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23 pages, 3933 KiB

Open AccessArticle

Digestive Profiles of Human Milk, Recombinant Human and Bovine Lactoferrin: Comparing the Retained Intact Protein and Peptide Release

by Bum Jin Kim, Russell F. Kuhfeld, Joanna L. Haas, Yanisa M. Anaya, Raysa Rosario Martinez, Baidya Nath P. Sah, Bella Breen, Kahler Newsham, Carrie-Anne Malinczak and David C. Dallas

Nutrients 2024, 16(14), 2360; https://doi.org/10.3390/nu16142360 - 21 Jul 2024

Viewed by 899

Abstract

Lactoferrin (LF) is a major component of human milk. LF supplementation (currently bovine) supports the immune system and helps maintain iron homeostasis in adults. No recombinant human lactoferrin (rhLF) is available for commercial food use. To determine the extent to which rhLF (Effera™) [...] Read more.

Lactoferrin (LF) is a major component of human milk. LF supplementation (currently bovine) supports the immune system and helps maintain iron homeostasis in adults. No recombinant human lactoferrin (rhLF) is available for commercial food use. To determine the extent to which rhLF (Effera™) produced by Komagataella phaffii digests similarly to hmLF, a validated in vitro digestion protocol was carried out. Bovine LF (bLF) was used as an additional control, as it is approved for use in various food categories. This study compared the extent of intact protein retention and the profile of peptides released in hmLF, bLF and rhLF (each with low and high iron saturation) across simulated adult gastric and intestinal digestion using gel electrophoresis, ELISA and LC-MS. Intact LF retention across digestion was similar across LF types, but the highest iron-saturated hmLF had greater retention in the simulated gastric fluid than all other sample types. Peptides identified in digested hmLF samples strongly correlated with digested rhLF samples (0.86 < r < 0.92 in the gastric phase and 0.63 < r < 0.70 in the intestinal phase), whereas digested bLF samples were significantly different. These findings support the potential for rhLF as a food ingredient for human consumption. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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12 pages, 2330 KiB

Open AccessArticle

Human Milk Protein-Derived Bioactive Peptides from In Vitro-Digested Colostrum Exert Antimicrobial Activities against Common Neonatal Pathogens

by Yang Lyu, Bum Jin Kim, Jagdish Suresh Patel, David C. Dallas and Yimin Chen

Nutrients 2024, 16(13), 2040; https://doi.org/10.3390/nu16132040 - 27 Jun 2024

Viewed by 840

Abstract

Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In [...] Read more.

Human milk reduces risk for necrotizing enterocolitis in preterm infants. Necrotizing enterocolitis occurs in the ileocecal region where thousands of milk protein-derived peptides have been released from digestion. Digestion-released peptides may exert bioactivity, such as antimicrobial and immunomodulatory activities, in the gut. In this study, we applied mass spectrometry-based peptidomics to characterize peptides present in colostrum before and after in vitro digestion. Sequence-based computational modeling was applied to predict peptides with antimicrobial activity. We identified more peptides in undigested samples, yet the abundances were much higher in the digested samples. Heatmapping demonstrated highly different peptide profiles between undigested and digested samples. Four peptides (α_S1-casein [157–163], α_S1-casein [157–165], β-casein [153–159] and plasminogen [591–597]) were selected, synthesized and tested against common pathogenic bacteria associated with necrotizing enterocolitis. All four exhibited bacteriostatic, though not bactericidal, activities against Klebsiella aerogenes, Citrobacter freundii and Serratia marcescens, but not Escherichia coli. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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Figure 1
Venn diagram and bar graph of (a) counts; (b) total chromatographic abundances (Log10) of human milk peptides identified from undigested and digested human milk samples; (c) Pie charts show the percentage of the total abundance of common (16% and 17%, respectively, in dark grey) and unique (84% and 83%, respectively) proteins identified in undigested (in white) and digested (in black) samples. * p = 0.005; difference in total peptide abundance between undigested and digested using related-samples Wilcoxon Signed Rank test. Full article ">Figure 2
Top 10 parent proteins based on highest number of peptides that were derived in both undigested (blue) and digested (red) human milk samples. Full article ">Figure 3
Heatmap and hierarchical clustering showing the similarity/differences of abundance pattern of human milk proteins that were identified using peptide profiles between undigested (U) and digested (D) samples. Full article ">Figure 4
Percent change in optical density at 600 nm from the growth of common pathogenic bacteria incubated with synthesized peptides SP1-SP4. (a) C. freundii; (b) E. coli; (c) K. aerogenes; (d) S. marcescens. Full article ">

17 pages, 3735 KiB

Open AccessArticle

The Immunomodulatory Effects of A2 β-Casein on Immunosuppressed Mice by Regulating Immune Responses and the Gut Microbiota

by Xiao Li, Xingru Lu, Ming Liu, Yu Zhang, Yujun Jiang, Xinyan Yang and Chaoxin Man

Nutrients 2024, 16(4), 519; https://doi.org/10.3390/nu16040519 - 13 Feb 2024

Cited by 2 | Viewed by 1328

Abstract

The aim of this study was to investigate the immunomodulatory effects of A2 β-casein (β-CN) in cyclophosphamide-induced immunosuppressed BALB/c mice. Experiments conducted in vitro revealed that A2 β-CN digestive products have potent immunostimulatory activities. Animal studies demonstrated that A2 β-CN improved the immunological [...] Read more.

