ABSTRACT
Purpose: Water freely diffuses across cell membranes, making it suitable for measuring absolute tissue perfusion. In this study, we introduce an imaging method for conducting coronary artery angiography and quantifying myocardial perfusion across the entire heart using hyperpolarized water. Methods:1H was hyperpolarized using dissolution dynamic nuclear polarization (dDNP) with UV-generated radicals. Submillimeter resolution coronary artery images were acquired as 2D projections using a spoiled GRE (SPGRE) sequence gated on diastole. Dynamic perfusion images were obtained with a multi-slice SPGRE with diastole gating, covering the entire heart. Perfusion values were analyzed through histograms, and the most frequent estimated perfusion value (the mode of the distribution), was compared with the average values for 15O water PET from the literature. Results: A liquid state polarization of 10% at the time of the injection and a 30 s T1 in D2O TRIS buffer were measured. Both coronary artery and dynamic perfusion images exhibited good quality. The main and small coronary artery branches were well resolved. The most frequent estimated perfusion value is around 0.6 mL/g/min, which is lower than the average values obtained from the literature for 15O-water PET (around 1.1 and 1.5 mL/g/min). Conclusions: The study successfully demonstrated the feasibility of achieving high-resolution, motion-free coronary artery angiography and 3D whole-heart quantitative myocardial perfusion using hyperpolarized water.
Subject(s)
Coronary Angiography , Coronary Vessels , Water , Humans , Coronary Angiography/methods , Coronary Vessels/diagnostic imaging , Myocardial Perfusion Imaging/methods , Male , Oxygen Radioisotopes , Heart/diagnostic imaging , Female , Coronary Circulation/physiologyABSTRACT
Hyperpolarized carbon-13 labeled compounds are increasingly being used in medical MR imaging (MRI) and MR imaging (MRI) and spectroscopy (MRS) research, due to its ability to monitor tissue and cell metabolism in real-time. Although radiological biomarkers are increasingly being considered as clinical indicators, biopsies are still considered the gold standard for a large variety of indications. Bioreactor systems can play an important role in biopsy examinations because of their ability to provide a physiochemical environment that is conducive for therapeutic response monitoring ex vivo. We demonstrate here a proof-of-concept bioreactor and microcoil receive array setup that allows for ex vivo preservation and metabolic NMR spectroscopy on up to three biopsy samples simultaneously, creating an easy-to-use and robust way to simultaneously run multisample carbon-13 hyperpolarization experiments. Experiments using hyperpolarized [1-13C]pyruvate on ML-1 leukemic cells in the bioreactor setup were performed and the kinetic pyruvate-to-lactate rate constants ( k PL ) extracted. The coefficient of variation of the experimentally found k PL s for five repeated experiments was C V = 35 % . With this statistical power, treatment effects of 30%-40% change in lactate production could be easily differentiable with only a few hyperpolarization dissolutions on this setup. Furthermore, longitudinal experiments showed preservation of ML-1 cells in the bioreactor setup for at least 6 h. Rat brain tissue slices were also seen to be preserved within the bioreactor for at least 1 h. This validation serves as the basis for further optimization and upscaling of the setup, which undoubtedly has huge potential in high-throughput studies with various biomarkers and tissue types.
Subject(s)
Metabolic Flux Analysis , Pyruvic Acid , Rats , Animals , Carbon Isotopes , Pyruvic Acid/metabolism , Lactic Acid/metabolism , Bioreactors , BiomarkersABSTRACT
We report dissolution Dynamic Nuclear Polarization (d-DNP) of [15 N3 ]metronidazole ([15 N3 ]MNZ) for the first time. Metronidazole is a clinically approved antibiotic, which can be potentially employed as a hypoxia-sensing molecular probe using 15 N hyperpolarized (HP) nucleus. The DNP process is very efficient for [15 N3 ]MNZ with an exponential build-up constant of 13.8â min using trityl radical. After dissolution and sample transfer to a nearby 4.7â T Magnetic Resonance Imaging scanner, HP [15 N3 ]MNZ lasted remarkably long with T1 values up to 343â s and 15 N polarizations up to 6.4 %. A time series of HP [15 N3 ]MNZ images was acquired in vitro using a steady state free precession sequence on the 15 NO2 peak. The signal lasted over 13â min with notably long T2 of 20.5â s. HP [15 N3 ]MNZ was injected in the tail vein of a healthy rat, and dynamic spectroscopy was performed over the rat brain. The in vivo HP 15 N signals persisted over 70â s, demonstrating an unprecedented opportunity for in vivo studies.
