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Article ; Online: CaMKII inhibition in human primary and pluripotent stem cell-derived chondrocytes modulates effects of TGFβ and BMP through SMAD signaling.

Saitta, B / Elphingstone, J / Limfat, S / Shkhyan, R / Evseenko, D

2018 Volume 27, Issue 1, Page(s) 158–171

Abstract: Objective: Upregulation of calcium/calmodulin-dependent kinase II (CaMKII) is implicated in the pathogenesis of osteoarthritis (OA) and reactivation of articular cartilage hypertrophy. However, direct inhibition of CaMKII unexpectedly augmented symptoms ...

Abstract	Objective: Upregulation of calcium/calmodulin-dependent kinase II (CaMKII) is implicated in the pathogenesis of osteoarthritis (OA) and reactivation of articular cartilage hypertrophy. However, direct inhibition of CaMKII unexpectedly augmented symptoms of OA in animal models. The role of CaMKII in OA remains unclear and requires further investigation. Methods: Analysis of CaMKII expression was performed in normal human and OA articular chondrocytes, and signaling mechanisms were assessed in articular, fetal and Pluripotent Stem Cell (PSC)-derived human chondrocytes using pharmacological (KN93), peptide (AC3-I) and small interfering RNA (siRNA) inhibitors of CaMKII. Results: Expression levels of phospho-CaMKII (pCaMKII) were significantly and consistently increased in human OA specimens. BMP2/4 activated expression of pCaMKII as well as COLII and COLX in human adult articular chondrocytes, and also increased the levels and nuclear localization of SMADs1/5/8, while TGFβ1 showed minimal or no activation of the chondrogenic program in adult chondrocytes. Targeted blockade of CaMKII with specific siRNAs decreased levels of pSMADs, COLII, COLX and proteoglycans in normal and OA adult articular chondrocytes in the presence of both BMP4 and TGFβ1. Both human fetal and PSC-derived chondrocytes also demonstrated a decrease of chondrogenic differentiation in the presence of small molecule and peptide inhibitors of CaMKII. Furthermore, immunoprecipitation for SMADs1/5/8 or 2/3 followed by western blotting for pCaMKII showed direct interaction between SMADs and pCaMKII in primary chondrocytes. Conclusion: Current study demonstrates a direct role for CaMKII in TGF-β and BMP-mediated responses in primary and PSC-derived chondrocytes. These findings have direct implications for tissue engineering of cartilage tissue from stem cells and therapeutic management of OA.
MeSH term(s)	Aged ; Bone Morphogenetic Protein 4/pharmacology ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/antagonists & inhibitors ; Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism ; Cartilage, Articular/cytology ; Cartilage, Articular/metabolism ; Cell Differentiation ; Cells, Cultured ; Chondrocytes/metabolism ; Collagen Type II/metabolism ; Collagen Type X/metabolism ; Extracellular Matrix Proteins/pharmacology ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis/metabolism ; Osteoarthritis/pathology ; Phosphorylation/drug effects ; Pluripotent Stem Cells/metabolism ; Signal Transduction/drug effects ; Smad Proteins, Receptor-Regulated/metabolism ; Smad Proteins, Receptor-Regulated/physiology ; Transforming Growth Factor beta/pharmacology ; Up-Regulation
Chemical Substances	BMP4 protein, human ; Bone Morphogenetic Protein 4 ; Collagen Type II ; Collagen Type X ; Extracellular Matrix Proteins ; Smad Proteins, Receptor-Regulated ; Transforming Growth Factor beta ; betaIG-H3 protein (148710-76-3) ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 (EC 2.7.11.17)
Language	English
Publishing date	2018-09-08
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1167809-4
ISSN	1522-9653 ; 1063-4584
ISSN (online)	1522-9653
ISSN	1063-4584
DOI	10.1016/j.joca.2018.08.017
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Zs.A 3828: Show issues

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Article ; Online: Modelling the relative benefits of using the measles vaccine outside cold chain for outbreak response.