The aim of this study was to investigate the immunomodulatory effects of A2 β-casein (β-CN) in cyclophosphamide-induced immunosuppressed BALB/c mice. Experiments conducted in vitro revealed that A2 β-CN digestive products have potent immunostimulatory activities. Animal studies demonstrated that A2 β-CN improved the immunological organ index reduction trend caused by cyclophosphamide, reduced the pathological damage to the spleen tissue in immunosuppressed mice, increased the release of IL-17A, IgG, and IgA, and reduced the production of IL-4. By regulating the relative abundance of advantageous bacteria like Oscillospira, Lactobacillus, and Bifidobacteria and harmful bacteria like Coprococcus and Desulfovibrionaceae, A2 β-CN improved gut microbiota disorders in immunosuppressed mice. Moreover, A2 β-CN promoted the production of short-chain fatty acids and increased the diversity of the gut microbiota. Therefore, ingestion of A2 β-CN is beneficial to the host’s immune system and gut health. These findings provide insights for the future application of A2 β-CN-related dairy products. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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Graphical abstract

Graphical abstract
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Effects of β-CN digests on three immune cells. (A) The proliferative capacity of splenic lymphocytes, (B) the phagocytic capacity of phagocytes, (C) NK cell activity. Comparison of different significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Full article ">Figure 2
Effect of A2 β-CN on organ indexes and immune cells in mice. (A) Spleen index, (B) thymus index, (C) proliferation of splenic lymphocytes, (D) phagocytosis percentage, (E) phagocytic index, (F) NK cell activity. NC: control group, M: model group, LA2: low-dose A2 β-CN group, MA2: medium-dose A2 β-CN group, HA2: high-dose A2 β-CN group, CN: high-dose A1 β-CN group. Compared with the normal group:, ## p < 0.01, ### p < 0.001; compared with the model group: * p < 0.05, ** p < 0.01, *** p < 0.001. Full article ">Figure 3
Effects of A2 β-CN on cytokines and immunoglobulins. (A) IL-4 content, (B) IL-17A content, (C) IgG content, (D) IgA content. Compared with the normal group: ## p < 0.01, ### p < 0.001; compared with the model group: * p < 0.05, ** p < 0.01, *** p < 0.001. Full article ">Figure 4
Effect of A2 β-casein on histopathology of spleen (H&E staining). Magnification: 200× and scale bar: 50 µm. Full article ">Figure 5
Effect of A2 β-CN on gut microbiota in immunosuppressed mice. (A) Dilution curve, (B) rank abundance curves, (C) Chao1 index, (D) Simpson index, (E) NMDS analysis. Full article ">Figure 6
Effect of A2 β-CN on gut microbiota and SCFAs in immunosuppressed mice. (A) Department level, (B) genus level, (C) heat map of species composition at the genus level, (D) content of SCFAs. Compared with the normal group: # p < 0.05, ## p < 0.01; compared with the model group: * p < 0.05, ** p < 0.01. Full article ">

15 pages, 1882 KiB

Open AccessArticle

Macrophage-Immunomodulatory Actions of Bovine Whey Protein Isolate, Glycomacropeptide, and Their In Vitro and In Vivo Digests

by Wyatt Olsen, Ningjian Liang and David C. Dallas

Nutrients 2023, 15(23), 4942; https://doi.org/10.3390/nu15234942 - 28 Nov 2023

Cited by 3 | Viewed by 1178

Abstract

Whey protein isolate (WPI) consists of an array of proteins and peptides obtained as a byproduct of the cheesemaking process. Research suggests that WPI, along with its peptides such as glycomacropeptide (GMP), possesses immunomodulatory properties. These properties hold potential for alleviating the adverse [...] Read more.

Whey protein isolate (WPI) consists of an array of proteins and peptides obtained as a byproduct of the cheesemaking process. Research suggests that WPI, along with its peptides such as glycomacropeptide (GMP), possesses immunomodulatory properties. These properties hold potential for alleviating the adverse effects of inflammatory conditions such as inflammatory bowel disease. Although promising, the immunoregulatory properties of the digested forms of WPI and GMP—those most likely to interact with the gut immune system—remain under-investigated. To address this knowledge gap, the current study examined the effects of in vitro-digested WPI and GMP, in vivo-digested WPI, and undigested WPI and GMP on the secretion of pro-inflammatory cytokines (TNF-α and IL-1β) in lipopolysaccharide-stimulated macrophage-like cells. Our results indicate that digested WPI and GMP reduced the expression of TNF-α and IL-1β, two pro-inflammatory cytokines. Whole WPI had no effect on TNF-α but reduced IL-1β levels. In contrast, in vivo-digested WPI reduced TNF-α but increased IL-1β. Undigested GMP, on the other hand, increased the secretion of both cytokines. These results demonstrate that digestion greatly modifies the effects of WPI and GMP on macrophages and suggest that digested WPI and GMP could help mitigate gastrointestinal inflammation. Further clinical studies are necessary to determine the biological relevance of WPI and GMP digestion products within the gut and their capacity to influence gut inflammation. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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14 pages, 1725 KiB