Subject(s)
Anti-Bacterial Agents , Metronidazole , Rats , Animals , Metronidazole/pharmacology , Anti-Bacterial Agents/pharmacology , Solubility , Magnetic Resonance Spectroscopy/methods , Magnetic Resonance ImagingABSTRACT
Shigellosis caused by Shigella is one of the most important foodborne illnesses in global health, but little is known about the metabolic cross talk between this bacterial pathogen and its host cells during the different stages of the infection process. A detailed understanding of the metabolism can potentially lead to new drug targets remedying the pressing problem of antibiotic resistance. Here, we use stable isotope-resolved metabolomics as an unbiased and fast method to investigate how Shigella metabolizes 13C-glucose in three different environments: inside the host cells, adhering to the host cells, and alone in suspension. We find that especially formate metabolism by bacteria is sensitive to these different environments. The role of formate in pathogen metabolism is sparsely described in the literature compared to the roles of acetate and butyrate. However, its metabolic pathway is regarded as a potential drug target due to its production in microorganisms and its absence in humans. Our study provides new knowledge about the regulatory effect of formate. Bacterial metabolism of formate is pH dependent when studied alone in culture medium, whereas this effect is less pronounced when the bacteria adhere to the host cells. Once the bacteria are inside the host cells, we find that formate accumulation is reduced. Formate also affects the host cells resulting in a reduced infection rate. This was correlated to an increased immune response. Thus, intriguingly formate plays a double role in pathogenesis by increasing the virulence of Shigella and at the same time stimulating the immune response of the host. IMPORTANCE Bacterial infection is a pressing societal concern due to development of resistance toward known antibiotics. Central carbon metabolism has been suggested as a potential new target for drug development, but metabolic changes upon infection remain incompletely understood. Here, we used a cellular infection model to study how the bacterial pathogen Shigella adapts its metabolism depending on the environment starting from the extracellular medium until Shigella successfully invaded and proliferated inside host cells. The mixed-acid fermentation of Shigella was the major metabolic pathway during the infectious process, and the glucose-derived metabolite formate surprisingly played a divergent role in the pathogen and in the host cell. Our data show reduced infection rate when both host cells and bacteria were treated with formate, which correlated with an upregulated immune response in the host cells. The formate metabolism in Shigella thus potentially provides a route toward alternative treatment strategies for Shigella prevention.
Subject(s)
Shigella flexneri , Shigella , Humans , HeLa Cells , Formates/metabolism , Formates/pharmacology , Glucose/metabolismABSTRACT
PURPOSE: There is a limit to the maximum achievable preamplifier decoupling. In many cases, this level is not enough. To overcome this limit, the preamplifier noise figure can be compromised for further decoupling increase. This is useful in flexible MRI arrays where ensuring coil insensitivity to changes in other array elements is a challenge. METHODS: This work establishes the relation between the preamplifier noise figure and preamplifier decoupling using closed-form equations. These equations allow the evaluation of preamplifier decoupling properties and benchmark different preamplifiers against each other. The method to design the corresponding decoupling networks is described. The derived generalized design equations, which are not limited to 50 Ω pre-matched preamplifiers, greatly improve design flexibility and enable use of new amplifiers in MRI detectors. RESULTS: Using the method, the decoupling properties of three preamplifiers are studied. For demonstration, the coil decoupling is further increased by 10.8 dB using one of the preamplifiers. The noise figure is sacrificed by 0.5 dB, which is predicted by equations and verified experimentally. Although examples are shown for 3 T systems at 32.13 MHz and 127.7 MHz, the approach and equations apply to any field strength and nucleus. CONCLUSION: Preamplifier decoupling can be improved beyond what is possible by traditional approaches. The derived design equations cover a wide range of cases, including inductive coils and self-resonant low-impedance and high-impedance coils.
