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Article ; Online: Effects of oxygen-prebreathing on tissue nitrogenation in normobaric and hyperbaric conditions.

Ashworth, Edward Tom / Ogawa, Ryotaro / Vera, David Robert / Lindholm, Peter

2024 Volume 19, Issue 1, Page(s) e0294611

Abstract: Background: Breathing pure oxygen causes nitrogen washout from tissues, a method commonly deployed to prevent decompression sickness from hypobaric exposure. Theoretically prebreathing oxygen increases the capacity for nitrogen uptake and potentially ... ...

Abstract	Background: Breathing pure oxygen causes nitrogen washout from tissues, a method commonly deployed to prevent decompression sickness from hypobaric exposure. Theoretically prebreathing oxygen increases the capacity for nitrogen uptake and potentially limits supersaturation during dives of short duration. We aimed to use 13N2, a radioactive nitrogen isotope, to quantify tissue nitrogen following normobaric and hyperbaric exposures. Methods: Twenty Sprague Dawley rats were divided in 4 conditions; normobaric prebreathe, normobaric control, hyperbaric prebreathe, hyperbaric control. Prebreathed rats breathed oxygen for 1 h prior to the experiment whilst controls breathed air. Normobaric rats breathed air containing 13N2 at 100 kPa for 30 min, whereas hyperbaric rats breathed 13N2 at 700 kPa before being decompressed and sedated using air-isoflurane (without 13N2 for a few minutes). After euthanization, blood, brain, liver, femur and thigh muscle were analyzed by gamma counting. Results: At normobaria prebreathing oxygen resulted in higher absolute nitrogen counts in blood (p = .034), as well as higher normalized counts in both the liver and muscle (p = .034). However, following hyperbaric exposure no differences were observed between conditions for any organ (p>.344). Both bone and muscle showed higher normalized counts after hyperbaria compared to normobaria. Conclusions: Oxygen prebreathing caused nitrogen elimination in normobaria that led to a larger "sink" and uptake of 13N2. The lack of difference between conditions in hyperbaria could be due to the duration and depth of the dive mitigating the effect of prebreathing. In the hyperbaric conditions the lower counts were likely due to off-gassing of nitrogen during the sedation procedure, suggest a few minutes was enough to off-gas in rodents. The higher normalized counts under hyperbaria in bone and muscle likely relate to these tissues being slower to on and off-gas nitrogen. Future experiments could include shorter dives and euthanization while breathing 13N2 to prevent off-gassing.
MeSH term(s)	Rats ; Animals ; Rats, Sprague-Dawley ; Oxygen ; Gases ; Muscles ; Nitrogen
Chemical Substances	Oxygen (S88TT14065) ; Gases ; Nitrogen (N762921K75)
Language	English
Publishing date	2024-01-22
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0294611
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Article ; Online: A novel method for tracking nitrogen kinetics in vivo under hyperbaric conditions using radioactive nitrogen-13 gas and positron emission tomography.

Ashworth, Edward T / Ogawa, Ryotaro / Nguyen, Juliana / Afif, Chloe / Sá, Rui C / Butts Pauly, Kim / Vera, David R / Lindholm, Peter

Journal of applied physiology (Bethesda, Md. : 1985)

2024 Volume 136, Issue 4, Page(s) 949–953

Abstract: Decompression sickness (DCS) is caused by gaseous nitrogen dissolved in tissues forming bubbles during decompression. To date, no method exists to identify nitrogen within tissues, but with advances in positron-emission tomography (PET) technology, it ... ...