Azam, James M / Saitta, Barbara / Bonner, Kimberly / Ferrari, Matthew J / Pulliam, Juliet R C

Vaccine

2021 Volume 39, Issue 40, Page(s) 5845–5853

Abstract: Introduction: Rapid outbreak response vaccination is a strategy for measles control and elimination. Measles vaccines must be stored and transported within a specified temperature range, but this can present significant challenges when targeting remote ... ...

Abstract	Introduction: Rapid outbreak response vaccination is a strategy for measles control and elimination. Measles vaccines must be stored and transported within a specified temperature range, but this can present significant challenges when targeting remote populations. Measles vaccine licensure for delivery outside cold chain (OCC) could provide more vaccine transport/storage space without ice packs, and a solution to shorten response times. However, due to vaccine safety and wastage considerations, the OCC strategy will require other operational changes, potentially including the use of 1-dose (monodose) instead of 10-dose vials, requiring larger transport/storage equipment currently achieved with 10-dose vials. These trade-offs require quantitative comparisons of vaccine delivery options to evaluate their relative benefits. Methods: We developed a modelling framework combining elements of the vaccine supply chain - cold chain, vial, team, and transport equipment types - with a measles transmission dynamics model to compare vaccine delivery options. We compared 10 strategies resulting from combinations of the vaccine supply elements and grouped into three main classes: OCC, partial cold chain (PCC), and full cold chain (FCC). For each strategy, we explored a campaign with 20 teams sequentially targeting 5 locations with 100,000 individuals each. We characterised the time needed to freeze ice packs and complete the campaign (campaign duration), vaccination coverage, and cases averted, assuming a fixed pre-deployment delay before campaign commencement. We performed sensitivity analyses of the pre-deployment delay, population sizes, and two team allocation schemes. Results: The OCC, PCC, and FCC strategies achieve campaign durations of 50, 51, and 52 days, respectively. Nine of the ten strategies can achieve a vaccination coverage of 80%, and OCC averts the most cases. Discussion: The OCC strategy, therefore, presents improved operational and epidemiological outcomes relative to current practice and the other options considered.
MeSH term(s)	Disease Outbreaks/prevention & control ; Drug Storage ; Humans ; Measles/epidemiology ; Measles/prevention & control ; Measles Vaccine ; Refrigeration
Chemical Substances	Measles Vaccine
Language	English
Publishing date	2021-09-01
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2021.08.053
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Zs.A 1930: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 458: Show issues

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Article ; Online: Metformin improves relevant disease parameters in an autosomal dominant polycystic kidney disease mouse model.

Pastor-Soler, Núria M / Li, Hui / Pham, Jessica / Rivera, Daniel / Ho, Pei-Yin / Mancino, Valeria / Saitta, Biagio / Hallows, Kenneth R

American journal of physiology. Renal physiology

2021 Volume 322, Issue 1, Page(s) F27–F41

Abstract: Autosomal dominant polycystic kidney disease (ADPKD), caused by mutations in the polycystin 1 ( ...

Abstract	Autosomal dominant polycystic kidney disease (ADPKD), caused by mutations in the polycystin 1 (
MeSH term(s)	Animals ; Cell Proliferation/drug effects ; Disease Models, Animal ; Disease Progression ; Female ; Genetic Predisposition to Disease ; Glomerular Filtration Rate/drug effects ; Inflammation Mediators/metabolism ; Kidney/drug effects ; Kidney/metabolism ; Kidney/pathology ; Kidney/physiopathology ; Kidney Failure, Chronic/metabolism ; Kidney Failure, Chronic/pathology ; Kidney Failure, Chronic/physiopathology ; Kidney Failure, Chronic/prevention & control ; Male ; Metformin/pharmacology ; Mice, 129 Strain ; Mice, Inbred C57BL ; Mice, Transgenic ; Mutation ; Polycystic Kidney, Autosomal Dominant/drug therapy ; Polycystic Kidney, Autosomal Dominant/metabolism ; Polycystic Kidney, Autosomal Dominant/pathology ; Polycystic Kidney, Autosomal Dominant/physiopathology ; Renal Agents/pharmacology ; TRPP Cation Channels/genetics ; Time Factors ; Mice
Chemical Substances	Inflammation Mediators ; Renal Agents ; TRPP Cation Channels ; polycystic kidney disease 1 protein ; Metformin (9100L32L2N)
Language	English
Publishing date	2021-11-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	603837-2
ISSN	1522-1466 ; 0363-6127
ISSN (online)	1522-1466
ISSN	0363-6127
DOI	10.1152/ajprenal.00298.2021
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Article: Application of an external fixator vascular compressor (EFVC) in the critically injured trauma patient: a novel damage control technique.