Subject(s)
Amplifiers, Electronic , Magnetic Resonance Imaging , Equipment DesignABSTRACT
PURPOSE: This article presents a novel 14-channel receive-only array for 13 C human head imaging at 3 T that explores the SNR gain by operating at cryogenic temperature cooled by liquid nitrogen. METHODS: Cryostats are developed to evaluate single-coil bench SNR performance and cool the 14-channel array with liquid nitrogen while having enough thermal insulation between the coils and the sample. The temperature distribution for the coil array is measured. Circuits are adapted to the -189°C environment and implemented in the 14-channel array. 13 C images are acquired with the array at cryogenic and room temperature in a 3T scanner. RESULTS: Compared with room temperature, the array at cryogenic temperature provides 27%-168% SNR improvement over all voxels and 47% SNR improvement near the image center. The measurements show a decrease of the element noise correlation at cryogenic temperature. CONCLUSION: It is demonstrated that higher SNR can be achieved by cryogenically cooling the 14-channel array. A cryogenic array suitable for clinical imaging can be further developed on the array proposed. The cryogenic coil array is most likely suited for scenarios in which high SNR deep in a head and decent SNR on the periphery are required.
Subject(s)
Cold Temperature , Magnetic Resonance Imaging , Humans , Magnetic Resonance Imaging/methods , Nitrogen , Phantoms, Imaging , Signal-To-Noise Ratio , Equipment DesignABSTRACT
PURPOSE: To develop a coil-based method to obtain accurate sensitivity profiles in 13 C MRI at 3T from the endogenous 23 Na. An eight-channel array is designed for 13 C MR acquisitions. As application examples, the array is used for two-fold accelerated acquisitions of both hyperpolarized 13 C metabolic imaging of pig kidneys and the human brain. METHODS: A flexible coil array was tuned optimally for 13 C at 3T (32.1 MHz), with the coil coupling coefficients matched to be nearly identical at the resonance frequency of 23 Na (33.8 MHz). This is done by enforcing a high decoupling (obtained through highly mismatched preamplifiers) and adjusting the coupling frequency response. The SNR performance is compared to reference coils. RESULTS: The measured sensitivity profiles on a phantom showed high spatial similarity for 13 C and 23 Na resonances, with average noise correlation of 9 and 11%, respectively. For acceleration factors 2, 3, and 4, the obtained maximum g-factors were 1.0, 1.1, and 2.6, respectively. The 23 Na profiles obtained in vivo could be used successfully to perform two-fold acceleration of hyperpolarized 13 C 3D acquisitions of both pig kidneys and a healthy human brain. CONCLUSION: A receive array has been developed in such a way that the 13 C sensitivity profiles could be accurately obtained from measurements at the 23 Na frequency. This technique facilitates accelerated acquisitions for hyperpolarized 13 C imaging. The SNR performance obtained at the 13 C frequency, compares well to other state-of-the-art coils for the same purpose, showing slightly better superficial and central SNR.
Subject(s)
Magnetic Resonance Imaging , Radio Waves , Animals , Brain/diagnostic imaging , Equipment Design , Magnetic Resonance Imaging/methods , Phantoms, Imaging , Signal-To-Noise Ratio , SwineABSTRACT
Polarisation transfer schemes and indirect detection are central to magnetic resonance. Using the trityl radical OX063 and a pulse electron paramagnetic resonance spectrometer operating in the Q-band (35â¯GHz, 1.2â¯T), we show here that it is possible to use pulsed dynamic nuclear polarisation (DNP) to transfer polarisation from electrons to protons and back. The latter is achieved by first saturating the electrons and then simply using a reverse DNP step. A variable mixing time between DNP and reverse DNP allows us to investigate the decay of polarisation on protons in the vicinity of the electrons. We qualitatively investigate the influence of solvent deuteration, temperature, and electron concentration. We expect reverse DNP to be useful in the investigation of nuclear spin diffusion and envisage its use in electron-nuclear double-resonance (ENDOR) experiments.