Abstract	Decompression sickness (DCS) is caused by gaseous nitrogen dissolved in tissues forming bubbles during decompression. To date, no method exists to identify nitrogen within tissues, but with advances in positron-emission tomography (PET) technology, it may be possible to track gaseous radionuclides into tissues. We aimed to develop a method to track nitrogen movement in vivo and under hyperbaric pressure that could then be used to further our understanding of DCS using nitrogen-13 (
MeSH term(s)	Humans ; Rats ; Animals ; Female ; Nitrogen ; Decompression Sickness/diagnostic imaging ; Diving/physiology ; Rats, Sprague-Dawley ; Decompression/adverse effects ; Gases ; Hyperbaric Oxygenation/methods ; Positron-Emission Tomography ; Mammals ; Nitrogen Radioisotopes
Chemical Substances	Nitrogen (N762921K75) ; Nitrogen-13 ; Gases ; Nitrogen Radioisotopes
Language	English
Publishing date	2024-02-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	219139-8
ISSN	1522-1601 ; 0021-8987 ; 0161-7567 ; 8750-7587
ISSN (online)	1522-1601
ISSN	0021-8987 ; 0161-7567 ; 8750-7587
DOI	10.1152/japplphysiol.00859.2023
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Article ; Online: Cranial-First Approach in Robot-Assisted Right Hemicolectomy.

Iwamoto, Masayoshi / Makutani, Yusuke / Ushijima, Hokuto / Ogawa, Ryotaro / Yoshioka, Yasumasa / Wada, Toshiaki / Ueda, Kazuki / Kawamura, Junichiro

Diseases of the colon and rectum

2024 Volume 67, Issue 6, Page(s) e358

MeSH term(s)	Humans ; Robotic Surgical Procedures/methods ; Colectomy/methods ; Colonic Neoplasms/surgery ; Colonic Neoplasms/pathology
Language	English
Publishing date	2024-02-13
Publishing country	United States
Document type	Journal Article ; Video-Audio Media ; Case Reports
ZDB-ID	212581-x
ISSN	1530-0358 ; 0012-3706
ISSN (online)	1530-0358
ISSN	0012-3706
DOI	10.1097/DCR.0000000000003114
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Article ; Online: Wells syndrome confined to the back of the hands mimicking contact dermatitis.

Amadearu, Abi / Kubota, Noriko / Ogawa, Ryotaro / Furuta, Junichi / Ishii, Yoshiyuki / Nomura, Toshifumi

The Journal of dermatology

2023

Language	English
Publishing date	2023-12-19
Publishing country	England
Document type	Letter
ZDB-ID	800103-0
ISSN	1346-8138 ; 0385-2407
ISSN (online)	1346-8138
ISSN	0385-2407
DOI	10.1111/1346-8138.17083
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Article ; Online: Fluorescent Guided Sentinel Lymph Mapping of the Oral Cavity with Fluorescent-Labeled Tilmanocept.

Guo, Theresa / Jang, Sophie S / Ogawa, Ryotaro / Davis, Morgan / Ashworth, Edward / Barback, Christopher V / Hall, David J / Vera, David R

The Laryngoscope

2023 Volume 134, Issue 3, Page(s) 1299–1307

Abstract: Objective: With the shift toward utilization of sentinel lymph node biopsy (SLNB) in oral cavity cancer, improved techniques for intraoperative sentinel node identification are needed. This study investigates the feasibility of fluorescently labeled ... ...

