Extensive Hydrolysate Research Articles

Glutamic acid intake by formula-fed infants: are acceptable daily intakes appropriate?

The 2017 European Food Safety Authority (EFSA) recommendation of an acceptable daily intake (ADI) of 30 mg glutamic acid/kg bw/day did not take into consideration the primary energy sources during infancy, including infant formulas. In the present study, we determined total daily intakes of glutamic acid in a contemporary cohort of healthy infants who were fed either cow milk formula (CMF) or extensive protein hydrolysate formula (EHF); the formulas differed substantially in glutamic acid content. The infants (n = 141) were randomized to be fed either CMF or EHF. Dietary intakes were determined from weighed bottle methods and/or prospective diet records, and body weights were measured on 14 occasions from 0.5 to 12.5 months. Secondary data analysis determined the glutamic acid content of the diet over time. The trial was registered at http://www.clinicaltrials.gov/ as NCT01700205, 3 October 2012. Glutamic acid intake from formula and other foods was significantly higher in infants fed EHF when compared to CMF. As glutamic acid intake from formula decreased, intake from other nutritional sources steadily increased from 5.5 months. Regardless of formula type, every infant exceeded the ADI of 30 mg/kg bw/day from 0.5 to 12.5 months. Conclusion: Given that the ADI recommendation was not based on actual intake data of primary energy sources during infancy, the present findings on the growing child’s ingestion of glutamic acid from infant formula and the complementary diet may be of interest when developing future guidelines and communications to parents, clinical care providers, and policy makers.What is Known:• The 2017 re-evaluation of the safety of glutamic acid-glutamates and the recommended acceptable daily intake (ADI) of 30 mg/kg bw/d by the European Food Safety Authority (EFSA) did not include actual intake data of the primary energy sources during infancy.What is New:• During the first year, glutamic acid intake from infant formula and other food sources exceeded the ADI of 30 mg/kg bw/day.

Soy Formula as An Alternative for Infants with Cow’s Milk Allergy: Perspective from Pediatricians and Midwives in Indonesia

The prevalence of food allergy in children is increasing globally, with cow's milk allergy beingthe most common in infants and increasing from 2% to 3% in developed countries. However, data oncow's milk allergy in infants is limited in Indonesia. This study used a mixed methods approach using anonline survey and in-depth interviews. The survey was conducted from September 2021 to March 2022.Qualitative analysis, all interviews were recorded and transcribed. Data were analyzed using simplethematic analysis. The results showed that a total of 93 pediatricians and 104 midwives participated inthe survey. However, among the 93 pediatricians, only 56 (60.2%) completed the survey. Meanwhile, 11midwives (10.6%) were excluded because they had no experience in handling CMA cases. The majorityof CMA cases occurred in children under two years old, and most were treated by pediatricians. In termsof diagnostic tests, 51.8% of pediatricians recommended all specific IgE tests, prick tests, and provocationtests to diagnose allergies. More than 80% (46/56) of pediatricians chose to avoid all types of cow's milkproducts in the mother and child's diet as the main treatment. Meanwhile, only 22.1% of midwives didthe same. Milk choice will differ depending on the severity of CMA. Soy milk formula is preferred bymidwives for patients with low-level and moderate to severe allergies. Meanwhile, pediatriciansrecommend extensive hydrolysate formula for low severity patients and amino acid-based formula formoderate to severe.

Peptide profiles and allergy-reactivity of extensive hydrolysates of milk protein

Milk protein concentrate (MPC) is one of the major allergens in food. This study aimed to analyze the peptide profiles and potential allergenicity of the extensive hydrolysates of MPC (EMPHs) using the peptidomics approach. Results demonstrated that when the hydrolysis time was 4 h, the degree of hydrolysis of the four EMPHs (AX, Alcalase-Protamex), (AD, Alcalase-Protease A 2SD), (AE, Alcalase-Flavourzyme) and (AH, Alcalase-ProteAXH) were 12.45 %, 18.48 %, 18.87 % and 16.77 %, respectively. The results of size exclusion chromatography showed no significant difference, when the hydrolysis time exceeded 3 h. A total of 16 allergic peptides were identified in the EMPHs by LC-MS/MS. The peptide profiles and the coverage of master protein of the four EMPHs were different. The results of the enzyme-linked immunoassay and KU812 cell model showed that the allergenicity of the EMPHs samples was significantly reduced. This study provided strong support for the application of EMPHs in hypoallergenic formula foods.

