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Article ; Online: The muscarinic M(4) receptor is the functionally predominant subtype in rat and mouse striatum as demonstrated using [(35)S] GTPγS binding.

Chapman, Kathryn L / Vaswani, Dina / Hendry, Nicola / Langmead, Christopher J / Kew, James N C / Watson, Jeannette M

2011 Volume 652, Issue 1-3, Page(s) 1–6

Abstract: We have used selective muscarinic receptor antagonists and M(2) and M(4) receptor knockout (KO ... of muscarinic acetylcholine receptors expressed are M(1) receptors. Radioligand binding studies suggest that the remaining ... muscarinic acetylcholine receptor population is largely M(4) with small levels of M(2). In agreement, carbachol-induced GTPγS ...

Abstract	We have used selective muscarinic receptor antagonists and M(2) and M(4) receptor knockout (KO) mouse tissue to define the functional muscarinic acetylcholine receptor populations in rodent striatum. [(3)H] NMS binding studies in rat and mouse striatum demonstrated that approximately 30% of muscarinic acetylcholine receptors expressed are M(1) receptors. Radioligand binding studies suggest that the remaining muscarinic acetylcholine receptor population is largely M(4) with small levels of M(2). In agreement, carbachol-induced GTPγS binding studies in M(2) and M(4) receptor KO mouse striatum implicated the M(4) receptor as the predominant functional receptor subtype. Based on these data we have developed a novel, native tissue M(4) receptor [(35)S] GTPγS binding assay. Pharmacological assessment of M(4) receptor agonist and positive 3modulators revealed clear differences in the potencies observed in a human recombinant CHO-M(4) receptor [(35)S] GTPγS binding assay as compared to the native tissue [(35)S] GTPγS binding assay. These differences are believed to reflect differences in receptor reserve between the assay systems as well as differences in compound pharmacology (relative contribution of compound affinity and efficacy to observed potency). These studies have demonstrated the importance of understanding the pharmacology of test compounds in a native environment when predicting in vivo response.
MeSH term(s)	Animals ; Cell Membrane/metabolism ; Corpus Striatum/metabolism ; Cricetinae ; Guanosine 5'-O-(3-Thiotriphosphate)/metabolism ; Humans ; Male ; Mice ; Mice, Knockout ; Muscarinic Antagonists/pharmacology ; Protein Binding/drug effects ; Radioligand Assay ; Rats ; Receptor, Muscarinic M4/agonists ; Receptor, Muscarinic M4/metabolism ; Receptors, Muscarinic/metabolism ; Sulfur Radioisotopes
Chemical Substances	Muscarinic Antagonists ; Receptor, Muscarinic M4 ; Receptors, Muscarinic ; Sulfur Radioisotopes ; Guanosine 5'-O-(3-Thiotriphosphate) (37589-80-3)
Language	English
Publishing date	2011-02-10
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	80121-5
ISSN	1879-0712 ; 0014-2999
ISSN (online)	1879-0712
ISSN	0014-2999
DOI	10.1016/j.ejphar.2010.10.079
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Article ; Online: IRAK-M regulates chromatin remodeling in lung macrophages during experimental sepsis.

Lyn-Kew, Kenneth / Rich, Eric / Zeng, Xianying / Wen, Haitao / Kunkel, Steven L / Newstead, Michael W / Bhan, Urvashi / Standiford, Theodore J

PloS one

2010 Volume 5, Issue 6, Page(s) e11145

Abstract: ... correlated with induction of IRAK-M mRNA and protein, which occurred in a MyD88-dependent fashion. PM ... isolated from IRAK-M(-/-) mice were largely refractory to CLP-induced impairment in cytokine expression ... gene expression in lung macrophages, and IRAK-M appears to be a critical mediator of this response. ...

