Use of 1H and 31P HRMAS to evaluate the relationship between quantitative alterations in metabolite concentrations and tissue features in human brain tumour biopsies

[EN] Quantitative multinuclear high-resolution magic angle spinning (HRMAS) was performed in order to determine the tissue pH values of and the absolute metabolite concentrations in 33 samples of human brain tumour tissue. Metabolite concentrations were quantified by 1D 1 H and 31P HRMAS using the electronic reference to in vivo concentrations (ERETIC) synthetic signal. 1 H–1 H homonuclear and 1 H–31P heteronuclear correlation experiments enabled the direct assessment of the 1 H–31P spin systems for signals that suffered from overlapping in the 1D 1 H spectra, and linked the information present in the 1D 1 H and 31P spectra. Afterwards, the main histological features were determined, and high heterogeneity in the tumour content, necrotic content and nonaffected tissue content was observed. The metabolite profiles obtained by HRMAS showed characteristics typical of tumour tissues: rather low levels of energetic molecules and increased concentrations of protective metabolites. Nevertheless, these characteristics were more strongly correlated with the total amount of living tissue than with the tumour cell contents of the samples alone, which could indicate that the sampling conditions make a significant contribution aside from the effect of tumour development in vivo. The use of methylene diphosphonic acid as a chemical shift and concentration reference for the 31P HRMAS spectra of tissues presented important drawbacks due to its interaction with the tissue. Moreover, the pH data obtained from 31P HRMAS enabled us to establish a correlation between the pH and the distance between the N(CH3)3 signals of phosphocholine and choline in 1 H spectra of the tissue in these tumour samples.The authors acknowledge the SCSIE-University of Valencia Microscopy Service for the histological preparations. They also acknowledge Martial Piotto (Bruker BioSpin, France) for providing the ERETIC synthetic signal. Furthermore, they acknowledge financial support from the Spanish Government project SAF2007-6547, the Generalitat Valenciana project GVACOMP2009-303, and the E.U.'s VI Framework Programme via the project "Web accessible MR decision support system for brain tumor diagnosis and prognosis, incorporating in vivo and ex vivo genomic and metabolomic data" (FP6-2002-LSH 503094). CIBER-BBN is an initiative funded by the VI National R&D&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.Esteve Moya, V.; Celda, B.; Martínez Bisbal, MC. (2012). Use of 1H and 31P HRMAS to evaluate the relationship between quantitative alterations in metabolite concentrations and tissue features in human brain tumour biopsies. Analytical and Bioanalytical Chemistry. 403:2611-2625. https://doi.org/10.1007/s00216-012-6001-zS26112625403Cheng LL, Chang IW, Louis DN, Gonzalez RG (1998) Cancer Res 58:1825–1832Opstad KS, Bell BA, Griffiths JR, Howe FA (2008) Magn Reson Med 60:1237–1242Sjobakk TE, Johansen R, Bathen TF, Sonnewald U, Juul R, Torp SH, Lundgren S, Gribbestad IS (2008) NMR Biomed 21:175–185Martinez-Bisbal MC, Marti-Bonmati L, Piquer J, Revert A, Ferrer P, Llacer JL, Piotto M, Assemat O, Celda B (2004) NMR Biomed 17:191–205Erb G, Elbayed K, Piotto M, Raya J, Neuville A, Mohr M, Maitrot D, Kehrli P, Namer IJ (2008) Magn Reson Med 59:959–965Wilson M, Davies NP, Brundler MA, McConville C, Grundy RG, Peet AC (2009) Mol Cancer 8:6Martinez-Bisbal MC, Monleon D, Assemat O, Piotto M, Piquer J, Llacer JL, Celda B (2009) NMR Biomed 22:199–206Martínez-Granados B, Monleón D, Martínez-Bisbal MC, Rodrigo JM, del Olmo J, Lluch P, Ferrández A, Martí-Bonmatí L, Celda B (2006) NMR Biomed 19:90–100Hubesch B, Sappey-Marinier D, Roth K, Meyerhoff DJ, Matson