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Article ; Online: Enzyme-activatable imaging probe reveals enhanced neutrophil elastase activity in tumors following photodynamic therapy.

Mitra, Soumya / Modi, Kshitij D / Foster, Thomas H

2013 Volume 18, Issue 10, Page(s) 101314

Abstract: We demonstrate the use of an enzyme-activatable fluorogenic probe, Neutrophil Elastase 680 FAST (NE680), for in vivo imaging of neutrophil elastase (NE) activity in tumors subjected to photodynamic therapy (PDT). NE protease activity was assayed in SCC ... ...

Abstract	We demonstrate the use of an enzyme-activatable fluorogenic probe, Neutrophil Elastase 680 FAST (NE680), for in vivo imaging of neutrophil elastase (NE) activity in tumors subjected to photodynamic therapy (PDT). NE protease activity was assayed in SCC VII and EMT6 tumors established in C3H and BALB/c mice, respectively. Four nanomoles of NE680 was injected intravenously immediately following PDT irradiation. 5 h following administration of NE680, whole-mouse fluorescence imaging was performed. At this time point, levels of NE680 fluorescence were at least threefold greater in irradiated versus unirradiated SCC VII and EMT6 tumors sensitized with Photofrin. To compare possible photosensitizer-specific differences in therapy-induced elastase activity, EMT6 tumors were also subjected to 2-(1-hexyloxyethyl)-2-devinyl pyropheophorbide-a (HPPH)-PDT. NE levels measured in HPPH-PDT-treated tumors were twofold higher than in unirradiated controls. Ex vivo labeling of host cells using fluorophore-conjugated antibodies and confocal imaging were used to visualize Gr1+ cells in Photofrin-PDT-treated EMT6 tumors. These data were compared with recently reported analysis of Gr1+ cell accumulation in EMT6 tumors subjected to HPPH-PDT. The population density of infiltrating Gr1+ cells in treated versus unirradiated drug-only control tumors suggests that the differential in NE680 fold enhancement observed in Photofrin versus HPPH treatment may be attributed to the significantly increased inflammatory response induced by Photofrin-PDT. The in vivo imaging of NE680, which is a fluorescent reporter of NE extracellular release caused by neutrophil activation, demonstrates that PDT results in increased NE levels in treated tumors, and the accumulation of the cleaved probe tracks qualitatively with the intratumor Gr1+ cell population.
MeSH term(s)	Animals ; Dihematoporphyrin Ether/pharmacology ; Female ; Fluorescent Dyes/analysis ; Fluorescent Dyes/chemistry ; Fluorescent Dyes/pharmacokinetics ; Leukocyte Elastase/analysis ; Leukocyte Elastase/metabolism ; Mammary Neoplasms, Experimental/drug therapy ; Mammary Neoplasms, Experimental/enzymology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Molecular Imaging/methods ; Optical Imaging/methods ; Photochemotherapy ; Photosensitizing Agents/pharmacology
Chemical Substances	Fluorescent Dyes ; Photosensitizing Agents ; Dihematoporphyrin Ether (97067-70-4) ; Leukocyte Elastase (EC 3.4.21.37)
Language	English
Publishing date	2013-07-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1309154-2
ISSN	1560-2281 ; 1083-3668
ISSN (online)	1560-2281
ISSN	1083-3668
DOI	10.1117/1.JBO.18.10.101314
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Article: In vivo bioluminescent imaging of mammary tumors using ivis spectrum

Lim, Ed / Modi, Kshitij D / Kim, JaeBeom

Journal of visualized experiments. 2009 Apr. 29, , no. 26

2009

Abstract: 4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system is valuable for the pre-clinical in vivo evaluation of putative antitumor compounds. The 4T1 cell ... ...

