scholarly journals Non-invasive detection of protein metabolism in vivo by n.m.r. spectroscopy. Application of a novel 19F-containing residualizing label

1989 ◽  
Vol 264 (3) ◽  
pp. 829-835 ◽  
Author(s):  
A Daugherty ◽  
N N Becker ◽  
L A Scherrer ◽  
B E Sobel ◽  
J J H Ackerman ◽  
...  
Keyword(s):  
Intravenous Administration ◽  
Half Life ◽  
Soft Tissues ◽  
Degradation Products ◽  
Chemical Shifts ◽  
Whole Body ◽  
Hepatic Tissue ◽  
Protein Accumulation ◽  
Non Invasive

Protein residualizing labels facilitate localization of tissue sites of protein catabolism and the quantification of protein accumulation because of their prolonged intracellular retention of protein accumulation because of their prolonged intracellular retention times. Radioiodinated residualizing labels have been used to define the metabolism of a wide variety of proteins, but this has necessitated destructive analysis. Here we describe the implementation and validation of a novel 19F-containing residualizing label for protein, NN-dilactitol-3,5-bis(trifluoromethyl)benzylamine (DLBA), that permits the non-invasive assessment of protein accumulation and catabolism by n.m.r. spectroscopy in vivo. DLBA comprises a reporter molecule containing six equivalent 19F atoms. 19F is strongly n.m.r.-active, has 100% natural abundance, and is present in minimal background concentrations in soft tissues. We validated the use of DLBA as a protein-labelling compound by coupling to asialofetuin (ASF), a protein that is recognized exclusively by hepatic tissue via a saturable receptor-mediated process. Coupling of DLBA to ASF by reductive amination had no effect on the physiological receptor-mediated uptake of the protein in rat liver in vivo. The 19F-n.m.r. spectrum of DLBA exhibited a single peak that was subject to a small chemical-shift change and broadening after coupling to ASF. Pronase digestion of DLBA-ASF was performed to simulate intracellular degradation products, and resulted in a narrower set of resonances, with chemical shifts intermediate between those of uncoupled DLBA and DLBA-ASF. Intravenous administration of DLBA-ASF to rats followed by quantification of 19F in homogenates of liver tissue indicated that the half-life of residence time of degradation products from DLBA-ASF in liver was approx. 2 days. This intracellular half-life was comparable with that described for similar residualizing labels that contain radioiodide as a reporter. Similar results for the half-life of retention were obtained non-destructively and non-invasively in situ with the use of a whole-body radio-frequency antenna to acquire sequential spectra over 80 h after intravenous administration of DLBA-ASF. Quantification of these spectra demonstrated an initial accumulation of DLBA-ASF in liver followed by an expected gradual loss of 19F-labelled degradation products. The approach developed offers promise for the sequential and longitudinal characterization of metabolism of specific proteins in individual experimental animals and ultimately in human subjects.

Turnover of Human Extrinsic (Tissue-Type) Plasminogen Activator in Rabbits

1981 ◽  
Vol 46 (03) ◽  
pp. 658-661 ◽  
Author(s):  
C Korninger ◽  
J M Stassen ◽  
D Collen
Keyword(s):  
Plasminogen Activator ◽  
Intravenous Injection ◽  
Active Site ◽  
Half Life ◽  
Degradation Products ◽  
Gel Filtration ◽  
Tissue Type ◽  
Organ Distribution ◽  
Small Rise

SummaryThe turnover of highly purified human extrinsic plasminogen activator (EPA) (one- and two-chain form) was studied in rabbits. Following intravenous injection, EPA-activity declined rapidly. The disappearance rate of EPA from the plasma could adequately be described by a single exponential term with a t ½ of approximately 2 min for both the one-chain and two-chain forms of EPA.The clearance and organ distribution of EPA was studied by using 125I-labeled preparations. Following intravenous injection of 125I-1abeled EPA the radioactivity disappeared rapidly from the plasma also with a t ½ of approximately 2 min down to a level of 15 to 20 percent, followed by a small rise of blood radioactivity. Gel filtration of serial samples revealed that the secondary increase of the radioactivity was due to the reappearance of radioactive breakdown products in the blood. Measurement of the organ distribution of 125I at different time intervals revealed that EPA was rapidly accumulated in the liver, followed by a release of degradation products in the blood.Experimental hepatectomy markedly prolonged the half-life of EPA in the blood. Blocking the active site histidine of EPA had no effect on the half-life of EPA in blood nor on the gel filtration patterns of 125I in serial plasma samples.It is concluded that human EPA is rapidly removed from the blood of rabbits by clearance and degradation in the liver. Recognition by the liver does not require a functional active site in the enzyme. Neutralization in plasma by protease inhibitors does not represent a significant pathway of EPA inactivation in vivo.

