scholarly journals Investigation of Mitochondrial Adaptations to Modulation of Carbohydrate Supply during Adipogenesis of 3T3-L1 Cells by Targeted 1H-NMR Spectroscopy

Biomolecules ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 662
Author(s):  
Manon Delcourt ◽  
Virginie Delsinne ◽  
Jean-Marie Colet ◽  
Anne-Emilie Declèves ◽  
Vanessa Tagliatti
Keyword(s):  
1H Nmr ◽  
Culture Medium ◽  
High Sensitivity ◽  
Proton Nuclear Magnetic Resonance ◽  
Partial Replacement ◽  
Resonance Spectroscopy ◽  
Low Glucose ◽  
And Function ◽  
Targeted Approach ◽  
Mitochondrial Adaptation

(1) Background: White adipose tissue (WAT) is a dynamic and plastic tissue showing high sensitivity to carbohydrate supply. In such a context, the WAT may accordingly modulate its mitochondrial metabolic activity. We previously demonstrated that a partial replacement of glucose by galactose in a culture medium of 3T3-L1 cells leads to a poorer adipogenic yield and improved global mitochondrial health. In the present study, we investigate key mitochondrial metabolic actors reflecting mitochondrial adaptation in response to different carbohydrate supplies. (2) Methods: The metabolome of 3T3-L1 cells was investigated during the differentiation process using different glucose/galactose ratios and by a targeted approach using 1H-NMR (Proton nuclear magnetic resonance) spectroscopy; (3) Results: Our findings indicate a reduction of adipogenic and metabolic overload markers under the low glucose/galactose condition. In addition, a remodeling of the mitochondrial function triggers the secretion of metabolites with signaling and systemic energetical homeostasis functions. Finally, this study also sheds light on a new way to consider the mitochondrial metabolic function by considering noncarbohydrates related pathways reflecting both healthier cellular and mitochondrial adaptation mechanisms; (4) Conclusions: Different carbohydrates supplies induce deep mitochondrial metabolic and function adaptations leading to overall adipocytes function and profile remodeling during the adipogenesis.

scholarly journals Neuroendocrine Neoplasms: Identification of Novel Metabolic Circuits of Potential Diagnostic Utility

Cancers ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 374
Author(s):  
Beatriz Jiménez ◽  
Mei Ran Abellona U ◽  
Panagiotis Drymousis ◽  
Michael Kyriakides ◽  
Ashley K. Clift ◽  
...  
Keyword(s):  
1H Nmr ◽  
Cancer Metabolism ◽  
Proton Nuclear Magnetic Resonance ◽  
Neuroendocrine Neoplasms ◽  
Resonance Spectroscopy ◽  
Diagnostic Utility ◽  
Signalling Molecules ◽  
Prognostic Accuracy ◽  
Diagnostic Potential ◽  
Healthy Control

The incidence of neuroendocrine neoplasms (NEN) is increasing, but established biomarkers have poor diagnostic and prognostic accuracy. Here, we aim to define the systemic metabolic consequences of NEN and to establish the diagnostic utility of proton nuclear magnetic resonance spectroscopy (1H-NMR) for NEN in a prospective cohort of patients through a single-centre, prospective controlled observational study. Urine samples of 34 treatment-naïve NEN patients (median age: 59.3 years, range: 36–85): 18 had pancreatic (Pan) NEN, of which seven were functioning; 16 had small bowel (SB) NEN; 20 age- and sex-matched healthy control individuals were analysed using a 600 MHz Bruker 1H-NMR spectrometer. Orthogonal partial-least-squares-discriminant analysis models were able to discriminate both PanNEN and SBNEN patients from healthy control (Healthy vs. PanNEN: AUC = 0.90, Healthy vs. SBNEN: AUC = 0.90). Secondary metabolites of tryptophan, such as trigonelline and a niacin-related metabolite were also identified to be universally decreased in NEN patients, while upstream metabolites, such as kynurenine, were elevated in SBNEN. Hippurate, a gut-derived metabolite, was reduced in all patients, whereas other gut microbial co-metabolites, trimethylamine-N-oxide, 4-hydroxyphenylacetate and phenylacetylglutamine, were elevated in those with SBNEN. These findings suggest the existence of a new systems-based neuroendocrine circuit, regulated in part by cancer metabolism, neuroendocrine signalling molecules and gut microbial co-metabolism. Metabonomic profiling of NEN has diagnostic potential and could be used for discovering biomarkers for these tumours. These preliminary data require confirmation in a larger cohort.