Abstract	4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system is valuable for the pre-clinical in vivo evaluation of putative antitumor compounds. The 4T1 cell line has been engineered to constitutively express the firefly luciferase gene (luc2). When mice carrying 4T1-luc2 tumors are injected with Luciferin the tumors emit a visual light signal that can be monitored using a sensitive optical imaging system like the IVIS Spectrum. The photon flux from the tumor is proportional to the number of light emitting cells and the signal can be measured to monitor tumor growth and development. IVIS is calibrated to enable absolute quantitation of the bioluminescent signal and longitudinal studies can be performed over many months and over several orders of signal magnitude without compromising the quantitative result. Tumor growth can be monitored for several days by bioluminescence before the tumor size becomes palpable or measurable by traditional physical means. This rapid monitoring can provide insight into early events in tumor development or lead to shorter experimental procedures. Tumor cell death and necrosis due to hypoxia or drug treatment is indicated early by a reduction in the bioluminescent signal. This cell death might not be accompanied by a reduction in tumor size as measured by physical means. The ability to see early events in tumor necrosis has significant impact on the selection and development of therapeutic agents. Quantitative imaging of tumor growth using IVIS provides precise quantitation and accelerates the experimental process to generate results.
Keywords	antineoplastic agents ; bioluminescence ; cell lines ; drug therapy ; genes ; growth and development ; hypoxia ; image analysis ; in vivo studies ; longitudinal studies ; luciferase ; luciferin ; mammary neoplasms (animal) ; mice ; models ; monitoring ; necrosis ; neoplasm cells ; photons
Language	English
Dates of publication	2009-0429
Size	p. e1210.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/1210
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: In vivo bioluminescent imaging of mammary tumors using IVIS spectrum.

Lim, Ed / Modi, Kshitij D / Kim, Jaebeom

Journal of visualized experiments : JoVE

2009 , Issue 26

Abstract	4T1 mouse mammary tumor cells can be implanted sub-cutaneously in nu/nu mice to form palpable tumors in 15 to 20 days. This xenograft tumor model system is valuable for the pre-clinical in vivo evaluation of putative antitumor compounds. The 4T1 cell line has been engineered to constitutively express the firefly luciferase gene (luc2). When mice carrying 4T1-luc2 tumors are injected with Luciferin the tumors emit a visual light signal that can be monitored using a sensitive optical imaging system like the IVIS Spectrum. The photon flux from the tumor is proportional to the number of light emitting cells and the signal can be measured to monitor tumor growth and development. IVIS is calibrated to enable absolute quantitation of the bioluminescent signal and longitudinal studies can be performed over many months and over several orders of signal magnitude without compromising the quantitative result. Tumor growth can be monitored for several days by bioluminescence before the tumor size becomes palpable or measurable by traditional physical means. This rapid monitoring can provide insight into early events in tumor development or lead to shorter experimental procedures. Tumor cell death and necrosis due to hypoxia or drug treatment is indicated early by a reduction in the bioluminescent signal. This cell death might not be accompanied by a reduction in tumor size as measured by physical means. The ability to see early events in tumor necrosis has significant impact on the selection and development of therapeutic agents. Quantitative imaging of tumor growth using IVIS provides precise quantitation and accelerates the experimental process to generate results.
MeSH term(s)	Animals ; Cell Line, Tumor ; Female ; Luciferases/biosynthesis ; Luciferases/genetics ; Luminescent Measurements/methods ; Mammary Neoplasms, Experimental/enzymology ; Mammary Neoplasms, Experimental/genetics ; Mammary Neoplasms, Experimental/pathology ; Mice ; Mice, Nude
Chemical Substances	Luciferases (EC 1.13.12.-)
Language	English
Publishing date	2009-04-29
Publishing country	United States
Document type	Journal Article ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/1210
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Monitoring tumor metastases and osteolytic lesions with bioluminescence and micro ct imaging

Lim, Ed / Modi, Kshitij / Christensen, Anna / Meganck, Jeff / Oldfield, Stephen / Zhang, Ning

Journal of visualized experiments. 2011 Apr. 14, , no. 50

2011

Abstract: Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells ... ...

Abstract	Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells following the delivery procedure including DLIT reconstruction to measure the tumor signal and its location. Development of metastatic lesions to the bone tissues triggers osteolytic activity and lesions to tibia and femur were evaluated longitudinally using micro CT. Imaging was performed using a Quantum FX micro CT system with fast imaging and low X-ray dose. The low radiation dose allows multiple imaging sessions to be performed with a cumulative X-ray dosage far below LD50. A mouse imaging shuttle device was used to sequentially image the mice with both IVIS Spectrum and Quantum FX achieving accurate animal positioning in both the bioluminescence and CT images. The optical and CT data sets were co-registered in 3-dimentions using the Living Image 4.1 software. This multi-mode approach allows close monitoring of tumor growth and development simultaneously with osteolytic activity.
Keywords	X-radiation ; bioluminescence ; computer software ; data collection ; femur ; growth and development ; image analysis ; lethal dose 50 ; metastasis ; mice ; monitoring ; neoplasm cells ; neoplasms ; tibia
Language	English
Dates of publication	2011-0414
Size	p. e2775.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/2775
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Monitoring tumor metastases and osteolytic lesions with bioluminescence and micro CT imaging.

