subjects.

There was a trend for higher eGFR and height SDS in subjects with adequate treatment compared to those without, but this was not statistically significant.

Conclusion

Currently available data demonstrate the difficulties in treating dRTA, with less than half of subjects achieving adequate control of their acidosis. By collecting long-term data, the registry will provide important information on the prognosis and complications of dRTA and to what degree these can be prevented with treatment. Enrollment of further, especially adult patients will contribute to our understanding of this rare disorder.
MO004PRIMARY BILIARY CHOLANGITIS PRESENTING WITH RENAL FANCONI SYNDROME: A FORGOTTEN PHENOTYPE
Authors: Er Chaoxui , Sayer John
Published: May 2021

Background and Aims

Primary biliary cholangitits (PBC) is an autoimmune liver disease, leading to liver fibrosis and cirrhosis. It is a rare disease affecting 1 in 3-4000 people and is more common in females. Symptoms may go unnoticed and include itch and fatigue. Most patients have anti-mitochondrial antibodies (AMA) as well as raised gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP) levels. First line treatment for PBC is ursodeoxycholic acid and is usually continued lifelong. It improves liver biochemistry, histological progression of liver disease and liver transplant-free survival. Renal complications of PBC include distal renal tubular acidosis (RTA), tubule-interstitial nephritis (TIN) and renal Fanconi syndrome.

Method

We reviewed a case of PBC presenting with renal Fanconi syndrome.

Results

A 48-year-old female was referred to the renal clinic due to progressive decline in renal function since she was diagnosed with type 2 diabetes in 2007. She was also known to have AMA (>1:640) and previously had transient transaminitis. She was clinically well with no major symptoms but reported that she had had a few episodes of urinary tract infection in the previous year. Her diabetes was managed with lifestyle modifications in the past until a few months ago when Metformin was introduced. However, her HbA1c level had never been greater than 55 mmol/mol. Urine dipstick in the clinic showed pH of 6, blood+, glucose+++, protein+++ and ketone trace. The severity of glycosuria was inconsistent with her glycaemic control. There was a disparity between her urine albumin/creatinine ration (ACR) of 18.8g/mol and protein/creatinine ratio (PCR) of 124mg/mmol. Myeloma screen was negative and further urine analysis showed generalised aminoaciduria. She also had hypouricaemia, intermittent hypophosphataemia and non-anion gap metabolic acidosis. These results are in keeping with renal Fanconi syndrome. Her eGFR was 48 ml/min/1.72m2 in 2007 and was 21 ml/min/1.72m at the time of review. A renal biopsy was undertaken, and the appearances were suggestive of mild tubulo-interstitial nephritis; the glomeruli were unremarkable; there was mild chronic tubulo-interstitial damage. She was started on oral steroid, sodium bicarbonate and ursodeoxycholic acid. The course of steroid had a slight transitory beneficial effect on the renal function.

Conclusion

Distal RTA is the usual renal feature of PBC, occurring in 1/3 of cases with advanced disease. In contrast, proximal RTA associating with renal Fanconi syndrome occurs rarely. Like our case, the cases that have been previously reported show that Fanconi syndrome occurred during the early phase of PBC in the absence of marked hepatic abnormalities, and were associated with CKD.

Fanconi syndrome and TIN are renal features of mitochondrial cytopathies and are perhaps a forgotten association of PBC. Antimitochondrial antibodies may play a role in the onset of tubulo-interstitial nephritis and Fanconi syndrome.
MO078DEEP LEARNING DIAGNOSIS OF ANTIBODY-MEDIATED REJECTION (AMR) ON GLOMERULAR TRANSECTIONS
Authors: Cicalese Pietro A , Rizvi Syed A , Roufosse Candice , Batal Ibrahim , Hellmich Martin , Ernst Angela , Moos Katharina , Clahsen-van Groningen Marian , Weidemann Alexander , Mohan Chandra , Nguyen Hien V , Becker Jan Ulrich
Published: May 2021

Background and Aims

Antibody-mediated rejection (AMR) is among the most common causes for kidney transplant loss. The histological diagnosis is hampered by significant intra- and interobserver variability. Training a deep learning classifier for the recognition of AMR on glomerular transections as the most decisive compartment could establish a reliable and perfectly reproducible diagnostic method.

