Breastmilk mineral composition among well-educated mothers from Central Poland - Associations with maternal dietary intake, dietary patterns and infant psychomotor development.

BACKGROUND: Maternal dietary habits could affect breastmilk mineral composition, which may influence infant development. Mineral dietary intake or supplementation slightly affects its breastmilk concentration. However, the intake of selected food groups or dietary patterns that reflect diet complexity could have a greater impact. Hence, the aim of the study was to assess breastmilk mineral composition at one, three, and six months of lactation among mothers living in urban area of Central Poland, as well as the evaluate maternal dietary determinants and associations with infant anthropometric and psychomotor development. METHODS: The study was conducted among 43 healthy and exclusively breastfeeding mothers. In the first, third, and sixth months of lactation, we collected breastmilk samples and assessed the concentration of Ca, P, Zn, Fe, Se, Ni, As, Pb, and Cd using the ICP-MS method. Maternal dietary habits were evaluated by a food frequency questionnaire in the first month of lactation, whereas in the third and sixth by the three-day food record. Based on the collected data adherence to the Polish-adapted Mediterranean (Pl-aMED; 1 month) and the DASH diet (Mellen's Index; 3 and 6 months) was assessed. In the third and sixth months of lactation infant anthropometric parameters and the sixth month of lactation psychomotor development were evaluated. RESULTS: Breastmilk Se, Ni, As, Pb, and Cd levels were under the LOQ in all the breastmilk samples at all study visits. Median breastmilk mineral concentrations of Ca, P, Zn, and Fe in the first, third, and sixth months of lactation varied from 381.9 to 332.7 mg/L, 161.6 to 139.1 mg/L, 2.2 to 0.8 mg/L, and 0.26 to 0.17 mg/L, respectively. Maternal dietary intake and supplementation did not affect breastmilk Ca, P, Zn, and Fe. Pl-aMED scores were associated with breastmilk Ca (ß = 0.489, 95% CI 0.180 - 0.799, p = 0.003) and Zn (ß = 0.499, 95% CI 0.199 - 0.798, p = 0.002) in the first month of lactation, whereas no association with the DASH diet were observed in the third and sixth month of lactation. Breastmilk Fe in the third month was associated with infant motor development (ß = 0.420, 95% CI 0.113 - 0.727, p = 0.009) in the sixth month of life, but no other associations with anthropometric or psychomotor development were observed. Moreover, we estimated that few infants meet their adequate intake (AI) requirements for P, Zn, and Fe. CONCLUSION: Our study showed that maternal adherence to Pl-aMED is a significant predictor of breastmilk Ca and Zn in the first month of lactation, which may be especially important considering that more than 75% of infants had inadequate Zn intake. Moreover, we found that breastmilk Fe positively influenced infant motor development, despite the majority of infants having inadequate intake. On the other hand, no infant had deficiency symptoms, which emphasizes the necessity to evaluate of AI norms for infants.

Fractionation of selenium isotopes during biofortification of Saccharomyces cerevisiae and the influence of metabolic labeling with 15N.

Isotope fractionation of metals/metalloids in biological systems is an emerging research area that demands the application of state-of-the-art analytical chemistry tools and provides data of relevance to life sciences. In this work, Se uptake and Se isotope fractionation were measured during the biofortification of baker's yeast (Saccharomyces cerevisiae)-a product widely used in dietary Se supplementation and in cancer prevention. On the other hand, metabolic labeling with 15N is a valuable tool in mass spectrometry-based comparative proteomics. For Se-yeast, such labeling would facilitate the assessment of Se impact on yeast proteome; however, the question arises whether the presence of 15N in the microorganisms affects Se uptake and its isotope fractionation. To address the above-mentioned aspects, extracellularly reduced and cell-incorporated Se fractions were analyzed by hydride generation-multi-collector inductively coupled plasma-mass spectrometry (HG MC ICP-MS). It was found that extracellularly reduced Se was enriched in light isotopes; for cell-incorporated Se, the change was even more pronounced, which provides new evidence of mass fractionation during biological selenite reduction. In the presence of 15N, a weaker preference for light isotopes was observed in both, extracellular and cell-incorporated Se. Furthermore, a significant increase in Se uptake for 15N compared to 14N biomass was found, with good agreement between hydride generation microwave plasma-atomic emission spectrometry (HG MP-AES) and quadrupole ICP-MS results. Biological effects observed for heavy nitrogen suggest 15N-driven alteration at the proteome level, which facilitated Se access to cells with decreased preference for light isotopes.