scholarly journals Differential Effects of ReplacingEscherichiacoli Ribosomal Protein L27 with Its Homologue from Aquifexaeolicus

2001 ◽  
Vol 183 (22) ◽  
pp. 6565-6572 ◽  
Author(s):  
Bruce A. Maguire ◽  
Anton V. Manuilov ◽  
Robert A. Zimmermann
Keyword(s):  
Growth Rate ◽  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Ribosomal Subunit ◽  
Proton Nuclear Magnetic Resonance ◽  
Resonance Spectroscopy ◽  
Extreme Thermophile ◽  
Ribosomal Subunit Protein ◽  
And Function ◽  
Structure In Solution

ABSTRACT The rpmA gene, which encodes 50S ribosomal subunit protein L27, was cloned from the extreme thermophileAquifex aeolicus, and the protein was overexpressed and purified. Comparison of the A.aeolicus protein with its homologue fromEscherichia coli by circular dichroism analysis and proton nuclear magnetic resonance spectroscopy showed that it readily adopts some structure in solution that is very stable, whereas the E. coli protein is unstructured under the same conditions. A mutant of E.coli that lacks L27 was found earlier to be impaired in the assembly and function of the 50S subunit; both defects could be corrected by expression of E. coliL27 from an extrachromosomal copy of the rpmA gene. WhenA. aeolicus L27 was expressed in the same mutant, an increase in the growth rate occurred and the “foreign” L27 protein was incorporated into E. coliribosomes. However, the presence of A.aeolicus L27 did not promote 50S subunit assembly. Thus, while the A. aeolicus protein can apparently replace its E. coli homologue functionally in completed ribosomes, it does not assist in the assembly of E. coli ribosomes that otherwise lack L27. Possible explanations for this paradoxical behavior are discussed.

scholarly journals ISOLATION OF 2-CHLOROBENZIMIDAZOLE FROM MELIA DUBIA LEAF EXTRACT AND ITS STRUCTURAL CHARACTERISATION

2017 ◽  
Vol 9 (10) ◽  
pp. 67 ◽  
Author(s):  
G. Dayana Jeyaleela ◽  
S. Irudaya Monisha ◽  
J. Rosaline Vimala ◽  
A. Anitha Immaculate
Keyword(s):  
Mass Spectrometry ◽  
1H Nmr ◽  
Research Work ◽  
Proton Nuclear Magnetic Resonance ◽  
Resonance Spectroscopy ◽  
Ft Ir ◽  
Uv Visible ◽  
Isolated Compound ◽  
Promising Source

Objective: Natural products from medicinal plants, either as isolated compounds or as standardized plant extracts exhibit promising source of medicinal activity against various diseases. The aim of the present work was to make an attempt of isolation of bioactive principle and characterization of the isolated compound, from the medicinal plant Melia dubaiMethods: The extraction was done by a cold percolation method and the compound was separated and isolated by chromatography technique such as a thin layer chromatography (TLC), column chromatography and high-performance liquid chromatography (HPLC). The isolated compound was crystallized and the structural characterization of the isolated compound was made using UV-Visible, FT-IR, 1H-NMR, GC-MS and MS techniques which confirmed the structure of the isolated compound.Results: The separated and isolated compound was characterized by both physical and spectral methods like Ultraviolet-Visible spectroscopy (UV-Visible), Fourier transform infrared spectroscopy (FT-IR), Proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR), Gas chromatography-mass spectrometry (GC-MS), and Mass spectrometry(MS). Based on the studies, organizational characteristics of one bioactive principle were deciphered. The results revealed that the isolated species is 2-chlorobenzimidazole and it agreed well with the reported value and spectra for 2-chlorobenzimidazole.Conclusion: The above results obtained in this research work clearly indicated the promising occurrence of 2-chlorobenzimidazole in Media dubia plant leaves. The future scope of these studies may guide us to view the biological activity of the isolated compound.