Method

We identified 48 biopsies with AMR (all positive for donor-specific antibody) and 38 biopsies without AMR according to Banff 2017 from our archive. Photographs were taken from all non-globally sclerosed glomeruli on two PAS-stained level sections, yielding a total of 1,655 images as a training set. 1,503 images could be labeled by three experienced nephropathologists conclusively as AMR or non-AMR in a blinded fashion. We trained a DenseNet-121 classifier (pre-trained on ImageNet) with basic online augmentation. In addition, we implemented StyPath++, a data augmentation algorithm that leverages a style transfer mechanism, addressing significant domain shifts in histopathology. Each sample was assigned a consensus label generated by the pathologists.

Results

Five-fold cross validation schemes produced a weighted glomerular level performance of 88.1%, exceeding the baseline performance by 5%. The improved generalization ability of the StyPath++ augmented model shows that it is possible to construct reliable glomerular classification algorithms with scarce datasets.

Conclusion

We created a deep learning classifier with excellent performance and reproducibility for the diagnosis of AMR on glomerular transections. We plan to expand the training set, including challenging cases of differential diagnoses like glomerulonephritis or other glomerulopathies. We are also interested in external clinicopathological datasets to further validate our results.
MO082THE EFFECTS OF DIFFERENT MICROMOLAR CONCENTRATIONS OF CADMIUM IONS ON THE PERITUBULAR MEMBRANE POTENTIAL OF PROXIMAL TUBULAR CELLS IN PERFUSED FROG KIDNEYS
Authors: Kovacevic Sanjin , Spasic Svetolik , Lopicic Srdjan , Todorovic Jasna , Dincic Marko , Stanojevic Marija , Nesovic - Ostojic Jelena
Published: May 2021

Background and Aims

Cadmium (Cd2+) is toxic metal and environmental pollutant. Accumulation of cadmium in the kidney results initially in proximal tubule dysfunction. Although Cd2+ toxicity is well documented, all mechanisms that are involved in the early stages of nephrotoxicity, especially considering low micromolar concentrations of Cd2+ ions are still unknown. The Aim of this study was to investigate the effects acute exposure to different peritubular micromolar concentrations of cadmium (0.25, 0.50, 1.0, 2.0, 3.0, 5.0 μmol/L) on the peritubular cell membrane potential in proximal tubular cells of frog kidney.

Method

The experiments were performed on isolated, doubly perfused kidneys of Rana esculenta of both sexes. Aortic and portal vein were cannulated in order to perfusate luminal and peritubular cell membraine. In controled conditions, Ringer solution was simultaneously used to perfusate both cell membraines. Cadmium chloride (different concentrations: 0.25, 0.50, 1.0, 2.0, 3.0, 5.0 μmol/L) were added to the peritubular perfusate separately, by switching the peritubular perfusate from the control Ringer solution to Ringer solution with addition of cadmium ions. Peritubular cell membrane potentials (PD) were measured with conventional 3 mol/L KCl microelectrodes.

Results

The peritubular application of different micromolar Cd2+ concentrations led to a rapid, sustained, reversible hyperpolarization of the peritubular cell membrane: 0.25 µmol/L, by −3.3±0.4 mV (n=8, p<0.001); 0.50 µmol/L, by −3.0±0.5 mV (n=11, p<0.001); 1.0 µmol/L, by −2.9±0.6 mV (n=8, p<0.01); 2.0 µmol/L, by −4.2±0.4 mV (n=13, p<0.01); 3.0 µmol/L, by −3.4±0.3 mV (n=14, p<0.001); 5.0 µmol/L, by −3.0±0.4 mV (n=10, p<0.001). After switching the perfusion from Ringer solution with addition of cadmium ions to control Ringer, the peritubular membraine potential returned to the average values that were maintained before the peritubular Cd2+ application (p>0.05). Comparing the effect of different Cd2+ concentrations, there was no difference in the hyperpolarization of the peritubular cell membrane (p>0.05).Each cell served as its own control.