Peptide profiles and antioxidant capacity of extensive hydrolysates of milk protein concentrate.

Milk protein concentrate was hydrolyzed using one-step enzymatic hydrolysis. Both the peptide profiles and antioxidant activities of the resulting extensive hydrolysates of milk protein concentrate (EMPH) were analyzed using a peptidomics approach based on liquid chromatography-tandem mass spectrometry. The results demonstrated that the degrees of hydrolysis of the 4 EMPH by Alcalase-Protamex, Alcalase-Protease A 2SD, Alcalase-Flavorzyme, and Alcalase-ProteAXH were 12.02%, 16.85%, 15.87%, and 15.77%, respectively. Using size exclusion chromatography, 99.85% of the peptides in the Alcalase-Protease A 2SD hydrolysate were shown to have a molecular weight of <3 kDa. A total of 33 common peptides were identified in the EMPH by liquid chromatography-tandem mass spectrometry, 16 of which were identified as bioactive peptides using bioinformatics. The peptide profiles and the coverage of master proteins of the 4 EMPH were different. The EMPH also exhibited strong free radical scavenging capacity, as indicated by the results of the 1,1-diphenyl-2-picrylhydrazyl radical, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), hydroxyl radical, and reducing power assays. The results of this study provided useful information on the peptide profiles and antioxidant activity of EMPH.

Differences between peptide profiles of extensive hydrolysates and their influence on functionality for the management of cow's milk allergy: A short review.

Extensively hydrolyzed formulas (eHFs) are recommended for the dietary management of cow's milk protein allergy (CMPA) in non-exclusively breastfed infants. Studies show that peptide profiles differ between eHFs. This short review aims to highlight the variability in peptides and their ability to influence allergenicity and possibly the induction of tolerance by different eHFs. The differences between eHFs are determined by the source of the protein fraction (casein or whey), peptide size-distribution profile and residual β-lactoglobulin which is the most immunogenic and allergenic protein in bovine milk for human infants as it is not present in human breastmilk. These differences occur from the hydrolyzation process which result in variable IgE reactivity against cow's milk allergen epitopes by subjects with CMPA and differences in the Th1, Th2 and pro-inflammatory cytokine responses elicited. They also have different effects on gut barrier integrity. Results suggest that one particular eHF-casein had the least allergenic potential due to its low residual allergenic epitope content and demonstrated the greatest effect on restoring gut barrier integrity by its effects on mucin 5AC, occludin and Zona Occludens-1 in human enterocytes. It also increased the production of the tolerogenic cytokines Il-10 and IFN-γ. In addition, recent studies documented promising effects of optional functional ingredients such as pre-, pro- and synbiotics on the management of cow's milk allergy and induction of tolerance, in part via the induction of the production of short chain fatty acids. This review highlights differences in the residual allergenicity, peptide size distribution, presence of optional functional ingredients and overall functionality of several well-characterized eHFs which can impact the management of CMPA and the ability to induce immune tolerance to cow's milk protein.

An In Vitro and In Vivo Translational Research Approach for the Assessment of Sensitization Capacity and Residual Allergenicity of an Extensive Whey Hydrolysate for Cow's Milk-Allergic Infants.

Introduction: Hypoallergenic formulas prepared from hydrolyzed cow’s milk proteins are often used for the management of cow’s milk allergy (CMA) in infants. In this study, both in vitro assays and an in vivo mouse model for CMA were used to assess the sensitizing and allergenic potential of a newly developed, extensive whey hydrolysate (eWH). Methods: Gel permeation chromatography was used to characterize the molecular weight distribution of the peptides. Residual antigenicity was measured using a beta-lactoglobulin ELISA as well as with immunoblotting using anti-beta-lactoglobulin (BLG) and anti-alpha-lactalbumin antibodies. In vitro residual allergenicity was assessed using huFcεRIα-RBL-2H3 cells sensitized with anti-bovine BLG human IgE. In vivo sensitizing and allergenic potential was assessed in a CMA mouse model by measuring the acute allergic skin response, anaphylactic shock score, body temperature, serum mMCP-1, whey-specific IgE, and cytokines. Results: There was no in vitro residual antigenicity and allergenicity observed of the eWH. Mice sensitized with eWH showed no acute allergic skin reaction after challenge with whey, confirmed by an absence of whey-specific IgE and anaphylactic symptoms and decrease in body temperature and mMCP-1 levels. Conclusions: Results from our in vitro and in vivo translational approach to assess sensitization capacity and residual allergenicity indicate that the newly developed eWH is safe for use in CMA infants. This was subsequently confirmed in a clinical study in which this eWH was tolerated by more than 90% (with 95% confidence) of infants or children with confirmed CMA.