Lim, Ed / Modi, Kshitij / Christensen, Anna / Meganck, Jeff / Oldfield, Stephen / Zhang, Ning

Journal of visualized experiments : JoVE

2011 , Issue 50

Abstract	Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells following the delivery procedure including DLIT reconstruction to measure the tumor signal and its location. Development of metastatic lesions to the bone tissues triggers osteolytic activity and lesions to tibia and femur were evaluated longitudinally using micro CT. Imaging was performed using a Quantum FX micro CT system with fast imaging and low X-ray dose. The low radiation dose allows multiple imaging sessions to be performed with a cumulative X-ray dosage far below LD50. A mouse imaging shuttle device was used to sequentially image the mice with both IVIS Spectrum and Quantum FX achieving accurate animal positioning in both the bioluminescence and CT images. The optical and CT data sets were co-registered in 3-dimentions using the Living Image 4.1 software. This multi-mode approach allows close monitoring of tumor growth and development simultaneously with osteolytic activity.
MeSH term(s)	Animals ; Bone Neoplasms/secondary ; Breast Neoplasms/pathology ; Cell Line, Tumor ; Female ; Humans ; Luminescent Measurements/methods ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Tomography, X-Ray Computed/methods ; Transplantation, Heterologous
Language	English
Publishing date	2011-04-14
Publishing country	United States
Document type	Journal Article ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/2775
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Noninvasive biophotonic imaging for monitoring of catheter-associated urinary tract infections and therapy in mice.

Kadurugamuwa, Jagath L / Modi, Kshitij / Yu, Jun / Francis, Kevin P / Purchio, Tony / Contag, Pamela R

Infection and immunity

2005 Volume 73, Issue 7, Page(s) 3878–3887

Abstract: Urinary tract infections (UTIs) are among the most common bacterial infections acquired by humans, particularly in catheterized patients. A major problem with catheterization is the formation of bacterial biofilms on catheter material and the risk of ... ...

Abstract	Urinary tract infections (UTIs) are among the most common bacterial infections acquired by humans, particularly in catheterized patients. A major problem with catheterization is the formation of bacterial biofilms on catheter material and the risk of developing persistent UTIs that are difficult to monitor and eradicate. To better understand the course of UTIs and allow more accurate studies of in vivo antibiotic efficacy, we developed a catheter-based biofilm infection model with mice, using bioluminescently engineered bacteria. Two important urinary tract pathogens, Pseudomonas aeruginosa and Proteus mirabilis, were made bioluminescent by stable insertion of a complete lux operon. Segments of catheter material (precolonized or postimplant infected) with either pathogen were placed transurethrally in the lumen of the bladder by using a metal stylet without surgical manipulation. The bioluminescent strains were sufficiently bright to be readily monitored from the outside of infected animals, using a low-light optical imaging system, including the ability to trace the ascending pattern of light-emitting bacteria through ureters to the kidneys. Placement of the catheter in the bladder not only resulted in the development of strong cystitis that persisted significantly longer than in mice challenged with bacterial suspensions alone but also required prolonged antibiotic treatment to reduce the level of infection. Treatment of infected mice for 4 days with ciprofloxacin at 30 mg/kg of body weight twice a day cured cystitis and renal infection in noncatheterized mice. Similarly, ciprofloxacin reduced the bacterial burden to undetectable levels in catheterized mice but did not inhibit rebound of the infection upon cessation of antibiotic therapy. This methodology easily allows spatial information to be monitored sequentially throughout the entire disease process, including ascending UTI, treatment efficacy, and relapse, all without exogenous sampling, which is not possible with conventional methods.
MeSH term(s)	Animals ; Diagnostic Imaging/methods ; Female ; Mice ; Microscopy, Electron, Scanning ; Photons ; Proteus Infections/diagnosis ; Proteus mirabilis/drug effects ; Proteus mirabilis/isolation & purification ; Pseudomonas Infections/diagnosis ; Pseudomonas aeruginosa/drug effects ; Pseudomonas aeruginosa/isolation & purification ; Urinary Catheterization/adverse effects ; Urinary Tract Infections/diagnosis
Language	English
Publishing date	2005-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.73.7.3878-3887.2005
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Article: Noninvasive Biophotonic Imaging for Monitoring of Catheter-Associated Urinary Tract Infections and Therapy in Mice

Kadurugamuwa, Jagath L / Modi, Kshitij / Yu, Jun / Francis, Kevin P / Purchio, Tony / Contag, Pamela R