Saitta, Bradley / Edgington, Jonathan / Hart, Theodore / Wilson, Kenneth / An, Gary / Daccarett, Miguel / Strelzow, Jason

European journal of orthopaedic surgery & traumatology : orthopedie traumatologie

2019 Volume 29, Issue 6, Page(s) 1337–1345

Abstract: Methods of controlling hemorrhage in penetrating abdominal injuries are varied, ranging from electrocautery, ligation, laparotomy sponge packing, angiography, hemostatic agents, and direct manual pressure. Unfortunately, traditional methods are sometimes ...

Abstract	Methods of controlling hemorrhage in penetrating abdominal injuries are varied, ranging from electrocautery, ligation, laparotomy sponge packing, angiography, hemostatic agents, and direct manual pressure. Unfortunately, traditional methods are sometimes unsuccessful due to the location or nature of the hemorrhage, and manual pressure cannot be held indefinitely. We describe a novel damage control technique for hemorrhage control in these situations, followed by three cases where an external fixator vascular compressor (EFVC) was used to hold continual pressure. Three patients are presented to a Level 1 trauma center following multiple ballistic injuries, all requiring emergent exploratory laparotomy. The first had a two-pin iliac crest EFVC placed during repeat exploratory laparotomy to control bleeding. The second patient had a supra-acetabular EFVC placed during initial exploratory laparotomy after emergent embolization failed to control bleeding from the L3 vertebral body. The third patient had a two-pin iliac crest EFVC placed at initial exploratory laparotomy due to uncontrollable bleeding from the sacral venous plexus and internal iliac veins. Of the three patients, two stabilized and survived, while one passed away due to multi-organ failure. We describe a novel damage control technique that may be a useful means of temporarily stemming intraabdominal bleeding that is otherwise recalcitrant to traditional hemostatic methods. Additionally, we provided a limited case series of patients who have undergone this technique to illustrate its utility and versatility. This technique is simple, fast, effective, and adaptable to a variety of circumstances that may be encountered in patients with intraabdominal bleeding recalcitrant to conventional hemorrhage control.
MeSH term(s)	Abdominal Injuries/complications ; Abdominal Injuries/surgery ; Adolescent ; Adult ; Equipment Design ; External Fixators ; Hemorrhage/etiology ; Hemorrhage/therapy ; Hemostasis, Surgical/instrumentation ; Hemostasis, Surgical/methods ; Humans ; Injury Severity Score ; Laparotomy/methods ; Male ; Multiple Trauma/complications ; Multiple Trauma/surgery ; Treatment Outcome ; Wounds, Penetrating/complications
Language	English
Publishing date	2019-04-16
Publishing country	France
Document type	Case Reports ; Journal Article ; Review
ZDB-ID	1231084-0
ISSN	1432-1068 ; 1633-8065 ; 0948-4817 ; 0940-3264
ISSN (online)	1432-1068
ISSN	1633-8065 ; 0948-4817 ; 0940-3264
DOI	10.1007/s00590-019-02439-x
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Zs.A 3479: Show issues

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Article ; Online: Association of Longitudinal Urinary Metabolic Biomarkers With ADPKD Severity and Response to Metformin in TAME-PKD Clinical Trial Participants.