Conclusion

Different low micromolar concentrations of Cd2+ provoked rapid and sustained hyperpolarization of peritubular membrane potential that did not show concentration-dependent response.
MO087PLASMALEMMAL VESICLE-ASSOCIATED PROTEIN-1 (PLVAP) INDICATES THE FORMATION OF DIAPHRAGM-BRIDGED FENESTRATIONS OF GLOMERULAR ENDOTHELIAL CELLS IN KIDNEY DISEASE
Authors: Tampe Björn , Schridde Laura , Hakroush Samy
Published: May 2021

Background and Aims

Plasmalemmal vesicle-associated protein-1 (PLVAP or PV-1) is a major protein of diaphragm-bridged fenestrated endothelial cells found in capillaries of neuroendocrine glands and peritubular capillaries. In contrast to peritubular capillaries, the glomerulus is known for its unique fenestrated endothelium without any diaphragm formation thereby ensuring free filtration. Here we aimed to investigate whether PLVAP is expressed in glomerular endothelial cells in various glomerular diseases and whether PLVAP expression is associated with the formation of diaphragm-bridged endothelial cells.

Method

A total number of 114 biopsy samples of glomerular diseases including diabetic nephropathy, FSGS, IgA-Nephritis, ANCA-GN and Lupus–Nephritis were analyzed immunohistochemistically for glomerular PLVAP expression. A fraction of PLVAP positive cases was subsequently investigated ultrastrucurally for the formation of diaphragm-bridged glomerular endothelial cells.

Results

One third of all cases showed at least one glomerulus with one single circumferential PLVAP staining. Interestingly, the most prominent staining, affecting the entire glomerular tuft, was observed in diabetic nephropathy and ANCA-GN. Ultrastructurally, such cases exhibited injured endothelium with focal detachment from the glomerular basement membrane, loss of pore formation and frequently diaphragm-bridged fenestrations reminiscent of peritubular capillaries.

Conclusion

Our data show that injured glomerular endothelium is capable of forming true diaphragm-bridged fenestrations, suggesting a possible role in preventing glomerular protein leakage and limiting its detachment from the GBM.
MO067DELETION AND BLOCKAGE OF KININ B1 RECEPTOR EXACERBATES CISPLATIN-INDUCED CKD
Authors: Estrela Gabriel , Budu Alexandre , Freitas-Lima Leandro , Arruda Adriano , Perilhão Mauro , Barrera-Chimal Jonatan , Araujo Ronaldo
Published: May 2021

Background and Aims

Kinins plays a major role in immune response, where kinin B2 receptor is constitutively expressed and kinin B1 receptor is induced under inflammatory stimuli. Kinin B1 receptor deletion and blockage has been shown to have beneficial effects in some models of renal diseases. Multiple acute renal insults, even if followed by renal recovery, is a risk factor for the future development of chronic kidney disease (CKD) and end-stage renal disease (ESRD). Our main objective was to determine the importance of kinin B1 receptor in tubulointerstitial fibrosis induced by ongoing cisplatin treatment.

Method

Male C57/Bl6 mice were divided in 3 groups, vehicle, cisplatin, cisplatin + R715 (kinin B1 receptor antagonist). Animals has been treated with multiple doses of cisplatin (7mg/kg i.p) once a week during 4 weeks and R715 (0.8mg/kg i.p) 48, 24 and 1 hour prior to cisplatin injections. We also used B1KO mice (C57Bl6 background) and WT mice (littermates). Mice were euthanized after 30 days of last cisplatin injection. Renal parameters, histology, real time PCR were performed to investigate renal injury, inflammation and fibrosis.