scholarly journals Quantitative Analysis of Apoptotic Cell Death Using Proton Nuclear Magnetic Resonance Spectroscopy

Blood ◽  
1997 ◽  
Vol 89 (10) ◽  
pp. 3778-3786 ◽  
Author(s):  
Francis G. Blankenberg ◽  
Peter D. Katsikis ◽  
Richard W. Storrs ◽  
Christian Beaulieu ◽  
Daniel Spielman ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Cell Death ◽  
Magnetic Resonance ◽  
1H Nmr ◽  
Signal Intensity ◽  
Intensity Ratio ◽  
Apoptotic Cell ◽  
Proton Nuclear Magnetic Resonance ◽  
Resonance Spectroscopy ◽  
Signal Intensity Ratio

Abstract Quantification of apoptotic cell death in vivo has become an important area of investigation in patients with acute lymphoblastic leukemia (ALL). We have devised a noninvasive analytical method to estimate the percentage of apoptotic lymphoblasts in doxorubicin-treated Jurkat T-cell ALL cultures, using proton nuclear magnetic resonance spectroscopy (1H NMR). We have found that the ratio of the methylene (CH2 ) resonance (at 1.3 ppm) to the methyl (CH3 ) resonance (at 0.9 ppm) signal intensity, as observed by 1H NMR, is directly proportional to the percentage of apoptotic lymphoblasts in vitro. The correlation between the CH2/CH3 signal intensity ratio and the percentage of apoptotic lymphoblasts was optimal 24 to 28 hours after doxorubicin treatment (r2 = .947, N = 27 samples). There was also a direct temporal relationship between an increase in the CH2/CH3 signal intensity ratio and the onset of apoptosis as detected by nuclear morphologic analysis, fluorescein-annexin V flow cytometry, and DNA gel electrophoresis. Thin-layer chromatography confirmed that a dynamic and/or compositional change of the plasma membrane, rather than increases in lipase activity or fatty acid production, appears to account for the increase in the CH2/CH3 signal intensity ratio during apoptosis. 1H NMR may have clinical utility for the early noninvasive assessment of chemotherapeutic efficacy in patients with ALL.

scholarly journals Changes in hepatic metabolic profile during the evolution of STZ-induced diabetic rats via an 1H NMR-based metabonomic investigation

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Minjiang Chen ◽  
Hong Zheng ◽  
Min Xu ◽  
Liangcai Zhao ◽  
Qianqian Zhang ◽  
...  
Keyword(s):  
Amino Acids ◽  
1H Nmr ◽  
Metabolic Pathways ◽  
Metabolic Profile ◽  
Diabetic Rats ◽  
Proton Nuclear Magnetic Resonance ◽  
Control Group ◽  
Resonance Spectroscopy ◽  
Discriminate Analysis ◽  
Protective Function

Abstract Background: The present study aimed to explore the changes in the hepatic metabolic profile during the evolution of diabetes mellitus (DM) and verify the key metabolic pathways. Methods: Liver samples were collected from diabetic rats induced by streptozotocin (STZ) and rats in the control group at 1, 5, and 9 weeks after STZ administration. Proton nuclear magnetic resonance spectroscopy (1H NMR)-based metabolomics was used to examine the metabolic changes during the evolution of DM, and partial least squares-discriminate analysis (PLS-DA) was performed to identify the key metabolites. Results: We identified 40 metabolites in the 1H NMR spectra, and 11 metabolites were further selected by PLS-DA model. The levels of α-glucose and β-glucose, which are two energy-related metabolites, gradually increased over time in the DM rats, and were significantly greater than those of the control rats at the three-time points. The levels of choline, betaine, and methionine decreased in the DM livers, indicating that the protective function in response to liver injury may be undermined by hyperglycemia. The levels of the other amino acids (leucine, alanine, glycine, tyrosine, and phenylalanine) were significantly less than those of the control group during DM development. Conclusions: Our results suggested that the hepatic metabolic pathways of glucose, choline-betaine-methionine, and amino acids were disturbed during the evolution of diabetes, and that choline-betaine-methionine metabolism may play a key role.