Abstract	Objective: With the shift toward utilization of sentinel lymph node biopsy (SLNB) in oral cavity cancer, improved techniques for intraoperative sentinel node identification are needed. This study investigates the feasibility of fluorescently labeled tilmanoscept in SLNB in an oral cancer rabbit model. Methods: An animal study was designed using 21 healthy male New Zealand rabbits. Gallium-68-labeled tilmanocept labeled with IRDye800CW was injected submucosally into the buccal mucosa (n = 6) or lateral tongue (n = 7) followed by PET imaging. One hour after injection, SLNB was performed using fluorescence imaging followed by a bilateral neck dissection and sampling of non-nodal surrounding tissue. All tissues were measured for radioactivity and fluorescence. In addition, eight rabbits were injected with delayed SLNB performed 48 h after injection. Results: Buccal injections all had ipsilateral SLN drainage and tongue injections exhibited 18.2% contralateral drainage. An average of 1.9 ± 1.0 SLN (range 1-5) were identified. In addition, an average of 16.9 ± 3.3 non-sentinel lymph nodes were removed per animal. SLNs had an average of 0.69 ± 0.60 percent-of-injected dose (%ID) compared with non-sentinel nodes with 0.012 ± 0.025 %ID and surrounding tissue with 0.0067 ± 0.015 %ID. There was 98.0% agreement between sentinel lymph nodes identified using fluorescence compared to radioactivity with Cohen's kappa coefficient of 0.879. In 48-h delayed SLNB, results were consistent with 97.8% agreement with radioactivity and Cohen's Kappa coefficient of 0.884. Fluorescence identified additional lymph nodes that were not identified by radioactivity, and with one false negative. Conclusion: Fluorescent-labeled Tc-99 m-tilmanocept represents a highly accurate adjunct to enhance SLNB for oral cavity cancer. Level of evidence: N/A Laryngoscope, 134:1299-1307, 2024.
MeSH term(s)	Male ; Animals ; Rabbits ; Sentinel Lymph Node Biopsy/methods ; Lymph Nodes/pathology ; Sentinel Lymph Node/diagnostic imaging ; Sentinel Lymph Node/surgery ; Sentinel Lymph Node/pathology ; Mouth Neoplasms/diagnostic imaging ; Mouth Neoplasms/surgery ; Mouth Neoplasms/pathology
Language	English
Publishing date	2023-09-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	80180-x
ISSN	1531-4995 ; 0023-852X
ISSN (online)	1531-4995
ISSN	0023-852X
DOI	10.1002/lary.31014
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Article ; Online: The Role of Tumor-Associated Neutrophils in Colorectal Cancer.

Mizuno, Rei / Kawada, Kenji / Itatani, Yoshiro / Ogawa, Ryotaro / Kiyasu, Yoshiyuki / Sakai, Yoshiharu

International journal of molecular sciences

2019 Volume 20, Issue 3

Abstract: Colorectal cancer (CRC) is one of the most common causes of cancer deaths worldwide and the number of CRC patients is increasing progressively. Despite the improvement of the surgical techniques and chemotherapy, we have not completely overcome this ... ...

Abstract	Colorectal cancer (CRC) is one of the most common causes of cancer deaths worldwide and the number of CRC patients is increasing progressively. Despite the improvement of the surgical techniques and chemotherapy, we have not completely overcome this disease yet due to the metastases. Therefore, understanding the mechanisms through which metastasis occurs is important for overcoming CRC. Normal host cells in the tumor microenvironment, such as macrophages and fibroblasts, have been reported to promote the growth of CRCs. Although neutrophils were originally considered to have defensive functions against tumor cells, it has been revealed that some populations of neutrophils, called as tumor-associated neutrophils (TANs), have tumor-supportive functions. The plasticity between tumor-suppressive and -supportive neutrophils are regulated by transforming growth factor (TGF)-β and Interferon-β signaling. Some studies have demonstrated that TANs promote the spread of cancer cells to distant organs. TANs contribute to the tumor invasion and angiogenesis through the production of matrix metalloproteinase-9 (MMP9), vascular endothelial growth factor (VEGF), and hepatocyte growth factor (HGF) in the primary and metastatic sites. Neutrophils also promotes tumor cell dissemination by capturing circulating tumor cells using neutrophil extracellular traps and promote their migration to distant sites. The neutrophil-to-lymphocyte ratio is a well-defined predictive marker for CRC patients. In this review, we highlight the molecular signaling between TANs and CRC cells and the possibility of TANs as a potential target for cancer therapy.
MeSH term(s)	Colorectal Neoplasms/immunology ; Colorectal Neoplasms/pathology ; Hepatocyte Growth Factor/metabolism ; Humans ; Interferon-beta/metabolism ; Matrix Metalloproteinase 9/metabolism ; Neoplasm Invasiveness ; Neutrophils/immunology ; Neutrophils/pathology ; Signal Transduction ; Transforming Growth Factor beta/metabolism ; Tumor Microenvironment ; Vascular Endothelial Growth Factor A/metabolism
Chemical Substances	HGF protein, human ; Transforming Growth Factor beta ; VEGFA protein, human ; Vascular Endothelial Growth Factor A ; Hepatocyte Growth Factor (67256-21-7) ; Interferon-beta (77238-31-4) ; MMP9 protein, human (EC 3.4.24.35) ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2019-01-27
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms20030529
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Article ; Online: Downregulation of osteoprotegerin in colorectal cancer cells promotes liver metastasis via activating tumor-associated macrophage.