Open AccessArticle

Evaluating the Potential of Casein Glycomacropeptide in Adult Irritable Bowel Syndrome Management: A Pilot Study

by Yunyao Qu, Si Hong Park and David C. Dallas

Nutrients 2023, 15(19), 4174; https://doi.org/10.3390/nu15194174 - 27 Sep 2023

Cited by 3 | Viewed by 1606

Abstract

Irritable bowel syndrome (IBS) is a common gastrointestinal disorder that affects 10–15% of the global population and presents symptoms such as abdominal discomfort, bloating and altered bowel habits. IBS is believed to be influenced by gut microbiota alterations and low-grade inflammation. Bovine kappa-casein [...] Read more.

Irritable bowel syndrome (IBS) is a common gastrointestinal disorder that affects 10–15% of the global population and presents symptoms such as abdominal discomfort, bloating and altered bowel habits. IBS is believed to be influenced by gut microbiota alterations and low-grade inflammation. Bovine kappa-casein glycomacropeptide (GMP), a bioactive dairy-derived peptide, possesses anti-adhesive, prebiotic and immunomodulatory properties that could potentially benefit IBS patients. This pilot study investigated the effects of daily supplementation with 30 g of GMP for three weeks on gut health in five people with IBS. We assessed alterations in gut microbiota composition, fecal and blood inflammatory makers, and gut-related symptoms before, during and after the GMP feeding period. The results revealed no changes in fecal microbiota, subtle effects on systemic and intestinal immune makers, and no changes in gut-related symptoms during and after the GMP supplementation. Further research is needed to assess the potential benefits of GMP in IBS patients, including the examination of dosage and form of GMP supplementation. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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22 pages, 6183 KiB

Open AccessArticle

Fingerprinting of Proteases, Protease Inhibitors and Indigenous Peptides in Human Milk

by Martin Nørmark Thesbjerg, Søren Drud-Heydary Nielsen, Ulrik Kræmer Sundekilde, Nina Aagaard Poulsen and Lotte Bach Larsen

Nutrients 2023, 15(19), 4169; https://doi.org/10.3390/nu15194169 - 27 Sep 2023

Cited by 3 | Viewed by 1616

Abstract

The presence of proteases and their resulting level of activity on human milk (HM) proteins may aid in the generation of indigenous peptides as part of a pre-digestion process, of which some have potential bioactivity for the infant. The present study investigated the [...] Read more.