Use of high-resolution proton nuclear magnetic resonance spectroscopy for rapid multi-component analysis of urine.

1984 ◽  
Vol 30 (3) ◽  
pp. 426-432 ◽  
Author(s):  
J R Bales ◽  
D P Higham ◽  
I Howe ◽  
J K Nicholson ◽  
P J Sadler
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Nmr Spectroscopy ◽  
Magnetic Resonance ◽  
High Resolution ◽  
1H Nmr ◽  
1H Nmr Spectroscopy ◽  
Proton Nuclear Magnetic Resonance ◽  
Resonance Spectroscopy ◽  
Simple Method ◽  
Nuclear Magnetic

Abstract Numerous low-Mr metabolites--including creatinine, citrate, hippurate, glucose, ketone bodies, and various amino acids--have been identified in 400- and 500-MHz proton nuclear magnetic resonance (1H NMR) spectra of intact human urine. The presence of many of these was related to the specific condition of the donors: humans in different physiological states (resting, fasting, or post-exercise) and pathological conditions (e.g., diabetes mellitus, cadmium-induced renal dysfunction). We have also monitored the metabolism of simple nonendogenous compounds (methanol and ethanol) and of acetaminophen. The pH-dependencies of the NMR chemical shifts of some urine components are reported. Our studies show that high-resolution 1H NMR spectroscopy provides a fast, simple method for "fingerprint" identification of urinary compounds. In some cases, analytes can be quantified by standard additions or by comparing integrated peak areas for the metabolites with those for creatinine. Determinations of creatinine by 1H NMR spectroscopy compared well with those by an independent chemical assay based on the Jaffé reaction.

scholarly journals The First Application of 1H NMR Spectroscopy for the Assessment of the Authenticity of Perfumes

2019 ◽  
Author(s):  
Grzegorz Ciepielowski ◽  
Barbara Pacholczyk-Sienicka ◽  
Łukasz Albrecht
Keyword(s):  
Nmr Spectroscopy ◽  
1H Nmr ◽  
1H Nmr Spectroscopy ◽  
Negative Impact ◽  
Poor Quality ◽  
Proton Nuclear Magnetic Resonance ◽  
Resonance Spectroscopy ◽  
Simple Method ◽  
Counterfeit Goods ◽  
Safety Issues

The industry of the counterfeit goods is one of the largest underground business in the world and it is rapidly growing. Counterfeits can lead not only to loss of profit for honest producers but also have a negative impact on consumers who receive poor quality goods at an excessive price and may be exposed to health damages and safety issues. Perfume industry is constantly exposed to the problem of counterfeits with the fast developing parallel market of inspired perfumes being an important issue. It prompts for the identification of methods that classify the quality of this type of products. In this paper the application of proton nuclear magnetic resonance spectroscopy is employed for the authentication of perfumery products for the first time. Molecular composition of several types of authentic brand fragrances for women were compared with their cheaper inspired equivalents and fake products. Our approach offers the prospect of a fast and simple method for discrimination and counterfeit detection of perfumes using 1H NMR spectroscopy.

scholarly journals An Optimization of Liquid–Liquid Extraction of Urinary Volatile and Semi-Volatile Compounds and Its Application for Gas Chromatography-Mass Spectrometry and Proton Nuclear Magnetic Resonance Spectroscopy

Molecules ◽  
2020 ◽  
Vol 25 (16) ◽  
pp. 3651
Author(s):  
Natalia Drabińska ◽  
Piotr Młynarz ◽  
Ben de Lacy Costello ◽  
Peter Jones ◽  
Karolina Mielko ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Nuclear Magnetic Resonance ◽  
Gas Chromatography ◽  
1H Nmr ◽  
Biomarker Discovery ◽  
Liquid Extraction ◽  
Proton Nuclear Magnetic Resonance ◽  
Gas Chromatography Mass Spectrometry ◽  
Resonance Spectroscopy ◽  
Liquid Liquid Extraction