Infection and immunity. 2005 July, v. 73, no. 7

2005

Abstract	Urinary tract infections (UTIs) are among the most common bacterial infections acquired by humans, particularly in catheterized patients. A major problem with catheterization is the formation of bacterial biofilms on catheter material and the risk of developing persistent UTIs that are difficult to monitor and eradicate. To better understand the course of UTIs and allow more accurate studies of in vivo antibiotic efficacy, we developed a catheter-based biofilm infection model with mice, using bioluminescently engineered bacteria. Two important urinary tract pathogens, Pseudomonas aeruginosa and Proteus mirabilis, were made bioluminescent by stable insertion of a complete lux operon. Segments of catheter material (precolonized or postimplant infected) with either pathogen were placed transurethrally in the lumen of the bladder by using a metal stylet without surgical manipulation. The bioluminescent strains were sufficiently bright to be readily monitored from the outside of infected animals, using a low-light optical imaging system, including the ability to trace the ascending pattern of light-emitting bacteria through ureters to the kidneys. Placement of the catheter in the bladder not only resulted in the development of strong cystitis that persisted significantly longer than in mice challenged with bacterial suspensions alone but also required prolonged antibiotic treatment to reduce the level of infection. Treatment of infected mice for 4 days with ciprofloxacin at 30 mg/kg of body weight twice a day cured cystitis and renal infection in noncatheterized mice. Similarly, ciprofloxacin reduced the bacterial burden to undetectable levels in catheterized mice but did not inhibit rebound of the infection upon cessation of antibiotic therapy. This methodology easily allows spatial information to be monitored sequentially throughout the entire disease process, including ascending UTI, treatment efficacy, and relapse, all without exogenous sampling, which is not possible with conventional methods.
Keywords	Proteus mirabilis ; Pseudomonas aeruginosa ; antibiotics ; bacteria ; bacterial infections ; biofilm ; bladder ; body weight ; ciprofloxacin ; cystitis ; genetically engineered microorganisms ; humans ; image analysis ; in vivo studies ; kidneys ; mice ; models ; monitoring ; operon ; pathogens ; patients ; relapse ; risk ; stylets ; therapeutics
Language	English
Size	p. 3878-3887.
Publishing place	American Society for Microbiology
Document type	Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
Database	NAL-Catalogue (AGRICOLA)

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Article: Reduction of Astrogliosis by Early Treatment of Pneumococcal Meningitis Measured by Simultaneous Imaging, In Vivo, of the Pathogen and Host Response

Kadurugamuwa, Jagath L / Modi, Kshitij / Coquoz, Olivier / Rice, Brad / Smith, Steven / Contag, Pamela R / Purchio, Tony

Infection and immunity. 2005 Dec., v. 73, no. 12

2005

Abstract: We developed a method for simultaneous in vivo biophotonic monitoring of pneumococcal meningitis and the accompanying neuronal injury in live transgenic mice. Streptococcus pneumoniae engineered for bioluminescence (lux) was used for direct visualization ...

Abstract	We developed a method for simultaneous in vivo biophotonic monitoring of pneumococcal meningitis and the accompanying neuronal injury in live transgenic mice. Streptococcus pneumoniae engineered for bioluminescence (lux) was used for direct visualization of disease progression and antibiotic treatment in a mouse model of meningitis. The host response was monitored in transgenic mice containing an inducible firefly luciferase (luc) reporter gene under transcriptional control of the mouse glial fibrillary acidic protein (GFAP) promoter. Based on the different spectra of light emission and substrate requirements for lux and luc, we were able to separately monitor the two reporters using a highly sensitive in vivo imaging system. The level of neuronal damage and recovery following antibiotic treatment was dependent on the time of treatment. This model has potential for simultaneous multiparameter monitoring and testing of therapies that target the pathogen or host response to prevent neuronal injury and recovery.
Keywords	Streptococcus pneumoniae ; animal models ; antibiotics ; bioluminescence ; disease course ; image analysis ; luciferase ; meningitis ; mice ; monitoring ; pathogens ; reporter genes ; transcription (genetics)
Language	English
Size	p. 7836-7843.
Publishing place	American Society for Microbiology
Document type	Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
Database	NAL-Catalogue (AGRICOLA)

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Article: Reduction of astrogliosis by early treatment of pneumococcal meningitis measured by simultaneous imaging, in vivo, of the pathogen and host response.