Hallows, Kenneth R / Abebe, Kaleab Z / Li, Hui / Saitta, Biagio / Althouse, Andrew D / Bae, Kyongtae T / Lalama, Christina M / Miskulin, Dana C / Perrone, Ronald D / Seliger, Stephen L / Watnick, Terry J

Kidney international reports

2022 Volume 8, Issue 3, Page(s) 467–477

Abstract: Introduction: Dysregulated cellular metabolism contributes to autosomal dominant polycystic kidney disease (ADPKD) pathogenesis. The Trial of Administration of Metformin in Polycystic Kidney Disease (TAME-PKD) tested the effects of metformin treatment ... ...

Abstract	Introduction: Dysregulated cellular metabolism contributes to autosomal dominant polycystic kidney disease (ADPKD) pathogenesis. The Trial of Administration of Metformin in Polycystic Kidney Disease (TAME-PKD) tested the effects of metformin treatment over 2 years in adult ADPKD patients with mild-moderate disease severity. Metformin was found to be safe and tolerable with an insignificant trend toward reduced estimated glomerular filtration rate (eGFR) decline compared to placebo. Here we tested whether targeted urinary metabolic biomarkers measured in TAME-PKD participants correlated with disease progression, severity, and metformin treatment in cross-sectional and longitudinal analyses. Methods: Concentrations of total protein, targeted metabolites (lactate, pyruvate, and succinate), and glycolytic enzymes (pyruvate kinase-M2, lactate dehydrogenase-A, and pyruvate dehydrogenase kinase-1) were measured and normalized by creatinine or osmolality in urine specimens and compared with height-adjusted total kidney volume (htTKV) and eGFR at the different study timepoints. Results: In cross-sectional analyses utilizing placebo group data, urinary succinate normalized by creatinine negatively correlated with ln (htTKV), whereas protein excretion strongly positively correlated with ln (htTKV), and negatively correlated with eGFR. Significant time-varying negative associations occurred with eGFR and the lactate/pyruvate ratio and with urine protein normalized by osmolality, indicating correlations of these biomarkers with disease progression. In secondary analyses, urinary pyruvate normalized by osmolality was preserved in metformin-treated participants but declined in placebo over the 2-year study period with a significant between-arm difference, suggesting time-dependent urinary pyruvate changes may serve as a discriminator for metformin treatment effects in this study population. Conclusion: Proteinuria with enhanced glycolytic and reduced oxidative metabolic markers generally correlated with disease severity and risk of progression in the TAME-PKD study population.
Language	English
Publishing date	2022-12-05
Publishing country	United States
Document type	Journal Article
ISSN	2468-0249
ISSN (online)	2468-0249
DOI	10.1016/j.ekir.2022.11.019
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Article: Beneficial effects of bempedoic acid treatment in polycystic kidney disease cells and mice.

Hallows, Kenneth R / Li, Hui / Saitta, Biagio / Sepehr, Saman / Huang, Polly / Pham, Jessica / Wang, Jonathan / Mancino, Valeria / Chung, Eun Ji / Pinkosky, Stephen L / Pastor-Soler, Núria M

Frontiers in molecular biosciences

2022 Volume 9, Page(s) 1001941

Abstract: ADPKD has few therapeutic options. Tolvaptan slows disease but has side effects limiting its tolerability. Bempedoic acid (BA), an ATP citrate-lyase (ACLY) inhibitor FDA-approved for hypercholesterolemia, catalyzes a key step in fatty acid/sterol ... ...

Abstract	ADPKD has few therapeutic options. Tolvaptan slows disease but has side effects limiting its tolerability. Bempedoic acid (BA), an ATP citrate-lyase (ACLY) inhibitor FDA-approved for hypercholesterolemia, catalyzes a key step in fatty acid/sterol synthesis important for cell proliferation. BA is activated by very long-chain acyl-CoA synthetase (FATP2) expressed primarily in kidney and liver. BA also activates AMPK. We hypothesized that BA could be a novel ADPKD therapy by inhibiting cyst growth, proliferation, injury, and metabolic dysregulation
Language	English
Publishing date	2022-11-25
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2814330-9
ISSN	2296-889X
ISSN	2296-889X
DOI	10.3389/fmolb.2022.1001941
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Article ; Online: Association of Baseline Urinary Metabolic Biomarkers with ADPKD Severity in TAME-PKD Clinical Trial Participants.

