Institut für Ernährungswissenschaften

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Anthropometrics, hemoglobin status and dietary micronutrient intake among Tanzanian and Mozambican pigeon pea farmers
(2022) Eleraky, Laila; Issa, Ramula; Maciel, Sónia; Mbwana, Hadijah; Rybak, Constance; Frank, Jan; Stuetz, Wolfgang
Inadequate consumption of micronutrient-dense and protein-rich foods such as vegetables, legumes and meat is an important contributing cause for anemia and deficiencies of vitamin A and iron in rural communities of Tanzania and Mozambique. A cross-sectional study was conducted to assess the nutritional status (anthropometrics and hemoglobin) and diets in particular micronutrient intake of female and male pigeon pea farmers from Lindi, Tanzania, and Gurué, the Zambézia province of Mozambique. A total of 1526 farmers (669 from Tanzania, 857 from Mozambique) were studied, of whom 16% were overweight and 35% were anemic. The highest prevalence of overweight and anemia, at 35% and 48%, was observed in Tanzanian and Mozambican women, respectively. Overall, only a small proportion of women and men reached the recommended daily dietary intake of vitamin A (10%), iron (51%) and zinc (44%). Multiple regression models revealed that dark green leafy vegetables (DGLVs) highly predicted vitamin A intake, whereas legumes in Tanzania and starchy plants in Mozambique were actually the dominant sources of vitamin A. Cereals covered over half of the iron and the zinc intake in both countries. An increased consumption of micronutrient-rich DGLVs and legumes, while reducing the high amounts of refined maize or polished rice, is suggested to counteract the high prevalence of anemia and overweight among smallholder farmers in East and South Eastern Africa.
Heritable variation of foliar spectral reflectance enhances genomic prediction of hydrogen cyanide in a genetically structured population of eucalyptus
(2022) Ballesta, Paulina; Ahmar, Sunny; Lobos, Gustavo A.; Mieres-Castro, Daniel; Jiménez-Aspee, Felipe; Mora-Poblete, Freddy
Plants produce a wide diversity of specialized metabolites, which fulfill a wide range of biological functions, helping plants to interact with biotic and abiotic factors. In this study, an integrated approach based on high-throughput plant phenotyping, genome-wide haplotypes, and pedigree information was performed to examine the extent of heritable variation of foliar spectral reflectance and to predict the leaf hydrogen cyanide content in a genetically structured population of a cyanogenic eucalyptus (Eucalyptus cladocalyx F. Muell). In addition, the heritable variation (based on pedigree and genomic data) of more of 100 common spectral reflectance indices was examined. The first profile of heritable variation along the spectral reflectance curve indicated the highest estimate of genomic heritability (hg2=0.41) within the visible region of the spectrum, suggesting that several physiological and biological responses of trees to environmental stimuli (ex., light) are under moderate genetic control. The spectral reflectance index with the highest genomic-based heritability was leaf rust disease severity index 1 (hg2=0.58), followed by the anthocyanin reflectance index and the Browning reflectance index (hg2=0.54). Among the Bayesian prediction models based on spectral reflectance data, Bayes B had a better goodness of fit than the Bayes-C and Bayesian ridge regression models (in terms of the deviance information criterion). All models that included spectral reflectance data outperformed conventional genomic prediction models in their predictive ability and goodness-of-fit measures. Finally, we confirmed the proposed hypothesis that high-throughput phenotyping indirectly capture endophenotypic variants related to specialized metabolites (defense chemistry), and therefore, generally more accurate predictions can be made integrating phenomics and genomics.
Gellan gum is a suitable biomaterial for manual and bioprinted setup of long-term stable, functional 3D-adipose tissue models
(2022) Albrecht, Franziska B.; Dolderer, Vera; Nellinger, Svenja; Schmidt, Freia F.; Kluger, Petra J.
Due to its wide-ranging endocrine functions, adipose tissue influences the whole body’s metabolism. Engineering long-term stable and functional human adipose tissue is still challenging due to the limited availability of suitable biomaterials and adequate cell maturation. We used gellan gum (GG) to create manual and bioprinted adipose tissue models because of its similarities to the native extracellular matrix and its easily tunable properties. Gellan gum itself was neither toxic nor monocyte activating. The resulting hydrogels exhibited suitable viscoelastic properties for soft tissues and were stable for 98 days in vitro. Encapsulated human primary adipose-derived stem cells (ASCs) were adipogenically differentiated for 14 days and matured for an additional 84 days. Live-dead staining showed that encapsulated cells stayed viable until day 98, while intracellular lipid staining showed an increase over time and a differentiation rate of 76% between days 28 and 56. After 4 weeks of culture, adipocytes had a univacuolar morphology, expressed perilipin A, and secreted up to 73% more leptin. After bioprinting establishment, we demonstrated that the cells in printed hydrogels had high cell viability and exhibited an adipogenic phenotype and function. In summary, GG-based adipose tissue models show long-term stability and allow ASCs maturation into functional, univacuolar adipocytes.
