Simultaneous Recording of the Uptake and Conversion of Glucose and Choline in Tumors by Deuterium Metabolic Imaging (DMI)

: Increased glucose and choline uptake are hallmarks of cancer. We investigated if the uptake and conversion of [2H9]choline alone and together with that of [6,6’ 2H2]glucose can be assessed in subcutaneous tumors by deuterium metabolic imaging (DMI) after bolus administration of these compounds. Therefore tumors with human renal carcinoma cells were grown subcutaneously in mice up to ~0.5 cm3. Mice were anesthetized with isoflurane and IV infused in the MR magnet for ~20 sec with ~0.2 ml solutions containing either [2H9]choline (0.05g/kg) alone or together with [6,6’ 2H2]glucose (1.3g/kg). 2H MR was performed on a 11.7T MR system with a home-built 2H/1H coil using a 900 excitation pulse and 400ms repetition time. 3D DMI was recorded at high resolution (2x2x2mm) in 37 min or at low resolution (3.7x3.7x3.7mm) in 2:24 min. Absolute tissue concentrations were calculate assuming initial [HOD]=13.7mM. Within 5 minutes after [2H9]choline infusion its signal appeared in tumor spectra representing concentration increasing up to 0.3–1.2 mM and then slowly decreased or remained constant over 100 minutes. In plasma [2H9]choline disappeared within 15 minutes post-infusion implying that its tumor signal arises from tissue and not blood. After infusing a mixture of [2H9]choline and [6,6’ 2H2]glucose their signals were observed separately in tumor 2H spectra. Over time the [2H9]choline signal broadened, possibly due to conversion to other choline compounds, [[6,6’ 2H2]glucose] declined, [HOD] increased and a lactate signal appeared, reflecting glycolysis. Metabolic maps of all 2H compounds were reconstructed from high resolution DMIs. As choline infusion and glucose DMI is feasible in patients, their simultaneous detection has clinical potential for tumor characterization.

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Simultaneous Recording of the Uptake and Conversion of Glucose and Choline in Tumors by Deuterium Metabolic Imaging

Cancers ◽

10.3390/cancers13164034 ◽

2021 ◽

Vol 13 (16) ◽

pp. 4034

Author(s):

Andor Veltien ◽

Jack van Asten ◽

Niveditha Ravichandran ◽

Robin A. de Graaf ◽

Henk M. De Feyter ◽

...

Keyword(s):

High Resolution ◽

Simultaneous Detection ◽

Simultaneous Recording ◽

Metabolic Imaging ◽

Choline Uptake ◽

Deuterated Water ◽

Post Infusion ◽

Repetition Time ◽

Tumor Accumulation ◽

Clinical Potential

Increased glucose and choline uptake are hallmarks of cancer. We investigated whether the uptake and conversion of [2H9]choline alone and together with that of [6,6′-2H2]glucose can be assessed in tumors via deuterium metabolic imaging (DMI) after administering these compounds. Therefore, tumors with human renal carcinoma cells were grown subcutaneously in mice. Isoflurane anesthetized mice were IV infused in the MR magnet for ~20 s with ~0.2 mL solutions containing either [2H9]choline (0.05 g/kg) alone or together with [6,6′-2H2]glucose (1.3 g/kg). 2H MR was performed on a 11.7T MR system with a home-built 2H/1H coil using a 90° excitation pulse and 400 ms repetition time. 3D DMI was recorded at high resolution (2 × 2 × 2 mm) in 37 min or at low resolution (3.7 × 3.7 × 3.7 mm) in 2:24 min. Absolute tissue concentrations were calculated assuming natural deuterated water [HOD] = 13.7 mM. Within 5 min after [2H9]choline infusion, its signal appeared in tumor spectra representing a concentration increase to 0.3–1.2 mM, which then slowly decreased or remained constant over 100 min. In plasma, [2H9]choline disappeared within 15 min post-infusion, implying that its signal arises from tumor tissue and not from blood. After infusing a mixture of [2H9]choline and [6,6′-2H2]glucose, their signals were observed separately in tumor 2H spectra. Over time, the [2H9]choline signal broadened, possibly due to conversion to other choline compounds, [[6,6′-2H2]glucose] declined, [HOD] increased and a lactate signal appeared, reflecting glycolysis. Metabolic maps of 2H compounds, reconstructed from high resolution DMIs, showed their spatial tumor accumulation. As choline infusion and glucose DMI is feasible in patients, their simultaneous detection has clinical potential for tumor characterization.

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BIMG-04. MAPPING HETEROGENEITY OF HIGH-GRADE GLIOMA METABOLISM USING HIGH RESOLUTION 7T MRSI

Neuro-Oncology Advances ◽

10.1093/noajnl/vdab024.003 ◽

2021 ◽

Vol 3 (Supplement_1) ◽

pp. i1-i1

Author(s):

Gilbert Hangel ◽

Cornelius Cadrien ◽

Philipp Lazen ◽

Sukrit Sharma ◽

Julia Furtner ◽

...