Results

Cisplatin treatment increases most of renal parameters, renal injury and fibrosis markers. Deletion of B1 receptor exacerbates significantly creatinine (WT CIS 0,60+0,03 B1KO 0,76+0,01 mg/dL), urea (WT CIS 111,8+5,638 B1KO CIS 240,8+28,60 mg/dL), and protein excretion (WT CIS 0,0058+0,0011 B1KO CIS 0,0121+0,0007 mg/24h). Association of cisplatin with R715 increased creatinine levels (veh 0,43+0,03 cis+R715 0,64+0,04 mg/dL), exacerbates urea (cis 95,51 + 3,926 cis+R715 158,9+14,40 mg/dL) and protein excretion (cis 0,012+0,002 cis+R715 0,018+0,003 mg/24h). Renal injury markers such as KIM-1 and TNF-a showed no significant differences. NGAL expression exacerbates (cis 2,53+0,44 cis+R715 5,66+1,34) and tubular injury score (cis 0,130+0,021 cis+R715 0,191+0,020) is higher in cisplatin+R715 group. Fibrosis markers a-SMA (cis 2,56+0,43 cis+R715 4,67+0,99), Col4 (cis 2,57+0,39 cis+R715 5,14+1,01) and Vimentin (2,69+0,31 cis+R715 4,62+0,98) were exacerbated in cisplatin treatment associated with R715. Picrosirius red staining were used to asses tubulointerstitial fibrosis, and we confirmed that R715 treatment here also exacerbates fibrosis (cis 0,225+0,025 cis+R715 0,345+0,042).

Conclusion

Here we show that both deletion and blockage of kinin B1 receptor has deleterious effects in renal injury and fibrosis induced by ongoing cisplatin treatment.
MO014DANHONG INJECTION INHIBITS LIPOPOLYSACCHARIDE-ENHANCED CELL PROLIFERATION OF RAT RENAL MESANGIAL CELLS VIA NF-ΚB SIGNALING PATHWAY
Authors: Hua Liu , Chen Lei , Wei Limin , Jiang Hongli
Published: May 2021

Background and Aims

Mesangioproliferative glomerulonephritis (MsPGN) is well known as one of the leading causes of end-stage renal failure (ESRD), especially in the Asian population. The principal characteristic of MsPGN is the over proliferation of glomerular mesangial cells (MCs) accompanied by expansion of the extracellular matrix (ECM). Our previous studies demonstrated that Danhong injection (DHI), is a traditional Chinese medicine injection, could improve the renal function and inhibit the MCs proliferation and ECM expansion in rats with MsPGN. However, the molecular mechanisms have not been fully elucidated. To explore the potential mechanisms of DHI in the treatment of MsPGN, we investigated the effects of DHI on lipopolysaccharide (LPS)-induced nuclear factor-kappa B (NF-κB) activation and its downstream inflammatory mediators, such as intercellular adhesion molecule-1 (ICAM-1), transforming growth factor-beta 1 (TGF-β1), inducible nitric oxide synthase (iNOS) and fibronectin (FN) protein expression in rat MCs.

Method

The rat MCs treated with different concentrations of DHI (0, 50, 100, 200, 500, 1000, and 2000 uL/L) for 12 h, then incubated with or without 100 ng/ml LPS for another 24 h. Subsequently, cell proliferation was determined by Cell Counting Kit-8 (CCK8). Furthermore, the rat MCs treated with low-dose DHI (250 uL/L), median-dose DHI (500 uL/L) and high-dose DHI (1000 uL/L) for 12 h or Ammonium pyrrolidine dithiocarbamate (PDTC) for 30 min before 24h treatment of LPS. Then the activation of NF-κB was detected by Western blot and immunofluorescence. The protein levels of ICAM-1, TGF-β1, iNOS and FN in rat MCs were detected by Western blot.

Results

The results of CCK-8 revealed that DHI significantly suppressed LPS-induced cell proliferation (shown in Fig.1). LPS stimulation resulted in a significant increment of p65 contents in nucleus and a decrement of p65 contents in cytoplasm in rat MCs compared with NC. PDTC and DHI exerted potent inhibitory effect on increasing expression of p65 in nucleus and decreasing in cytoplasm compared with LPS treatment group. The inhibitory effect on NF-κB nuclear translocation of DHI was in a dose- dependent manner (shown in Fig.2). The Western blot assay showed that the protein level of IκB-α in cytoplasm treated by LPS decreased significantly compared with that in control (shown in Fig. 3) and this decrement was significantly reversed by PDTC and DHI. In addition, the protein expression of ICAM-1, TGF-β1, iNOS and FN was also inhibited by PDTC and DHI (shown in Fig. 4).