Scientific reports

2023 Volume 13, Issue 1, Page(s) 22217

Abstract: Osteoprotegerin (OPG) is a secreted cytokine that functions as a decoy receptor for receptor activator of nuclear factor kappa-B (RANK) ligand (RANKL). Anti-RANKL treatment for bone metastasis has been widely accepted for solid tumors. However, the ... ...

Abstract	Osteoprotegerin (OPG) is a secreted cytokine that functions as a decoy receptor for receptor activator of nuclear factor kappa-B (RANK) ligand (RANKL). Anti-RANKL treatment for bone metastasis has been widely accepted for solid tumors. However, the mechanism of OPG-RANKL-RANK signaling in systemic colorectal cancer (CRC) metastasis remains unclear. In this study, we investigated the relevance and function of OPG expression in CRC liver metastasis. First, we performed in silico analysis using The Cancer Genome Atlas public database and found that lower OPG expression in CRC was associated with poor overall survival. Immunohistochemistry analyses using resected specimen from patients with CRC in our institute confirmed the result. Patient-matched primary CRC and liver metastases showed a significant downregulation of OPG expression in metastatic lesions. In CRC cell lines, OPG expression did not suppress cell proliferation and migration. However, OPG expression inhibited macrophage migration by suppressing the RANKL-RANK pathway. Moreover, in vivo mouse liver metastasis models showed that OPG expression in CRC cells suppressed liver metastases. In addition, treatment with an anti-RANKL neutralizing antibody also suppressed liver metastases. These results showed that downregulation of OPG expression in CRC cells promotes liver metastasis by activating tumor-associated macrophage, which can become a candidate for targeted therapy with anti-RANKL neutralizing antibody for CRC liver metastasis.
MeSH term(s)	Animals ; Humans ; Mice ; Antibodies, Neutralizing/metabolism ; Colorectal Neoplasms/genetics ; Down-Regulation ; Liver Neoplasms/genetics ; Osteoprotegerin/genetics ; Osteoprotegerin/metabolism ; RANK Ligand/genetics ; RANK Ligand/metabolism ; Receptor Activator of Nuclear Factor-kappa B/genetics ; Receptor Activator of Nuclear Factor-kappa B/metabolism ; Tumor-Associated Macrophages/metabolism
Chemical Substances	Antibodies, Neutralizing ; Osteoprotegerin ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Tnfrsf11b protein, mouse ; TNFRSF11B protein, human
Language	English
Publishing date	2023-12-14
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-49312-w
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Article ; Online: Neuronal Ndst1 depletion accelerates prion protein clearance and slows neurodegeneration in prion infection.

PLoS pathogens

2023 Volume 19, Issue 9, Page(s) e1011487

Abstract: Select prion diseases are characterized by widespread cerebral plaque-like deposits of amyloid fibrils enriched in heparan sulfate (HS), a abundant extracellular matrix component. HS facilitates fibril formation in vitro, yet how HS impacts fibrillar ... ...