The presence of proteases and their resulting level of activity on human milk (HM) proteins may aid in the generation of indigenous peptides as part of a pre-digestion process, of which some have potential bioactivity for the infant. The present study investigated the relative abundance of indigenous peptides and their cleavage products in relation to the abundance of observed proteases and protease inhibitors. The proteomes and peptidomes in twelve HM samples, representing six donors at lactation months 1 and 3, were profiled. In the proteome, 39 proteases and 29 protease inhibitors were identified in 2/3 of the samples. Cathepsin D was found to be present in higher abundance in the proteome compared with plasmin, while peptides originating from plasmin cleavage were more abundant than peptides from cathepsin D cleavage. As both proteases are present as a system of pro- and active- forms, their activation indexes were calculated. Plasmin was more active in lactation month 3 than month 1, which correlated with the total relative abundance of the cleavage product ascribed to plasmin. By searching the identified indigenous peptides in the milk bioactive peptide database, 283 peptides were ascribed to 10 groups of bioactivities. Antimicrobial peptides were significantly more abundant in month 1 than month 3; this group comprised 103 peptides, originating from the β-CN C-terminal region. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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Figure 1
The top 25 most abundant proteins identified in the proteome. Bars to the left represent the relative abundance of the protein in the proteome, while the bars to the right represent the relative abundance of the protein in the peptidome. Relative abundancies (in% of total protein) are displayed on the x-axis with bar plots, where the bar represents the median relative abundance and the inner and outer whiskers represent the 1st and 3rd quartiles, respectively. The y-axis represents the gene names for the identified proteins according to uniprot.org. Full article ">Figure 2
Relationship between the degree of activation (y-axis) for plasmin and cathepsin D enzyme systems and relative abundance of cleavage products (peptides in peptidome) ascribed to either enzyme system based on cleavage specificities. The x-axis indicates the relative abundance of the enzyme products, i.e., peptides in the peptidome. The y-axis shows the activation degree (enzyme/(enzyme + proenzyme)) for plasminogen and procathepsin D, based on the relative abundance of peptides originating from the activation peptide regions and peptides from the mature enzyme. Full article ">Figure 3
Average relative abundance (y-axis) of the 39 proteases (x-axis) identified in the proteome (white bars, right y-axis) or giving rise to peptides found in the peptidome, based on enzyme specificities (grey bars, right y-axis), and as represented in at least 2/3 of the HM samples. The top and bottom whiskers represent the 1st and 3rd quartiles, respectively. Full article ">Figure 4
Residues at P1 (top panel) and P1′ (bottom panel) positions of the peptides in the peptidome of the indigenous peptides. The amino acid residues are represented by one-letter codes on the x-axis. The bars represent the median count of the total observations within each of the samples. The top and bottom whiskers represent the 1st and 3rd quartiles, respectively. The line represents the average summarized peptide intensities representing each residue across each sample. The left y-axis shows the peptide counts; the right y-axis shows the peptide intensities. Full article ">Figure 5
Shows a peptide intensity map of the caseins in the peptidome (graphs) with cleavage sites for the four most active proteases (vertical lines). The x-axis shows the amino acid position, including signal peptides. The y-axis shows the ion intensity (does not apply to the vertical lines). The summarized intensities were calculated using PepEx and plotted as graphs for the individual donors (lines). The color code applies to all three plots. Full article ">Figure 6
Shows a peptide intensity map of the caseins in the peptidome (graphs) with cleavage sites for the four most active proteases (vertical lines). The x-axis shows the amino acid position, including signal peptides. The y-axis shows the ion intensity (does not apply to the vertical lines). The summarized intensities were calculated using PepEx and plotted as graphs for the individual donors (lines). The color code applies to both plots. Full article ">Figure 7
Shows the relative abundance of the groups of bioactive peptides in the peptidome. Light grey bars represent lactation month 1, while dark grey bars represent lactation month 3. The bar represents the average, while the top and bottom whiskers represent the 1st and 3rd quartiles, respectively. Of the ten groups of bioactive peptides, the relative abundances of the antimicrobial and antioxidant groups were found to be significantly different between lactation months 1 and 3. The break in the y-axis is from 0.25 to 2. Asterisks denote the level of significance between the lactation groups: *, p ≤ 0.05; **, p ≤ 0.01. Full article ">

12 pages, 763 KiB

Open AccessArticle

Effects of Whey Protein Supplementation on Inflammatory Marker Concentrations in Older Adults

by Samuel Adler, Wyatt Olsen, Bryna Rackerby, Rachel Spencer and David C. Dallas

Nutrients 2023, 15(18), 4081; https://doi.org/10.3390/nu15184081 - 21 Sep 2023

Viewed by 3931

Abstract

Although whey protein isolate (WPI) has been shown to be immunomodulatory, its ability to modulate production of a broad array of inflammatory markers has not previously been investigated in healthy adults. We investigated the effects of daily supplementation with 35 g of WPI [...] Read more.

Although whey protein isolate (WPI) has been shown to be immunomodulatory, its ability to modulate production of a broad array of inflammatory markers has not previously been investigated in healthy adults. We investigated the effects of daily supplementation with 35 g of WPI for 3 weeks on inflammatory marker concentrations in the blood serum and feces of 14 older adult subjects (mean age: 59). Serum was analyzed using a multiplex assay to quantify the cytokines IFN-γ, IL-1β, IL-1RA, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, IL-13, IL-17A and TNF-α. Fecal samples were analyzed using an ELISA for the inflammatory markers calprotectin and lactoferrin. Our results yielded high inter-subject variability and a significant proportion of cytokine concentrations that were below our method’s limit of quantification. We observed decreases in serum IL-12p70 in the washout phase compared with baseline, as well as the washout stage for fecal lactoferrin relative to the intervention stage. Serum IL-13 was also significantly reduced during the intervention and washout stages. Our data suggest that whey protein supplementation did not significantly alter most inflammatory markers measured but can alter concentrations of some inflammatory markers in healthy older adults. However, our study power of 35% suggests the number of participants was too low to draw strong conclusions from our data. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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11 pages, 976 KiB

Open AccessArticle

Effects of Lactoferrin on Oral and Throat Conditions under Low Humidity Environments: A Randomized, Double-Blind, and Placebo-Controlled Crossover Trial

by Shutaro Kubo, Hirotsugu Oda, Miyuki Tanaka, Takashi Koikeda and Shinichi Tomita

Nutrients 2023, 15(18), 4033; https://doi.org/10.3390/nu15184033 - 18 Sep 2023

Viewed by 1598

Abstract

To evaluate the effects of a single ingestion of bovine lactoferrin (bLF) on oral and throat conditions under a low-humidity environment. A randomized, double-blind, 2-sequence, 2-treatment, and 2-period placebo-controlled crossover trial was conducted. Healthy adult subjects orally ingested bLF dissolved in water, or [...] Read more.