GB, Weiner MW (1990) Radiology 174:401–409Albers MJ, Krieger MD, Gonzalez-Gomez I, Gilles FH, McComb JG, Nelson MD Jr, Bluml S (2005) Magn Reson Med 53:22–29Wijnen JP, Scheenen TW, Klomp DW, Heerschap A (2010) NMR Biomed 23:968–976Podo F (1999) NMR Biomed 12:413–439Griffiths JR, Cady E, Edwards RH, McCready VR, Wilkie DR, Wiltshaw E (1983) Lancet 1:1435–1436Robitaille PL, Robitaille PA, Gordon Brown G, Brown GG (1991) J Magn Reson 92:73–84, 1969Griffiths JR (1991) Br J Cancer 64:425–427Payne GS, Troy H, Vaidya SJ, Griffiths JR, Leach MO, Chung YL (2006) NMR Biomed 19:593–598De Silva SS, Payne GS, Thomas V, Carter PG, Ind TE, deSouza NM (2009) NMR Biomed 22:191–198Wang Y, Cloarec O, Tang H, Lindon JC, Holmes E, Kochhar S, Nicholson JK (2008) Anal Chem 80:1058–1066Lehnhardt FG, Rohn G, Ernestus RI, Grune M, Hoehn M (2001) NMR Biomed 14:307–317Srivastava NK, Pradhan S, Gowda GA, Kumar R (2010) NMR Biomed 23:113–122Akoka S, Barantin L, Trierweiler M (1999) Anal Chem 71:2554–2557Albers MJ, Butler TN, Rahwa I, Bao N, Keshari KR, Swanson MG, Kurhanewicz J (2009) Magn Reson Med 61:525–532Ben Sellem D, Elbayed K, Neuville A, Moussallieh FM, Lang-Averous G, Piotto M, Bellocq JP, Namer IJ (2011) J Oncol 2011:174019Bourne R, Dzendrowskyj T, Mountford C (2003) NMR Biomed 16:96–101Martinez-Bisbal MC, Esteve V, Martinez-Granados B, Celda B (2011) J Biomed Biotechnol 2011:763684, Epub 2010 Sep 5Celda B, Montelione GT (1993) J Magn Reson B 101:189–193Esteve V, Celda B (2008) Magn Reson Mater Phys MAGMA 21:484–484Collins TJ (2007) Biotechniques 43:25–30Govindaraju V, Young K, Maudsley AA (2000) NMR Biomed 13:129–153Fan TW-M (1996) Prog Nucl Magn Reson Spectrosc 28:161–219Ulrich EL, Akutsu H, Doreleijers JF, Harano Y, Ioannidis YE, Lin J, Livny M, Mading S, Maziuk D, Miller Z, Nakatani E, Schulte CF, Tolmie DE, Kent Wenger R, Yao H, Markley JL (2008) Nucleic Acids Res 36:D402–D408Kriat M, Vion-Dury J, Confort-Gouny S, Favre R, Viout P, Sciaky M, Sari H, Cozzone PJ (1993) J Lipid Res 34:1009–1019Subramanian A, Shankar Joshi B, Roy AD, Roy R, Gupta V, Dang RS (2008) NMR Biomed 21:272–288Daykin CA, Corcoran O, Hansen SH, Bjornsdottir I, Cornett C, Connor SC, Lindon JC, Nicholson JK (2001) Anal Chem 73:1084–1090Griffin JL, Lehtimaki KK, Valonen PK, Grohn OH, Kettunen MI, Yla-Herttuala S, Pitkanen A, Nicholson JK, Kauppinen RA (2003) Cancer Res 63:3195–3201Petroff OAC, Prichard JW (1995) In: Kraicer J, Dixon SJ (eds) Methods in neurosciences. Academic, San DiegoBarton S, Howe F, Tomlins A, Cudlip S, Nicholson J, Anthony Bell B, Griffiths J (1999) Magn Reson Mater Phys Biol Med 8:121–128Sitter B, Sonnewald U, Spraul M, Fjosne HE, Gribbestad IS (2002) NMR Biomed 15:327–337Coen M, Hong YS, Cloarec O, Rhode CM, Reily MD, Robertson DG, Holmes E, Lindon JC, Nicholson JK (2007) Anal Chem 79:8956–8966Russell D, Rubinstein LJ (1998) Russel and Rubinstein's pathology of tumors of the nervous system. Arnold, LondonTynkkynen T, Tiainen M, Soininen P, Laatikainen R (2009) Anal Chim Acta 648:105–112Kjaergaard M, Brander S, Poulsen F (2011) J Biomol NMR 49:139–149Robert O, Sabatier J, Desoubzdanne D, Lalande J, Balayssac S, Gilard V, Martino R, Malet-Martino M (2011) Anal Bioanal Chem 399:987–999Chadzynski GL, Bender B, Groeger A, Erb M, Klose U (2011) J Magn Reson 212:55–63Weljie AM, Jirik FR (2011) Int J Biochem Cell Biol 43:981–989Barba I, Cabanas ME, Arus C (1999) Cancer Res 59:1861–1868Liimatainen T, Hakumaki JM, Kauppinen RA, Ala-Korpela M (2009) NMR Biomed 22:272–279Opstad KS, Bell BA, Griffiths JR, Howe FA (2008) NMR Biomed 21:677–685Schmitz JE, Kettunen MI, Hu D, Brindle KM (2005) Magn Reson Med 54:43–50Glunde K, Artemov D, Penet MF, Jacobs MA, Bhujwalla ZM (2010) Chem Rev 110:3043–3059Hertz L (2008) Neuropharmacology 55:289–309Takahashi T, Otsuguro K, Ohta T, Ito S (2010) Br J Pharmacol 161:1806–181