ABSTRACT
The use of hyperpolarised 13 C pyruvate for nononcological neurological applications has not been widespread so far, possibly due to delivery issues limiting the visibility of metabolites. First proof-of-concept results have indicated that metabolism can be detected in human brain, and this may supersede the results obtained in preclinical settings. One major difference between the experimental setups is that preclinical MRI/MRS routinely uses anaesthesia, which alters both haemodynamics and metabolism. Here, we used hyperpolarised [1-13 C]pyruvate to compare brain metabolism in awake rats and under isoflurane, urethane or medetomidine anaesthesia. Spectroscopic [1-13 C]pyruvate time courses measured sequentially showed that pyruvate-to-bicarbonate and pyruvate-to-lactate labelling rates were lower in isoflurane animals than awake animals. An increased bicarbonate-to-lactate ratio was observed in the medetomidine group compared with other groups. The study shows that hyperpolarised [1-13 C]pyruvate experiments can be performed in awake rats, thus avoiding anaesthesia-related issues. The results suggest that haemodynamics probably dominate the observed pyruvate-to-metabolite labelling rates and area-under-time course ratios of referenced to pyruvate. On the other hand, the results obtained with medetomidine suggest that the ratios are also modulated by the underlying cerebral metabolism. However, the ratios between intracellular metabolites were unchanged in awake compared with isoflurane-anaesthetised rats.
Subject(s)
Brain/metabolism , Isoflurane/pharmacology , Pyruvic Acid/metabolism , Anesthesia , Animals , Carbon Isotopes , Female , Rats , Rats, Sprague-Dawley , Rats, Wistar , WakefulnessABSTRACT
Hyperpolarized 13C isotope resolved spectroscopy boosts NMR signal intensity, which improves signal detection and allows metabolic fluxes to be analyzed. Such hyperpolarized flux data may offer new approaches to tissue classification and biomarker identification that could be translated in vivo. Here we used hyperpolarized stable isotope resolved analysis (SIRA) to measure metabolite specific 13C isotopic enrichments in the central carbon metabolism of mouse prostate. Prostate and tumor tissue samples were acquired from transgenic adenocarcinomas of the mouse prostate (TRAMP) mice. Before euthanasia, mice were injected with [U-13C]glucose intraperitoneally (i.p.). Polar metabolite extracts were prepared, and hyperpolarized 1D-13C NMR spectra were obtained from normal prostate (n = 19) and cancer tissue (n = 19) samples. Binary classification and feature analysis was performed to make a separation model and to investigate differences between samples originating from normal and cancerous prostate tissue, respectively. Hyperpolarized experiments were carried out according to a standardized protocol, which showed a high repeatability (CV = 15%) and an average linewidth in the 1D-13C NMR spectra of 2 ± 0.5 Hz. The resolution of the hyperpolarized 1D-13C spectra was high with little signal overlap in the carbonyl region and metabolite identification was easily accomplished. A discrimination with 95% success rate could be made between samples originating from TRAMP mice prostate and tumor tissue based on isotopomers from uniquely identified metabolites. Hyperpolarized 13C-SIRA allowed detailed metabolic information to be obtained from tissue specimens. The positional information of 13C isotopic enrichments lead to easily interpreted features responsible for high predictive classification of tissue types. This analytical approach has matured, and the robust experimental protocols currently available allow systematic tracking of metabolite flux ex vivo.