To evaluate the effects of a single ingestion of bovine lactoferrin (bLF) on oral and throat conditions under a low-humidity environment. A randomized, double-blind, 2-sequence, 2-treatment, and 2-period placebo-controlled crossover trial was conducted. Healthy adult subjects orally ingested bLF dissolved in water, or placebo water, followed by exposure to low humidity (20 °C, 20% relative humidity (RH)) for 2 h. The primary endpoint was subjective oral and throat discomfort assessed by a visual analog scale (VAS), which positively correlated with the discomfort. Secondary endpoints were unstimulated whole salivary flow rate (UWSFR) and salivary immunoglobulin A (IgA) secretion rate. Overall, 40 subjects were randomly assigned to two sequences (20 each) and 34 were analyzed. The VAS values for oral and throat discomfort in the bLF treatment were significantly lower than in the placebo treatment, whereas UWSFR and IgA secretion rates were comparable between the two treatments. Adverse drug reactions were not observed. Subjective oral and throat discomfort associated with low humidity is suppressed by a single ingestion of bLF. Our findings demonstrate the novel use of bLF in a clinical situation that leverages its unique characteristics. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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9 pages, 586 KiB

Open AccessArticle

Effects of Bovine Lactoferrin on the Maintenance of Respiratory and Systemic Physical Conditions in Healthy Adults—A Randomized, Double-Blind, Placebo-Controlled Trial

by Hirotsugu Oda, Shutaro Kubo, Asuka Tada, Takumi Yago, Chihiro Sugita, Hiroki Yoshida, Tatsunori Toida, Miyuki Tanaka and Masahiko Kurokawa

Nutrients 2023, 15(18), 3959; https://doi.org/10.3390/nu15183959 - 13 Sep 2023

Viewed by 2086

Abstract

Objectives: We investigated the effects of bovine lactoferrin (LF) on the maintenance of the respiratory and systemic physical conditions. Methods: A randomized, double-blind, placebo-controlled trial was conducted. Healthy adults at Kyushu University of Health and Welfare ingested a placebo or bovine LF (200 [...] Read more.

Objectives: We investigated the effects of bovine lactoferrin (LF) on the maintenance of the respiratory and systemic physical conditions. Methods: A randomized, double-blind, placebo-controlled trial was conducted. Healthy adults at Kyushu University of Health and Welfare ingested a placebo or bovine LF (200 mg/day) for 12 weeks. The primary endpoints were the total respiratory and systemic symptom scores. The secondary endpoint was the activity of plasmacytoid dendritic cells (pDCs) in peripheral blood. Results: A total of 157 subjects were randomized (placebo, n = 79; LF, n = 78), of whom, 12 dropped out. The remaining 145 participants were included in the full analysis set (placebo group, n = 77; LF group, n = 68). The total scores for respiratory and systemic symptoms during the intervention were significantly lower in the LF group than in the placebo group. The expression of CD86 and HLA-DR on pDCs was significantly higher in the LF group than in the placebo group at week 12. Adverse events were comparable between the groups, and no adverse drug reactions were observed. Conclusions: These results suggest that orally ingested LF supports the normal immune system via maintaining pDC activity, and maintains respiratory and systemic physical conditions in healthy adults. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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12 pages, 1821 KiB

Open AccessArticle

Effects of Whey Protein Isolate on Body Composition, Muscle Mass, and Strength of Chronic Heart Failure Patients: A Randomized Clinical Trial

by Elisa M. dos Santos, Annie S. B. Moreira, Grazielle V. B. Huguenin, Eduardo Tibiriça and Andrea De Lorenzo

Nutrients 2023, 15(10), 2320; https://doi.org/10.3390/nu15102320 - 16 May 2023

Cited by 1 | Viewed by 4226

Abstract

Heart failure (HF) is associated with a reduction of skeletal muscle mass. Whey protein isolate (WPI) has been beneficial in increasing muscle mass and strength, in addition to improving body composition. The goal of this research was to evaluate the effect of WPI [...] Read more.

Heart failure (HF) is associated with a reduction of skeletal muscle mass. Whey protein isolate (WPI) has been beneficial in increasing muscle mass and strength, in addition to improving body composition. The goal of this research was to evaluate the effect of WPI on the body composition, muscle mass, and strength of chronic HF patients. For this purpose, twenty-five patients of both genders with predominantly NYHA I functional class and a median age of 65.5 (60.5–71.0) years were used to conduct a randomized, single-blind, placebo-controlled clinical trial and received 30 g per day of WPI for 12 weeks. Anthropometric measurements, body composition analysis, and biochemical exams were performed at the beginning and end of the study. An increase in skeletal muscle mass was observed in the intervention group after 12 weeks. A reduction in waist circumference, body fat percentage, and an increase in skeletal muscle index was observed when compared to the placebo group. No significant effect on muscle strength was observed after 12 weeks of intervention. These data demonstrate that WPI consumption contributed to the increase of skeletal muscle mass, strength, and reduction of body fat in HF patients. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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12 pages, 296 KiB