Maternal diet during breastfeeding in correlation to calcium and phosphorus concentrations in human milk.

BACKGROUND: The impact of maternal diet on mineral concentration in human milk (HM) remains unclear. The main aim of this study was to investigate the relationship between maternal dietary intake and calcium and phosphorus concentrations in HM. Furthermore, we aimed to evaluate the intake of both minerals by exclusively breastfed infants. METHODS: HM samples were obtained from 30 mothers at 6-8 weeks postpartum. Each mother was asked to express pre- and postfeeding milk four times during a 24-h period (6.00-12.00, 12.00-18.00, 18.00-24.00, 24.00-6.00). Maternal dietary assessment was based on a food frequency questionnaire and 3-day dietary records. Analysed minerals were determined using an inductively coupled plasma mass spectrometer (NexION 300D ICP mass spectrometer, Perkin Elmer SCIEX). RESULTS: The mean concentrations of calcium and phosphorus in HM samples were 278.7 ± 61.0 and 137.1 ± 21.9 mg/L, respectively, maintaining 2:1 ratio by weight. The concentration of both minerals was correlated with each other (r = 0.632, p = <0.001). The infants' mean calcium intake was 149.53 ± 36.41 mg/L, and their mean phosphorus intake was 74.62 ± 19.41 mg/L. The risk of insufficient intake of calcium was reported in 60% of infants (n = 18). Spearman's/Pearson's correlation coefficients did not reveal any correlations between HM calcium concentration and maternal diet, contrary to HM phosphorus concentration, which was positively correlated with energy (r = 0.369, p = 0.045), total protein (r = 0.464, p = 0.01), calcium (r = 385, p = 0.036), phosphorus (r = 501, p = 0.005), niacin (p < 0.001) and pyridoxine (r = 382, 0.037) intake. However, in multivariable analysis we observed that maternal dietary intake of both minerals had a positive influence on their concentration in HM. CONCLUSIONS: Maternal calcium and phosphorus intake influenced the concentration of both minerals in HM; however, the relationship was rather weak. In addition, we observed that calcium intake by most of the exclusively breastfed infants was insufficient to meet the recommended daily intake.

Label-Free Mass Spectrometry-Based Proteomic Analysis in Lamb Tissues after Fish Oil, Carnosic Acid, and Inorganic Selenium Supplementation.

Selenium is an essential nutrient, building twenty five identified selenoproteins in humans known to perform several important biological functions. The small amount of selenium in the earth's crust in certain regions along with the risk of deficiency in organisms have resulted in increasingly popular dietary supplementation in animals, implemented via, e.g., inorganic selenium compounds. Even though selenium is included in selenoproteins in the form of selenocysteine, the dietary effect of selenium may result in the expression of other proteins or genes. Very little is known about the expression effects modulated by selenium. The present study aimed to examine the significance of protein expression in lamb tissues obtained after dietary supplementation with selenium (sodium selenate) and two other feed additives, fish oil and carnosic acid. Label-free mass spectrometry-based proteomic analysis was successfully applied to examine the animal tissues. Protein-protein interaction network analysis of forty differently-expressed proteins following inorganic selenium supplementation indicated two significant clusters which are involved in cell adhesion, heart development, actin filament-based movement, plasma membrane repair, and establishment of organelle localization.

Cancer Influences the Elemental Composition of the Myocardium More Strongly than Conjugated Linoleic Acids-Chemometric Approach to Cardio-Oncological Studies.

