All posts by Pau

Metabolomic fingerprinting of pig seminal plasma identifies in vivo fertility biomarkers

PREPRINT VERSION AT Journal of Animal Science and Biotechnology

Yentel Mateo-Otero, Pol Fernández-López, Ariadna Delgado-Bermúdez, Pau Nolis, Jordi Roca, Jordi Miró, Isabel Barranco, Marc Yeste


Metabolomic approaches, which include the study of low molecular weight molecules, is an emerging -omics technology useful for the identification of biomarkers. In this field, nuclear magnetic resonance (NMR) spectroscopy approach has already been used to uncover (in)fertility biomarkers in the seminal plasma (SP) of several mammalian species. However, NMR studies profiling SP metabolome to uncover in vivo fertility biomarkers are yet to be carried out in pigs. Thus, this study aimed to evaluate the putative relationship between the presence/concentration of SP-metabolites and in vivo fertility outcomes. To this end, 24 entire ejaculates (three ejaculates per boar) were collected from artificial insemination (AI)-boars throughout a year (one ejaculate every four months). Immediately after collection, ejaculates were centrifuged (1,500×g for 10 min twice) to obtain SP-samples and were stored (− 80°C) for subsequent metabolomic analysis by NMR spectroscopy. Fertility outcomes from 1,525 inseminations were recorded over a year, including farrowing rate, litter size, stillbirths per litter and the duration of pregnancy. These data were corrected to isolate the direct boar effect on each in vivo fertility parameter using a multivariate statistical model.


A total of 24 metabolites were identified and quantified in all SP-samples. ROC curve analysis showed that lactate levels in SP had discriminative capacity for farrowing rate (area under the curve (AUC) = 0.764; P < 0.05) while carnitine (AUC = 0.847), hypotaurine (AUC = 0.819), sn-glycero-3-phosphocholine (AUC = 0.833), glutamate (AUC = 0.799) and glucose (AUC = 0.750) had it for litter size (P < 0.05). Similarly, citrate (AUC = 0.743), creatine (AUC = 0.812), phenylalanine (AUC = 0.750), tyrosine (AUC = 0.753) and malonate (AUC = 0.868) levels had discriminative capacity for stillbirths per litter (P < 0.05); and malonate (AUC = 0.767) and fumarate (AUC = 0.868) concentrations for gestation length (P < 0.05).


Considering these results, the assessment of selected SP-metabolites in ejaculates through NMR spectroscopy could be considered as a promising non-invasive tool to predict in vivo fertility outcomes in pigs. Moreover, supplementing AI-doses with specific metabolites should also be contemplated as a way to improve their fertility potential.

Early socialization and environmental enrichment of lactating piglets affects the caecal microbiota and metabolomic response after weaning

M. Saladrigas-García, M. D’Angelo, H. L. Ko, S. Traserra, P. Nolis, Y. Ramayo-Caldas, J. M. Folch, P. Vergara, P. Llonch, J. F. Pérez & S. M. Martín-Orúe

Scientific Reports volume 11, Article number: 6113 (2021)


The aim of this study was to determine the possible impact of early socialization and an enriched neonatal environment to improve adaptation of piglets to weaning. We hypothesized that changes in the microbiota colonization process and in their metabolic response and intestinal functionality could help the animals face weaning stress. A total of 48 sows and their litters were allotted into a control (CTR) or an enriched treatment (ENR), in which piglets from two adjacent pens were combined and enriched with toys. The pattern of caecal microbial colonization, the jejunal gene expression, the serum metabolome and the intestinal physiology of the piglets were assessed before (-2 d) and after weaning (+ 3d). A differential ordination of caecal microbiota was observed after weaning. Serum metabolome suggested a reduced energetic metabolism in ENR animals, as evidenced by shifts in triglycerides and fatty acids, VLDL/LDL and creatine regions. The TLR2 gene showed to be downregulated in the jejunum of ENR pigs after weaning. The integration of gene expression, metabolome and microbiota datasets confirmed that differences between barren and enriched neonatal environments were evident only after weaning. Our results suggest that improvements in adaptation to weaning could be mediated by a better response to the post-weaning stress.

Measuring Long-Range Heteronuclear NMR Correlations in Natural Products

LR-HSQMBC versus LR-selHSQMBC: Enhancing the Observation ofTiny Long-Range Heteronuclear NMR Correlations

Kumar Motiram-Corral, Pau Nolis, Josep Saurí, and Teodor Parella


ABSTRACT:The detection of ultra-long-range (4JCHand higher) heteronuclear connectivities can complement the conventionaluse of HMBC/HSQMBC data in structure elucidation NMR studies of proton-deficient natural products, where two-bond andthree-bond correlations are usually observed. The performance of the selHSQMBC experiment with respect to its broadbandHSQMBC counterpart is evaluated. Despite its frequency-selectivity nature, selHSQMBC efficiently prevents any unwanted signalphase and intensity modulations due to passive proton−proton coupling constants typically involved in HSQMBC. As a result,selHSQMBC offers a significant sensitivity enhancement and provides pure in-phase multiplets, improving the detection levels forshort- and long-range cross-peaks corresponding to small heteronuclear coupling values. This is particularly relevant for experimentsoptimized to smallnJCHvalues (2−3 Hz), referred to as LR-selHSQMBC, where key cross-peaks that are not visible in the equivalentbroadband LR-HSQMBC spectrum can become observable in optimum conditions.

Removal of the synthetic hormone methyltestosterone from aqueous solution using a β-cyclodextrin/silica composite

Lucas Bragança Carvalho, Pricila Maria BatistaChagas, Tamara Rezende Marques, Angelina Razafitianamaharavo, Manuel Pelletier, Pau Nolis, Carlos Jaime, Sérgio Scherrer Thomasi, Lucianade Matos Alves Pinto.