Abstract	Sepsis results in a profound state of immunosuppression, which is temporally associated with impaired leukocyte function. The mechanism of leukocyte reprogramming in sepsis is incompletely understood. In this study, we explored mechanisms contributing to dysregulated inflammatory cytokine expression by pulmonary macrophages during experimental sepsis. Pulmonary macrophages (PM) recovered from the lungs of mice undergoing cecal ligation and puncture (CLP) display transiently reduced expression of some, but not all innate genes in response to LPS. Impaired expression of TNF-alpha and iNOS was associated with reduced acetylation and methylation of specific histones (AcH4 and H3K4me3) and reduced binding of RNA polymerase II to the promoters of these genes. Transient impairment in LPS-induced cytokine responses in septic PM temporally correlated with induction of IRAK-M mRNA and protein, which occurred in a MyD88-dependent fashion. PM isolated from IRAK-M(-/-) mice were largely refractory to CLP-induced impairment in cytokine expression, chromatin remodeling, recruitment of RNA polymerase II, and induction of histone deacetylase-2 observed during sepsis. Our findings indicate that systemic sepsis induces epigenetic silencing of cytokine gene expression in lung macrophages, and IRAK-M appears to be a critical mediator of this response.
MeSH term(s)	Animals ; Base Sequence ; Chromatin Assembly and Disassembly/physiology ; Chromatin Immunoprecipitation ; DNA Primers ; Interleukin-1 Receptor-Associated Kinases/genetics ; Interleukin-1 Receptor-Associated Kinases/physiology ; Lung/pathology ; Macrophages/pathology ; Mice ; Mice, Knockout ; Microscopy, Fluorescence ; Reverse Transcriptase Polymerase Chain Reaction ; Sepsis/pathology ; Sepsis/physiopathology
Chemical Substances	DNA Primers ; Interleukin-1 Receptor-Associated Kinases (EC 2.7.11.1) ; Irak3 protein, mouse (EC 2.7.11.1)
Language	English
Publishing date	2010-06-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0011145
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Article: The muscarinic M₄ receptor is the functionally predominant subtype in rat and mouse striatum as demonstrated using [³⁵S] GTPγS binding

Chapman, Kathryn L / Vaswani, Dina / Hendry, Nicola / Langmead, Christopher J / Kew, James N.C / Watson, Jeannette M

European journal of pharmacology. 2011 Feb. 10, v. 652, no. 1-3

2011

Abstract: We have used selective muscarinic receptor antagonists and M₂ and M₄ receptor knockout (KO) mouse ... of muscarinic acetylcholine receptors expressed are M₁ receptors. Radioligand binding studies suggest that the remaining ... muscarinic acetylcholine receptor population is largely M₄ with small levels of M₂. In agreement, carbachol-induced GTPγS binding ...

Abstract	We have used selective muscarinic receptor antagonists and M₂ and M₄ receptor knockout (KO) mouse tissue to define the functional muscarinic acetylcholine receptor populations in rodent striatum. [³H] NMS binding studies in rat and mouse striatum demonstrated that approximately 30% of muscarinic acetylcholine receptors expressed are M₁ receptors. Radioligand binding studies suggest that the remaining muscarinic acetylcholine receptor population is largely M₄ with small levels of M₂. In agreement, carbachol-induced GTPγS binding studies in M₂ and M₄ receptor KO mouse striatum implicated the M₄ receptor as the predominant functional receptor subtype. Based on these data we have developed a novel, native tissue M₄ receptor [³⁵S] GTPγS binding assay. Pharmacological assessment of M₄ receptor agonist and positive 3modulators revealed clear differences in the potencies observed in a human recombinant CHO-M₄ receptor [³⁵S] GTPγS binding assay as compared to the native tissue [³⁵S] GTPγS binding assay. These differences are believed to reflect differences in receptor reserve between the assay systems as well as differences in compound pharmacology (relative contribution of compound affinity and efficacy to observed potency). These studies have demonstrated the importance of understanding the pharmacology of test compounds in a native environment when predicting in vivo response.
Keywords	acetylcholine ; agonists ; antagonists ; cholinergic receptors ; humans ; mice ; prediction ; rats
Language	English
Dates of publication	2011-0210
Size	p. 1-6.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	80121-5
ISSN	1879-0712 ; 0014-2999
ISSN (online)	1879-0712
ISSN	0014-2999
DOI	10.1016/j.ejphar.2010.10.079
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Article ; Online: IRAK-M regulates chromatin remodeling in lung macrophages during experimental sepsis.