Conclusion

DHI significantly repressed LPS-induced cell proliferation and FN expression in rat MCs through inhibiting the activation of NF-κB signaling pathway and protein expression of its downstream inflammatory mediators. The ameliorative effects of DHI on MsPGN might be associated with this inhibition effect on NF-B signaling pathway.
MO015ANALYZING THE EFFECT OF MICROPLASTIC PARTICLES ON HUMAN PODOCYTES
Authors: Yarbakht Melina , Sarau George , Kling Lasse , Müller-Deile Janina , Kotb Ahmed , Christiansen Silke , Schiffer Mario
Published: May 2021

Background and Aims

Despite increasing use of plastic based products and their potential health risks on the humans, very little is known about their possible accumulation in the food chain and their further long-term effect on the human health. Recently, there are increasing reports related to the potential risk of polystyrene microplastics to the human respiratory system and human intestinal epithelia cell line. In this study, we assayed the primary effect of microplastic particles on the human kidney cells. To that aim, we used human podocytes cells and four different types of plastic particles including; polyvinyl chloride (PVC), polypropylene (PP), polyamide (PA) and tyre wear particles to evaluate the effects of microplastics on the viability and morphology of human podocytes in vitro.

Method

In this study, we applied different biological methods such as, cell viability test and phalloidin staining, to assay the toxicity of particles and their further effects on the actin cytoskeleton organization in human podocytes, respectively. Furthermore, Raman imaging is used to track particle attachment on the cells and to evaluate the possible changes in the cell compartment following the particle treatment. The particle uptake by the cells and changes in cellular biological features were visualized with the use of scanning electron microscopy (SEM).

Results

As a primary result, the cytotoxicity response of particle treatment was found to be dependent on the polymer type. As an example higher concentration of PP particle as compared to PVC, PA, and tyre wear caused a similar rate of cell mortality. Furthermore, the degree of particle attachment on the cells depended on their adhesion properties, which was higher in PA, PVC and tyre wear in comparison to PP particles. These particles remained attached to the cell surface even after two-three times of washing with PBS. Based on the phalloidin staining results, particle treatment induced cytoskeleton reorganization in podocytes in vitro. With the use of Raman imaging particle attachment was confirmed based on the fingerprint spectra related to each particle.

Conclusion

This study suggests that exposure duration and particle concentrations are two of the key factors to evaluate the toxicological effect of particles on podocytes as a highly-specialized epithelial cells in the kidney. It is supposed that two mechanisms can be related to the harmful effects of plastic particles on podocytes. First, particle attachment on the cell surface leading to limitation of nutrient uptake by the cells. Second, uptake of smaller size particles into the cells through phagocytosis. More studies are necessary to determine the direct effect of microplastics on human kidney cells.
MO018QUORUM SENSING MOLECULE INDUCES THE RECEPTOR-INTERACTING PROTEIN KINASE 1-DEPENDENT APOPTOSIS SYNERGISTICALLY WITH LIPOPOLYSACCHARIDE IN ENDOTHELIAL CELLS
Authors: Shin Jungho , Ahn Sun Hee , Oh Dong-Jin
Published: May 2021

Background and Aims

Endothelial dysfunction is an initial step for sepsis-associated organ failure. Bacterial quorum sensing molecules play as pathogen-associated molecular patterns, however, the impact of quorum sensing molecules on endothelial cells remains uncertain. This study investigated the molecular mechanism of quorum sensing molecule-induced cell death and the interaction with lipopolysaccharide (LPS) in human umbilical vein endothelial cells.

Method

Endothelial cells were treated with varying concentrations of N-3-oxododecanoyl-HSL (3OC12-HSL) and LPS derived from Pseudomonas aeruginosa.

Results

Treatment of 3OC12-HSL reduced cell viability in dose-dependent manners, and the combination of this molecule and LPS exacerbated cell deaths in compared to single treatment of LPS. The terminal deoxynucleotidyl transferase dUTP nick end labeling assay revealed that there were clear increases in apoptotic death following the 3OC12-HSL treatment. Treatment of 3CO12-HSL activated the receptor-interacting protein kinase 1 (RIPK1) pathway, resulting in increases in expression of caspase 8 and caspase 3. Moreover, the combination treatment of LPS and 3OC12-HSL amplified these expressions.