Subject(s)
Prostatic Neoplasms , Animals , Biomarkers, Tumor , Carbon Isotopes , Humans , Magnetic Resonance Spectroscopy , Male , MiceABSTRACT
Drastic sensitivity enhancement of dynamic nuclear polarization is becoming an increasingly critical methodology to monitor real-time metabolic and physiological information in chemistry, biochemistry, and biomedicine. However, the limited number of available hyperpolarized 13C probes, which can effectively interrogate crucial metabolic activities, remains one of the major bottlenecks in this growing field. Here, we demonstrate [1-13C] N-acetyl cysteine (NAC) as a novel probe for hyperpolarized 13C MRI to monitor glutathione redox chemistry, which plays a central part of metabolic chemistry and strongly influences various therapies. NAC forms a disulfide bond in the presence of reduced glutathione, which generates a spectroscopically detectable product that is separated from the main peak by a 1.5 ppm shift. In vivo hyperpolarized MRI in mice revealed that NAC was broadly distributed throughout the body including the brain. Its biochemical transformation in two human pancreatic tumor cells in vitro and as xenografts differed depending on the individual cellular biochemical profile and microenvironment in vivo. Hyperpolarized NAC can be a promising non-invasive biomarker to monitor in vivo redox status and can be potentially translatable to clinical diagnosis.
Subject(s)
Acetylcysteine/metabolism , Brain/metabolism , Carbon Isotopes/analysis , Glutathione/metabolism , Pancreatic Neoplasms/pathology , Animals , Apoptosis , Cell Proliferation , Humans , Magnetic Resonance Imaging , Mice , Oxidation-Reduction , Pancreatic Neoplasms/metabolism , Tumor Cells, Cultured , Xenograft Model Antitumor AssaysABSTRACT
Acute ischemic stroke patients benefit from reperfusion in a short time-window after debut. Later treatment may be indicated if viable brain tissue is demonstrated and this outweighs the inherent risks of late reperfusion. Magnetic resonance imaging (MRI) with hyperpolarized [1-13C]pyruvate is an emerging technology that directly images metabolism. Here, we investigated its potential to detect viable tissue in ischemic stroke. Stroke was induced in pigs by intracerebral injection of endothelin 1. During ischemia, the rate constant of pyruvate-to-lactate conversion, kPL, was 52% larger in penumbra and 85% larger in the infarct compared to the contralateral hemisphere (P = 0.0001). Within the penumbra, the kPL was 50% higher in the regions that later infarcted compared to non-progressing regions (P = 0.026). After reperfusion, measures of pyruvate-to-lactate conversion were slightly decreased in the infarct compared to contralateral. In addition to metabolic imaging, we used hyperpolarized pyruvate for perfusion-weighted imaging. This was consistent with conventional imaging for assessment of infarct size and blood flow. Lastly, we confirmed the translatability of simultaneous assessment of metabolism and perfusion with hyperpolarized MRI in healthy volunteers. In conclusion, hyperpolarized [1-13C]pyruvate may aid penumbral characterization and increase access to reperfusion therapy for late presenting patients.
Subject(s)
Brain Infarction/diagnostic imaging , Ischemic Stroke/diagnostic imaging , Magnetic Resonance Imaging/methods , Pyruvic Acid/metabolism , Reperfusion/methods , Animals , Brain/metabolism , Brain Infarction/metabolism , Brain Infarction/pathology , Case-Control Studies , Cerebrovascular Circulation/physiology , Endothelin-1/metabolism , Female , Humans , Ischemic Stroke/metabolism , Models, Animal , Perfusion Imaging/methods , SwineABSTRACT
Magnetic Resonance Imaging combined with hyperpolarized 13C-labelled metabolic contrast agents produced via dissolution Dynamic Nuclear Polarization can, non-invasively and in real-time, report on tissue specific aberrant metabolism. However, hyperpolarization equipment is expensive, technically demanding and needs to be installed on-site for the end-user. In this work, we provide a robust methodology that allows remote production of the hyperpolarized 13C-labelled metabolic contrast agents. The methodology, built on photo-induced thermally labile radicals, allows solid sample extraction from the hyperpolarization equipment and several hours' lifetime of the 13C-labelled metabolic contrast agents at appropriate storage/transport conditions. Exemplified with [U-13C, d7]-D-glucose, we remotely produce hyperpolarized 13C-labelled metabolic contrast agents and generate above 10,000-fold liquid-state Magnetic Resonance signal enhancement at 9.4 T, keeping on-site only a simple dissolution device.