Kadurugamuwa, Jagath L / Modi, Kshitij / Coquoz, Olivier / Rice, Brad / Smith, Steven / Contag, Pamela R / Purchio, Tony

Infection and immunity

2005 Volume 73, Issue 12, Page(s) 7836–7843

Abstract	We developed a method for simultaneous in vivo biophotonic monitoring of pneumococcal meningitis and the accompanying neuronal injury in live transgenic mice. Streptococcus pneumoniae engineered for bioluminescence (lux) was used for direct visualization of disease progression and antibiotic treatment in a mouse model of meningitis. The host response was monitored in transgenic mice containing an inducible firefly luciferase (luc) reporter gene under transcriptional control of the mouse glial fibrillary acidic protein (GFAP) promoter. Based on the different spectra of light emission and substrate requirements for lux and luc, we were able to separately monitor the two reporters using a highly sensitive in vivo imaging system. The level of neuronal damage and recovery following antibiotic treatment was dependent on the time of treatment. This model has potential for simultaneous multiparameter monitoring and testing of therapies that target the pathogen or host response to prevent neuronal injury and recovery.
MeSH term(s)	Animals ; Anti-Bacterial Agents/therapeutic use ; Astrocytes/pathology ; Brain/microbiology ; Brain/pathology ; Disease Models, Animal ; Genes, Reporter ; Glial Fibrillary Acidic Protein/genetics ; Gliosis/drug therapy ; Gliosis/pathology ; Luciferases, Firefly/analysis ; Luciferases, Firefly/genetics ; Luminescent Agents/analysis ; Luminescent Measurements ; Meningitis, Pneumococcal/drug therapy ; Meningitis, Pneumococcal/microbiology ; Meningitis, Pneumococcal/pathology ; Mice ; Mice, Transgenic ; Promoter Regions, Genetic/genetics ; Streptococcus pneumoniae/genetics
Chemical Substances	Anti-Bacterial Agents ; Glial Fibrillary Acidic Protein ; Luminescent Agents ; Luciferases, Firefly (EC 1.13.12.7)
Language	English
Publishing date	2005-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.73.12.7836-7843.2005
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Article: Noninvasive monitoring of pneumococcal meningitis and evaluation of treatment efficacy in an experimental mouse model.

Kadurugamuwa, Jagath L / Modi, Kshitij / Yu, Jun / Francis, Kevin P / Orihuela, Carlos / Tuomanen, Elaine / Purchio, Anthony F / Contag, Pamela R

Molecular imaging

2005 Volume 4, Issue 2, Page(s) 137–142

Abstract: Noninvasive real-time in vivo bioluminescent imaging was used to assess the spread of Streptococcus pneumoniae throughout the spinal cord and brain during the acute stages of bacterial meningitis. A mouse model was established by lumbar (LP) or ... ...

Abstract	Noninvasive real-time in vivo bioluminescent imaging was used to assess the spread of Streptococcus pneumoniae throughout the spinal cord and brain during the acute stages of bacterial meningitis. A mouse model was established by lumbar (LP) or intracisternal (IC) injection of bioluminescent S. pneumoniae into the subarachnoid space. Bacteria replicated initially at the site of inoculation and spread progressively from the spinal cord to the brain or from the brain down to the cervical part of the spinal column and to the lower vertebral levels. After 24 hr, animals showed strong bioluminescent signals throughout the spinal canal, indicating acute meningitis of the intracranial and intraspinal meninges. A decline in bacterial cell viability, as judged by a reduction in the bioluminescent signal, was observed over time in animals treated with ceftriaxone, but not in untreated groups. Mice treated with the antibiotic survived infection, whereas all mice in untreated groups became moribund, first in the IC group then in the LP group. No untreated animal survived beyond 48 hr after induction of infection. Colony counts of infected cerebrospinal fluid (CSF) correlated positively with bioluminescent signals. This methodology is especially appealing because it allows detecting infected mice as early as 3 hr after inoculation, provide temporal, sequential, and spatial distribution of bacteria within the brain and spinal cord throughout the entire disease process and the rapid monitoring of treatment efficacy in a nondestructive manner. Moreover, it avoids the need to sacrifice the animals for CSF sampling and the potential manipulative damage that can occur with other conventional methods.
MeSH term(s)	Animals ; Diagnostic Imaging ; Disease Models, Animal ; Female ; Luminescent Measurements/methods ; Meningitis, Pneumococcal/diagnostic imaging ; Meningitis, Pneumococcal/drug therapy ; Meningitis, Pneumococcal/microbiology ; Mice ; Mice, Inbred BALB C ; Radiography ; Streptococcus pneumoniae/drug effects
Language	English
Publishing date	2005-04
Publishing country	England
Document type	Journal Article
ZDB-ID	2137435-1
ISSN	1536-0121 ; 1535-3508
ISSN (online)	1536-0121
ISSN	1535-3508
DOI	10.1177/13505068211024891
Database	MEDical Literature Analysis and Retrieval System OnLINE

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