The aim of the study was to verify in a cardio-oncological model experiment if conjugated linoleic acids (CLA) fed to rats with mammary tumors affect the content of selected macro- and microelements in their myocardium. The diet of Sprague-Dawley females was supplemented either with CLA isomers or with safflower oil. In hearts of rats suffering from breast cancer, selected elements were analyzed with a quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). In order to better understand the data trends, cluster analysis, principal component analysis and linear discriminant analysis were applied. Mammary tumors influenced macro- and microelements content in the myocardium to a greater extent than applied diet supplementation. Significant influences of diet (p = 0.0192), mammary tumors (p = 0.0200) and interactions of both factors (p = 0.0151) were documented in terms of Fe content. CLA significantly decreased the contents of Cu and Mn (p = 0.0158 and p = 0.0265, respectively). The level of Ni was significantly higher (p = 0.0073), which was more pronounced in groups supplemented with CLA. The obtained results confirmed antioxidant properties of CLA and the relationship with Se deposition. Chemometric techniques distinctly showed that the coexisting pathological process induced differences to the greater extent than diet supplementation in the elemental content in the myocardium, which may impinge on cardiac tissue's susceptibility to injuries.

Evaluation of the influence of diet supplementation with conjugated linoleic acid isomers on elemental composition in the cardio-oncological nutritional programming rat' model.

BACKGROUND: The 'developmental origin of health and disease' (DOHaD) hypothesis assumes that due to the action of some stimuli during fetal life the long-term physiological changes occurs and may affects the risk of various diseases. The main aim of this study was to assess impact of supplementation of maternal' and early postnatal diet with conjugated linoleic acids (CLA) isomers on selected elements content in hearts of their female offspring with chemically induced breast cancer. METHODS: Elemental composition was determined by quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). RESULTS: The effect of maternal' diet on the elements content was more pronounced than the progeny diet modifications. Significant correlations among determined elements, especially macroelements, and lipidomic parameters, in the experimental factors dependent manner were observed. It can be concluded that supplementation of maternal and progeny diets with CLA isomers may significantly influence the contents of both macro- and microelements in cardiac tissue of newborns. CONCLUSION: Our results also indicate, that dynamic and intricate balance among various elements in body may be affected by the lipid dietary supplements also in the pathological state. Utility of cardio-oncological approach in developmental programming study was confirmed.

The matrix metalloproteinase inhibitor marimastat inhibits seizures in a model of kainic acid-induced status epilepticus.

An intra-hippocampus injection of kainic acid serves as a model of status epilepticus and the subsequent development of temporal lobe epilepsy. Matrix metalloproteinase-9 (MMP-9) is an enzyme that controls remodeling of the extracellular milieu under physiological and pathological conditions. In response to brain insult, MMP-9 contributes to pathological synaptic plasticity that may play a role in the progression of an epileptic condition. Marimastat is a metalloproteinase inhibitor that was tested in clinical trials of cancer. The present study assessed whether marimastat can impair the development of epilepsy. The inhibitory efficacy of marimastat was initially tested in neuronal cultures in vitro. As a marker substrate, we used nectin-3. Next, we investigated the blood-brain barrier penetration of marimastat using mass spectrometry and evaluated the therapeutic potential of marimastat against seizure outcomes. We found that marimastat inhibited the cleavage of nectin-3 in hippocampal neuronal cell cultures. Marimastat penetrated the blood-brain barrier and exerted an inhibitory effect on metalloproteinase activity in the brain. Finally, marimastat decreased some seizure parameters, such as seizure score and number, but did not directly affect status epilepticus. The long-term effects of marimastat were evident up to 6 weeks after kainic acid administration, in which marimastat still inhibited seizure duration.

Searching for Low Molecular Weight Seleno-Compounds in Sprouts by Mass Spectrometry.