In vitro anti-inflammatory and antioxidant activity of chickpea (Cicer arietinum L.) proteins hydrolysate fractions

Chickpea is a legume that has exhibited various biological activities. The anti-inflammatory and antioxidant activities of chickpea proteins hydrolysate fractions were evaluated. The extensive protein hydrolysate (greater than 50 %) obtained by enzymatic hydrolysis was fractionated and 5-10 kDa, 3-5 kDa, 1-3 kDa, and ≤1 kDa fractions were obtained. The anti-inflammatory activity was determined in murine peritoneal macrophages stimulated with lipopolysaccharide (LPS). Fractions of F5-10 kDa, F3-5 kDa, F1-3 kDa presented the highest inhibition of nitric oxide (76 %), Tumoral Necrosis Factor-α (93 %), and Interleukin-1β (90 %) synthesis. Antioxidant activity was evaluated by ABTS, DPPH methods, Fe3+ reducing power and Cu2+ chelating capacity. The fraction of F1-3 kDa showed higher inhibition of ABTS (78 %) and DPPH (7.76 %) radicals, besides showed the highest Fe3+ reducing capacity (0.508 Abs700nm), for Cu2+ chelating capacity F≤1 kDa showed higher inhibition (52.60 %). Chickpea protein shows biological activities such as anti-inflammatory and antioxidant, so it can be considered a source of low molecular weight bioactive peptides that can be used as alternative therapies for certain diseases related to chronic inflammation.

The Macronutrient Composition of Infant Formula Produces Differences in Gut Microbiota Maturation That Associate with Weight Gain Velocity and Weight Status.

This proof-of-principle study analyzed fecal samples from 30 infants who participated in a randomized controlled trial on the effects of the macronutrient composition of infant formula on growth and energy balance. In that study, infants randomized to be fed cow milk formula (CMF) had faster weight-gain velocity during the first 4 months and higher weight-for-length Z scores up to 11.5 months than those randomized to an isocaloric extensive protein hydrolysate formula (EHF). Here we examined associations among infant formula composition, gut microbial composition and maturation, and children’s weight status. Fecal samples collected before and monthly up to 4.5 months after randomization were analyzed by shotgun metagenomic sequencing and targeted metabolomics. The EHF group had faster maturation of gut microbiota than the CMF group, and increased alpha diversity driven by Clostridia taxa. Abundance of Ruminococcus gnavus distinguished the two groups after exclusive feeding of the assigned formula for 3 months. Abundance of Clostridia at 3–4 months negatively correlated with prior weight-gain velocity and body weight phenotypes when they became toddlers. Macronutrient differences between the formulas likely led to the observed divergence in gut microbiota composition that was associated with differences in transient rapid weight gain, a well-established predictor of childhood obesity and other comorbidities.

β Cyclodextrin Nanocomplexes with Biologically Active Peptides from Hydrolysed Bovine Whey and Colostrum