Nutrition and health aspects of mothers and their infants in Bukavu region, Democratic Republic of the Congo : a follow-up study with cross-sectional analyses and nutrition interventions
(2024) Beitze, Damaris Elisabeth; Scherbaum, Veronika
Hintergrund: Mangelernährung in verschiedenen Ausprägungen ist eine Herausforderung für die Bevölkerungsgesundheit in der Demokratischen Republik Kongo, einem Land, das von politischer Instabilität, Gewalt, Nahrungsmittelunsicherheit und Armut geprägt ist. Vor allem verborgener Hunger und Anämie machen einen großen Teil davon aus. Unzureichende Ernährung während der ersten 1000 Lebenstage beeinträchtigt die Gesundheit und Entwicklung eines Kindes kurz- und langfristig. Das Ernährungsverhalten der Mütter spielt sowohl für ihren eigenen Ernährungszustand als auch für den ihrer Kinder eine wichtige Rolle, insbesondere während dieser sensiblen Phase. Der Ernährungszustand und das Ernährungsverhalten können durch vielfältige Faktoren auf verschiedenen Ebenen beeinflusst werden. Diese müssen untersucht und berücksichtigt werden, um zielführende Interventionen zu gestalten. Zielsetzung: Diese Dissertation hatte zum Ziel, den Ernährungsstatus, insbesondere Anämie, stillender Frauen und ihrer Säuglinge zu untersuchen, ebenso wie damit assoziierte Faktoren. Des Weiteren sollte der Einfluss von Ernährungsschulungen und Lipid-basierten Nährstoffsupplementen auf Anämie, Ernährungswissen und Ernährungsverhalten von unter- und übergewichtigen Müttern ermittelt werden. Methoden: Eine Follow-up-Studie mit Querschnittserhebungen und inkludierter Interventionsstudie wurde mit 471 stillenden Müttern und ihren Säuglingen in der Region von Bukavu, Demokratische Republik Kongo von Dezember 2017 bis Juni 2019 durchgeführt. Mutter-Kind-Paare wurden nach der Entbindung in einem von drei Krankenhäusern der Studie (zwei davon semi-urban und eines im ländlichen Gebiet) in der ersten Woche postpartum rekrutiert. Die erste Datenerhebung beinhaltete anthropometrische Messungen der Mütter und Neugeborenen sowie ein strukturiertes Interview zu sozio-demographischen Faktoren und Ernährungsaspekten während der Schwangerschaft. Folgeerhebungen, die 3-6 und 6-9 Monate nach der Geburt stattfanden, umfassten die Messung von anthropometrischen Parametern und der Hämoglobin-Konzentration (Hb) von Müttern und ihren Säuglingen, ein 24-Stunden Ernährungsprotokoll der Mütter sowie ein strukturiertes Interview zu Ernährungswissen und -verhalten. Die randomisierte kontrollierte Interventionsstudie fand zwischen diesen beiden Folgeerhebungen statt. Vor der Intervention wurden die Mütter anhand ihres mittleren Oberarmumfangs (MUAC) in niedrigen (≥ 21 und < 25 cm), normalen (≥ 25 und < 28 cm) und hohen MUAC (≥ 28 cm) eingestuft. Anschließend wurden sie zufällig den folgenden Interventions- und Kontrollgruppen zugeteilt: niedriger MUAC – Ernährungsschulung; niedriger MUAC – Supplement; niedriger MUAC – Kontrolle; mittlerer MUAC – Kontrolle; hoher MUAC – Ernährungsschulung; hoher MUAC – Kontrolle. Über drei Monate erhielten Mütter mit Ernährungsschulungen vier theoretische und zwei praktische Einheiten zu ausgewogener Ernährung, Eisen, Vitamin A und Nährstoffmängeln. Mütter in der Supplement-Gruppe erhielten ein tägliches Lipid-basiertes Nährstoffsupplement. Für 416 Mütter und Säuglinge waren Daten vor und nach der Intervention vorhanden. Ergebnisse: Die Studie zeigte die Koexistenz verschiedener Formen von Mangelernährung in der Studienpopulation. Ernährungs- und Gesundheitsaspekte unterschieden sich zwischen den semiurbanen und ländlichen Gegenden. Das Erhalten von Leistungen während der Schwangerschaftsbetreuung (Malaria-Medikamente, Entwurmung, Supplemente, Ernährungsinformationen durch Gesundheitspersonal) und das Wissen zu sowie die Umsetzung von Familienplanung waren im ländlichen Gebiet geringer als im semi-urbanen Gebiet, ebenso anthropometrische Parameter wie der mütterliche MUAC und das Geburtsgewicht. Eine multiple lineare Regression ergab zunehmenden mütterlichen MUAC, Alter und Gestationsalter als positive Einflussfaktoren auf das Geburtsgewicht, während die ländliche Lage, eine erstgebärende Mutter, eine Tätigkeit der Mutter in der Landwirtschaft und das weibliche Geschlecht des Neugeborenen negative Prädiktoren des Geburtsgewichts waren. Die Anämieraten 3-6 Monate nach der Geburt waren mit 28,2 % bei den Müttern von mittlerer Relevanz für Bevölkerungsgesundheit, lagen bei den Säuglingen mit 74,3 % jedoch im schwerwiegenden Bereich. Sowohl das mütterliche Ernährungswissen als auch ihre Ernährungsvielfalt waren gering ausgeprägt. Im Median hatten die Mütter ein Wissen von 0,39 in einem möglichen Bereich von 0 bis 1 und eine Ernährungsvielfalt von 3,0 (möglicher Bereich 0-10). Ernährungswissen und mütterlicher Hb waren leicht positiv korreliert, aber es bestanden keine signifikanten Korrelationen der Ernährungsvielfalt mit einem dieser beiden Parameter. Geschmack (68,1 %), Aussehen (42,5 %), die Wirkung von Lebensmitteln auf die Gesundheit (29,0 %) und Verfügbarkeit (25,6 %) waren häufige Kriterien der Lebensmittelauswahl. Nach der Intervention hatten Mütter, die Ernährungsschulungen erhalten hatten, ein signifikant höheres Ernährungswissen als diejenigen ohne Schulungen, aber keine verbesserte Ernährungsvielfalt. Außerdem modifizierten mehr Mütter mit Schulungen die Beikost für ihr Kind als die anderen Mütter, häufig im Zusammenhang mit Inhalten der Ernährungsschulungen. Weder der mütterliche Hb nach der Intervention noch die Hb-Entwicklung über die Interventionsphase unterschieden sich signifikant zwischen den sechs Interventions- und Kontrollgruppen. Allerdings war der Anteil an Müttern mit einer Verbesserung des Hb in den Gruppen mit Ernährungsschulungen größer, wenn auch nicht statistisch signifikant. Bei Betrachtung ausschließlich der Frauen, die zu Beginn der Intervention an Anämie litten, zeigte sich eine größere Zunahme des Hb in den Gruppen, die entweder Schulungen oder Supplemente erhielten, allerdings nicht statistisch signifikant. Schlussfolgerungen: Der Ernährungsstatus stillender Frauen und ihrer Säuglinge im Studiengebiet war mangelhaft. Verschiedene Faktoren, die die Mangelernährung befördern könnten, wurden identifiziert: Unzureichende Nahrungsaufnahme und eingeschränkte Leistungen in der primären Gesundheitsversorgung können den Ernährungsstatus beeinflussen. Zu ihren möglichen, vielschichtigen Determinanten gehören geringes Ernährungswissen, die ländliche Gegend und Nahrungsmittelunsicherheit, aber auch persönliche Präferenzen. Um den Ernährungszustand effektiv zu verbessern, müssen Ernährungsinterventionen diese verschiedenen Einflussfaktoren der Nahrungsaufnahme adressieren. Ernährungsschulungen und Lipid-basierte Nährstoffsupplemente könnten einen positiven Effekt haben, vor allem bei Müttern mit Anämie. Sie sollten weiter untersucht werden. Es wird empfohlen, das Bewusstsein und die Qualifikation von Gesundheitspersonal in der primären Gesundheitsversorgung hinsichtlich Ernährung zu stärken und praktische Elemente in Maßnahmen mit Ernährungsschulungen zu integrieren.