Keyword(s):

High Resolution ◽

Imaging Techniques ◽

Basis Set ◽

Metabolic Imaging ◽

Tumor Segmentation ◽

Low Grade ◽

High Grade ◽

Isotropic Resolution ◽

Contrast Enhanced ◽

Definition Of

Abstract OBJECTIVES Neurosurgical resection in gliomas depends on the precise preoperative definition of the tumor and its margins to realize a safe maximum resection that translates into a better patient outcome. New metabolic imaging techniques could improve this delineation as well as designate targets for biopsies. We validated the performance of our fast high-resolution whole-brain 3D-magnetic resonance spectroscopic imaging (MRSI) method at 7T in high-grade gliomas (HGGs) as first step to this regard. METHODS We measured 23 patients with HGGs at 7T with MRSI covering the whole cerebrum with 3.4mm isotropic resolution in 15 min. Quantification used a basis-set of 17 neurochemical components. They were evaluated for their reliability/quality and compared to neuroradiologically segmented tumor regions-of-interest (necrosis, contrast-enhanced, non-contrast-enhanced+edema, peritumoral) and histopathology (e.g., grade, IDH-status). RESULTS We found 18/23 measurements to be usable and ten neurochemicals quantified with acceptable quality. The most common denominators were increases of glutamine, glycine, and total choline as well as decreases of N-acetyl-aspartate and total creatine over most tumor regions. Other metabolites like taurine and serine showed mixed behavior. We further found that heterogeneity in the metabolic images often continued into the peritumoral region. While 2-hydroxy-glutarate could not be satisfyingly quantified, we found a tendency for a decrease of glutamate in IDH1-mutant HGGs. DISCUSSION Our findings corresponded well to clinical tumor segmentation but were more heterogeneous and often extended into the peritumoral region. Our results corresponded to previous knowledge, but with previously not feasible resolution. Apart from glycine/glutamine and their role in glioma progression, more research on the connection of glutamate and others to specific mutations is necessary. The addition of low-grade gliomas and statistical ROI analysis in a larger cohort will be the next important steps to define the benefits of our 7T MRSI approach for the definition of spatial metabolic tumor profiles.

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Ultrastructural visualization of cytoskeletal mRNAs and their associated proteins using double-label in situ hybridization.

The Journal of Cell Biology ◽

10.1083/jcb.108.6.2343 ◽

1989 ◽

Vol 108 (6) ◽

pp. 2343-2353 ◽

Cited By ~ 105

Author(s):

R H Singer ◽

G L Langevin ◽

J B Lawrence

Keyword(s):

In Situ Hybridization ◽

High Resolution ◽

Colloidal Gold ◽

Messenger Rna ◽

Signal To Noise Ratio ◽

Simultaneous Detection ◽

Gold Particles ◽

Rna Molecules ◽

Nucleic Acid Molecule

We have been able to visualize cytoskeletal messenger RNA molecules at high resolution using nonisotopic in situ hybridization followed by whole-mount electron microscopy. Biotinated cDNA probes for actin, tubulin, or vimentin mRNAs were hybridized to Triton-extracted chicken embryo fibroblasts and myoblasts. The cells were then exposed to antibodies against biotin followed by colloidal gold-conjugated antibodies and then critical-point dried. Identification of mRNA was possible using a probe fragmented to small sizes such that hybridization of several probe fragments along the mRNA was detected as a string of colloidal gold particles qualitatively and quantitatively distinguishable from nonspecific background. Extensive analysis showed that when eight gold particles were seen in this iterated array, the signal to noise ratio was greater than 30:1. Furthermore, these gold particles were colinear, often spiral, or circular suggesting detection of a single nucleic acid molecule. Antibodies against actin, vimentin, or tubulin proteins were used after in situ hybridization, allowing simultaneous detection of the protein and its cognate message on the same sample. This revealed that cytoskeletal mRNAs are likely to be extremely close to actin protein (5 nm or less) and unlikely to be within 20 nm of vimentin or tubulin filaments. Actin mRNA was found to be more predominant in lamellipodia of motile cells, confirming previous results. These results indicate that this high resolution in situ hybridization approach is a powerful tool by which to investigate the association of mRNA with the cytoskeleton.

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A novel pentaplex real time (RT)- PCR high resolution melt curve assay for simultaneous detection of emetic and enterotoxin producing Bacillus cereus in food

Food Control ◽

10.1016/j.foodcont.2015.08.030 ◽

2016 ◽

Vol 60 ◽

pp. 560-568 ◽

Cited By ~ 10

Author(s):

Fereidoun Forghani ◽

Prashant Singh ◽

Kun-Ho Seo ◽

Deog-Hwan Oh

Keyword(s):

High Resolution ◽

Bacillus Cereus ◽

Real Time ◽

Simultaneous Detection ◽

Rt Pcr ◽

High Resolution Melt

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Large-Scale Production of Bioactive Terrein by Aspergillus terreus Strain S020 Isolated from the Saudi Coast of the Red Sea

Biomolecules ◽

10.3390/biom9090480 ◽

2019 ◽

Vol 9 (9) ◽

pp. 480 ◽

Cited By ~ 5

Author(s):

Hani Z. Asfour ◽

Zuhier A. Awan ◽

Alaa A. Bagalagel ◽

Mahmoud A. Elfaky ◽

Reda F. A. Abdelhameed ◽

...

