title: Analysis of Nucleosides and Nucleotides in Milk and Infant Formula

authors: Gill, Brendon D.

subjects: Nucleotide

abstract: Nucleotides have been routinely supplemented to infant formulas due to the important roles they play in metabolism and to replicate the higher concentrations typically found in human milk.  A method utilising anion exchange solid-phase extraction clean-up and liquid chromatography was developed for the rapid, routine determination of supplemented cytidine 5′ monophosphate, uridine 5′ monophosphate, inosine 5′ monophosphate, guanosine 5′ monophosphate, and adenosine 5′ monophosphate in bovine milk-based infant formula.  Chromatographic analyses were performed using a C18 stationary phase with gradient elution, UV detection, and quantitation by an internal standard technique.  A single-laboratory validation was performed, with recoveries of 92–101% and repeatability of 1.0–2.3%.  An extension study demonstrated the expansion in scope to a wider range of different infant formula products including milk protein and hydrolysate-based products, low and high fat products, soy protein-based and elemental products, adult nutritional and infant formulations, in both ready-to-feed and powder forms.
The development of a method to measure the total potentially available nucleosides (TPAN) in human milk has made an important contribution to further understanding the distribution of nucleosides and nucleotides.  This method was applied in a lactation study of bovine milk with colostrum and milk samples collected from two herds over the course of the first month post-partum, pooled within each herd by stage of lactation and the TPAN concentrations were determined.  Sample analysis consisted of parallel enzymatic treatments, phenylboronate affinity gel extraction, and liquid chromatography to quantify contributions of nucleosides, monomeric nucleotides, nucleotide adducts, and polymeric nucleotides to the nutritionally available nucleoside pool.  Bovine colostrum contained high levels of nucleosides and monomeric nucleotides, which rapidly decreased as lactation progressed into transitional milk.  Mature milk was relatively consistent in nucleoside and monomeric nucleotide concentrations from approximately the tenth day post-partum.  Differences in concentrations between summer-milk and winter-milk herds were largely attributable to variability in uridine and monomeric nucleotide concentrations.
The TPAN method was subsequently applied to the analysis of mature bovine, caprine, and ovine milk.  The contributions to TPAN from polymeric nucleotides, monomeric nucleotides, and nucleotide adducts were then calculated.  Ovine milk contained the highest concentration of TPAN (374.1 µmol dL-1), with lower concentrations in caprine milk (97.4 µmol dL-1) and bovine milk (7.9 µmol dL-1).  Ovine milk contained the highest concentrations of each of the different nucleoside and nucleotide forms, and bovine milk contained the lowest.
A method for the simultaneous analysis of nucleosides and nucleotides in infant formula using reversed-phase liquid chromatography-tandem mass spectrometry was developed.  Following sample dissolution, protein was removed by centrifugal ultrafiltration.  Chromatographic analyses were performed using a C18 stationary phase and gradient elution, with mass spectrometric detection, and quantitation by stable isotope labelled internal standard technique.  A single laboratory validation study was performed with recoveries of 80.1–112.9% and repeatability relative standard deviations of 1.9–7.2%.  The method was validated for the analysis of bovine milk-based, soy-based, caprine milk-based and hydrolysate-based infant formula.

text: Analysis of Nucleosides and Nucleotides in Milk and Infant Formula Nucleotides have been routinely supplemented to infant formulas due to the important roles they play in metabolism and to replicate the higher concentrations typically found in human milk.  A method utilising anion exchange solid-phase extraction clean-up and liquid chromatography was developed for the rapid, routine determination of supplemented cytidine 5′ monophosphate, uridine 5′ monophosphate, inosine 5′ monophosphate, guanosine 5′ monophosphate, and adenosine 5′ monophosphate in bovine milk-based infant formula.  Chromatographic analyses were performed using a C18 stationary phase with gradient elution, UV detection, and quantitation by an internal standard technique.  A single-laboratory validation was performed, with recoveries of 92–101% and repeatability of 1.0–2.3%.  An extension study demonstrated the expansion in scope to a wider range of different infant formula products including milk protein and hydrolysate-based products, low and high fat products, soy protein-based and elemental products, adult nutritional and infant formulations, in both ready-to-feed and powder forms.
The development of a method to measure the total potentially available nucleosides (TPAN) in human milk has made an important contribution to further understanding the distribution of nucleosides and nucleotides.  This method was applied in a lactation study of bovine milk with colostrum and milk samples collected from two herds over the course of the first month post-partum, pooled within each herd by stage of lactation and the TPAN concentrations were determined.  Sample analysis consisted of parallel enzymatic treatments, phenylboronate affinity gel extraction, and liquid chromatography to quantify contributions of nucleosides, monomeric nucleotides, nucleotide adducts, and polymeric nucleotides to the nutritionally available nucleoside pool.  Bovine colostrum contained high levels of nucleosides and monomeric nucleotides, which rapidly decreased as lactation progressed into transitional milk.  Mature milk was relatively consistent in nucleoside and monomeric nucleotide concentrations from approximately the tenth day post-partum.  Differences in concentrations between summer-milk and winter-milk herds were largely attributable to variability in uridine and monomeric nucleotide concentrations.
The TPAN method was subsequently applied to the analysis of mature bovine, caprine, and ovine milk.  The contributions to TPAN from polymeric nucleotides, monomeric nucleotides, and nucleotide adducts were then calculated.  Ovine milk contained the highest concentration of TPAN (374.1 µmol dL-1), with lower concentrations in caprine milk (97.4 µmol dL-1) and bovine milk (7.9 µmol dL-1).  Ovine milk contained the highest concentrations of each of the different nucleoside and nucleotide forms, and bovine milk contained the lowest.
A method for the simultaneous analysis of nucleosides and nucleotides in infant formula using reversed-phase liquid chromatography-tandem mass spectrometry was developed.  Following sample dissolution, protein was removed by centrifugal ultrafiltration.  Chromatographic analyses were performed using a C18 stationary phase and gradient elution, with mass spectrometric detection, and quantitation by stable isotope labelled internal standard technique.  A single laboratory validation study was performed with recoveries of 80.1–112.9% and repeatability relative standard deviations of 1.9–7.2%.  The method was validated for the analysis of bovine milk-based, soy-based, caprine milk-based and hydrolysate-based infant formula.