Effect of a diet rich in galactose or fructose, with or without fructooligosaccharides, on gut microbiota composition in rats
(2022) Mhd Omar, Nor Adila; Dicksved, Johan; Kruger, Johanita; Zamaratskaia, Galia; Michaëlsson, Karl; Wolk, Alicja; Frank, Jan; Landberg, Rikard
Recent studies suggest that a diet rich in sugars significantly affects the gut microbiota. Adverse metabolic effects of sugars may partly be mediated by alterations of gut microbiota and gut health parameters, but experimental evidence is lacking. Therefore, we investigated the effects of high intake of fructose or galactose, with/without fructooligosaccharides (FOS), on gut microbiota composition in rats and explored the association between gut microbiota and low-grade systemic inflammation. Sprague–Dawley rats (n = 6/group) were fed the following isocaloric diets for 12 weeks (% of the dry weight of the sugars or FOS): (1) starch (control), (2) fructose (50%), (3) galactose (50%), (4) starch+FOS (15%) (FOS control), (5) fructose (50%)+FOS (15%), (6) galactose (50%)+FOS (15%), and (7) starch+olive (negative control). Microbiota composition in the large intestinal content was determined by sequencing amplicons from the 16S rRNA gene; 341F and 805R primers were used to generate amplicons from the V3 and V4 regions. Actinobacteria, Verrucomicrobia, Tenericutes, and Cyanobacteria composition differed between diets. Bifidobacterium was significantly higher in all diet groups where FOS was included. Modest associations between gut microbiota and metabolic factors as well as with gut permeability markers were observed, but no associations between gut microbiota and inflammation markers were observed. We found no coherent effect of galactose or fructose on gut microbiota composition. Added FOS increased Bifidobacterium but did not mitigate potential adverse metabolic effects induced by the sugars. However, gut microbiota composition was associated with several metabolic factors and gut permeability markers which warrant further investigations.
Antioxidants Attenuate Heat Shock Induced Premature Senescence of Bovine Mesenchymal Stem Cells
(2022) Nir, Dana; Ribarski-Chorev, Ivana; Shimoni, Chen; Strauss, Carmit; Frank, Jan; Schlesinger, Sharon
Mesenchymal stem cells (MSC) have many roles that are important for the body’s proper functioning. When the MSC pool is damaged, it is often correlated with impaired development or health of the organism. MSC are known for their anti-inflammatory, immunomodulatory and trophic characteristics that play an important role in the physiological homeostasis of many tissues. Heat shock impairs MSC capacity by inducing the generation of reactive oxygen species and mitochondrial dysfunction, which, in turn, send the cells into a state of premature senescence. Here, we pre-exposed MSC to melatonin, resveratrol, or curcumin, which are natural antioxidative compounds, and tested the protective effects of these substances from oxidative stress and aging. Our data showed that pre-exposure of MSC to antioxidants decreased reactive oxygen species while mitochondrial damage remained high. Additionally, although the proliferation of the cells was slow, antioxidants protected the cells from premature senescence, and subsequent cytokine release was prevented. We conclude that while elevated temperatures directly cause mitochondrial damage, senescence is induced by elevated ROS levels. We suggest that heat shock alters cell and tissue homeostasis by several independent mechanisms; however, reducing tissue senescence will reduce damage and provide a pathway to overcome physiological challenges in animals.
Uptake and time‐dependent subcellular localization of native and micellar curcumin in intestinal cells
(2022) Flory, Sandra; Benz, Ann‐Kathrin; Frank, Jan
Uptake into intestinal cells and intracellular distribution into metabolically competent organelles, such as the endoplasmic reticulum, are important processes potentially limiting the bioavailability of xenobiotics. The incorporation of curcumin into polysorbate 80 micelles improves its naturally low oral bioavailability in humans. Here, we investigated uptake and time‐dependent localization of curcumin in intestinal cells when administered as native or micellar formulation. Differentiated Caco‐2 cells were incubated with 200 μmol/L native or micellar curcumin for up to 180 min and cellular uptake was quantified. Intracellular curcumin was detected already after 30 min and did not differ significantly between formulations or over time. Subcellular localization of native and micellar curcumin in Caco‐2 cells was studied by density gradient centrifugation. After 30 min, curcumin from both formulations was mainly associated with mitochondria and lysosomes, after 180 min native curcumin was associated with mitochondria and peroxisomes, micellar curcumin with peroxisomes only. Uptake and localization of native and micellar curcumin in intestinal cells do not differ significantly and consequently do not explain differences in bioavailability in humans. The temporary co‐localization with lysosomes is in agreement with the previously proposed role of endocytosis in cellular uptake of curcumin and warrants further investigation.
Vitamin E and carotenoid profiles in leaves, stems, petioles and flowers of stinging nettle (Urtica leptophylla Kunth) from Costa Rica
(2022) Montoya‐Arroyo, Alexander; Toro‐González, Camilo; Sus, Nadine; Warner, Jorge; Esquivel, Patricia; Jiménez, Víctor M; Frank, Jan
BACKGROUND Local leafy vegetables are gaining attention as affordable sources of micronutrients, including vitamins, pro-vitamin carotenoids and other bioactive compounds. Stinging nettles (Urtica spp.) are used as source of fibers, herbal medicine and food. However, despite the relatively wide geographical spread of Urtica leptophylla on the American continent, little is known about its content of vitamin E congeners and carotenoids. We therefore investigated the particular nutritional potential of different plant structures of wild Costa Rican U. leptophylla by focusing on their vitamin E and carotenoid profiles. RESULTS Young, mature and herbivore-damaged leaves, flowers, stems and petioles were collected and freeze-dried. Vitamin E and carotenoids were determined by high-performance liquid chromatography after liquid/liquid extraction with hexane. α-Tocopherol was the major vitamin E congener in all structures. Flowers had a high content of γ-tocopherol. Herbivore-damaged leaves had higher contents of vitamin E than undamaged leaves. Lutein was the major and β-carotene the second most abundant carotenoid in U. leptophylla. No differences in carotenoid profiles were observed between damaged and undamaged leaves. CONCLUSION The leaves of U. leptophylla had the highest nutritional value of all analyzed structures; therefore, they might represent a potential source of α-tocopherol, lutein and β-carotene.