Kenneth Lyn-Kew / Eric Rich / Xianying Zeng / Haitao Wen / Steven L Kunkel / Michael W Newstead / Urvashi Bhan / Theodore J Standiford

PLoS ONE, Vol 5, Iss 6, p e

2010 Volume 11145

Abstract	Sepsis results in a profound state of immunosuppression, which is temporally associated with impaired leukocyte function. The mechanism of leukocyte reprogramming in sepsis is incompletely understood. In this study, we explored mechanisms contributing to dysregulated inflammatory cytokine expression by pulmonary macrophages during experimental sepsis. Pulmonary macrophages (PM) recovered from the lungs of mice undergoing cecal ligation and puncture (CLP) display transiently reduced expression of some, but not all innate genes in response to LPS. Impaired expression of TNF-alpha and iNOS was associated with reduced acetylation and methylation of specific histones (AcH4 and H3K4me3) and reduced binding of RNA polymerase II to the promoters of these genes. Transient impairment in LPS-induced cytokine responses in septic PM temporally correlated with induction of IRAK-M mRNA and protein, which occurred in a MyD88-dependent fashion. PM isolated from IRAK-M(-/-) mice were largely refractory to CLP-induced impairment in cytokine expression, chromatin remodeling, recruitment of RNA polymerase II, and induction of histone deacetylase-2 observed during sepsis. Our findings indicate that systemic sepsis induces epigenetic silencing of cytokine gene expression in lung macrophages, and IRAK-M appears to be a critical mediator of this response.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2010-06-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Dynamics of tuberculosis infection in various populations during the 19th and 20th century: The impact of conservative and pharmaceutical treatments.

Holloway-Kew, K L / Henneberg, M

Tuberculosis (Edinburgh, Scotland)

2023 Volume 143S, Page(s) 102389

Abstract: Humans and Mycobacterium tuberculosis have co-evolved together for thousands of years. Many individuals are infected with the bacterium, but few show signs and symptoms of tuberculosis (TB). Pharmacotherapy to treat those who develop disease is useful, ... ...

Abstract	Humans and Mycobacterium tuberculosis have co-evolved together for thousands of years. Many individuals are infected with the bacterium, but few show signs and symptoms of tuberculosis (TB). Pharmacotherapy to treat those who develop disease is useful, but drug resistance and non-adherence significantly impact the efficacy of these treatments. Prior to the introduction of antibiotic therapies, public health strategies were used to reduce TB mortality. This work shows how these strategies were able to reduce TB mortality in 19th and 20th century populations, compared with antibiotic treatments. Previously published mortality data from historical records for several populations (Switzerland, Germany, England and Wales, Scotland, USA, Japan, Brazil and South Africa) were used. Curvilinear regression was used to examine the reduction in mortality before and after the introduction of antibiotic treatments (1946). A strong decline in TB mortality was already occurring in Switzerland, Germany, England and Wales, Scotland and the USA prior to the introduction of antibiotic treatment. This occurred following many public health interventions including improved sanitation, compulsory reporting of TB cases, diagnostic techniques and sanatoria treatments. Following the introduction of antibiotics, mortality rates declined further, however, this had a smaller effect than the previously employed strategies. In Japan, Brazil and South Africa, reductions in mortality rates were largely driven by antibiotic treatments that caused rapid decline of mortality, with a smaller contribution from public health strategies. For the development of active disease, immune status is important. Individuals infected with the bacterium are more likely to develop signs and symptoms if their immune function is reduced. Effective strategies against TB can therefore include enhancing immune function of the population by improving nutrition, as well as reducing transmission by improving living conditions and public health. This has been effective in the past. Improving immunity may be an important strategy against drug resistant TB.
MeSH term(s)	Humans ; Mycobacterium tuberculosis ; Tuberculosis/diagnosis ; Tuberculosis/drug therapy ; Tuberculosis/epidemiology ; Antitubercular Agents/therapeutic use ; Antitubercular Agents/pharmacology ; Tuberculosis, Multidrug-Resistant/diagnosis ; Tuberculosis, Multidrug-Resistant/drug therapy ; Tuberculosis, Multidrug-Resistant/epidemiology ; Latent Tuberculosis/drug therapy ; Pharmaceutical Preparations
Chemical Substances	Antitubercular Agents ; Pharmaceutical Preparations
Language	English
Publishing date	2023-11-25
Publishing country	Scotland
Document type	Journal Article ; Review
ZDB-ID	2046804-0
ISSN	1873-281X ; 1472-9792
ISSN (online)	1873-281X
ISSN	1472-9792
DOI	10.1016/j.tube.2023.102389
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Article ; Online: 193 nm Ultraviolet Photodissociation for the Characterization of Singly Charged Proteoforms Generated by MALDI.