Conclusion

Treatment of 3OC12-HSL reduces endothelial cell viability, and this effect is synergistical with LPS. 3OC12-HSL induces apoptotic cell death, through the activation of the RIPK1 pathway, moreover, it is amplified in combination with LPS.
MO007ANALYSIS OF CALCIUM SIGNALING IN AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE (ADPKD)
Authors: Van Giel Dorien , Decuypere Jean-Paul , Mekahli Djalila , Vennekens Rudi
Published: May 2021

Background and Aims

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is an inheritable kidney disease characterized by the development of fluid-filled cysts in all nephron segments, leading to loss of renal function. Mutations in PKD1 or PKD2, which encode polycystin-1 and polycystin-2, are the most common cause of ADPKD. The molecular mechanisms underlying cystogenesis are poorly characterized but it is postulated that disturbed calcium homeostasis is a primary event in cystogenesis. The precise molecular players that cause this disturbance are still a poorly explored area, especially in relevant human cell types. We therefore aim to characterize the profile of calcium-coupled receptors and channels in a human renal epithelial cell model, to identify which receptors and channels are present and whether their function is affected in ADPKD.

Method

Human urine-derived conditionally immortalized proximal tubule epithelial cells (ciPTECs) of ADPKD patients and healthy controls were screened for calcium-coupled GPCRs, using a GPCR agonist library on Fura-2 loaded cell populations seeded in 96-well format using the Flexstation3 (Molecular Devices). Validation of specific hits was done using single-cell measurements with a fluorescence microscope and built-in perfusion system. The expression of TRP channels and STIM/Orai proteins was determined via qPCR.

Results

From a library of 418 GPCR agonists a selective amount of calcium-coupled GPCRs was found functionally active in ciPTECs. ciPTECs from both healthy controls and ADPKD patients were found to functionally express purinergic -, histamine -, serotonin and dopamine receptors. Through qPCR we found expression of various TRP channels, including TRPML1, TRPC1/3, TRPM3/4/7, TRPV4 and TRPA1, as well as high expression of STIM1/2 and Orai1/2/3.

Conclusion

We describe the first thorough characterization of molecular players involved in calcium signalling mechanisms in human renal epithelial cells, including the profile of calcium-coupled GPCRs and the expression of TRP channels and STIM/Orai proteins, of further interest to investigate disturbed calcium dynamics in ADPKD.
MO012ACE2 AND SARS-COV-2 INFECTION RISK: INSIGHTS FROM PATIENTS WITH TWO RARE GENETIC TUBULOPATHIES, GITELMAN‘S AND BARTTER’S SYNDROMES
Authors: Bertoldi Giovanni , Rigato Matteo , Sgarabotto Luca , Gianesello Lisa , Ravarotto Verdiana , Davis Paul A , Calò Lorenzo
Published: May 2021

Background and Aims

COVID-19 is spreading globally with Angiotensin Converting Enzyme (ACE)-2 serving as the entry point of SARS-CoV-2 virus. This raised concerns how ACE2 and Renin-Angiotensin (Ang)-System (RAS) are to be dealt with given their involvement in COVID-19’s morbidity and mortality. Specifically, increased ACE2 expression in response to treatment with ACE inhibitors (ACEi) and Ang II receptor blockers (ARBs) might theoretically increase COVID-19 risk by increasing SARS-CoV-2 binding sites. However, ACE2 is part of the protective counter-regulatory ACE2-Ang1-7-MasR axis, which opposes the classical ACE-AngII-AT1R regulatory axis. We used Gitelman’s and Bartter’s syndromes (GS/BS) patients, rare genetic tubulopathies, who have endogenously increased levels of ACE2, to provide more insight on these issues.

Method

128 genetically confirmed GS/BS patients, living in Lombardia, Emilia Romagna and Veneto, the Northern Italy hot spots for COVID-19, were surveyed via telephone survey regarding COVID-19.

Results

The survey found no COVID-19 infection and absence of COVID-19 symptoms in any patient. Comparison analysis with the prevalence of COVID-19 in those Regions [8.96% (95% CI 8.96-8.99% vs 0.00% (95% IC 0.00-3.62%)] showed statistical significance (p<0.01).