year: 2013

title: The Reduced Effect of International Conservation Agreements: A New Zealand Case Study

authors: Wallace, Philippa Jane

subjects: Science & Technology

abstract: Despite proliferation of law and policy directed at halting global biodiversity decline, it is a common concern that decline continues. Using a case study of six New Zealand birds, this article demonstrates reasons for the reduced effect of the law in the context of three international agreements: the Convention on Wetlands of International Importance; the Convention on Biological Diversity; and the Convention on Migratory Species of Wild Animals. The article concludes that the agreements are deficient in terms of obligation and consistency, and that these deficiencies are compounded by insufficient implementation and siloed approaches at the national level. Success is also compromised by the ways in which the law privileges resource use to the detriment of species due to insufficient environment standards, sectoral defences, and widespread externalities. Ironically for birds, mobility which in evolutionary terms has been a survival strategy, may become a liability in the Anthropocene.

text: The Reduced Effect of International Conservation Agreements: A New Zealand Case Study Despite proliferation of law and policy directed at halting global biodiversity decline, it is a common concern that decline continues. Using a case study of six New Zealand birds, this article demonstrates reasons for the reduced effect of the law in the context of three international agreements: the Convention on Wetlands of International Importance; the Convention on Biological Diversity; and the Convention on Migratory Species of Wild Animals. The article concludes that the agreements are deficient in terms of obligation and consistency, and that these deficiencies are compounded by insufficient implementation and siloed approaches at the national level. Success is also compromised by the ways in which the law privileges resource use to the detriment of species due to insufficient environment standards, sectoral defences, and widespread externalities. Ironically for birds, mobility which in evolutionary terms has been a survival strategy, may become a liability in the Anthropocene.

year: 2015-11-01

title: Harnessing Indigenous Tweets: The Reo Maori Twitter corpus

authors: Trye, David

subjects: Science & Technology

abstract: Te reo Māori, the Indigenous language of Aotearoa New Zealand, is a distinctive feature of the nation’s cultural heritage. This paper documents our efforts to build a corpus of 79,000 Māori-language tweets using computational methods. The Reo Māori Twitter (RMT) Corpus was created by targeting Māori-language users identified by the Indigenous Tweets website, pre-processing their data and filtering out non-Māori tweets, together with other sources of noise. Our motivation for creating such a resource is three-fold: (1) it serves as a rich and unique dataset for linguistic analysis of te reo Māori on social media; (2) it can be used as training data to develop and augment Natural Language Processing (NLP) tools with robust, real-world Māori-language applications; and (3) it will potentially promote awareness of, and encourage positive interaction with, the growing community of Māori tweeters, thereby increasing the use and visibility of te reo Māori in an online environment. While the corpus captures data from 2007 to 2020, our analysis shows that the number of tweets in the RMT Corpus peaked in 2014, and the number of active tweeters peaked in 2017, although at least 600 users were still active in 2020. To the best of our knowledge, the RMT Corpus is the largest publicly-available collection of social media data containing (almost) exclusively Māori text, making it a useful resource for language experts, NLP developers and Indigenous researchers alike.

text: Harnessing Indigenous Tweets: The Reo Maori Twitter corpus Te reo Māori, the Indigenous language of Aotearoa New Zealand, is a distinctive feature of the nation’s cultural heritage. This paper documents our efforts to build a corpus of 79,000 Māori-language tweets using computational methods. The Reo Māori Twitter (RMT) Corpus was created by targeting Māori-language users identified by the Indigenous Tweets website, pre-processing their data and filtering out non-Māori tweets, together with other sources of noise. Our motivation for creating such a resource is three-fold: (1) it serves as a rich and unique dataset for linguistic analysis of te reo Māori on social media; (2) it can be used as training data to develop and augment Natural Language Processing (NLP) tools with robust, real-world Māori-language applications; and (3) it will potentially promote awareness of, and encourage positive interaction with, the growing community of Māori tweeters, thereby increasing the use and visibility of te reo Māori in an online environment. While the corpus captures data from 2007 to 2020, our analysis shows that the number of tweets in the RMT Corpus peaked in 2014, and the number of active tweeters peaked in 2017, although at least 600 users were still active in 2020. To the best of our knowledge, the RMT Corpus is the largest publicly-available collection of social media data containing (almost) exclusively Māori text, making it a useful resource for language experts, NLP developers and Indigenous researchers alike.

year: 2022-02-14