β Cyclodextrin nanocomplexes with extensive whey and colostrum hydrolysates possessing acceptable flavor properties serve as potential sources of bioactive peptides. In this study, comparative characterization of dairy protein hydrolysates and their complexes with β cyclodextrin is presented. Antioxidant activity of studied samples was estimated by fluorometric method, the formation of clathrates with cyclic oligosaccharide was determined using thermogravimetric analysis. A significant decrease in bitterness of peptides included in cyclic oligosaccharides was established compared with samples of dairy hydrolysates. 2.1/1.3 fold increase in the antioxidant potential of β cyclodextrin clathrates with whey/colostrum hydrolysates was recorded versus unbound peptide fractions. According to toxicological tests on Tetrahymena pyriformis, the samples of whey hydrolysate and the resulting nanocomplex were referred to as non-toxic and slightly hazardous compounds, respectively. The dynamics of body weight gain and the relative weight coefficient of internal organs revealed no differences compared to the control group of Rattus norvegicus. The data on differentiation of blood cells, their death, and cytogenetic disorders demonstrated that a sample of cyclic oligosaccharides with whey peptides is non-toxic at the maximum dosages allowable for administration. β Cyclodextrin complexing with dairy peptides resulted in enhanced radical-reducing activity and improved flavor properties, making the clathrates promising and safe ingredients of special nutrition formulas.

Current Guidelines and Future Strategies for the Management of Cow's Milk Allergy.

Exclusive breast feeding is recommended in all guidelines as the first choice feeding. Cow milk allergy (CMA) can be diagnosed by a diagnostic elimination diet for 2 to 4 weeks with a hypo-allergenic formula, followed by a challenge test with intact cow milk protein. The most often used hypo-allergenic formula for the diagnostic elimination diet and the therapeutic diet is a CM based extensive hydrolysate. CM-based partial hydrolysates cannot be recommended in the management of CMA because of insufficient efficacy and possible reactions, but about half of the infants with CMA may tolerate a partial hydrolysate. The pros and cons of other dietary options are discussed in this paper. The use of an amino acid-based formula and/or rice based hydrolysate formula during the diagnostic elimination and therapeutic diet is debated. When available, there is sufficient evidence to consider rice hydrolysates as an adequate alternative to CM-based hydrolysates, since some infants will still react to the CM hydrolysate. The pros and cons of dietary options such as soy formula, buckwheat, almond, pea or other plant based dietary products are discussed. Although the majority of the plant-based beverages are nutritionally inadequate, some are nutritionally adapted for toddlers. However, accessibility and content vary by country and, thus far there is insufficient evidence on the efficacy and tolerance of these plant-based drinks (except for soy formula and rice hydrolysates) to provide an opinion on them.ConclusionA diagnostic elimination diet, followed by a challenge remains the diagnostic standard. The use of an awareness tool may result in a decrease of delayed diagnosis. Breastmilk remains the ideal source of nutrition and when not available a CM extensively hydrolyzed formula, rice hydrolysate or amino acid formula should be recommended. More evidence is needed regarding plant-based drinks.

Antihypertensive and Antioxidant Activity of Chia Protein Techno-Functional Extensive Hydrolysates.

Twelve high-quality chia protein hydrolysates (CPHs) were produced from chia protein isolate (CPI) in a pilot plant of vegetable proteins. To obtain functional hydrolysate, four CPHs were hydrolyzed by the action of Alcalase, an endoprotease, and the other eight CPHs were hydrolyzed by the action of Flavourzyme, an exoprotease. Alcalase-obtained CPHs showed significant antihypertensive properties particularly, the CPH obtained after 15 min of hydrolysis with Alcalase (CPH15A), which showed a 36.2% hydrolysis degree. In addition, CPH15A increased the antioxidant capacity compared to CPI. The CPH15A physicochemical composition was characterized and compared to chia defatted flour (CDF) and CPI, and its techno-functional properties were determined by in vitro experiments through the analysis of its oil absorption capacity, as well as the capacity and stability of foaming and emulsifying, resulting in an emulsifier and stabilizer better than the intact protein. Therefore, the present study revealed that CPH15A has potent antihypertensive and antioxidant properties and can constitute an effective alternative to other plant protein ingredients sources that are being used in the food industry.

Глубокие ферментативные гидролизаты белков сыворотки молока и молозива для продуктов специализированного питания

Проведено комплексное сравнительное исследование пептидного состава и биологических активностей глубоких ферментативных гидролизатов коровьего молока. Образцы гидролизованных сывороточных белков и молозива обладают подтвержденным антиоксидантным, антимутагенным и антибактериальным действием. A comprehensive comparative study of peptide profile and bioactivities of extensive enzymatic dairy hydrolysates was carried out. Samples of hydrolysed whey proteins and colostrum possess proven antioxidant, antimutagenic and antibacterial effects.