Conclusion

The results of the study contribute to suggest that increased ACE2 does not increase risk of COVID-19 and that ACEi and ARBs by blocking excessive AT1R-mediated Ang II activation, might favour the increase of ACE2-derived Ang 1-7. The GS/BS patients’ increased ACE2 and Ang 1-7 levels and their characteristic chronic metabolic alkalosis might suggest for SARS-COV-2 a mechanism similar to that of chloroquine/hydroxychloroquine effect altering ACE2 glycosylation which resulted, in previous studies, in SARS-COV binding inhibition and block/inhibition of viral entry. Studies from our laboratory are ongoing to explore in GS/BS ACE2 glycosylation.
MO029CLINICAL CHARACTERISTICS AND PATHOGENIC GENES OF CONGENITAL SOLITARY KIDNEY WITH REPRODUCTIVE SYSTEM MALFORMATION*
Authors: HU Rongrong , Xu Lubin , Chen Minting , Chen Na , Ma Tiantian , Ji Peili , Liu Zhenzhen , Zhu Lan , Chen Limeng
Published: May 2021

Background and Aims

Congenital solitary kidney – one category of congenital anomalies of the kidney and urinary tract (CAKUT) may combine with other system malformations such as reproductive, cardiac, skeletal system, and so on. Our study analysed the clinical characteristics among congenital solitary kidney patients and their reproductive system malformations. And further work about probable pathogenic genes was explored.

Method

The information of CAKUT patients who were indicated by Doppler ultrasound was collected. The clinical and imaging features including reproductive system abnormalities were retrospectively reviewed in patients with congenital solitary kidney. In patients with Mayer-Rokitansky- Küster-Hauser (MRKH) syndrome, a disorder of congenital agenesis of uterus and vagina, whole exome sequencing was performed. Rare variants in CAKUT-related genes were analysed. Trio analysis was conducted to identify de novo mutations.

Results

We identified 209 patients with congenital solitary kidney from July 20, 2017 to July 19, 2020 among 1160 CAKUT patients in Peking Union Medical College Hospital. There were 152 females. The average age of congenital solitary kidney patients was 35.26±18.42 years when they were diagnosed. 53.2% showed different degrees of proteinuria and hematuria. Serum creatinine elevating was proved 13.1% and 40% in women and men separately. Among 81 females who also had a gynecological ultrasound report, 88.9% combined with genital malformation, oblique vaginal septum syndrome 48.7%, the Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome 22.2%, malformed uterus 23.5%, vaginal atresia and other genital malformation 7%. Congenital heart disease, complete transposition of viscera, and scoliosis were also found in some patients with congenital solitary kidney. Furthermore, based on the whole exome data of 443 patients with MRKH syndrome, seven function-lost mutations were confirmed. And also two De nove mutations (NOTCH2 (NM_024408.3: c.703A>T(p.Thr235Ser), ESRRG (NM_001243512.1:c.-169-8delT)), one homozygous patients with parents heterozygous (NM_133433.3:c.8084C>T(p.Thr2695Me) were identified as possible pathogenic genes caused CAKUT.

Conclusion

We should be aware of reproductive system malformations in CAKUT patients. Whole exome sequencing may suggest common pathogenic genes between the two kinds of diseases.
MO042PREVALENCE OF FABRY DISEASE CAUSING VARIANTS IN THE UK BIOBANK
Authors: Gilchrist Mark , Casanova Francesco , Tyrrell Jessica , Fife Nicole , Young Katie , Oram Richard , Weedon Michael
Published: May 2021

Background and Aims

Fabry disease (FD) is an X-linked lysosomal storage disorder resulting from deficiency of the alpha-galactosidase A enzyme. This leads to the accumulation of globotriaosylceramide in multiple organ sites with prominent renal involvement, cardio and cerebrovascular disease, as well as neurological, dermatological, and gastrointestinal pathology in affected individuals. Clinical manifestations are highly heterogeneous with considerable variation depending on the specific variant and even within families with the same variant. Prevalence estimates from studies of case control cohorts range from 1 in 40,000 to 1 in 140,000. We sought to determine the prevalence of Fabry disease causing mutations in an unbiased adult population sample using the UK Biobank.