A fit for purpose analytical protocol was designed towards searching for low molecular weight seleno-compounds in sprouts. Complementary analytical techniques were used to collect information enabling the characterization of selenium speciation. Conceiving the overall characterization of the behavior of selenium, inductively plasma optical mass spectrometry (ICP-MS) was used to determine the total selenium content in entire sprouts as well as in selected extracts or chromatographic fractions. Then, high-performance liquid chromatography combined with ICP-MS (HPLC-ICP-MS) was used to evaluate the presence of inorganic and organic seleno-compounds, with the advantages of being very sensitive towards selenium, but limited by available selenium standard compounds. Finally, ultra-high performance liquid chromatography electrospray ionization triple quadrupole mass spectrometry (UHPLC-ESI-QqQ-MS/MS) and UHPLC-ESI-Orbitrap-MS/MS were used for the confirmation of the identity of selected compounds and identification of several unknown compounds of selenium in vegetable sprouts (sunflower, onion, radish), respectively. Cultivation of plants was designed to supplement sprouts with selenium by using solutions of selenium (IV) at the concentration of 10, 20, 40, and 60 mg/L. The applied methodology allowed to justify that vegetable sprouts metabolize inorganic selenium to a number of organic derivatives, such as seleno-methylselenocysteine (SeMetSeCys), selenomethionine (SeMet), 5'-seleno-adenosine, 2,3-DHP-selenolanthionine, Se-S conjugate of cysteine-selenoglutathione, 2,3-DHP-selenocysteine-cysteine, 2,3-DHP-selenocysteine-cysteinealanine, glutathione-2,3-DHP-selenocysteine, gamma-Glu-MetSeCys or glutamyl-glycinyl-N-2,3-DHP-selenocysteine.

Determination of Selenium Species in Muscle, Heart, and Liver Tissues of Lambs Using Mass Spectrometry Methods.

Identification and quantification of the selenium species in biological tissues is imperative, considering the need to properly understand its metabolism and its importance in various field of sciences, especially nutrition science. Although a number of studies deals with the speciation of selenium, speciation analysis is still far from being a routine task, and so far strongly depends on the type of the samples. We present a study aimed to examine speciation analysis of Se in tissues of livers, muscles, and hearts obtained from lambs, namely in liver, muscle, and heart. The studied lambs were fed with the diet enriched with an inorganic (as sodium selenate) and organic chemical form of Se (as Se-enriched yeast) compounds with simultaneous addition of fish oil (FO) and carnosic acid (CA). The first part of the work was focused on the optimization of the extraction procedure of selenium compounds from tissues. Next, hyphenated high performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) was used for the identification of five seleno-compounds-Se-methionine (SeMet), Se-cystine (SeCys2), Se-methyl-Se-cysteine (SeMetSeCys), and Se(IV) and Se(VI). Verification of the identified seleno-compounds was achieved using triple-quadrupole mass spectrometer coupled to high performance liquid chromatography (HPLC-ESI-MS/MS). The applied procedure allowed for quantitative analysis of SeMet, SeCys2, and SeMetSeCys, in biological tissues. The developed analytical protocol is feasible for speciation analysis of small molecular seleno-compounds in animals samples.

Magnesium-Isotope Fractionation in Chlorophyll-a Extracted from Two Plants with Different Pathways of Carbon Fixation (C3, C4).

Relatively few studies have been focused so far on magnesium-isotope fractionation during plant growth, element uptake from soil, root-to-leaves transport and during chlorophylls biosynthesis. In this work, maize and garden cress were hydroponically grown in identical conditions in order to examine if the carbon fixation pathway (C4, C3, respectively) might have impact on Mg-isotope fractionation in chlorophyll-a. The pigment was purified from plants extracts by preparative reversed phase chromatography, and its identity was confirmed by high-resolution mass spectrometry. The green parts of plants and chlorophyll-a fractions were acid-digested and submitted to ion chromatography coupled through desolvation system to multiple collector inductively coupled plasma-mass spectrometry. Clear preference for heavy Mg-isotopes was found in maize green parts (∆26Mgplant-nutrient 0.65, 0.74 for two biological replicates, respectively) and in chlorophyll-a (∆26Mgchlorophyll-plant 1.51, 2.19). In garden cress, heavy isotopes were depleted in green parts (∆26Mgplant-nutrient (-0.87)-(-0.92)) and the preference for heavy isotopes in chlorophyll-a was less marked relative to maize (∆26Mgchlorophyll-plant 0.55-0.52). The observed effect might be ascribed to overall higher production of energy in form of adenosine triphosphate (ATP), required for carbon fixation in C4 compared to C3, which could reduce kinetic barrier and make equilibrium fractionation prevailing during magnesium incorporation to protoporphyrin ring.