Zemaitis, Kevin J / Zhou, Mowei / Kew, William / Paša-Tolić, Ljiljana

Journal of the American Society for Mass Spectrometry

2023 Volume 34, Issue 2, Page(s) 328–332

Abstract: MALDI imaging allows for the near-cellular profiling of proteoforms directly from microbial, plant, and mammalian samples. Despite detecting hundreds of proteoforms, identification of unknowns with only intact mass information remains a distinct ... ...

Abstract	MALDI imaging allows for the near-cellular profiling of proteoforms directly from microbial, plant, and mammalian samples. Despite detecting hundreds of proteoforms, identification of unknowns with only intact mass information remains a distinct challenge, even with high mass resolving power and mass accuracy. To this end, many supplementary methods have been used to create experimental databases for accurate mass matching, including bulk or spatially resolved bottom-up and/or top-down proteomics. Herein, we describe the application of 193 nm ultraviolet photodissociation (UVPD) for fragmentation of quadrupole isolated singly charged ubiquitin (
MeSH term(s)	Animals ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods ; Tandem Mass Spectrometry/methods ; Proteins/analysis ; Ubiquitin ; Proteomics/methods ; Ultraviolet Rays ; Mammals
Chemical Substances	Proteins ; Ubiquitin
Language	English
Publishing date	2023-01-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	1073671-2
ISSN	1879-1123 ; 1044-0305
ISSN (online)	1879-1123
ISSN	1044-0305
DOI	10.1021/jasms.2c00302
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Article ; Online: Primary fungal laryngitis mimicking recurrent laryngeal carcinoma.

Thomas, Reuben Abraham / Kew, Thean Yean / Mat Baki, Marina

BMJ case reports

2022 Volume 15, Issue 2

Abstract: A 79-year-old smoker with a background history of a treated glottic carcinoma and chronic obstructive pulmonary disease presented with progressive hoarseness, symptoms of aspiration and shortness of breath for 6 months. Examination revealed an ulcero- ... ...

Abstract	A 79-year-old smoker with a background history of a treated glottic carcinoma and chronic obstructive pulmonary disease presented with progressive hoarseness, symptoms of aspiration and shortness of breath for 6 months. Examination revealed an ulcero-fungating mass over the posterior commissure of the larynx. A tracheostomy, direct laryngoscopy and biopsy of the mass was performed to secure his airway and to exclude recurrent glottic carcinoma. Reassuringly, a histopathological examination of the mass revealed numerous fungal yeast bodies. He was then treated with itraconazole for 4 weeks and was followed up as and outpatient with complete resolution and no recurrence of the disease.
MeSH term(s)	Aged ; Carcinoma ; Humans ; Laryngitis/diagnosis ; Laryngitis/drug therapy ; Laryngoscopy ; Larynx ; Male ; Neoplasm Recurrence, Local
Language	English
Publishing date	2022-02-09
Publishing country	England
Document type	Case Reports ; Journal Article
ISSN	1757-790X
ISSN (online)	1757-790X
DOI	10.1136/bcr-2021-245678
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Article ; Online: Natural Abundance Isotope Ratio Measurements of Organic Molecules Using 21 T FTICR MS

Kew, William / Boiteau, Rene M. / Eiler, John M. / Paša-Tolić, Ljiljana / Moran, James J.

Analytical Chemistry. 2023 Nov. 15, v. 95, no. 47 p.17203-17211

2023

Abstract: Subtle variations in stable isotope ratios at natural abundance are challenging to measure but can yield critical insights into biological, physical, and geochemical processes. Well-established methods, particularly multicollector, gas-source, or plasma ... ...