Urinary volatile compounds (VCs) have been recently assessed for disease diagnoses. They belong to very diverse chemical classes, and they are characterized by different volatilities, polarities and concentrations, complicating their analysis via a single analytical procedure. There remains a need for better, lower-cost methods for VC biomarker discovery. Thus, there is a strong need for alternative methods, enabling the detection of a broader range of VCs. Therefore, the main aim of this study was to optimize a simple and reliable liquid–liquid extraction (LLE) procedure for the analysis of VCs in urine using gas chromatography-mass spectrometry (GC-MS), in order to obtain the maximum number of responses. Extraction parameters such as pH, type of solvent and ionic strength were optimized. Moreover, the same extracts were analyzed using Proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR), to evaluate the applicability of a single urine extraction for multiplatform purposes. After the evaluation of experimental conditions, an LLE protocol using 2 mL of urine in the presence of 2 mL of 1 M sulfuric acid and sodium sulphate extracted with dichloromethane was found to be optimal. The optimized method was validated with the external standards and was found to be precise and linear, and allowed for detection of >400 peaks in a single run present in at least 50% of six samples—considerably more than the number of peaks detected by solid-phase microextracton fiber pre-concentration-GC-MS (328 ± 6 vs. 234 ± 4). 1H-NMR spectroscopy of the polar and non-polar extracts extended the range to >40 more (mainly low volatility compounds) metabolites (non-destructively), the majority of which were different from GC-MS. The more peaks detectable, the greater the opportunity of assessing a fingerprint of several compounds to aid biomarker discovery. In summary, we have successfully demonstrated the potential of LLE as a cheap and simple alternative for the analysis of VCs in urine, and for the first time the applicability of a single urine solvent extraction procedure for detecting a wide range of analytes using both GC-MS and 1H-NMR analysis to enhance putative biomarker detection. The proposed method will simplify the transport between laboratories and storage of samples, as compared to intact urine samples.

scholarly journals PSIX-7 1H NMR-based plasma metabolomics reveals a potential biomarker of aflatoxin ingestion in dairy cows

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 395-396
Author(s):  
Ibukun M Ogunade ◽  
Yun Jiang
Keyword(s):  
Dairy Cows ◽  
1H Nmr ◽  
Fixed Effects ◽  
Plasma Concentrations ◽  
Latin Square ◽  
Proton Nuclear Magnetic Resonance ◽  
Control Treatment ◽  
Resonance Spectroscopy ◽  
Potential Biomarker ◽  
Biomarker Analysis

Abstract The study applied high-resolution proton nuclear magnetic resonance spectroscopy (1H NMR)-based plasma metabolomics to identify candidate biomarkers of aflatoxin B1 (AFB1) ingestion in dairy cows fed no sequestering agents and evaluate the effect of supplementing clay and/or a Saccharomyces cerevisiae fermentation product (SCFP) on such biomarkers. Eight lactating cows were randomly assigned to 1 of 4 treatments in a balanced 4 × 4 Latin square design with 2 squares. Treatments were control, toxin (T; 1725 µg AFB1/head/day), T with clay (CL; 200 g/head/day), and CL with SCFP (CL+SCFP; 35 g of SCFP/head/day). Cows in T, CL, and CL+SCFP were dosed with AFB1 from d 26 to 30. The sequestering agents were top-dressed from d 1 to 33. On d 30 of each period, 15 mL of blood was taken from the coccygeal vessels and plasma samples were prepared by centrifugation. Plasma sample (200 µL) was subjected to spectral analysis using a 700 MHz Avance III spectrometer. The metabolite data were subjected to statistical analysis using the GLIMMIX procedure of SAS. Biomarker analysis was done using metaboanalyst 4.0. The model included the fixed effects of treatment, period, interaction of treatment and period, and random effects of cow and square. Significance was declared at P ≤ 0.05. Compared to the control, T decreased (P ≤ 0.05) plasma concentrations of alanine, acetic acid, leucine, arginine, and valine. In contrast, T increased plasma ethanol concentration 3.56-fold compared to control. Treatment with CL+SCFP increased concentrations of mannose and 12 amino acids. Based on size of the area under the curve (AUC) of receiver operating characteristic and fold change (FC) analyses, ethanol was the most significantly altered metabolite in T (AUC = 0.88; FC = 3.56); hence, it was chosen as the candidate biomarker of aflatoxin ingestion in dairy cows fed no sequestering agent.

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