Open AccessArticle

Comparison of 30 Cytokines in Human Breast Milk between 1989 and 2013 in Japan

by Tomoki Takahashi, Hiroshi M. Ueno, Fumiya Yamaide, Taiji Nakano, Yuki Shiko, Yohei Kawasaki, Chisako Mitsuishi and Naoki Shimojo

Nutrients 2023, 15(7), 1735; https://doi.org/10.3390/nu15071735 - 1 Apr 2023

Cited by 4 | Viewed by 2382

Abstract

Milk cytokines play a vital role in mucosal immunity during infancy by supporting immune development and functions. Although the maternal background characteristics influence milk cytokines, changes in cytokine levels across generations remain unclear. Colostrum (C, n = 48) and mature milk (MM, n [...] Read more.

Milk cytokines play a vital role in mucosal immunity during infancy by supporting immune development and functions. Although the maternal background characteristics influence milk cytokines, changes in cytokine levels across generations remain unclear. Colostrum (C, n = 48) and mature milk (MM, n = 49) samples were collected from lactating Japanese women in 1989 (2727 samples) and 2013 (1408 samples). Milk cytokines were comprehensively measured using a suspension array and immunosorbent assays. The positive rates and cytokine concentrations were compared between the two generations using logistic and multiple regression analyses. Twenty-eight cytokines tested positive in all sample groups (1989-C, 1989-MM, 2013-C, and 2013-MM). The median osteopontin (OPN) level was significantly higher in the 1989-C group than in the 2013-C group (318.1 vs. 137.5 μg/mL; p = 0.0016) but did not differ between the MM groups. The median TGF-β1 level was significantly lower in the 1989-MM group than in the 2013-MM group (1056.2 vs. 1330.8 pg/mL; p = 0.008) but did not differ between the C groups. Most cytokines were comparable between generations, except for potential variation in the C-OPN and TGF-β1 levels. Milk cytokine secretion may reflect temporal changes in maternal background characteristics; however, the results from the analysis of 30-year-old samples may have influenced the milk cytokine levels. Further studies are needed with a larger number of milk samples collected from the same individuals at multiple time points over a wide lactation period, with detailed data on the maternal and infant background characteristics and diets. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

11 pages, 1041 KiB

Open AccessFeature PaperArticle

The Effect of Human and Bovine Milk Osteopontin on Intestinal Caco-2 Cells: A Transcriptome Comparison

by Brian Christensen, Albert J. Buitenhuis, Lotte N. Jacobsen, Marie S. Ostenfeld and Esben S. Sørensen

Nutrients 2023, 15(5), 1166; https://doi.org/10.3390/nu15051166 - 25 Feb 2023

Cited by 4 | Viewed by 2077

Abstract

Osteopontin (OPN) is a multifunctional protein abundantly present in human milk, whereas the concentration is significantly lower in bovine milk. Human and bovine milk OPN are structurally similar and both proteins resist gastric digestion and reach the intestines in a bioactive form. Intervention [...] Read more.

Osteopontin (OPN) is a multifunctional protein abundantly present in human milk, whereas the concentration is significantly lower in bovine milk. Human and bovine milk OPN are structurally similar and both proteins resist gastric digestion and reach the intestines in a bioactive form. Intervention studies have indicated the beneficial effects of supplementing infant formula with bovine milk OPN and several in vivo and in vitro studies have shown that bovine milk OPN positively influences intestinal development. To investigate the functional relationship, we compared the effect of simulated gastrointestinal digested human and bovine milk OPN on gene expression in Caco-2 cells. After incubation, total RNA was extracted and sequenced and transcripts were mapped to the human genome. Human and bovine milk OPN regulated the expression of 239 and 322 genes, respectively. A total of 131 genes were similarly regulated by the OPNs. As a control, a whey protein fraction with a high content of alpha-lactalbumin had a very limited transcriptional impact on the cells. Enrichment data analysis showed that biological processes related to the ubiquitin system, DNA binding, and genes associated with transcription and transcription control pathways were affected by the OPNs. Collectively, this study shows that human and bovine milk OPN have a significant and highly comparable effect on the intestinal transcriptome. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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Review

Jump to: Research

32 pages, 3227 KiB

Open AccessReview

Science and Faith to Understand Milk Bioactivity for Infants

by Per T. Sangild

Nutrients 2024, 16(11), 1676; https://doi.org/10.3390/nu16111676 - 29 May 2024

Viewed by 1198

Abstract

Milk bioactivity refers to the specific health effects of milk components beyond nutrition. The science of milk bioactivity involves the systematic study of these components and their health effects, as verified by empirical data, controlled experiments, and logical arguments. Conversely, ’faith in milk [...] Read more.