Anemia among Syrian refugee children aged 6 to 23 months living in Greater Beirut, Lebanon, including the Voices of Mothers’ and Local Healthcare Staff: A mixed-methods study
(2023) Jeremias, Theresa; Abou-Rizk, Joana; Burgard, Leonie; Entenmann, Isa; Nasreddine, Lara; Jomaa, Lamis; Hwalla, Nahla; Frank, Jan; Scherbaum, Veronika
Globally, the prevalence of anemia among children during the period of complementary feeding is high. A cross-sectional, mixed-methods study was conducted to examine the main determinants of anemia among Syrian refugee children aged 6 to 23 months (n = 215) and to illuminate the knowledge, attitudes and perceptions of their mothers and Lebanese healthcare staff on its causes and available treatment options. 42% of the children and 20% of their mothers were anemic. Determinants of child anemia were the mother having anemia or not knowing that fish/seafood is a source of iron; the child having been acutely ill the last two weeks or receiving cow’s milk, but not consuming iron-rich infant formula, added fats/oils, or fruits in the previous 24 hours. Several Syrian mothers knew some causes of anemia and named dizziness as a leading symptom but did not mention flesh foods as a key source of heme iron. They reported financial constraints in accessing iron-rich foods and supplements. Lebanese doctors largely gave appropriate dietary advice and prescribed iron supplements as treatment. Multisectoral interventions are needed that combine medical and financial support with nutrition counseling for mothers to reduce the high burden of anemia among young children living in a multiple crises situation.
Optimization of supercritical carbon dioxide extraction of polyphenols from black rosehip and their bioaccessibility using an In Vitro digestion/Caco-2 cell model
(2023) Kasapoğlu, Kadriye Nur; Kruger, Johanita; Barla-Demirkoz, Aslı; Gültekin-Özgüven, Mine; Frank, Jan; Özçelik, Beraat
The fruits of Rosa pimpinellifolia are rich sources of (poly)phenols, however they are underutilized due to the limited information available. The influence of the pressure, temperature, and co-solvent concentration (aqueous ethanol) of the supercritical carbon dioxide extraction (SCO2-aqEtOH) on the extraction yield, total phenolic-, total anthocyanin-, catechin-, cyanidin-3-O-glucoside contents, and total antioxidant activity of black rosehip was investigated simultaneously. The maximum obtained total phenolic and total anthocyanin contents under the optimized extraction conditions (280 bar, 60 °C and 25% ethanol, v/v) were 76.58 ± 4.25 mg gallic acid equivalent and 10.89 ± 1.56 mg cyanidin-3-O-glucoside equivalent per g of the dry fruits, respectively. The optimal extract obtained by SCO2-aqEtOH was compared to two other extraction procedures: ultrasonication using ethanol as solvent (UA-EtOH) and pressurized hot water extraction (PH-H2O). The bioaccessibility and cellular metabolism of the phenolic compounds in the different black rosehip extracts were assessed using an in vitro digestion coupled with a human intestinal Caco-2 cell model. The in vitro digestive stability and cellular uptake of the phenolic compounds had no significant difference among the different extraction methods. The results of this study confirm the efficiency of SCO2-aqEtOH extraction for phenolic compounds and, in particular, for anthocyanins, and could be used to produce new functional food ingredients from black rosehip with high antioxidant power containing both hydrophilic and lipophilic compounds.
Combined effects of drought and soil fertility on the synthesis of vitamins in green leafy vegetables
(2023) Park, Taewan; Fischer, Sahrah; Lambert, Christine; Hilger, Thomas; Jordan, Irmgard; Cadisch, Georg
Green leafy vegetables, such as Vigna unguiculata, Brassica oleraceae, and Solanum scabrum, are important sources of vitamins A, B1, and C. Although vitamin deficiencies considerably affect human health, not much is known about the effects of changing soil and climate conditions on vegetable vitamin concentrations. The effects of high or low soil fertility and three drought intensities (75%, 50%, and 25% pot capacity) on three plant species were analysed (n = 48 pots) in a greenhouse trial. The fresh yield was reduced in all the vegetables as a result of lower soil fertility during a severe drought. The vitamin concentrations increased with increasing drought stress in some species. Regardless, the total vitamin yields showed a net decrease due to the significant biomass loss. Changes in vitamin concentrations as a result of a degrading environment and increasing climate change events are an important factor to be considered for food composition calculations and nutrient balances, particularly due to the consequences on human health, and should therefore be considered in agricultural trials.
Bioactive compounds and antioxidant activity in the fruit of rosehip (Rosa canina L. and Rosa rubiginosa L.)
(2023) Peña, Fabiola; Valencia, Sebastián; Tereucán, Gonzalo; Nahuelcura, Javiera; Jiménez-Aspee, Felipe; Cornejo, Pablo; Ruiz, Antonieta
Rosehips (Rosa spp., Rosaceae) are wild rose bushes with more than 100 species. Its fruits vary in colour and size, depending on the species, and are recognised for their nutritional characteristics. Ten samples of Rosa canina L. and Rosa rubiginosa L. fruits were collected at different geographical points from Southern Chile. Nutrients such as crude protein and minerals and functional properties such as phenolic compounds, ascorbic acid, and also antioxidant activities were evaluated by HPLC-DAD-ESI-MS/MS. The results revealed a high content of bioactive compounds, primarily ascorbic acid (6.0 to 8.2 mg g−1 fresh weight (FW)), flavonols (427.9 ± 0.4 μg g−1 FW) and antioxidant activity. We established a relationship between the antioxidant activity using Trolox equivalent antioxidant capacity (TEAC), cupric reducing antioxidant capacity (CUPRAC) and 2,2-diphenyl radical (DPPH) methods and the concentration of uncoloured compounds, such as flavonols and catechin. This antioxidant activity was primarily associated with the samples from Gorbea, Lonquimay, Loncoche, and Villarrica localities, and all of them were of the species Rosa rubiginosa L. The results here obtained represent novel information of rosehip fruits. In this sense, the reported information about compounds and antioxidant activities in rosehip fruits allowed us to continue new lines of research in relation to the potential formulation of new functional foods and also in the treatment and/or prevention of some diseases.