Method

UK Biobank comprises approximately 500,000 participants with extensive phenotyping and genetic data linked to clinical care records. Exome sequencing data from 200643 individuals from the UK Biobank were examined for variants in the GLA gene. Likely mutations were searched for in ClinVar and HGMD with subsequent review of supporting literature. We used ACMG guidelines to classify the pathogenicity of variants.

Results

Eighty one coding variants were identified in GLA. Nine of these variants were rare (<1 in 10,000 individuals) and either protein truncating variants or previously reported to cause Fabry’s disease. Thirty seven individuals, (14 males, 22 females, 1 missing data) carried one of these GLA variants. The most common variant was the cardiac predominant phenotype causing N215S variant (18/37). 6/37 had the R363C variant, 4/37 had R356Q. The R112H, R301Q, I198T variants were each identified in 2 individuals. There were single individuals with I232T, K240fs, and E66G variants.

Two male individuals (with I232T, K240fs variants) had HES codes compatible with a known diagnosis of Fabry disease. The remaining 12/14 males did not have evidence of an existing diagnosis of Fabry disease. No female participants had HES codes indicating an existing diagnosis of Fabry disease.

The overall prevalence of Fabry disease causing variants in the UK Biobank is 1 in 5,422. The N215 variant alone has a prevalence of 1 in 11,147.

Conclusion

Reported Fabry disease causing GLA variants are far more prevalent in an unselected population sample than would be expected from the reported prevalence of Fabry disease. This may be because some of the reported variants are not pathogenic, or are predominantly later-onset causing variants with reduced penetrance. The true prevalence of Fabry disease may be substantially higher than current estimates.
MO045THE APPLICATION OF A NGS KIDNEY PANEL REVEALED KEY CHALLENGES OF PKD1-2 ANALYSIS: INTERPRETATION OF MISSENSE VARIANTS, SIGNIFICANCE OF VARIANTS IN DUPLICATED REGIONS AND HIGH ALLELIC HETEROGENEITY
Authors: Blasio Giancarlo , Zacchia Miriam , Del Vecchio Blanco Francesca , Capolongo Giovanna , Perna Alessandra , Nigro Vincenzo , Capasso Giovambattista
Published: May 2021

Background and Aims

Genetic testing has changed the clinical management of inherited kidney diseases patients, improving prognosis, surveillance and therapy. On the other hand, it has put geneticists and clinicians in front of new challenges, as the heterogeneity of these disorders and the high number of variants, with no clear genotype-phenotype correlation.

Method

108 patients underwent genetic analysis through a kidney focused NGS panel, named Nephroplex, containing 119 genetic loci associated with inherited kidney disorders. The study aimed to addressed the genetic landscape of cystic individuals and to analyze PKD1 and PKD2 variants in non-cystic individuals.

Results

Following diagnostic criteria, patients were divided as cystic kidney diseases (n=36) and non-cystic kidney diseases (n=72). Among the group of cystic patients, a causative mutation was detected in 51% of cases. We found thirty-seven PKD1 and PKD2 variants in 26 out of 35 individuals. In particular, 12 variants were shown to be damaging and nine of that were reported in public database, as CLINVAR and Mayo Clinic databases. Among pathogenic variants, twelve were truncating and the remaining were missense variants. Of note, 7 out of 12 damaging PKD1 mutations were located in duplicated regions. Moreover, in three cystic patients, we found a (i) a frameshift hemizygote OFD1 mutation (ii) compound heterozygote PKHD1 variants and (iii) a frameshift MUC1 variant, framing the diagnosis of oro-facio-digital type 1, autosomal recessive polycystic kidney disease and autosomal dominant tubulointerstitial disease, respectively. Interestingly, we detected 28 PKD1-2 rare variants in 21 out of 75 adult non cystic patients (28%). The most were observed in PKD1 genes (82% vs 18% in PKD2). Eighteen of 28 variants were described in the literature as likely benign or as mutations of uncertain significance, while we found 10 novel variants. In silico analysis revealed as pathogenic a frameshift mutation located in exon 15. Of note, the great part of these variations reside into the duplicated PKD1 regions.

Conclusion

Our data showed that genetic analysis of ADPKD retains unique challenges, given the high degree of homology of PKD1 with his pseudogenes and the high allelic heterogeneity in non-cystic individuals.