Keyword(s):

Mass Spectrometry ◽

High Resolution ◽

Crude Extract ◽

Large Scale ◽

Aspergillus Terreus ◽

Antimicrobial Activities ◽

Carcinoma Cells ◽

Small Scale ◽

Scale Production ◽

Fungal Strains

The diversity of symbiotic fungi derived from two marine sponges and sediment collected off Obhur, Jeddah (Saudi Arabia), was investigated in the current study. A total of 23 isolates were purified using a culture-dependent approach. Using the morphological properties combined with internal transcribed spacer-rDNA (ITS-rDNA) sequences, 23 fungal strains (in the majority Penicillium and Aspergillus) were identified from these samples. The biological screening (cytotoxic and antimicrobial activities) of small-scale cultures of these fungi yielded several target fungal strains which produced bioactive secondary metabolites. Amongst these isolates, the crude extract of Aspergillus terreus strain S020, which was cultured in fermentation static broth, 21 L, for 40 days at room temperature on potato dextrose broth, displayed strong antimicrobial activities against Pseudomonas aeruginosa and Staphylococcus aureus and significant antiproliferative effects on human carcinoma cells. Chromatographic separation of the crude extract by silica gel column chromatography indicated that the S020 isolate could produce a series of chemical compounds. Among these, pure crystalline terrein was separated with a high yield of 537.26 ± 23.42 g/kg extract, which represents the highest fermentation production of terrein to date. Its chemical structure was elucidated on the basis of high-resolution electrospray ionization mass spectrometry (HRESIMS) or high-resolution mass spectrometry (HRMS), 1D, and 2D NMR spectroscopic analyses and by comparison with reported data. The compound showed strong cytotoxic activity against colorectal carcinoma cells (HCT-116) and hepatocellular carcinoma cells (HepG2), with IC50 values of 12.13 and 22.53 µM, respectively. Our study highlights the potential of A. terreus strain S020 for the industrial production of bioactive terrein on a large scale and the importance of future investigations of these strains to identify the bioactive leads in these fungal extracts.

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Label-free, High-Resolution Optical Metabolic Imaging of Human Cervical Precancers Reveals Potential for Intraepithelial Neoplasia Diagnosis

Cell Reports Medicine ◽

10.1016/j.xcrm.2020.100017 ◽

2020 ◽

Vol 1 (2) ◽

pp. 100017

Author(s):

Dimitra Pouli ◽

Hong-Thao Thieu ◽

Elizabeth M. Genega ◽

Laura Baecher-Lind ◽

Michael House ◽

...

Keyword(s):

High Resolution ◽

Intraepithelial Neoplasia ◽

Metabolic Imaging ◽

Label Free

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A two-days full-scanning of the CFTR gene for simultaneous detection of point mutations and large rearrangements using High Resolution Melting analysis (HRM) in combination with quantitative real-time PCR

Journal of Cystic Fibrosis ◽

10.1016/s1569-1993(09)60064-3 ◽

2009 ◽

Vol 8 ◽

pp. S15

Author(s):

C. Thèze ◽

V. Gaston ◽

G. Carriere ◽

M. Claustres ◽

M. des Georges ◽

...

Keyword(s):

High Resolution ◽

Real Time ◽

Real Time Pcr ◽

High Resolution Melting ◽

Simultaneous Detection ◽

Point Mutations ◽

Cftr Gene ◽

Quantitative Real Time Pcr ◽

Melting Analysis ◽

Large Rearrangements

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Clostridium Spore Attachment to Human Cells

Microscopy and Microanalysis ◽

10.1017/s1431927600013143 ◽

1997 ◽

Vol 3 (S2) ◽

pp. 1251-1252

Author(s):

B. Panessa-Warren ◽

G. Tortora ◽

J. Warren

Keyword(s):

High Resolution ◽

Cell Surface ◽

Human Cells ◽

Carcinoma Cells ◽

Embryonic Fibroblasts ◽

Clostridium Sporogenes ◽

Colon Carcinoma Cells ◽

Spore Attachment ◽

Cooked Meat ◽

Commercial Field

This paper uses high resolution scanning electron microscopy (SEM) with a LaB6 gun and the newest commercial field emission guns, to obtain high magnification images of intact clostridial spores throughout the activation/germination/outgrowth process. by high resolution SEM, the clostridial exosporial membrane can be seen to produce numerous delicate projections (following activation), that extend from the exosporial surface to a nutritive substrate (agar), or cell surface when anaerobically incubated in the presence of human cells (embryonic fibroblasts and colon carcinoma cells). Magnifications of 20,000 to 200,000Xs at accelerating voltages low enough to minimize or eliminate specimen damage (1-5 kV) have permitted the entire surface of C.sporogenes and C.difficile endospores to be examined during all stages of germination. The relationships between the spore and the agar or human cell surface were also clearly visible.Clostridium sporogenes ATCC 3584 and C.difficile ATCC 43594 and 9689 were grown in cooked meat media, filtered and cleaned.

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