Hallows, Kenneth R / Althouse, Andrew D / Li, Hui / Saitta, Biagio / Abebe, Kaleab Z / Bae, Kyongtae T / Miskulin, Dana C / Perrone, Ronald D / Seliger, Stephen L / Watnick, Terry J

Kidney360

2021 Volume 2, Issue 5, Page(s) 795–808

Abstract: Background: Recent work suggests that dysregulated cellular metabolism may play a key role in the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD). The TAME-PKD clinical trial is testing the safety, tolerability, and efficacy of ... ...

Abstract	Background: Recent work suggests that dysregulated cellular metabolism may play a key role in the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD). The TAME-PKD clinical trial is testing the safety, tolerability, and efficacy of metformin, a regulator of cell metabolism, in patients with ADPKD. This study investigates the cross-sectional association of urinary metabolic biomarkers with ADPKD severity among TAME-PKD trial participants at baseline. Methods: Concentrations of total protein, targeted metabolites (lactate, pyruvate, succinate, and cAMP), and key glycolytic enzymes (pyruvate kinase M2 [PKM2], lactate dehydrogenase A [LDHA], and pyruvate dehydrogenase kinase 1 [PDK1]) were measured by ELISA, enzymatic assays, and immunoblotting in baseline urine specimens of 95 TAME-PKD participants. These analytes, normalized by urinary creatinine or osmolality to estimate excretion, were correlated with patients' baseline height-adjusted total kidney volumes (htTKVs) by MRI and eGFR. Additional analyses were performed, adjusting for participants' age and sex, using multivariable linear regression. Results: Greater htTKV correlated with lower eGFR ( Conclusions: Proteinuria correlated with ADPKD severity, and urinary excretion of PKM2 and LDHA correlated with ADPKD severity at baseline in the TAME-PKD study population. These findings are the first to provide evidence in human urine samples that upregulated glycolytic flux is a feature of ADPKD severity. Future analysis may reveal if metformin treatment affects both disease progression and the various urinary metabolic biomarkers in patients throughout the study.
MeSH term(s)	Adult ; Biomarkers/metabolism ; Cross-Sectional Studies ; Glomerular Filtration Rate ; Humans ; Kidney/metabolism ; Polycystic Kidney, Autosomal Dominant/complications
Chemical Substances	Biomarkers
Language	English
Publishing date	2021-07-26
Publishing country	United States
Document type	Clinical Trial ; Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	2641-7650
ISSN (online)	2641-7650
DOI	10.34067/KID.0005962020
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Article ; Online: Lentiviral Gene Therapy for Bone Repair Using Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells.

Bougioukli, Sofia / Saitta, Biagio / Sugiyama, Osamu / Tang, Amy H / Elphingstone, Joseph / Evseenko, Denis / Lieberman, Jay R

Human gene therapy

2019 Volume 30, Issue 7, Page(s) 906–917

Abstract: Umbilical cord blood (UCB) has been increasingly explored as an alternative source of stem cells for use in regenerative medicine due to several advantages over other stem-cell sources, including the need for less stringent human leukocyte antigen ... ...