Computerized Tomography: A Perspective in the Pediatric Patient

PEDIATRICS ◽  
1977 ◽  
Vol 59 (2) ◽  
pp. 305-308
Author(s):  
Derek Harwood-Nash ◽  
Herman Grossman ◽  
Alvin Felman ◽  
John Kirkpatrick ◽  
Leonard Swischuk
Keyword(s):  
Computerized Tomography ◽  
Soft Tissues ◽  
Conventional Radiography ◽  
The United States ◽  
The Body ◽  
Whole Body ◽  
Central Research ◽  
X Ray ◽  
Whole Body Ct

Computerized tomography (CT), a technique conceptualized by Oldendorf in 19611 and developed by Hounsfield2 of EMI-Tronics Inc. (EMI) Central Research Laboratories, has proven to be a successful innovation in neuroradiology. Reviews by Ambrose3 in England and by Baker et al.4 and by New et al.5 in the United States have clearly demonstrated the value of this new modality in neuroradiological diagnosis. In 1975 Houser et al.6 and Harwood-Nash et al.7 provided the initial clinical and radiological data about CT in infants and children. More recently this technique has been extended to the study of tissues and organs in the body other than those in the head. This has been accomplished by modification of the original machine into a whole-body CT system. Early reviews by Ledley et al.8 and by Alfidi et al.9 suggest a significant potential for diagnosis of lesions in the abdomen, pelvis, and thorax. The advantages of CT are that it is less invasive than standard special diagnostic radiological procedures and that for the first time it provides in vivo information regarding the content and the characteristics of tissue composing organs and masses. DESCRIPTION OF EQUIPMENT In conventional radiography an image is made on radiographic film by an attenuated X-ray beam. In passing through a core of tissue, each ray of the beam is attenuated as it is absorbed and scattered by the tissue in its path. The intensity of the transmitted ray depends on the sum total of X-ray attenuation by all the different soft tissues in its path.

scholarly journals Quantitative PET imaging of PD-L1 expression in xenograft and syngeneic tumour models using a site-specifically labelled PD-L1 antibody

2019 ◽  
Vol 47 (5) ◽  
pp. 1302-1313 ◽  
Author(s):  
Camilla Christensen ◽  
Lotte K. Kristensen ◽  
Maria Z. Alfsen ◽  
Carsten H. Nielsen ◽  
Andreas Kjaer
Keyword(s):  
Pet Imaging ◽  
Predictive Value ◽  
Ex Vivo ◽  
Response To Therapy ◽  
Whole Body ◽  
Therapeutic Monitoring ◽  
Non Invasive ◽  
Tumour Bearing ◽  
Quantitative Pet