Seleno-compounds and Carnosic Acid Added to Diets with Rapeseed and Fish Oils Affect Concentrations of Selected Elements and Chemical Composition in the Liver, Heart and Muscles of Lambs.

The objective of our studies was to investigate effects of carnosic acid (CA), selenized yeast (SeY) and selenate (SeVI) added to the diet including rapeseed oil (RO) and fish oil (FO) on concentrations of elements, fatty acids (FAs), tocopherols, cholesterol, and malondialdehyde in the liver, heart, musculus longissimus dorsi (MLD), and musculus biceps femoris (MBF) of lambs. Lambs were fed diets: group I-the basal diet (BD) with RO; group II-BD with RO and FO; group III-BD with RO, FO, and CA; group IV-BD with RO, FO, CA, and SeY; group V-BD with RO, FO, CA, and SeVI. The diets with Se compounds increased Se concentrations in all tissues compared with other diets. The diet with SeVI increased Cd, Sb, and Pb concentrations in the liver compared to groups I, II, and IV. The diets containing Se compounds increased Sb and Pb concentrations in MBF compared to groups I and II. All diets with CA reduced As, Sb, and Pb concentrations in MLD compared to groups I and II. All diets with FO increased concentrations of FAs and malondialdehyde in the liver compared to group I. All diets with FO decreased FAs concentrations in MBF compared to group I. The diets containing CA with/without Se compounds increased malondialdehyde concentrations in MBF compared to groups I and II. The diet with Se compounds reduced malondialdehyde concentrations in MLD compared to group II. All diets with FO changed concentrations of tocopherols and cholesterol in all tissues compared to group I. Our study showed that the addition of SeY or SeVI to the experimental diet increased the concentration of Se in all assayed tissues of lambs without adversely influencing performance or causing physiological disorders in internal organs. Both, SeY or SeVI added to the experimental diet decreased the oxidative stress and the concentrations of As, Sb, and Pb in MLD compared with the diets containing RO, irrespective of the presence of FO (groups I and II). Our study provides useful knowledge for nutritionists carrying out further investigations aimed at improving farm animal health, performance, and the nutritional quality of animal products for humans.

Allium cepa L. Response to Sodium Selenite (Se(IV)) Studied in Plant Roots by a LC-MS-Based Proteomic Approach.

Liquid chromatography-high-resolution mass spectrometry was used for the first time to investigate the impact of Se(IV) (10 mgSe L-1 as sodium selenite) on Allium cepa L. root proteome. Using MaxQuant platform, more than 600 proteins were found; 42 were identified based on at least 2 razor + unique peptides, score > 25, and were found to be differentially expressed in the exposed versus control roots with t-test difference > ±0.70 (p < 0.05, Perseus). Se(IV) caused growth inhibition and the decrease of total RNA in roots. Different abundances of proteins involved in transcriptional regulation, protein folding/assembly, cell cycle, energy/carbohydrate metabolism, stress response, and antioxidant defense were found in the exposed vs nonexposed roots. New evidence was obtained on the alteration of sulfur metabolism due to S-Se competition in A. cepa L. which, together with the original analytical approach, is the main scientific contribution of this study. Specifically, proteins participating in assimilation and transformation of both elements were affected; formation of volatile Se compounds seemed to be favored. Changes observed in methionine cycle suggested that Se(IV) stress might repress methylation capability in A. cepa L., potentially limiting accumulation of Se in the form of nonprotein methylated species and affecting adversely transmethylation-dependent signaling pathways.

Bio-transformation of selenium in Se-enriched bacterial strains of Lactobacillus casei.