Milk bioactivity refers to the specific health effects of milk components beyond nutrition. The science of milk bioactivity involves the systematic study of these components and their health effects, as verified by empirical data, controlled experiments, and logical arguments. Conversely, ’faith in milk bioactivity’ can be defined as personal opinion, meaning, value, trust, and hope for health effects that are beyond investigation by natural, social, or human sciences. Faith can be strictly secular, but also influenced by spirituality or religion. The aim of this paper is to show that scientific knowledge is frequently supplemented with faith convictions to establish personal and public understanding of milk bioactivity. Mammalian milk is an immensely complex fluid containing myriad proteins, carbohydrates, lipids, and micronutrients with multiple functions across species, genetics, ages, environments, and cultures. Human health includes not only physical health, but also social, mental, and spiritual health, requiring widely different fields of science to prove the relevance, safety, and efficacy of milk interventions. These complex relationships between milk feeding and health outcomes prevent firm conclusions based on science and logic alone. Current beliefs in and understanding of the value of breast milk, colostrum, infant formula, or isolated milk proteins (e.g., immunoglobulins, α-lactalbumin, lactoferrin, and growth factors) show that both science and faith contribute to understand, stimulate, or restrict the use of milk bioactivity. The benefits of breastfeeding for infants are beyond doubt, but the strong beliefs in its health effects rely not only on science, and mechanisms are unclear. Likewise, fear of, or trust in, infant formula may rely on both science and faith. Knowledge from science safeguards individuals and society against ‘milk bioactivity superstition’. Conversely, wisdom from faith-based convictions may protect science from unrealistic ‘milk bioactivity scientism’. Honesty and transparency about the potentials and limitations of both scientific knowledge and faith convictions are important when informing individuals and society about the nutritious and bioactive qualities of milk. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Schematic overview of how both science and faith may form beliefs in milk bioactivity. Here, ‘sciences’ represent all academic disciplines at universities. ‘Faith’ covers secular but also spiritual or religious views and attitudes to human meaning and existence, including health effects of milk (milk bioactivity) for infants. Examples of scientific study fields and faith convictions related to milk bioactivity are listed. Scientific knowledge helps to avoid superstition related to milk bioactivity. Faith elements help to avoid ‘scientism’ related to milk bioactivity, thus preventing unrealistic and exclusive reliance on scientific theories and analyses. Together, science and faith may synergize to form beliefs that determine how to understand and implement milk bioactivity for infant health. Full article ">Figure 2
Illustration of some selected milk proteins and their roles as nutrients (indicated by the green ellipse), bioactives (indicated by the red ellipse), or a mix of the two (brown overlapping area). Nutrition effects of milk proteins (e.g., tissue building blocks and energy) interact with roles to regulate body functions and health. Those listed represent only a small fraction of the thousands of known proteins in mammalian milk. Whether a protein is considered a nutrient or a bioactive factor also depends on its digestibility and concentration; both are high for nutrients. In addition to proteins, mammalian milk contains numerous other nutrients and bioactive components, categorized as carbohydrates (e.g., lactose and oligosaccharides), lipids (e.g., glycerides and fatty acids), minerals, vitamins, or other biological categories, interacting with milk proteins and peptides. Full article ">Figure 3
An illustration of the fields of study in milk bioactivity research across natural, social, and human (humanity) sciences. The differently colored ellipses denote the natural sciences, social sciences, and human sciences, as well as their overlaps. The gray circles denote the more specific fields of study relating to milk bioactivity within these different sciences. The text below the circles shows the spectrum of methodologies across the different sciences. Note the overlaps among scientific domains, specific topics, and methodologies, despite their unique characteristics. By research target, and especially by research methodology, social science can be seen as being intermediate between the natural and human sciences. Social science is engaged with studies on both nature and human society and relationships. Social sciences use both qualitative and quantitative research methods. For further information, see <a href="#app1-nutrients-16-01676" class="html-app">Supplementary Figures S1 and S2</a>. Full article ">Figure 4
Illustration of the three domains of human health (physical, social, and mental health shown by green, yellow and red ‘leaves’), as defined by the World Health Organization (WHO) [<a href="#B74-nutrients-16-01676" class="html-bibr">74</a>]. A fourth dimension, spiritual health, has been suggested to be an important addition to the 3-fold WHO health definition [<a href="#B77-nutrients-16-01676" class="html-bibr">77</a>,<a href="#B78-nutrients-16-01676" class="html-bibr">78</a>] and is added to the illustration (the grey ‘leaf’). Spiritual health reflects aspects of personal existence, meaning, hope, love, and trust in something greater than oneself, with or without involvement of religious faith. Embedded pictures obtained from Pixibay.com. Full article ">Figure 5
Overview of some milk bioactive proteins (left side) that may affect human health (right side). The blue arrows denote the relationship between these two aspects, influencing each other. The protein list is not complete and may include many others [<a href="#B48-nutrients-16-01676" class="html-bibr">48</a>,<a href="#B49-nutrients-16-01676" class="html-bibr">49</a>,<a href="#B107-nutrients-16-01676" class="html-bibr">107</a>], varying among species, stages of lactation, and health states. When health outcomes include not only physical health (investigated by natural science), but also social, mental, and spiritual health (<a href="#nutrients-16-01676-f004" class="html-fig">Figure 4</a>), the possible interactions with milk bioactivity are endless and highly complex. Milk bioactive proteins affect health outcomes, and each health state influences how milk bioactive proteins work in the body. Full article ">Figure 6
Scientific research into the bioactivity of breastfeeding and human milk for infants requires natural, social, and human sciences to answer all relevant scientific questions. The (left side) of the figure provides examples of such questions and the corresponding science field(s). Scientific evidence is incomplete, in part due to the multitudes of interacting variables and difficulties in performing randomized, controlled studies on mothers and infants. Physical, social, and personal benefits of breastfeeding for mothers and infants are well documented by clinical science, but cause–effect relationships and molecular mechanisms (natural science) are unclear. Faith elements, including existential, spiritual, and/or religious attitudes to breastfeeding and human milk may antagonize, complement, or synergize with knowledge from science to form the beliefs that are the basis for public understanding, guidelines, and practice. Examples of faith convictions indicating existentiality, spirituality, or religion, or a combination, are shown in the figure (right side). Looking into breastfeeding and human milk from both science and faith ‘windows’ facilitates a broader and more nuanced picture of breastfeeding than from the science perspective alone. Image from WikiMedia: ‘Madonna Litta’, attributed to Leonardo da Vinci (1452–1519), Italian scientist, naturalist, and artist. Full article ">