Effect of two-step enzymatic hydrolysis on the antioxidant properties and proteomics of hydrolysates of milk protein concentrate

Food protein and peptides are generally considered a source of dietary antioxidants. The antioxidant activity and peptide profiles of four extensive hydrolysates of milk protein concentrate (MPC) were examined using the two-step enzymatic method. The hydrolysis combinations were Alcalase-Flavourzyme (AE), Alcalase-ProteAXH (AH), Alcalase-Protamex (AX) and Alcalase-Protease A 2SD (AD). The results showed that highest degree of hydrolysis corresponded to the AE sample (20.41%). High-efficiency gel-filtration chromatography results indicated that the relative proportions of extensive hydrolysates with molecular weights < 3 kDa were 99.89%, 99.57%, 99.93%, and 99.89% for AX, AE, AD and AH, respectively. The hydrolysates of the MPC exhibited increased radical-scavenging capacity, as evidenced through an analysis with 1,1-diphenyl-2-pycryl-hydrazyl (DPPH), 2,2-azinobis (3-ethylbenzothiazo-line-6-sulfonic acid) diammonium salt (ABTS), reducing power and hydroxyl-radical scavenging activity testing. The main bioactive peptides were identified through EASY-nLC-orbitrap MS/MS and bioinformatics. The study may provide useful information regarding the antioxidant properties of extensive hydrolysates of MPC.

Strategies and Future Opportunities for the Prevention, Diagnosis, and Management of Cow Milk Allergy.

The prevalence of food allergy has increased over the last 20-30 years, including cow milk allergy (CMA) which is one of the most common causes of infant food allergy. International allergy experts met in 2019 to discuss broad topics in allergy prevention and management of CMA including current challenges and future opportunities. The highlights of the meeting combined with recently published developments are presented here. Primary prevention of CMA should start from pre-pregnancy with a focus on a healthy lifestyle and food diversity to ensure adequate transfer of inhibitory IgG- allergen immune complexes across the placenta especially in mothers with a history of allergic diseases and planned c-section delivery. For non-breastfed infants, there is controversy about the preventive role of partially hydrolyzed formulae (pHF) despite some evidence of health economic benefits among those with a family history of allergy. Clinical management of CMA consists of secondary prevention with a focus on the development of early oral tolerance. The use of extensive Hydrolysate Formulae (eHF) is the nutrition of choice for the majority of non-breastfed infants with CMA; potentially with pre-, probiotics and LCPUFA to support early oral tolerance induction. Future opportunities are, among others, pre- and probiotics supplementation for mothers and high-risk infants for the primary prevention of CMA. A controlled prospective study implementing a step-down milk formulae ladder with various degrees of hydrolysate is proposed for food challenges and early development of oral tolerance. This provides a more precise gradation of milk protein exposure than those currently recommended.

Structural and antioxidant properties of cowpea protein hydrolysates

Cowpea protein isolates extracted at pH 8.0 (I8) and pH 10.0 (I10) were hydrolysed by alcalase to obtain limited (I8LH and I10LH) and extensive (I8HH and I10HH) hydrolysates. Hydrolysis degree was 2.3% for I8LH, 4.6% for I10LH, 25.8% for I8HH and 24.1% for I10HH. The effect of hydrolysis was assessed by using solubility-pH profile assay, exclusion chromatography, gel electrophoresis, infrared and fluorescence spectroscopy and surface hydrophobicity. Subsequently, ABTS.+, ORAC and HORAC methods were applied to measure antioxidant activity in all samples. Electrophoresis, chromatography and surface hydrophobicity showed different profiles for I8LH and I10LH, presumably, due to the initial extraction pH of protein isolates. Limited hydrolysis increased antioxidant activity in both I8LH and I10LH with values higher than 70% for ABTS.+, 60% for ORAC and 45% for HORAC when compared to their respective isolates. Concerning extensive hydrolysis, I8HH and I10HH yielded similar effects when considering physicochemical, functional and antioxidant properties. Both I8HH and I10HH showed increments above 80% for ABTS.+, 80% for ORAC and 70% for HORAC in comparison to their respective isolates. These results indicate that limited and extensive alcalase hydrolysis of both isolates increases antioxidant activity regardless of the pH for initial protein extraction. Finally, peptides with a molecular weight between 1.8 and 6.5 kDa (I8HH and I10HH from FPLC soluble fractions) exhibited the highest ORAC activity. Thus, cowpea alcalase hydrolysates emerge as viable ingredients to formulate foods with antioxidant properties.