Abstract	Umbilical cord blood (UCB) has been increasingly explored as an alternative source of stem cells for use in regenerative medicine due to several advantages over other stem-cell sources, including the need for less stringent human leukocyte antigen matching. Combined with an osteoinductive signal, UCB-derived mesenchymal stem cells (MSCs) could revolutionize the treatment of challenging bone defects. This study aimed to develop an
MeSH term(s)	Animals ; Biomarkers ; Bone Morphogenetic Protein 2/genetics ; Bone Regeneration ; Cell Differentiation/genetics ; Cells, Cultured ; Disease Models, Animal ; Fetal Blood/cytology ; Gene Expression ; Gene Order ; Gene Transfer Techniques ; Genetic Therapy/methods ; Genetic Vectors/genetics ; Humans ; Lentivirus/genetics ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stem Cells/metabolism ; Mice ; Phenotype ; Transduction, Genetic ; Transgenes
Chemical Substances	Biomarkers ; Bone Morphogenetic Protein 2
Language	English
Publishing date	2019-04-22
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1028152-6
ISSN	1557-7422 ; 1043-0342
ISSN (online)	1557-7422
ISSN	1043-0342
DOI	10.1089/hum.2018.054
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Zs.A 2988: Show issues

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Article ; Online: Ensuring access to affordable, timely vaccines in emergencies.

Elder, Kate / Saitta, Barbara / Ducomble, Tanja / Alia, Miriam / Close, Ryan / Scheele, Suzanne / Erickson, Elise / Scourse, Rosalind / Kahn, Patricia / Elder, Greg

Bulletin of the World Health Organization

2019 Volume 97, Issue 12, Page(s) 851–853

MeSH term(s)	Developing Countries ; Drug Industry/organization & administration ; Emergencies ; Global Health ; Humans ; Pneumococcal Vaccines/economics ; Pneumococcal Vaccines/supply & distribution ; Relief Work ; Vaccines/economics ; Vaccines/supply & distribution ; World Health Organization
Chemical Substances	Pneumococcal Vaccines ; Vaccines
Language	English
Publishing date	2019-10-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	80213-x
ISSN	1564-0604 ; 0042-9686 ; 0366-4996 ; 0510-8659
ISSN (online)	1564-0604
ISSN	0042-9686 ; 0366-4996 ; 0510-8659
DOI	10.2471/BLT.18.228585
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Zs.A 215: Show issues

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Article: Ex vivo kidney slice preparations as a model system to study signaling cascades in kidney epithelial cells.

Saitta, Biagio / Jalili, Michael F / Zohoorkari, Hamidreza / Rao, Renee / Hallows, Kenneth R / Baty, Catherine J / Pastor-Soler, Nuria M

Methods in cell biology

2019 Volume 153, Page(s) 185–203

Abstract: Several model systems have been used to study signaling cascades in kidney epithelial cells, including kidney histology after systemic treatments, ex vivo isolated tubule perfusion, epithelial cell lines in culture, kidney micropuncture, and ex vivo ... ...

Abstract	Several model systems have been used to study signaling cascades in kidney epithelial cells, including kidney histology after systemic treatments, ex vivo isolated tubule perfusion, epithelial cell lines in culture, kidney micropuncture, and ex vivo kidney slices. We and others have found the ex vivo kidney slice method useful to study the signaling cascades involved in the regulation of kidney transport proteins. In this chapter we describe our adaptations to this classic method for the study of the regulation of kinases and endocytosis in rodent kidney epithelial cells. Briefly, slices are obtained by sectioning of freshly harvested rat or mouse kidneys using a Stadie-Riggs tissue slicer. Alternatively, a vibratome can be used to obtain slices at a more consistent and finer thickness. The harvested kidney and kidney slices are kept viable in either cell culture media or in buffers that mimic physiological conditions equilibrated with 5% CO
MeSH term(s)	Animals ; Biological Assay/instrumentation ; Biological Assay/methods ; Epithelial Cells/metabolism ; Histocytological Preparation Techniques/instrumentation ; Histocytological Preparation Techniques/methods ; Kidney/cytology ; Kidney/metabolism ; Mice ; Microscopy, Confocal/instrumentation ; Microscopy, Confocal/methods ; Rats ; Signal Transduction
Language	English
Publishing date	2019-06-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	0091-679X
ISSN	0091-679X
DOI	10.1016/bs.mcb.2019.04.017
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