Abstract	Select prion diseases are characterized by widespread cerebral plaque-like deposits of amyloid fibrils enriched in heparan sulfate (HS), a abundant extracellular matrix component. HS facilitates fibril formation in vitro, yet how HS impacts fibrillar plaque growth within the brain is unclear. Here we found that prion-bound HS chains are highly sulfated, and that the sulfation is essential for accelerating prion conversion in vitro. Using conditional knockout mice to deplete the HS sulfation enzyme, Ndst1 (N-deacetylase / N-sulfotransferase) from neurons or astrocytes, we investigated how reducing HS sulfation impacts survival and prion aggregate distribution during a prion infection. Neuronal Ndst1-depleted mice survived longer and showed fewer and smaller parenchymal plaques, shorter fibrils, and increased vascular amyloid, consistent with enhanced aggregate transit toward perivascular drainage channels. The prolonged survival was strain-dependent, affecting mice infected with extracellular, plaque-forming, but not membrane bound, prions. Live PET imaging revealed rapid clearance of recombinant prion protein monomers into the CSF of neuronal Ndst1- deficient mice, neuronal, further suggesting that HS sulfate groups hinder transit of extracellular prion protein monomers. Our results directly show how a host cofactor slows the spread of prion protein through the extracellular space and identify an enzyme to target to facilitate aggregate clearance.
MeSH term(s)	Animals ; Mice ; Heparitin Sulfate/metabolism ; Mice, Knockout ; Neurons/enzymology ; Prion Diseases/metabolism ; Prion Proteins/genetics ; Prions/metabolism ; Sulfotransferases/genetics ; Sulfotransferases/metabolism
Chemical Substances	Heparitin Sulfate (9050-30-0) ; Prion Proteins ; Prions ; heparitin sulfotransferase (EC 2.8.2.8) ; Sulfotransferases (EC 2.8.2.-)
Language	English
Publishing date	2023-09-25
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2205412-1
ISSN	1553-7374 ; 1553-7374
ISSN (online)	1553-7374
ISSN	1553-7374
DOI	10.1371/journal.ppat.1011487
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Article ; Online: Bone marrow-derived mesenchymal stem cells promote colorectal cancer progression via CCR5.

Nishikawa, Gen / Kawada, Kenji / Nakagawa, Jun / Toda, Kosuke / Ogawa, Ryotaro / Inamoto, Susumu / Mizuno, Rei / Itatani, Yoshiro / Sakai, Yoshiharu

Cell death & disease

2019 Volume 10, Issue 4, Page(s) 264

Abstract: Mesenchymal stem cells (MSCs) are recruited from BM to the stroma of developing tumors, where they serve as critical components of the tumor microenvironment by secreting growth factors, cytokines, and chemokines. The role of MSCs in colorectal cancer ( ... ...

Abstract	Mesenchymal stem cells (MSCs) are recruited from BM to the stroma of developing tumors, where they serve as critical components of the tumor microenvironment by secreting growth factors, cytokines, and chemokines. The role of MSCs in colorectal cancer (CRC) progression was controversial. In this study, we found that C-C chemokine receptor type 5 (CCR5) ligands (i.e., C-C motif chemokine ligand 3 (CCL3), CCL4, and CCL5) were highly produced from MSCs using a chemokine array screening with conditioned media from the cultured human MSCs. A relatively strong CCR5 expression could be detected within the cytoplasm of several CRC cell lines. Regarding the effect of MSC, we found that the xenografts in which CCR5-overexpressing HCT116 cells were inoculated into immunocompromised mice were highly promoted in vivo by a mixture with MSCs. Notably, the CCR5 inhibitor, maraviroc, significantly abolished the MSC-induced tumor growth in vivo. In human clinical specimens (n = 89), 20 cases (29%) were high for CCR5, whereas 69 cases (71%) were low. Statistical analyses indicated that CCR5 expression in primary CRC was associated with CRC patients' prognosis. Especially, stage III/IV patients with CCR5-high CRCs exhibited a significantly poorer prognosis than those with CCR5-low CRCs. Furthermore, we investigated the effects of preoperative serum CCR5 ligands on patients' prognosis (n = 114), and found that CRC patients with high serum levels of CCL3 and CCL4 exhibited a poorer prognosis compared to those with low levels of CCL3 and CCL4, while there was no association between CCL5 and prognosis. These results suggest that the inhibition of MSC-CRC interaction by a CCR5 inhibitor could provide the possibility of a novel therapeutic strategy for CRC, and that serum levels of CCL3 and CCL4 could be predictive biomarkers for the prognosis of CRC patients.
MeSH term(s)	Aged ; Animals ; Bone Marrow/metabolism ; Chemokine CCL3/blood ; Chemokine CCL3/metabolism ; Chemokine CCL4/blood ; Chemokine CCL4/metabolism ; Chemokine CCL5/blood ; Chemokine CCL5/metabolism ; Colonic Neoplasms/genetics ; Colonic Neoplasms/metabolism ; Colonic Neoplasms/pathology ; Colorectal Neoplasms/genetics ; Colorectal Neoplasms/metabolism ; Colorectal Neoplasms/pathology ; Culture Media, Conditioned ; Disease Progression ; Female ; HCT116 Cells ; HT29 Cells ; Humans ; Mesenchymal Stem Cells/metabolism ; Mice ; Mice, Nude ; Prognosis ; Receptors, CCR5/blood ; Receptors, CCR5/genetics ; Receptors, CCR5/metabolism ; Signal Transduction/genetics ; Transplantation, Heterologous
Chemical Substances	CCR5 protein, human ; CCR5 protein, mouse ; Chemokine CCL3 ; Chemokine CCL4 ; Chemokine CCL5 ; Culture Media, Conditioned ; Receptors, CCR5
Language	English
Publishing date	2019-03-19
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2541626-1
ISSN	2041-4889 ; 2041-4889
ISSN (online)	2041-4889
ISSN	2041-4889
DOI	10.1038/s41419-019-1508-2
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Article ; Online: Disruption of CCR1-mediated myeloid cell accumulation suppresses colorectal cancer progression in mice.