Abstract Purpose Despite remarkable clinical responses and prolonged survival across several cancers, not all patients benefit from PD-1/PD-L1 immune checkpoint blockade. Accordingly, assessment of tumour PD-L1 expression by immunohistochemistry (IHC) is increasingly applied to guide patient selection, therapeutic monitoring, and improve overall response rates. However, tissue-based methods are invasive and prone to sampling error. We therefore developed a PET radiotracer to specifically detect PD-L1 expression in a non-invasive manner, which could be of diagnostic and predictive value. Methods Anti-PD-L1 (clone 6E11, Genentech) was site-specifically conjugated with DIBO-DFO and radiolabelled with 89Zr (89Zr-DFO-6E11). 89Zr-DFO-6E11 was optimized in vivo by longitudinal PET imaging and dose escalation with excess unlabelled 6E11 in HCC827 tumour-bearing mice. Specificity of 89Zr-DFO-6E11 was evaluated in NSCLC xenografts and syngeneic tumour models with different levels of PD-L1 expression. In vivo imaging data was supported by ex vivo biodistribution, flow cytometry, and IHC. To evaluate the predictive value of 89Zr-DFO-6E11 PET imaging, CT26 tumour-bearing mice were subjected to external radiation therapy (XRT) in combination with PD-L1 blockade. Results 89Zr-DFO-6E11 was successfully labelled with a high radiochemical purity. The HCC827 tumours and lymphoid tissue were identified by 89Zr-DFO-6E11 PET imaging, and co-injection with 6E11 increased the relative tumour uptake and decreased the splenic uptake. 89Zr-DFO-6E11 detected the differences in PD-L1 expression among tumour models as evaluated by ex vivo methods. 89Zr-DFO-6E11 quantified the increase in PD-L1 expression in tumours and spleens of irradiated mice. XRT and anti-PD-L1 therapy effectively inhibited tumour growth in CT26 tumour-bearing mice (p < 0.01), and the maximum 89Zr-DFO-6E11 tumour-to-muscle ratio correlated with response to therapy (p = 0.0252). Conclusion PET imaging with 89Zr-DFO-6E11 is an attractive approach for specific, non-invasive, whole-body visualization of PD-L1 expression. PD-L1 expression can be modulated by radiotherapy regimens and 89Zr-DFO-6E11 PET is able to monitor these changes and predict the response to therapy in an immunocompetent tumour model.

scholarly journals A fluorescent residualizing label for studies on protein uptake and catabolism in vivo and in vitro

1990 ◽  
Vol 267 (1) ◽  
pp. 155-162 ◽  
Author(s):  
J L Maxwell ◽  
L Terracio ◽  
T K Borg ◽  
J W Baynes ◽  
S R Thorpe
Keyword(s):  
Half Life ◽  
Normal Tissue ◽  
Degradation Products ◽  
Rat Serum ◽  
Peripheral Tissues ◽  
Protein Uptake ◽  
Kinetics Of ◽  
Rat Serum Albumin

Residualizing labels are tracers which remain in lysosomes after uptake and catabolism of the carrier protein and have been especially useful for studies on the sites of plasma protein degradation. Thus far these labels have contained radioactive reporters such as 3H or 125I. In the present paper we describe a fluorescent residualizing label, NN-dilactitol-N′-fluoresceinylethylenediamine (DLF). Modification of asialofetuin (ASF) or rat serum albumin (RSA) with DLF affected neither their normal kinetics of clearance from the rat circulation nor their normal tissue sites of uptake and degradation. After injection of DLF-ASF, fluorescent degradation products were recovered nearly quantitatively in liver and retained with a half-life of about 2 days. Fluorescent degradation products from DLF-RSA were recovered in skin and muscle, and were localized in fibroblasts by fluorescence microscopy. These results confirm previous studies with radioactive residualizing labels in which fibroblasts in peripheral tissues were identified as primary sites of albumin degradation. Fluorescent catabolites also accumulated in fibroblasts incubated with DLF-RSA in vitro, and residualized with a half-life of about 2 days. Overall, the data establish that DLF functions efficiently as a fluorescent residualizing label both in vivo and in vitro. The advantages of fluorescent, compared with radioactive, residualizing labels should make them valuable tools for studies on protein uptake and catabolism in biological systems.

Non-Invasive Measurement of In-Vivo Elasticity of Skeletal Muscles with MR-Elastography

2007 ◽  
Vol 342-343 ◽  
pp. 901-904
Author(s):  
Yu Bong Kang ◽  
T. Oida ◽  
Duk Young Jung ◽  
A. Fukuma ◽  
T. Azuma ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Shear Modulus ◽  
Skeletal Muscles ◽  
Soft Tissues ◽  
Viscosity Coefficient ◽  
Non Invasive ◽  
Novel Method ◽  
Human Calf ◽  
Invasive Measurement

In order to evaluate the mechanical properties of the human skeletal muscles, the elasticity and viscosity of the human calf muscles were measured with Magnetic Resonance Elastography (MRE). MRE is a novel method to measure the mechanical properties of living soft tissues in vivo quantitatively by observing the strain waves propagated in the object. In this study, the shear modulus and viscosity coefficient were measured with MRE. The shear modulus was 3.7 kPa in relaxed state, and increased with increasing the muscle forces. Interestingly, the viscosity was changed with the vibration frequency applied to the muscles, that was 4.5 Pa·s at 100Hz vibration and 2.4 Pa·s at 200Hz vibration. This shows clearly the visco-elastic property.

scholarly journals Why Are Viscosity and Nonlinearity Bound to Make an Impact in Clinical Elastographic Diagnosis?