BACKGROUND: Selenium is an element of very great importance for the proper functioning of the human body, mainly due to its antioxidant properties. Selenium exhibits a preventive effect in the case of cardiovascular disease, the immune system, male infertility and inhibits the toxic action of other agents. Selenium is important for Hashimoto's disease. Intake of selenium in the diet slows the aging process. The biological and toxicological effects of selenium strongly depend on its chemical form. Some organisms for example: plant, yeast, are capable of metabolizing low bioavailable selenium compounds (inorganic selenium) into its high bioavailable forms (organic selenium). OBJECTIVE: The aim of this study was to investigate the bio-transformation of selenium by Lactobacillus bacteria towards the characterisation of selenium metabolites. MATERIAL AND METHODS: The speciation of selenium was evaluated by high performance liquid chromatography with inductively coupled plasma mass spectrometry detector. The extraction of selenium species from lyophilized bacteria was executed with water, the mixture of lipase and protease, as well as lisozyme and sodium dodecyl sulphate. RESULTS: All investigated bacteria strains cultivated in the presence of Na2SeO3 effectively uptake selenium. Surprisingly, none of the applied extraction media exhibited a strong power to release the majority of the uptaken selenium compounds. Thus a maximum of 10% of the selenium was extracted from bacteria exposed to the enzymes. However, it was found that Lactobacillus bacteria are able to metabolize inorganic ions of selenium (IV) into Se-methionine, Se-methyloselenocysteine and other unidentified forms. CONCLUSIONS: The study confirmed the ability of probiotic bacteria to biotransform inorganic selenium into its organic derivatives. Therefore, Se-enriched bacteria can be considered as an addition to the functional food. KEY WORDS: selenium speciation, extraction procedure, Lactobacillus casei bacteria, Lactic acid bacteria (LAB), HPLC ICP-MS, functional food.

In vivo investigation of the distribution and the local speciation of selenium in Allium cepa L. by means of microscopic X-ray absorption near-edge structure spectroscopy and confocal microscopic X-ray fluorescence analysis.

In this work, microscopic X-ray absorption near-edge structure spectroscopy (mu-XANES) and confocal microscopic X-ray fluorescence analysis (mu-XRF) were used for the in vivo determination of the distribution of total selenium and for the local speciation of selenium in roots and leaves of onion. Selected Allium cepa L. plants were grown hydroponically in a standard medium containing inorganic selenium compounds (selenite or selenate). The measurements were performed in vivo, that is, on living plants without the need for any form of sampling or sample pretreatment and without the necessity for cutting plant tissues into pieces. Distinct energy differences of the XANES spectra of various selenium reference compounds having different oxidation states allow adjusting the excitation energies used for mu-XRF mapping in such a manner that the distribution of selenium in various oxidation states is obtained with a spatial resolution of a few tens of micrometers within the virtual cross section of the onion tissues. We find that the ratio of inorganic to amino acid selenium compounds differs in various subparts of the plant. Detailed in vivo investigation of the distribution of various selenium species in virtual cross sections of root tips and green leaf shows that the selenium transport takes place via different mechanisms, depending on the nature of the selenium compounds originally taken up.

[Low zinc and high copper levels in mothers of children with isolated cleft lip and palate]. / Niskie stezenia cynku i wysokie stezenia miedzi w surowicy matek dzieci z izolowanym rozszczepem wargi i podniebienia.

UNLABELLED: Changes in orofacial development have been reported in offspring of animals treated with copper injections or low zinc diet during pregnancy. The aim of our study was to determine serum levels of zinc and copper in mothers of children with non-syndromic orofacial clefts and in women who gave birth to healthy children. Ninety six healthy women participated in the study: 35 mothers of children with cleft lip and cleft palate (CLP), 30 mothers of children with cleft palate (CP) and 31 control mothers of healthy children. Serum zinc and copper concentrations were determined by the use of inductively coupled plasma-mass spectrometry (ICP-MS). RESULTS: Between CLP and controls there were significant differences in mean zinc levels (511 +/- 121 microg/L vs 572 +/- 76 microg/L, p = 0.012) and copper levels (991 +/- 388 microg/L vs 810 +/- 201 microg/L, p = 0.02). In CP mean serum zinc and copper levels were 540 +/- 130 microg/L and 846 +/- 291 microg/L. The ratio of serum zinc to copper levels was significantly lower in CLP comparing to CP and controls (0.57 +/- 0.19 vs 0.67 +/- 0.18 and 0.74 +/- 0.19, p = 0.003). CONCLUSION: Our results suggest that trace elements may play a role in facial clefting in humans.