Uncovering the relationship between selenium status, age, health, and dietary habits: Insights from a large population study including nonagenarian offspring from the MARK-AGE project
(2023) Giacconi, Robertina; Piacenza, Francesco; Aversano, Valentina; Zampieri, Michele; Bürkle, Alexander; Villanueva, María Moreno; Dollé, Martijn E.T.; Jansen, Eugène; Grune, Tilman; Gonos, Efstathios S.; Franceschi, Claudio; Capri, Miriam; Weinberger, Birgit; Sikora, Ewa; Toussaint, Olivier; Debacq-Chainiaux, Florence; Stuetz, Wolfgang; Slagboom, Pieternella Eline; Bernhardt, Jürgen; Fernández-Sánchez, Maria Luisa; Provinciali, Mauro; Malavolta, Marco
An inadequate selenium (Se) status can accelerate the aging process, increasing the vulnerability to age-related diseases. The study aimed to investigate plasma Se and Se species in a large population, including 2200 older adults from the general population (RASIG), 514 nonagenarian offspring (GO), and 293 GO Spouses (SGO). Plasma Se levels in women exhibit an inverted U-shaped pattern, increasing with age until the post-menopausal period and then declining. Conversely, men exhibit a linear decline in plasma Se levels with age. Subjects from Finland had the highest plasma Se values, while those from Poland had the lowest ones. Plasma Se was influenced by fish and vitamin consumption, but there were no significant differences between RASIG, GO, and SGO. Plasma Se was positively associated with albumin, HDL, total cholesterol, fibrinogen, and triglycerides and negatively associated with homocysteine. Fractionation analysis showed that Se distribution among plasma selenoproteins is affected by age, glucometabolic and inflammatory factors, and being GO or SGO. These findings show that sex-specific, nutritional, and inflammatory factors play a crucial role in the regulation of Se plasma levels throughout the aging process and that the shared environment of GO and SGO plays a role in their distinctive Se fractionation.
Association of Torquetenovirus Viremia with physical frailty and cognitive impairment in three independent European cohorts
(2023) Giacconi, Robertina; Laffon, Blanca; Costa, Solange; Teixeira-Gomes, Armanda; Maggi, Fabrizio; Macera, Lisa; Spezia, Pietro Giorgio; Piacenza, Francesco; Bürkle, Alexander; Moreno-Villanueva, María; Bonassi, Stefano; Valdiglesias, Vanessa; Teixeira, Joao Paulo; Dollé, Martijn E.T.; Rietman, M. Liset; Jansen, Eugène; Grune, Tilman; Gonos, Efstathios S.; Franceschi, Claudio; Capri, Miriam; Weinberger, Birgit; Sikora, Ewa; Stuetz, Wolfgang; Toussaint, Olivier; Debacq-Chainiaux, Florence; Hervonen, Antti; Hurme, Mikko; Slagboom, P. Eline; Schön, Christiane; Bernhardt, Jürgen; Breusing, Nicolle; Pásaro, Eduardo; Maseda, Ana; Lorenzo-López, Laura; Millán-Calenti, José Carlos; Provinciali, Mauro; Malavolta, Marco
Introduction: Immunosenescence and inflammaging have been implicated in the pathophysiology of frailty. Torquetenovirus (TTV), a single-stranded DNA anellovirus, the major component of the human blood virome, shows an increased replication rate with advancing age. An elevated TTV viremia has been associated with an impaired immune function and an increased risk of mortality in the older population. The objective of this study was to analyze the relation between TTV viremia, physical frailty, and cognitive impairment. Methods: TTV viremia was measured in 1,131 nonfrail, 45 physically frail, and 113 cognitively impaired older adults recruited in the MARK-AGE study (overall mean age 64.7 ± 5.9 years), and then the results were checked in two other independent cohorts from Spain and Portugal, including 126 frail, 252 prefrail, and 141 nonfrail individuals (overall mean age: 77.5 ± 8.3 years). Results: TTV viremia ≥4log was associated with physical frailty (OR: 4.69; 95% CI: 2.06–10.67, p < 0.0001) and cognitive impairment (OR: 3.49, 95% CI: 2.14–5.69, p < 0.0001) in the MARK-AGE population. The association between TTV DNA load and frailty status was confirmed in the Spanish cohort, while a slight association with cognitive impairment was observed (OR: 1.33; 95% CI: 1.000–1.773), only in the unadjusted model. No association between TTV load and frailty or cognitive impairment was found in the Portuguese sample, although a negative association between TTV viremia and MMSE score was observed in Spanish and Portuguese females. Conclusions: These findings demonstrate an association between TTV viremia and physical frailty, while the association with cognitive impairment was observed only in the younger population from the MARK-AGE study. Further research is necessary to clarify TTV’s clinical relevance in the onset and progression of frailty and cognitive decline in older individuals.
Demethoxycurcumin and Bisdemethoxycurcumin are more bioavailable than Curcumin: A meta-analysis of randomized cross-over trials in healthy umans and an In Vitro mechanistic exploration
(2023) Desmarchelier, Charles; Sus, Nadine; Marconot, Grégory; Gillet, Guillian; Resseguier, Noémie; Frank, Jan
Background: Curcuminoids are secondary plant metabolites found in turmeric and many dietary supplements. They usually consist of a mixture of curcumin (CUR), demethoxycurcumin (dCUR) and bisdemethoxycurcumin (bdCUR). CUR, the main curcuminoid, has been intensely investigated for its putative effects against, e.g., inflammation, oxidative stress and cancer. However, CUR displays very poor bioavailability. We have previously shown that, when brought by turmeric, dCUR and bdCUR, which can also exert health effects, display greater in vitro bioaccessibility than CUR (PMID: 37073511). However, their bioavailability relative to that of CUR has not been thoroughly investigated. Objective: We aimed to compare the bioavailability of dCUR and bdCUR to that of CUR in a meta-analysis of clinical trials in healthy humans and to compare their in vitro bioaccessibility and enterocyte uptake efficiency. Methods and Results: Studies published until 2022 were searched for using Medline and Scopus. The included studies were randomized trials that measured the bioavailability of CUR, dCUR and bdCUR in healthy participants. Estimates were calculated using a random-effects model. Fifteen trials were included in the study, representing a total of 50 interventions, i.e., each trial investigated several curcuminoid formulations, in 762 participants. The relative bioavailabilities were calculated using the inverse variance method. dCUR was 2.32 (95% CI:1.70, 3.13) times more bioavailable than CUR, while bdCUR was 2.57 (95% CI: 1.58, 4.16) times more bioavailable than CUR, with some heterogeneity across the formulations used. Using an in vitro gastro-intestinal digestion model with pure curcuminoids, we showed that dCUR solubilization efficiency was 4.8 and 5.3 times higher than that of CUR and bdCUR, respectively (p < 0.001), while its micellization efficiency was 10.3 and 5.1 times higher than that of CUR and bdCUR, respectively (p < 0.001). Conclusions: bdCUR and dCUR display greater bioavailability in humans compared to CUR. A subgroup analysis by formulation is undergoing investigation and will be presented. For dCUR, this difference is partly explained by higher in vitro bioaccessibility. Uptake efficiency measurements of pure curcuminoids and of curcuminoids from in vitro digestion fluids are undergoing investigation and will be presented. bdCUR and dCUR might therefore represent relevant alternatives to CUR for the systematic delivery of curcuminoids.