Sensors ◽  
2020 ◽  
Vol 20 (8) ◽  
pp. 2379 ◽  
Author(s):  
Guillermo Rus ◽  
Inas H. Faris ◽  
Jorge Torres ◽  
Antonio Callejas ◽  
Juan Melchor
Keyword(s):  
Soft Tissues ◽  
Elastic Parameters ◽  
Tissue Microstructure ◽  
Non Invasive ◽  
Recent Developments ◽  
Mechanical Biomarkers ◽  
Nonlinear Elastic

The adoption of multiscale approaches by the biomechanical community has caused a major improvement in quality in the mechanical characterization of soft tissues. The recent developments in elastography techniques are enabling in vivo and non-invasive quantification of tissues’ mechanical properties. Elastic changes in a tissue are associated with a broad spectrum of pathologies, which stems from the tissue microstructure, histology and biochemistry. This knowledge is combined with research evidence to provide a powerful diagnostic range of highly prevalent pathologies, from birth and labor disorders (prematurity, induction failures, etc.), to solid tumors (e.g., prostate, cervix, breast, melanoma) and liver fibrosis, just to name a few. This review aims to elucidate the potential of viscous and nonlinear elastic parameters as conceivable diagnostic mechanical biomarkers. First, by providing an insight into the classic role of soft tissue microstructure in linear elasticity; secondly, by understanding how viscosity and nonlinearity could enhance the current diagnosis in elastography; and finally, by compounding preliminary investigations of those elastography parameters within different technologies. In conclusion, evidence of the diagnostic capability of elastic parameters beyond linear stiffness is gaining momentum as a result of the technological and imaging developments in the field of biomechanics.

High-resolution magnetic resonance imaging tracks changes in organ and tissue mass in obese and aging rats

2002 ◽  
Vol 282 (3) ◽  
pp. R890-R899 ◽  
Author(s):  
Haiying Tang ◽  
Joseph R. Vasselli ◽  
Ed X. Wu ◽  
Carol N. Boozer ◽  
Dympna Gallagher
Keyword(s):  
Magnetic Resonance Imaging ◽  
Adipose Tissue ◽  
Magnetic Resonance ◽  
High Resolution ◽  
Soft Tissues ◽  
Whole Body ◽  
Resonance Imaging ◽  
Sprague Dawley ◽  
Tissue Mass

Magnetic resonance imaging (MRI) has the ability to discriminate between various soft tissues in vivo. Whole body, specific organ, total adipose tissue (TAT), intra-abdominal adipose tissue (IAAT), and skeletal muscle (SM) weights determined by MRI were compared with weights determined by dissection and chemical analysis in two studies with male Sprague-Dawley rats. A 4.2-T MRI machine acquired high-resolution, in vivo, longitudinal whole body images of rats as they developed obesity or aged. Weights of the whole body and specific tissues were determined using computer image analysis software, including semiautomatic segmentation algorithms for volume calculations. High correlations were found for body weight ( r = 0.98), TAT ( r = 0.99), and IAAT ( r = 0.98) between MRI and dissection and chemical analyses. MRI estimated the weight of the brain, kidneys, and spleen with high accuracy ( r > 0.9), but overestimated IAAT, SM, and liver volumes. No differences were detected in organ weights using MRI and dissection measurements. Longitudinal MRI measurements made during the development of obesity and aging accurately represented changes in organ and tissue mass.

scholarly journals Non-invasive synchronous monitoring of neutrophil migration using whole body near-infrared fluorescence-based imaging

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jack Leslie ◽  
Stuart M. Robinson ◽  
Fiona Oakley ◽  
Saimir Luli
Keyword(s):  
Real Time ◽  
Fluorescence Imaging ◽  
Near Infrared ◽  
Spectral Unmixing ◽  
Whole Body ◽  
Spectral Signature ◽  
Cell Labelling ◽  
Cellular Interactions ◽  
Non Invasive