ABSTRACT
PURPOSE: Preamplifier decoupling is useful for minimizing interaction between MRI array elements. The purpose of this work is to propose a general approach to designing networks for preamplifier decoupling while keeping the number of elements to a minimum. The approach is applicable to arbitrary impedance preamplifiers and arbitrary coil impedances. METHODS: Closed form design equations for decoupling networks are derived based on maximum decoupling and minimum preamplifier noise conditions. The analytical solutions are verified using numerical simulations. Design examples at 32.1, 64, 128, and 298 MHz are shown. One of the examples is realized on a test bench. The fabricated circuit is tested for decoupling and minimum noise properties. RESULTS: The design equations are verified numerically and experimentally. The fabricated network demonstrates 30.7 dB of decoupling and minimum output noise at the design frequency. CONCLUSION: The design equations lead to four alternative network solutions. Each network is realized as a T-shape or Π-shape three elements circuit topology. All four networks are identical in performance providing minimum amplifier noise and maximum decoupling for a given preamplifier and coil combination. An MRI array designer can choose any solution out of four. The considerations for choosing the most practical solution are given. The presented method enables the use of arbitrary impedance preamplifiers or transistors (not necessary 50 Ω) and provides the most compact design possible (with the least number of components), which is particularly useful in multi-element systems.
Subject(s)
Amplifiers, Electronic , Magnetic Resonance Imaging , Electric Impedance , Equipment DesignABSTRACT
Dissolution-DNP is a method to boost liquid-state NMR sensitivity by several orders of magnitude. The technique consists in hyperpolarizing samples by solid-state dynamic nuclear polarization at low temperature and moderate magnetic field, followed by an instantaneous melting and dilution of the sample happening inside the polarizer. Although the technique is well established and the outstanding signal enhancement paved the way towards many applications precluded to conventional NMR, the race to develop new methods allowing higher throughput, faster and higher polarization, and longer exploitation of the signal is still vivid. In this work, we review the most recent advances on dissolution-DNP methods trying to overcome the original technique's shortcomings. The review describes some of the new approaches in the field, first, in terms of sample formulation and properties, and second, in terms of instrumentation.
Subject(s)
Magnetic Resonance Imaging , Cold Temperature , Magnetic Fields , Magnetic Resonance Spectroscopy , SolubilityABSTRACT
PURPOSE: Cancer has a multitude of phenotypic expressions and identifying these are important for correct diagnosis and treatment selection. Clinical molecular imaging such as positron emission tomography can access several of these hallmarks of cancer non-invasively. Recently, hyperpolarized magnetic resonance spectroscopy with [1-13C] pyruvate has shown great potential to probe metabolic pathways. Here, we investigate simultaneous dual modality clinical molecular imaging of angiogenesis and deregulated energy metabolism in canine cancer patients. METHODS: Canine cancer patients (n = 11) underwent simultaneous [68Ga]Ga-NODAGA-E[(cRGDyK)]2 (RGD) PET and hyperpolarized [1-13C]pyruvate-MRSI (hyperPET). Standardized uptake values and [1-13C]lactate to total 13C ratio were quantified and compared generally and voxel-wise. RESULTS: Ten out of 11 patients showed clear tumor uptake of [68Ga]Ga-NODAGA-RGD at both 20 and 60 min after injection, with an average SUVmean of 1.36 ± 0.23 g/mL and 1.13 ± 0.21 g/mL, respectively. A similar pattern was seen for SUVmax values, which were 2.74 ± 0.41 g/mL and 2.37 ± 0.45 g/mL. The [1-13C]lactate generation followed patterns previously reported. We found no obvious pattern or consistent correlation between the two modalities. Voxel-wise tumor values of RGD uptake and lactate generation analysis revealed a tendency for each canine cancer patient to cluster in separated groups. CONCLUSION: We demonstrated combined imaging of [68Ga]Ga-NODAGA-RGD-PET for angiogenesis and hyperpolarized [1-13C]pyruvate-MRSI for probing energy metabolism. The results suggest that [68Ga]Ga-NODAGA-RGD-PET and [1-13C]pyruvate-MRSI may provide complementary information, indicating that hyperPET imaging of angiogenesis and energy metabolism is able to aid in cancer phenotyping, leading to improved therapy planning.