Nutrition-related knowledge, attitudes, practices, and anemia status of lactating mothers in Bukavu, Democratic Republic of the Congo - A cross-sectional analysis
(2024) Beitze, Damaris Elisabeth; Kavira Malengera, Céline; Barhwamire Kabesha, Theophile; Scherbaum, Veronika
Maternal nutrition is impacted by personal and environmental factors including dietary intake, knowledge, food availability, and affordability. This cross-sectional analysis aimed to evaluate nutrition-related knowledge, attitudes, practices, and associations with hemoglobin concentration among lactating mothers in the Bukavu region, Democratic Republic of the Congo. In 444 lactating mothers, nutrition-related knowledge and practice were assessed by questionnaires and translated into knowledge and practice scores ranging from 0 to 1, attitudes and drivers of food choice were assessed, the Dietary Diversity Score (DDS) was assessed with 24 h dietary recalls in a potential range from 0 to 10, and hemoglobin (Hb) was measured in mothers and their infants. Anemia prevalence was 28.2% among mothers and 74.3% among infants aged 3–8 months. Nutritional knowledge and practice were limited (the median total knowledge score was 0.39, median DDS was 3.0). While there were slight positive correlations between knowledge and maternal Hb, DDS did not significantly correlate with either knowledge or Hb. Although half of the mothers stated a perception about their own susceptibility to anemia or vitamin A deficiency (56.4%, 47.4%), less than half of those could justify their estimation (40.9%, 44.2%). Taste (68.1%), appearance (42.5%), availability (29.0%), and health effects (25.6%) were important drivers of food choice. In conclusion, interventions on the different influencing factors including nutrition education strategies are needed.
Dietary intake of fructooligosaccharides protects against metabolic derangements evoked by chronic exposure to fructose or galactose in rats
(2023) Almasri, Fidèle; Collotta, Debora; Aimaretti, Eleonora; Sus, Nadine; Aragno, Manuela; Dal Bello, Federica; Eva, Carola; Mastrocola, Raffaella; Landberg, Rikard; Frank, Jan; Collino, Massimo
Scope: Diets rich in fat and sugars evoke chronic low-grade inflammation, leading to metabolic derangements. This study investigates the impact of fructose and galactose, two commonly consumed simple sugars, on exacerbation of the harmful effects caused by high fat intake. Additionally, the potential efficacy of fructooligosaccharides (FOS), a fermentable dietary fiber, in counteracting these effects is examined. Methods and results: Male Sprague-Dawley rats (six/group) are fed 8 weeks as follows: control 5% fat diet (CNT), 20% fat diet (FAT), FAT+10% FOS diet (FAT+FOS), FAT+25% galactose diet (FAT+GAL), FAT+GAL+10% FOS diet (FAT+GAL+FOS), FAT+25% fructose diet (FAT+FRU), FAT+FRU+10% FOS diet (FAT+FRU+FOS). The dietary manipulations tested do not affect body weight gain, blood glucose, or markers of systemic inflammation whereas significant increases in plasma concentrations of triacylglycerols, cholesterol, aspartate aminotransferase, and alanine aminotrasferase are detected in both FAT+FRU and FAT+GAL compared to CNT. In the liver and skeletal muscle, both sugars induce significant accumulation of lipids and advanced glycation end-products (AGEs). FOS supplementation prevents these impairments. Conclusion: This study extends the understanding of the deleterious effects of a chronic intake of simple sugars and demonstrates the beneficial role of the prebiotic FOS in dampening the sugar-induced metabolic impairments by prevention of lipid and AGEs accumulation.
Stability of bioactive compounds and antioxidant activity in rosehip juice (Rosa spp.)
(2024) Peña, Fabiola; González, Felipe; Jiménez-Aspee, Felipe; Bustamante, Luis; Ruiz, Antonieta
Rosehip fruits, characterized by their high concentrations of bioactive compounds and antioxidant activity (AA), have been traditionally used to make jams, infusions, and juices. Thus, the objective of this research was to evaluate the stability of rosehip juice by determining the concentrations of bioactive compounds and total phenols and the AA using chromatographic and spectroscopic methods. The stability of the juice was evaluated with three treatments and different storage conditions, namely, unpasteurized–refrigerated, pasteurized–room temperature, and pasteurized–refrigerated, and measurements were taken for eight months. Individual and total phenolic compounds, evaluated by chromatographic methods, reported differences until the end of this study. The total phenolic compounds by Folin–Ciocalteu method presented an average decrease of 57% in the three treatments in relation to the initial conditions. On the other hand, the ascorbic acid content decreased considerably, disappearing at week six. Furthermore, for the unpasteurized–refrigerated and pasteurized–refrigerated samples, a correlation was found between flavonols, total phenols, ascorbic acid, and antioxidant activity determined by the TEAC method. For the pasteurized–room temperature samples, correlations were found between the levels of several flavonols, hydroxycinnamic acid, total phenols, and ascorbic acid and the antioxidant activity determined by the CUPRAC method. The stability of the compounds was mainly correlated with the storage conditions of the juice and not with pasteurization. The highest stability was observed for the unpasteurized–refrigerated and pasteurized–refrigerated samples. Although the concentrations of the compounds evaluated decreased during this study, significant levels of AA persisted, providing beneficial characteristics for consumer health.