Kiyasu, Yoshiyuki / Kawada, Kenji / Hirai, Hideyo / Ogawa, Ryotaro / Hanada, Keita / Masui, Hideyuki / Nishikawa, Gen / Yamamoto, Takamasa / Mizuno, Rei / Itatani, Yoshiro / Kai, Masayuki / Taketo, Makoto Mark / Sakai, Yoshiharu

Cancer letters

2020 Volume 487, Page(s) 53–62

Abstract: Tumor-stromal interaction is implicated in tumor progression. Although CCR1 expression in myeloid cells could be associated with pro-tumor activity, it remains elusive whether disruption of CCR1-mediated myeloid cell accumulation can suppress tumor ... ...

Abstract	Tumor-stromal interaction is implicated in tumor progression. Although CCR1 expression in myeloid cells could be associated with pro-tumor activity, it remains elusive whether disruption of CCR1-mediated myeloid cell accumulation can suppress tumor progression. Here, we investigated the role of CCR1 depletion in myeloid cells in two syngeneic colorectal cancer mouse models: MC38, a transplanted tumor model and CMT93, a liver metastasis model. Both cells induced tumor accumulation of CCR1
MeSH term(s)	Animals ; Bone Marrow/metabolism ; Bone Marrow/pathology ; Cell Line, Tumor ; Cell- and Tissue-Based Therapy ; Colorectal Neoplasms/genetics ; Colorectal Neoplasms/pathology ; Colorectal Neoplasms/therapy ; Disease Models, Animal ; Disease Progression ; Gene Expression Regulation, Neoplastic/genetics ; Heterografts ; Humans ; Liver Neoplasms/genetics ; Liver Neoplasms/pathology ; Liver Neoplasms/secondary ; Liver Neoplasms/therapy ; Matrix Metalloproteinase 2/genetics ; Matrix Metalloproteinase 9/genetics ; Mice ; Myeloid Cells/metabolism ; Myeloid Cells/pathology ; Neoplasm Metastasis ; Nitric Oxide Synthase Type II/genetics ; Protein-Serine-Threonine Kinases/genetics ; Protein-Serine-Threonine Kinases/therapeutic use ; Vascular Endothelial Growth Factor A/genetics
Chemical Substances	Vascular Endothelial Growth Factor A ; Nitric Oxide Synthase Type II (EC 1.14.13.39) ; SNF1-related protein kinases (EC 2.7.1.-) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1) ; Matrix Metalloproteinase 2 (EC 3.4.24.24) ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2020-05-27
Publishing country	Ireland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	195674-7
ISSN	1872-7980 ; 0304-3835
ISSN (online)	1872-7980
ISSN	0304-3835
DOI	10.1016/j.canlet.2020.05.028
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