Journal of Environmental Chemical Engineering – Available online 20 October 2019

ABSTRACT Contamination of water with steroid residues can cause a number of environmental damages, affecting exposed organisms including man. The development of technologies for treatment or removal of this type of micropollutant from water is of paramount importance. In this study, citric acid was used to functionalize β-cyclodextrin (bCD) on the silica surface generating an organic-inorganic hybrid composite for application in molecular sequestration. The functionalization percentage was high, with about 62.6% of the composite mass corresponding to the organic part of the material. 13C NMR and infrared spectroscopic analysis indicate that the functionalization mechanism occurs by an esterification reaction between the citric acid with the silanol groups from silica and the primary hydroxyls of the bCDs. Fast adsorption of the methyltestosterone steroid was observed at acid pH, with a high adsorption capacity of 11 mg g-1. The best kinetic and isotherm models fit indicated that the adsorption occurred by a physical mechanism at independent sites with the steroid molecule possibly captured by two bCDs. The removal process was spontaneous and exothermic, with the existence of weak interactions between the hormone and the composite, and its regeneration is quite fast efficient with the displacement of the complexation equilibrium. The results obtained in this study demonstrate the considerable potential of the composite for use in the treatment of wastewater containing the steroid studied, and its efficacy should be evaluated for other steroid molecules. 

Measuring Long-Range Heteronuclear Coupling Constants with Sel-HSQMBC Experiments: A Tutorial

How to measure long‐range proton‐carbon coupling constants from 1H‐selective HSQMBC experiments, by Josep Saurí, Pau Nolis and Teodor Parella. Magn. Reson. Chem. 2019. Early View, DOI:

Heteronuclear long‐range scalar coupling constants (nJCH) are a valuable tool for solving problems in organic chemistry and are especially suited for stereochemical and configurational analyses of small molecules and natural products. This tutorial will focus on the step‐by‐step implementation of several 2D 1H frequency selective HSQMBC experiments for the easy and accurate measurement of either the magnitude or both the magnitude and the sign of long‐range nJCH couplings. The performance of these experiments will be showcased with several scenarios in a range of different experimental conditions.

Bruker pulse program code for selHSQMBC experiments available here.

Bruker pulse program code for selHSQMBC-TOCSY experiment available here.

Strychnine dataset examples available here.

Rethinking COSY and TOCSY acquisition

Nolis, Pau & Teodor Parella. 2019. Practical aspects of the simultaneous collection of COSY and TOCSY spectra. Magnetic Resonance in Chemistry. DOI: 10.1002/mrc.4835

The practical aspects of some NMR experiments designed for the simultaneous acquisition of 2D COSY and 2D TOCSY spectra are presented and discussed. Several techniques involving afterglow‐based, CTP‐based and NOAH‐based strategies for the collection of different FIDs within the same scan are evaluated and compared. These methods offer a faster recording of these spectra in small‐molecule NMR when sensitivity is not a limiting factor, with a reduction in spectrometer time about 45%‐60% when compared to the conventional sequential acquisition of the parent experiments. It is also shown how the optimized design of an extended three‐FID approach yields one COSY and two TOCSY spectra simultaneously by combining CTP and NOAH principles in the same experiment, affording substantial sensitivity enhancements per time unit.

New methodology to obtain High Resolution aliased HSQC

Nolis, Pau, Kumar Motiram-Corral, Míriam Pérez-Trujillo & Teodor Parella. 2019. Simultaneous acquisition of two 2D HSQC spectra with different 13C spectral widths. Journal of Magnetic Resonance. DOI: 10.1016/j.jmr.2019.01.004

ABSTRACT: A time-efficient NMR strategy that involves the interleaved acquisition of two 2D HSQC spectra having different spectral widths in the indirect 13C dimension is presented. We show how the two equivalent coherence transfer pathways involved in sensitivity-enhanced HSQC experiments are managed selectively and detected separately in different FID periods within the same scan. The feasibility of this new SADA-HSQC (Spectral Aliasing in Dually Acquired HSQC) technique is demonstrated by recording simultaneously two complementary datasets, conventional and highly-resolved spectral-aliased 2D HSQC spectra, in a single NMR experiment. Combining the information from both datasets, accurate chemical shift determination and excellent signal dispersion is achieved in a unique measurement using only few t1 increments.

Solid State NMR supports a new study on hydride composite for hydrogen storage

Bergemann, N., C. Pistidda, C. Milanese, M. Aramini, S. Huotari, P. Nolis, A. Santoru, M. R. Chierotti, A.-L. Chaudhary, M. D. Baro, T. Klassen & M. Dornheim. 2018. A hydride composite featuring mutual destabilisation and reversible boron exchange: Ca(BH4)2–Mg2NiH4. Journal of Materials Chemistry A 6(37). 17929–17946. DOI: 10.1039/c8ta04748k

The system Ca(BH4)2–Mg2NiH4 is used as a model to prove the unique possibility to fully reverse the borohydride decomposition process even in cases where the decomposition reaction leads to undesired stable boron containing species (boron sinks). The formation of MgNi2.5B2 directly from Ca(BH4)2 or from CaB12H12 and amorphous boron allows an unexpectedly easy transfer of the boron atoms to reversibly form Ca(BH4)2 during rehydrogenation. In addition, to the best of our knowledge, the mutual destabilisation of the starting reactants is observed for the first time in Ca(BH4)2 based Reactive Hydride Composite (RHC) systems. A detailed account of dehydrogenation and rehydrogenation reaction mechanisms as the function of applied experimental conditions is given.