Entwicklung und Charakterisierung verschiedener humaner 3D Fettgewebemodelle mit und ohne Hydrogelmatrix als in vivo nahe Alternative
(2023) Albrecht, Franziska Brigitte; Kluger, Petra
Humanes Fettgewebe sekretiert hunderte regulatorisch aktive Hormone, die bei der Entstehung und Manifestierung schwerwiegender Krankheiten wie Diabetes oder kardiovaskulären Erkrankungen beteiligt sind. Um die Vorhersagekraft von in vitro Modellen zu verbessern und die Komplexität des nativen Fettgewebes besser nachzubilden, werden dringend dreidimensionale (3D) in vivo nahe Fettgewebemodelle benötigt. Um solch ein flexibel anwendbares möglichst physiologisches Modell aufzubauen wurde in dieser Arbeit der Aufbau und die Evaluierung verschiedener 3D Fettgewebemodelle teils mit artifizieller Matrix angestrebt und im Anschluss mit dem nativen Zustand vergleichen. Beginnend wurden aus primärem humanem Fett Lobuli in verschiedenen Größenbereichen isoliert, kultiviert und der Zelltod sowie Zeichen der Lipolyse bestimmt. Es hat sich gezeigt, dass Lobuli mit einem Gewicht von 27 – 70 mg über 15 Tagen die stabilsten Ergebnisse geliefert haben und wurden somit tiefergehend analysiert. Bei der Analyse der Viabilität und Zellfunktionalität in Form der Lipidakkumulation und der Expression von Perilipin A konnte bewiesen werden, dass Lobuli als Fettgewebemodell genutzt werden können. Als weiteres Modell erfolgte die Etablierung von Sphäroiden aus humanen primären Stammzellen aus dem Fettgewebe (adipose-derived stem cells, ASC). Die Sphäroide wurden für 14 Tage adipogen differenziert und währenddessen die Veränderung des Volumens und der Rundheit sowie die Expression von Zelltodmarkern und das adipogene Differenzierungspotenzial betrachtet. Dabei haben Sphäroide mit mehr als 100.000 Zellen ihr Volumen über die Zeit verkleinert und Apoptose- und Nekrosemarker an den Tagen 0 und 14 gezeigt. Sphäroide bis 100.000 Zellen haben ihr Volumen während der Differenzierung leicht vergrößert und haben die meisten Lipide akkumuliert, Perilipin A und Kollagen Ⅵ exprimiert. Somit konnten ASC-basierte Sphäroide aus 10.000 Zellen, mit signifikanter Lipidzunahme, als weiteres Fettgewebemodell mit hohem Potenzial identifiziert werden. Im Anschluss erfolgte die Etablierung von methacrylierter Gelatine (GelMA) als Biomaterial für die Verkapselung von primären ASCs und reifen Adipozyten (adipocyte, AC) mit anschließendem extrusions-basierten 3D Druck. Es konnte gezeigt werden, dass die homogene Verteilung der lipidgefüllten ACs in eine wässrige Biotintenlösung nur durch schnelles Herunterkühlen auf Eis möglich war. Der anschließende 3D Druck hatte, weder auf die Viabilität noch Funktionalität bzw. adipogene Differenzierung der Zellen signifikante Einflüsse. Die additiv aufgebauten Modelle wiesen an Tag 1 und 8 bzw. 15 in der Lebend-Tot-Färbung keinen vermehrten Zelltod im Vergleich zu den manuellen Modellen auf. Die Färbung der intrazellulären Lipide und Perilipin A sowie die Glycerolfreisetzung als Funktionalitätsmarker, haben dies bestätigt. Im Vergleich zu nativem Gewebe haben differenzierte ASCs (diffASC) nach der Differenzierung signifikant weniger lipidpositive Zellen und freigesetztes Glycerol gezeigt. Auch morphologisch betrachtet, haben ACs in GelMA mehr Ähnlichkeiten zum nativen Gewebe als diffASCs. Aufgrund der begrenzten Langzeitstabilität von GelMA, des Ursprungs und der Vernetzung wurden Biomaterialien ohne tierischen Ursprung evaluiert. Dabei hat sich das niedrig acetylierte Polysaccharid Gellan Gum (GG) als vielversprechendes Material erwiesen. Es konnten stabile und transparente Hydrogele durch die Vernetzung mit divalenten Ionen im Zellkulturmedium aufgebaut werden. Die 1 %-igen Hydrogele waren weder bei indirekter noch direkter Testung zytotoxisch oder haben Monozyten aktiviert. Bestätigt wurde es durch die Laktatdehydrogenase (LDH)-Freisetzung, den Resazurinumsatz und eine Lebend-Tot-Färbung. Azelluläre Gele wurden über 98 Tagen ohne signifikante Veränderungen erhalten und zeigten für Fettgewebe geeignete Materialeigenschaften. Aufgrund dieser Langzeitstabilität konnten diffASCs für 98 Tage in GG kultiviert werden und zu univakuolären Fettzellen reifen. Dies wurde durch die Färbung von intrazellulären Lipiden und Perilipin A sowie der Leptinsekretion und Glycerolfreisetzung bewiesen. Verglichen mit nativem Gewebe wiesen die diffASCs eine vergleichbare, wenn auch kleinere Morphologie, ähnliche Anteile an lipidpositiven und univakuolären Zellen auf. Auf Basis von GG konnten ACs erfolgreich verkapselt und aufgebaut werden. Hierbei erwies sich eine GG Konzentration von 0,5 % als geeignet, da homogen gemischte Hydrogele mit hohem Resazurinumsatz und geringer Glycerolfreisetzung kultiviert werden konnten. Für den additive Aufbau der GG-basierten Modelle, fand eine Optimierung zur Biotinte statt, was durch die initiale Zugabe divalenter Ionen zur Erhöhung der Viskosität und damit Druckbarkeit ermöglicht wurde. Die additiv gefertigten Modelle zeigten keine signifikanten Unterschiede in Bezug auf Viabilität oder Funktionalität. Nach 32 Tagen in Kultur konnten univakuoläre und funktionale Zellen gezeigt werden. Durch die Erhöhung der GG Konzentration auf 1,5 % wurde ein erfolgreiches 6D Bioprintingverfahren etabliert. Auch hier zeigten die in GG verkapselten ASCs keine Viabilitäts-, Morphologie-, oder Differenzierungsunterschiede nach dem Druckprozess. Abschließend erfolgte ein Vergleich der etablierten Modelle wobei die Adipogenese der ASC-basierten Modelle und zum anderen der adipozytenspezifische Phänotyp evaluiert wurde. Hinsichtlich der Viabilität konnten keine Unterschiede festgestellt werden. Das adipogene Differenzierungspotenzial zeigte sich am stärksten im diffASC Hydrogel. Sowohl hier als auch in den Sphäroiden konnten eingelagerte Lipide ohne hormonelle Induktion mit Medium nachgewiesen werden. Die Betrachtung der adipozytenspezifischen Morphologie der ausgereiften Modelle erlaubte den Rückschluss, dass die Zellen in den Monolayern den unausgereiftesten Zustand in Form von multivakuolären, elongierten diffASCs mit Aktin-Stressfasern aufweisen. Die Zellen der anderen Modelle zeigen deutlich größere Lipidvakuolen mit einem abgerundeten Zytoskelett. Die quantitative Bestimmung der Lipid-, Leptin- und Glycerolmenge legt nahe, dass sich die Zellen, außer die im Monolayer, in einem Steady-State befinden, was die relative Genexpression adipozytenspezifischer Gene untermauert. Zusammengefasst wurden in dieser Arbeit sechs unterschiedliche Fettgewebemodelle entwickelt und mit nativem Gewebe verglichen. Jedes Modell weist individuelle Stärken auf, wodurch sich der Anwendungsbereich und die Forschungsfragen unterscheiden. Um ein möglichst in vivo-nahes Modell zu erreichen, sind diffASCs in GG Hydrogelen ein vielversprechendes, langzeitstabiles, ausgereiftes Fettgewebemodell und können daher als flexible Plattform für in vitro Testungen genutzt werden.