Multiproject–multicenter evaluation of automatic brain tumor classification by magnetic resonance spectroscopy

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Additive QTLs on three chromosomes control flowering time in woodland strawberry (Fragaria vesca L.)

Flowering time is an important trait that affects survival, reproduction and yield in both wild and cultivated plants. Therefore, many studies have focused on the identification of flowering time quantitative trait locus (QTLs) in different crops, and molecular control of this trait has been extensively investigated in model species. Here we report the mapping of QTLs for flowering time and vegetative traits in a large woodland strawberry mapping population that was phenotyped both under field conditions and in a greenhouse after flower induction in the field. The greenhouse experiment revealed additive QTLs in three linkage groups (LG), two on both LG4 and LG7, and one on LG6 that explain about half of the flowering time variance in the population. Three of the QTLs were newly identified in this study, and one co-localized with the previously characterized FvTFL1 gene. An additional strong QTL corresponding to previously mapped PFRU was detected in both field and greenhouse experiments indicating that gene(s) in this locus can control the timing of flowering in different environments in addition to the duration of flowering and axillary bud differentiation to runners and branch crowns. Several putative flowering time genes were identified in these QTL regions that await functional validation. Our results indicate that a few major QTLs may control flowering time and axillary bud differentiation in strawberries. We suggest that the identification of causal genes in the diploid strawberry may enable fine tuning of flowering time and vegetative growth in the closely related octoploid cultivated strawberry.Peer reviewe

In vivo magnetic resonance spectroscopy: basic methodology and clinical applications

The clinical use of in vivo magnetic resonance spectroscopy (MRS) has been limited for a long time, mainly due to its low sensitivity. However, with the advent of clinical MR systems with higher magnetic field strengths such as 3 Tesla, the development of better coils, and the design of optimized radio-frequency pulses, sensitivity has been considerably improved. Therefore, in vivo MRS has become a technique that is routinely used more and more in the clinic. In this review, the basic methodology of in vivo MRS is described—mainly focused on 1H MRS of the brain—with attention to hardware requirements, patient safety, acquisition methods, data post-processing, and quantification. Furthermore, examples of clinical applications of in vivo brain MRS in two interesting fields are described. First, together with a description of the major resonances present in brain MR spectra, several examples are presented of deviations from the normal spectral pattern associated with inborn errors of metabolism. Second, through examples of MR spectra of brain tumors, it is shown that MRS can play an important role in oncology

CRF-Like Diuretic Hormone Negatively Affects Both Feeding and Reproduction in the Desert Locust, Schistocerca gregaria

Diuretic hormones (DH) related to the vertebrate Corticotropin Releasing Factor (CRF) have been identified in diverse insect species. In the migratory locust, Locusta migratoria, the CRF-like DH (CRF/DH) is localized in the same neurosecretory cells as the Ovary Maturating Parsin (OMP), a neurohormone that stimulates oocyte growth, vitellogenesis and hemolymph ecdysteroid levels in adult female locusts. In this study, we investigated whether CRF-like DH can influence feeding and reproduction in the desert locust, Schistocerca gregaria. We identified two highly similar S. gregaria CRF-like DH precursor cDNAs, each of which also encodes an OMP isoform. Alignment with other insect CRF-like DH precursors shows relatively high conservation of the CRF/DH sequence while the precursor region corresponding to OMP is not well conserved. Quantitative real-time RT-PCR revealed that the precursor transcripts mainly occur in the central nervous system and their highest expression level was observed in the brain. Injection of locust CRF/DH caused a significantly reduced food intake, while RNAi knockdown stimulated food intake. Therefore, our data indicate that CRF-like DH induces satiety. Furthermore, injection of CRF/DH in adult females retarded oocyte growth and caused lower ecdysteroid titers in hemolymph and ovaries, while RNAi knockdown resulted in opposite effects. The observed effects of CRF/DH may be part of a wider repertoire of neurohormonal activities, constituting an integrating control system that affects food intake and excretion, as well as anabolic processes like oocyte growth and ecdysteroidogenesis, following a meal. Our discussion about the functional relationship between CRF/DH and OMP led to the hypothesis that OMP may possibly act as a monitoring peptide that can elicit negative feedback effects

Fluorescence-guided surgical sampling of glioblastoma identifies phenotypically distinct tumour-initiating cell populations in the tumour mass and margin.

BACKGROUND: Acquiring clinically annotated, spatially stratified tissue samples from human glioblastoma (GBM) is compromised by haemorrhage, brain shift and subjective identification of 'normal' brain. We tested the use of 5-aminolevulinic acid (5-ALA) fluorescence to objective tissue sampling and to derive tumour-initiating cells (TICs) from mass and margin. METHODS: The 5-ALA was administered to 30 GBM patients. Samples were taken from the non-fluorescent necrotic core, fluorescent tumour mass and non-fluorescent margin. We compared the efficiency of isolating TICs from these areas in 5-ALA versus control patients. HRMAS (1)H NMR was used to reveal metabolic alterations due to 5-ALA. We then characterised TICs for self-renewal in vitro and tumorigenicity in vivo. RESULTS: The derivation of TICs was not compromised by 5-ALA and the metabolic profile was similar between tumours from 5-ALA patients and controls. The TICs from the fluorescent mass were self-renewing in vitro and tumour-forming in vivo, whereas TICs from non-fluorescent margin did not self-renew in vitro but did form tumours in vivo. CONCLUSION: Our data show that 5-ALA does not compromise the derivation of TICs. It also reveals that the margin contains TICs, which are phenotypically different from those isolated from the corresponding mass