22 pages, 1263 KiB

Open AccessReview

The Role of Bovine Kappa-Casein Glycomacropeptide in Modulating the Microbiome and Inflammatory Responses of Irritable Bowel Syndrome

by Yunyao Qu, Si Hong Park and David C. Dallas

Nutrients 2023, 15(18), 3991; https://doi.org/10.3390/nu15183991 - 15 Sep 2023

Cited by 6 | Viewed by 2156

Abstract

Irritable bowel syndrome (IBS) is a common gastrointestinal disorder marked by chronic abdominal pain, bloating, and irregular bowel habits. Effective treatments are still actively sought. Kappa-casein glycomacropeptide (GMP), a milk-derived peptide, holds promise because it can modulate the gut microbiome, immune responses, gut [...] Read more.

Irritable bowel syndrome (IBS) is a common gastrointestinal disorder marked by chronic abdominal pain, bloating, and irregular bowel habits. Effective treatments are still actively sought. Kappa-casein glycomacropeptide (GMP), a milk-derived peptide, holds promise because it can modulate the gut microbiome, immune responses, gut motility, and barrier functions, as well as binding toxins. These properties align with the recognized pathophysiological aspects of IBS, including gut microbiota imbalances, immune system dysregulation, and altered gut barrier functions. This review delves into GMP’s role in regulating the gut microbiome, accentuating its influence on bacterial populations and its potential to promote beneficial bacteria while inhibiting pathogenic varieties. It further investigates the gut microbial shifts observed in IBS patients and contemplates GMP’s potential for restoring microbial equilibrium and overall gut health. The anti-inflammatory attributes of GMP, especially its impact on vital inflammatory markers and capacity to temper the low-grade inflammation present in IBS are also discussed. In addition, this review delves into current research on GMP’s effects on gut motility and barrier integrity and examines the changes in gut motility and barrier function observed in IBS sufferers. The overarching goal is to assess the potential clinical utility of GMP in IBS management. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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20 pages, 864 KiB

Open AccessReview

Milk Osteopontin and Human Health

by Esben S. Sørensen and Brian Christensen

Nutrients 2023, 15(11), 2423; https://doi.org/10.3390/nu15112423 - 23 May 2023

Cited by 9 | Viewed by 3541

Abstract

Osteopontin (OPN) is a multifunctional protein found in all vertebrates. OPN is expressed in many different cell types, and is consequently found in most tissues and physiological secretions. OPN is involved in a multitude of biological processes, such as activation and regulation of [...] Read more.

Osteopontin (OPN) is a multifunctional protein found in all vertebrates. OPN is expressed in many different cell types, and is consequently found in most tissues and physiological secretions. OPN is involved in a multitude of biological processes, such as activation and regulation of the immune system; biomineralization; tissue-transformative processes, including growth and development of the gut and brain; interaction with bacteria; and many more. OPN is found in the highest concentrations in milk, where it is believed to initiate and regulate developmental, immunological and physiological processes in infants who consume milk. Processes for the isolation of bovine OPN for use in infant formula have been developed, and in recent years, many studies have investigated the effects of the intake of milk OPN. The purpose of this article is to review and compare existing knowledge about the structure and function of milk OPN, with a particular focus on the effects of milk OPN on human health and disease. Full article

(This article belongs to the Special Issue Bioactive Milk Proteins and Human Health)

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