Epidemiology and social determinants of chronic diseases attributed adult mortality and its influence on maternal and young child nutrition in Tigray, 2009-2015: evidence from Kilte Awlaelo- Health and Demographic Surveillance Site
(2023) Abera, Semaw Ferede; Scherbaum, Veronika
In Ethiopia, the burden of disease related to communicable diseases has recently decreased significantly, while morbidity and mortality due to non-communicable diseases (NCDs) have increased. At the same, maternal and child malnutrition remained a major public health problem of Ethiopia. In developing countries, where health insurance is largely unavailable, individual medical conditions can also affect the overall and nutritional well-being of household members. In particular, the occurrence of disease and adult mortality in households can affect the nutritional well-being of the most vulnerable household members, especially lactating mothers and their young children. If the diseases are of chronic nature, which usually are costly and adult household members die from it in the long-term, this can be devastating for the family. The aim of this Ph.D. project was to investigate the epidemiology and social determinants of NCDs-attributed adult mortality, and to examine the association of chronic diseases attributed adult mortality with undernutrition of lactating mothers and their young children in rural population of Kilte Awlaelo-Health and Demographic Surveillance Site (KA-HDSS), Eastern Zone of Tigray, Ethiopia. During the data analysis, causes of death in adults were classified into chronic and non-chronic causes. The category of adult mortality due to chronic diseases refers to all causes that may be characterized by a long duration of illness. This group includes all deaths caused by NCDs and chronic communicable diseases such as tuberculosis and HIV/AIDS. The thesis has three articles, all published in peer-reviewed journals. The first article reports findings on the epidemiology and social-determinants of adult mortality caused by NCDs among 45,982 adult residents of KA-HDSS using population-based longitudinal data collected from 2009 to 2015. The second article tested whether the burden of undernutrition was higher among lactating mothers who were living in households with adult mortality from chronic diseases than among lactating mothers living in households with no adult mortality from chronic diseases. The third article examined whether there was an association between undernutrition of children and adult mortality from chronic diseases. Both longitudinal and cross-sectional data were used in the second and third articles. To our knowledge, this study showed for the first time that adult mortality caused by NCDs varied according household members’ relationship to their household head: extended family and non-family members of the household head had higher hazard of mortality compared to the household heads. In addition, this work can be considered as the first study from a low-income setting to examine whether mortality of an adult household member from chronic diseases is associated with undernutrition of lactating mothers and their young children. The results of the first study indicate a double mortality burden from both communicable diseases and NCDs in the study population. Between 2009 and 2015, the leading causes of NCDs-attributed adult mortality were cardiovascular diseases, cancer and renal failure. Compared to heads of households, extended family and non-family co-residents had an increased hazard of mortality from NCDs. Literacy and younger age were protective factors against adult mortality caused by NCDs. However, the protective role of literacy against adult mortality from NCDs decreased with increasing age. Next, we assessed the level of undernutrition among the lactating mothers and examined its association with household-level occurrence of adult mortality from chronic diseases by controlling the effect of a wide range of epi-demographic and agro-ecological variables. Nearly two-fifths (38%; 95% CI: 36.1, 40.1%) of the mothers were undernourished. We found an increased risk of maternal undernutrition for lactating mothers who were living in households which experienced adult mortality from chronic diseases. In addition, maternal undernutrition was strongly associated with recent history of household-level morbidity, poor health-seeking practice, lack of diverse food crops, and a low index score for housing and environmental factors. In the third article, we determined the burden of undernutrition among children of complementary feeding age (6 to 23 months) and its factors within the context of nutrition-specific and -sensitive drivers of young child undernutrition. Here, mortality from chronic diseases were constructed as a nutrition-specific factor. We found high prevalence of wasting (13.7%; 95% CI: 12.1, 15.5%) and inadequate child dietary diversity (81.3%; 95%CI: 79.2, 83.1%). Adult mortality history from chronic diseases was not associated with young child undernutrition and child dietary diversity. However, child undernutrition was strongly associated with recent history of household-level morbidity, maternal undernutrition, low child dietary diversity, poverty, larger family size, insecure employment of household heads, and living in highland areas. Poor household wealth status and lack of diverse food crops production, particularly in highland areas, were also strongly associated with lower child dietary diversity. Overall, this thesis has shown that an epidemiological transition is ongoing in the surveillance population. Population-based intervention measures are recommended that aim to reduce NCD-related adult mortality by targeting the leading causes of death and focusing on vulnerable population subgroups, such as the extended family and nonfamily household members. In this study, there was no association between the occurrence of chronic diseases attributed adult mortality and young child undernutrition. However, adult mortality from chronic diseases was associated with maternal undernutrition. Our findings appear to call for multi-sectoral interventions, mainly by the agriculture, nutrition and health sectors, to promote nutritional well-being of lactating mothers and their dyads in the long-term.

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