Administration of Extensive Hydrolysates From Caseins and Lactobacillus rhamnosus GG Probiotic Does Not Prevent Cow's Milk Proteins Allergy in a Mouse Model.

BackgroundEarly nutrition may influence the development of food allergies later in life. In the absence of breastfeeding, hydrolysates from cow’s milk proteins (CMP) were indicated as a prevention strategy in at risk infants, but their proof of effectiveness in clinical and pre-clinical studies is still insufficient. Thanks to a validated mouse model, we then assessed specific and nonspecific preventive effects of administration of extensive hydrolysates from caseins (eHC) on the development of food allergy to CMP. The additional nonspecific effect of the probiotic Lactobacillus GG (LGG), commonly used in infant formula, was also assessed.MethodsGroups of young BALB/cByJ female mice were pretreated by repeated gavage either with PBS (control mice), or with PBS solution containing non-hydrolyzed milk protein isolate (MPI), eHC or eHC+LGG (eq. of 10 mg of protein/gavage). All mice were then experimentally sensitized to CMP by gavage with whole CM mixed with the Th2 mucosal adjuvant Cholera toxin. All mice were further chronically exposed to cow’s milk. A group of mice was kept naïve. Sensitization to both caseins and to the non-related whey protein β-lactoglobulin (BLG) was evaluated by measuring specific antibodies in plasma and specific ex vivo Th2/Th1/Th17 cytokine secretion. Elicitation of the allergic reaction was assessed by measuring mMCP1 in plasma obtained after oral food challenge (OFC) with CMP. Th/Treg cell frequencies in gut-associated lymphoid tissue and spleen were analyzed by flow cytometry at the end of the protocol. Robust statistical procedure combining non-supervised and supervised multivariate analyses and univariate analyses, was conducted to reveal any effect of the pretreatments.ResultsPBS pretreated mice were efficiently sensitized and demonstrated elicitation of allergic reaction after OFC, whereas mice pretreated with MPI were durably protected from allergy to CMP. eHC+/-LGG pretreatments had no protective effect on sensitization to casein (specific) or BLG (non-specific), nor on CMP-induced allergic reactions. Surprisingly, eHC+LGG mice demonstrated significantly enhanced humoral and cellular immune responses after sensitization with CMP. Only some subtle changes were evidenced by flow cytometry.ConclusionNeither specific nor nonspecific preventive effects of administration of casein-derived peptides on the development of CMP food allergy were evidenced in our experimental setup. Further studies should be conducted to delineate the mechanisms involved in the immunostimulatory potential of LGG and to clarify its significance in clinical use.

Hydrolyzed Rice Protein-Based Formulas, a Vegetal Alternative in Cow’s Milk Allergy

Formulas adapted to infant feeding, although most of the time made from cow’s milk proteins, can be made from hydrolyzed rice protein but they must be classified as “formulas for specific medical needs”, according to European regulations. The nutritional quality of rice proteins is thus suitable to be used in infant formulas giving that it is supplemented by certain amino acids which can be lacking. Besides, hydrolysis is required to facilitate their water solubility and digestibility. Owing to a low allergenicity of rice and to the absence of the cross-allergy between milk proteins and rice proteins, these formulas are adapted to the diet of children with cow’s milk protein allergy (CMPA), which explains their growing use in some countries. However, CMPA, an expanding disorder, has consequences for growth, bone mineralization, and often has an association with allergy to other foods, including cow’s milk extensive hydrolysate, so that a surveillance of the adaption of hydrolyzed rice protein formulas (HRPF) to CMPA, the absence of unexpected side effects, and the appropriate response to its various health hazards seems mandatory. This paper analyses the health problem deriving from CMPA, the industrial development of hydrolyzed rice protein formulas, and the limited number of clinical studies, which confirms, at the moment, a good allergic tolerance and safety. The goal is to better advise heath care professionals on their use of HRPFs during CMPA.