Subject(s)
Neoplasms , Pyruvic Acid , Acetates , Animals , Dogs , Gallium Radioisotopes , Heterocyclic Compounds, 1-Ring , Humans , Neoplasms/diagnostic imaging , Positron-Emission TomographyABSTRACT
Pancreatic ß-cells become irreversibly damaged by long-term exposure to excessive glucose concentrations and lose their ability to carry out glucose stimulated insulin secretion (GSIS) upon damage. The ß-cells are not able to control glucose uptake and they are therefore left vulnerable for endogenous toxicity from metabolites produced in excess amounts upon increased glucose availability. In order to handle excess fuel, the ß-cells possess specific metabolic pathways, but little is known about these pathways. We present a study of ß-cell metabolism under increased fuel pressure using a stable isotope resolved NMR approach to investigate early metabolic events leading up to ß-cell dysfunction. The approach is based on a recently described combination of 13C metabolomics combined with signal enhanced NMR via dissolution dynamic nuclear polarization (dDNP). Glucose-responsive INS-1 ß-cells were incubated with increasing concentrations of [U-13C] glucose under conditions where GSIS was not affected (2-8 h). We find that pyruvate and DHAP were the metabolites that responded most strongly to increasing fuel pressure. The two major divergence pathways for fuel excess, the glycerolipid/fatty acid metabolism and the polyol pathway, were found not only to operate at unchanged rate but also with similar quantity.
Subject(s)
Insulin-Secreting Cells/metabolism , Insulin-Secreting Cells/physiology , Animals , Cell Line , Fatty Acids/metabolism , Glucose/metabolism , Insulin/metabolism , Insulin Secretion/physiology , Lipid Metabolism/physiology , Metabolic Networks and Pathways/physiology , Metabolomics/methods , Pressure , Pyruvic Acid/metabolism , Rats , Signal Transduction/physiologyABSTRACT
Seaweeds contain many valuable compounds that can be used in the food industry. Carrageenan is a polysaccharide which has been extracted from seaweed for centuries and is used as a texturizer in food and non-food products. However, seaweeds contain compounds other than carrageenan, such as proteins, which could also be extracted. This extraction should be done without compromising the industrial scale carrageenan extraction yield and quality. This study aimed at up-stream protein extraction from red seaweed Eucheuma denticulatum by using of an optimized enzyme-assisted extraction, including of an aqueous/enzymatic treatment followed by alkaline extraction, and then the commercial carrageenan extraction. The protein extraction efficiency of four enzymes was evaluated including Celluclast® 1.5L, Shearzyme® 500 L, Alcalase® 2.4 L FG and Viscozyme® L at a concentration of 0.0, 0.1, 0.2 and 0.4% (w/w). To avoid detrimental effects on carrageenan, all the experiments were performed at pH 7 at room temperature. The results showed that 0.2% w/w Alcalase® or Viscozyme® added individually achieved the highest protein extraction efficiencies (59 and 48%, respectively) at pH 7 and room temperature (p < 0.05). Determination of the most common carrageenan quality parameters indicated that using any of these enzymes had no negative effect on the carrageenan yield and quality.