Abstract	Subtle variations in stable isotope ratios at natural abundance are challenging to measure but can yield critical insights into biological, physical, and geochemical processes. Well-established methods, particularly multicollector, gas-source, or plasma isotope ratio mass spectrometry, are the gold standard for stable isotope measurement, but inherent limitations in these approaches make them ill-suited to determining site-specific and multiply substituted isotopic abundances of all but a few compounds or to characterizing larger intact molecules. Fourier transform mass spectrometry, namely, Orbitrap mass spectrometry, has recently demonstrated the ability to measure natural abundance isotope ratios with chemically informative accuracy and precision. Here, we report the first use of Fourier transform ion cyclotron resonance mass spectrometry for the accurate (<1‰) and precise (<1‰ standard error) simultaneous determination of δ¹³C and δ¹⁵N in caffeine isotopologues and provide a discussion of the critical instrumental parameters necessary to make such measurements. We further report the ability to make these measurements with online liquid chromatography, expanding the ability of this technique to explore mixtures in the future.
Keywords	analytical chemistry ; caffeine ; liquid chromatography ; mass spectrometry ; stable isotopes
Language	English
Dates of publication	2023-1115
Size	p. 17203-17211.
Publishing place	American Chemical Society
Document type	Article ; Online
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.3c01816
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Article ; Online: Natural Abundance Isotope Ratio Measurements of Organic Molecules Using 21 T FTICR MS.

Kew, William / Boiteau, Rene M / Eiler, John M / Paša-Tolić, Ljiljana / Moran, James J

Analytical chemistry

2023 Volume 95, Issue 47, Page(s) 17203–17211

Abstract	Subtle variations in stable isotope ratios at natural abundance are challenging to measure but can yield critical insights into biological, physical, and geochemical processes. Well-established methods, particularly multicollector, gas-source, or plasma isotope ratio mass spectrometry, are the gold standard for stable isotope measurement, but inherent limitations in these approaches make them ill-suited to determining site-specific and multiply substituted isotopic abundances of all but a few compounds or to characterizing larger intact molecules. Fourier transform mass spectrometry, namely, Orbitrap mass spectrometry, has recently demonstrated the ability to measure natural abundance isotope ratios with chemically informative accuracy and precision. Here, we report the first use of Fourier transform ion cyclotron resonance mass spectrometry for the accurate (<1‰) and precise (<1‰ standard error) simultaneous determination of δ
Language	English
Publishing date	2023-11-15
Publishing country	United States
Document type	Journal Article
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.3c01816
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Article ; Online: Relating Molecular Properties to the Persistence of Marine Dissolved Organic Matter with Liquid Chromatography-Ultrahigh-Resolution Mass Spectrometry.

Boiteau, Rene M / Corilo, Yuri E / Kew, William R / Dewey, Christian / Alvarez Rodriguez, Maria Cristina / Carlson, Craig A / Conway, Tim M

Environmental science & technology

2024

Abstract: Marine dissolved organic matter (DOM) contains a complex mixture of small molecules that eludes rapid biological degradation. Spatial and temporal variations in the abundance of DOM reflect the existence of fractions that are removed from the ocean over ... ...

Abstract	Marine dissolved organic matter (DOM) contains a complex mixture of small molecules that eludes rapid biological degradation. Spatial and temporal variations in the abundance of DOM reflect the existence of fractions that are removed from the ocean over different time scales, ranging from seconds to millennia. However, it remains unknown whether the intrinsic chemical properties of these organic components relate to their persistence. Here, we elucidate and compare the molecular compositions of distinct DOM fractions with different lability along a water column in the North Atlantic Gyre. Our analysis utilized ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry at 21 T coupled to liquid chromatography and a novel data pipeline developed in CoreMS that generates molecular formula assignments and metrics of isomeric complexity. Clustering analysis binned 14 857 distinct molecular components into groups that correspond to the depth distribution of semilabile, semirefractory, and refractory fractions of DOM. The more labile fractions were concentrated near the ocean surface and contained more aliphatic, hydrophobic, and reduced molecules than the refractory fraction, which occurred uniformly throughout the water column. These findings suggest that processes that selectively remove hydrophobic compounds, such as aggregation and particle sorption, contribute to variable removal rates of marine DOM.
Language	English
Publishing date	2024-02-09
Publishing country	United States
Document type	Journal Article
ISSN	1520-5851
ISSN (online)	1520-5851
DOI	10.1021/acs.est.3c08245
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