Antimutagenic and Antibacterial Activity of β‑Cyclodextrin Clathrates with Extensive Hydrolysates of Colostrum and Whey

In this study, experimental samples of extensive enzymatic hydrolysates of bovine colostrum and whey, inclusion complexes of β cyclodextrin with dairy peptides were derived. 1.6 fold increase in the degree of proteolysis was established for whey hydrolysate compared to cleaved first milk. A significant decline in the bitterness of peptide constituents of clathrates was recorded versus samples of initial extensive hydrolysates. The thermogravimetric analysis confirmed the formation of inclusion complexes of cyclic oligosaccharide with dairy peptides. The Ames test demonstrated comparable induced mutation rates for samples of hydrolysates and clathrates. Antimutagenic activity of the samples reached 23.9–28.3 % when tested on strain Salmonella typhimurium TA 98 and 19.4–21.8 % on strain TA 100. According to the impedimetric method, the antibacterial potential of extensive colostrum hydrolysate exceeded the action of cleaved whey against Escherichia coli ATCC 8739 and Staphylococcus aureus ATCC 6538. Enhanced antibacterial activity was found for whey and colostrum peptide fractions included in clathrates with a cyclic oligosaccharide. β cyclodextrin complexing preserved antimutagenic capacity, promoted antibacterial effect, and improved organoleptic properties of milk peptides. These advantages acquire maximum relevance during the clathrate upgrading of specialized nutrition formulas.

Identification of bitter constituents in milk-based infant formula with hydrolysed milk protein through a sensory-guided technique

Key bitter taste-active fractions were fractionated and identified from cows' milk-based infant formula (IF) containing hydrolysed (bovine) milk protein. A systematic and sensory-guided approach was applied to separate and identify taste-active bitter peptides. The degree of hydrolysis for the extensively hydrolysed (EH) and partially hydrolysed (PH) samples was >22% and <22%, respectively. The sample with EH milk protein had the highest sensory score for bitter taste, and more bitter peptides were identified from it. The proportions of micromolecular peptides (<500 Da) distributed in the EH sample and PH sample were approximately 100% and 80%, respectively. More importantly, a sequential difference in the peptides generated from milk-based IFs with PH and EH was observed, and total numbers of four and nine peptides were discovered in the PH and EH samples, respectively; Ser–Leu–Ala, Leu–Leu–Pro and Try–Try–Gln, were reported for the very first time.

Timing and Patterning of Primary Teeth Eruption as a Biomarker for Early Rapid Weight Gain (P11-080-19)

ObjectivesProspective and retrospective studies suggest that the timing and patterning of tooth eruption in infancy is a biomarker for later obesity. We recently discovered that although within the range of normal growth, healthy infants randomized to be fed cow milk formula (CMF) for the first year of life experienced greater gains in weight, but not length, during the first 4.5 months, when compared to those fed an extensive protein hydrolysate formula (EHF). In the present study, we conducted secondary analysis to determine whether the type of infant formula interacted with weight gain velocity on the timing and patterning of developmental milestones including tooth eruption. MethodsAt each monthly visit from 0.5 until 12.5 mos, infants (N = 113) were weight, measured and the number and location of each tooth, if any, erupted during the prior month was recorded as was the age at which they began crawling, sitting independently, standing and walking without assistance. Generalized estimating equations were conducted to determine whether infant formula had a direct impact on teething via effect on weight gain velocity independent of treatment group. ResultsWe found a significant time × treatment group effect and a significant treatment group effect on the number of teeth erupted such that CMF erupted more teeth than EHF infants by one year of age. The rate of tooth eruption was significantly greater during the first year among the CMF when compared to the EHF group (P < 0.05). Regardless of group, the greater the velocities in weight gain during the first 4 months (but not later), the earlier infants erupted their first tooth (P < 0.05). The formula-induced changes in weight gain velocity did not generalize to the other milestones measures. ConclusionsThe patterning of primary teeth eruption paralleled the formula-induced differences in early rapid weight gain, a risk factor for later obesity. Early and rapid eruption of primary teeth may be a biomarker for obesity as well as detrimental for dental health since caries lesions progress faster and are cavitated more quickly during childhood. Funding SourcesNIH Grants HD072307 and R03HD09408.