AbstractAdvances in fluorescence imaging coupled with the generation of near infrared probes have significantly improved the capabilities of non-invasive, real-time imaging in whole animals. In this study we were able to overcome a limitation of in vivo fluorescence imaging and have established a dual cell tracking method where two different cell types can be monitored according to the spectral signature of the cell labelling fluorophore. Using a mouse model of acute liver injury, we have characterised the in vivo migration patterns of wild type and transgenic neutrophils with impaired chemotaxis. Here, we were able to demonstrate that IVIS provides a sensitive multiplexing technology to differentiate two different cell populations based on the spectral signature of the cell labelling fluorophores. This spectral unmixing methodology has the potential to uncover multidimensional cellular interactions involved in many diseases such as fibrosis and cancer. In vivo spectral un-mixing provides a useful tool for monitoring multiple biological process in real-time in the same animal.

scholarly journals Thrombin-Antithrombin III and Plasm in-Antiplasmin Complexes as Indicators of in Vivo Activation of the Coagulation and/or Fibrinolytic Systems

1977 ◽  
Author(s):  
D. Collen ◽  
F.de Cock
Keyword(s):  
Human Blood ◽  
Half Life ◽  
Antithrombin Iii ◽  
Enzyme Inhibitor ◽  
Degradation Products ◽  
Clinical Value ◽  
Fibrinogen Degradation Products ◽  
Prethrombotic State ◽  
Immunochemical Methods

Both the thrombin-antithrombin III and plasmin-antiplasmin complexes contain neoantigenic structures, not present in the parent molecules, to which non cross-reacting precipitating antibodies can be produced (Thromb.Res. 5, 577, 1974). This phenomenon has been used to develop immunochemical methods for the quantitation of these complexes in human blood (Thromb. Res 7, 235, 1975). Patients with in vivo activation of the coagulation and/or fibrinolytic systems consistently showed increased levels of enzyme-inhibitor complexes (Eur.J.Clin.Invest, in press, 1977). The circulatory half-life of these complexes was estimated to be several hours (0.5 to 0.75 days).On several occasions, increased plasmin-antiplasmin titers were observed in the presence of a normal screening hemostasis analysis, suggesting that these assays may be more sensitive indicators of in vivo activation of the coagulation-fibrinolytic systems that assays for fibrinogen degradation products.Studies to determine the clinical value of the measurement of these enzyme-inhibitor complexes as indicators of a prethrombotic state are in progress and will be reported.

scholarly journals The stability of rat liver ribonucleic acid in vivo after methylation with methyl methanesulphonate or dimethylnitrosamine

1971 ◽  
Vol 125 (3) ◽  
pp. 821-827 ◽  
Author(s):  
Marilyn J. McElhone ◽  
P. J. O'Connor ◽  
A. W. Craig
Keyword(s):  
Rat Liver ◽  
Half Life ◽  
Orotic Acid ◽  
Degradation Products ◽  
Rna Degradation ◽  
Biological Half Life ◽  
Extraction And Purification ◽  
The Stability ◽  
Methylating Agents

1. RNA was isolated from rat liver at selected times after the intraperitoneal injection of either [14C]methyl methanesulphonate (50mg/kg) or [14C]dimethylnitrosamine (2mg/kg). These doses were chosen to minimize effects due to toxicity. 2. Two methods of extraction and purification of RNA were used and an analysis of the radioactivity present was made by column chromatography of acid hydrolysates of the purified RNA. 3. The extent of methylation of guanine, the principal site of alkylation in rat liver RNA, was determined at times up to 14 days after injection. Although dimethylnitrosamine is a potent liver carcinogen and methyl methanesulphonate is not carcinogenic to rat liver, the rate of disappearance of 7-methylguanine from RNA was similar for both compounds, with a half-life of about 3.5 days. 4. An estimate of the biological half-life of rRNA was made by using [3H]orotic acid. A half-life of 5 days was obtained and this was not affected by injecting animals with unlabelled methyl methanesulphonate at the same dosage of 50mg/kg used in the studies of RNA methylation. 5. After administration of labelled orotic acid, reutilization of labelled RNA degradation products probably results in an overestimation of the biological half-life for rRNA. It is suggested that non-toxic doses of methylating agents such as methyl methanesulphonate and dimethylnitrosamine may prove to be a more effective way of accurately estimating the biological turnover of RNA species.

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