BACKGROUND: About 70-80% of children with cow's milk allergy (CMA) become outgrown clinically by the age of 3 years. Casein, one of the three major cow's milk proteins (casein, beta-lactoglobulin (BLG), alpha-lactoalbumin (ALA) ) has been reported to play an important role in the persistence of CMA. The aim of this study was to determine different effects of causative milk proteins on the persistence of CMA between two age groups. METHODS: A total of 65 patients with CMA were enrolled in this study. Their cow's milk-specific IgEs were positive ( 0.7 U/ml by Pharmacia CAP). After dividing 65 patients into two age groups, under the age of 3 years and over 3 years (persistent CMA), we compared the levels of casein-, BLG- and ALA-specific IgE antibodies between the two groups. RESULTS: There were 44 patients in the group of less than 3 years of age and 21 patients in the group of more than 3 years of age. The concentrations of the specific IgE antibodies to casein, BLG and ALA were not significantly different between the two groups. However, although statistically insignificant, those more than 3 years of age had higher mean values of casein-specific IgE antibodies and lower mean values of whey protein (BLG and ALA) - specific IgE antibodies compared with those less than 3 years of age. A single dominant allergenic milk protein was not identified within either of the two age groups, but the con centrations of the casein-specific IgE antibodies in children with more than 3 years of age tended to be higher than those of whey protein-specific IgE antibodies. CONCLUSION: Although statistically insignificant, the concentrations of the casein-specific IgE antibodies were higher in the group of more than 3 years of age than in the younger group. Moreover, the concentrations of the casein-specific IgE antibodies in children more than 3 years of age tended to be higher than those of whey proteins. These findings implicate that casein plays a certain role in the persistence of CMA.
Antibodies ; Caseins ; Child* ; Humans ; Immunoglobulin E* ; Lactoglobulins ; Milk Hypersensitivity* ; Milk Proteins ; Milk* ; Whey Proteins

OBJECTIVETo examine the effects of the whey basic protein on bone metabolism of Sprague-Dawley (SD) rats and healthy mid-aged women.

METHODSForty-four female SD rats were randomized by weight into four groups of eleven rats each and fed 10 mg x kg BW(-1) x d(-1), 20 mg x kg BW(-1) x d(-1), 30 mg x kg BW(-1) x d(-1) of whey basic protein and control diet was given respectively by intragastrically injection for 90 days. Bone mineral density of femur was measured by dual-energy X-ray absorptiometry in vitro. Sixty-three health women [(37.9 +/- 4.3) years old] were randomly assigned to treatment with placebo, 30 mg whey basic protein per day or 60 mg whey basic protein per day for 24 weeks. The bone mineral density (BMD) of the lumbar vertebrae L2-LA, femoral neck and right calcaneus of each subject were measured by dual-energy X-ray absorptiometry (DXA) at 0 and the 24th week of treatment. Serum bone specific alkaline phosphatase and N-telopeptide (NTX) were measured at 0 and the 14th week.

RESULTSThe mean BMD value of the distal end of the femur in 10 mg x kg BW(-1) x d(-1) whey basic protein group was significantly higher than that of the control group at the end of the trail. But after treatment by doses of whey basic protein used in the study, there were no differences between the control group and others groups on bone mineral density in the human trail.

CONCLUSIONWhey basic protein should enhance the bone mineral density of the rats' femur and no obvious effect was detected in the human trail.

Adult ; Animals ; Bone Density ; drug effects ; Female ; Food, Formulated ; Humans ; Lactalbumin ; pharmacology ; Milk ; Milk Proteins ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Whey Proteins

Milk proteins are composed of casein, further classified into αS1-casein, αS2-casein, β-casein, and κ-casein, and whey protein, which is separated into α-lacatalbumin, β-lactoglobulin, serum albumin, and some minor proteins, such as lactoferrin and immunoglobulin. To reduce the allergenicity of protein, heat treatment and enzymatic protein hydrolysis by endopeptidase are necessarily required. Additionally, membrane technology should be applied to produce a protein hydrolyzate, which has consistent molecular weight of peptide and low in free amino acid without allergenic peptide or protein. Extensive casein hydrolyzate and whey protein hydrolyzate are used for protein source of mainly extensively hydrolyzed protein formula (eHF) intended for the treatment of cow's milk allergy. Also, partially hydrolyzed formula (pHF) is developed, which is using a single protein source e.g., whey protein hydrolyzate. The allergenicity of infant formula can be determined according to molecular weight profile and antigenicity reduction compared to intact protein. More than 90% peptides are present in eHF have a molecular weight of <3,000 Da. Peptide molecular weight profiles of pHF range mainly between 3,000 and 10,000 Da, but have a small percentage of >10,000 Da. Generally, antigenicity reduction in eHF and pHF is 10-6 and 10-3, respectively. Even if protein hydrolyzate is manufactured under strict quality control, there is still a risk of cross contamination of allergenic milk components through environmental conditions and the shared manufacturing process. Thus, quality assessment of protein hydrolyzate formula must be performed routinely.
Caseins ; Hot Temperature ; Humans ; Hydrolysis ; Immunoglobulins ; Infant Formula* ; Infant* ; Lactoferrin ; Membranes ; Milk ; Milk Hypersensitivity ; Milk Proteins ; Molecular Weight ; Peptides ; Quality Control ; Serum Albumin ; Whey Proteins

Introduction: It is important to wean mechanically-ventilated patients as early as possible to avoid complications such as ventilator-associated pneumonia. Supplementing the diet with additional protein may help to stimulate muscle protein synthesis which may enhance respiratory muscle function and ventilator drive. This study aims to determine the effect of whey protein supplementation on the duration of mechanical ventilation in intensive care unit patients of Ospital ng Makati. Methods: We enrolled forty eligible patients in this openlabel randomized controlled trial and were assigned into two groups: Group A (N=20): patients were given enteral feeding using commercial formula with added whey protein, given as one serving every eight hours (equivalent to 18 g of additional protein per day) and Group B (N=20): patients were given enteral feeding using commercial formula alone. Weaning was started as soon as the patient fulfilled the criteria of spontaneous breathing trial and was considered successful if the patient maintained these criteria for 48 hours after extubation. Results: Our study’s results showed that the mechanically intubated patients in Group A have lesser ventilatordependent days with an average of 5.4 days as compared to those in Group B with an average of 7.45 days (p=0.00). Patients in Group A were also noted to have statistically significant higher increase in serum albumin, mid-arm circumference and triceps skin fold from baseline. Twentyfive percent of patients in Group B developed ventilator acquired pneumonia and none in Group A. No mortality was noted in both groups. Conclusion Whey protein supplementation in mechanically ventilated patients can be recommended to facilitate early weaning because of its effect on early muscle protein synthesis leading to improvement of lung function and ventilator drive. It can also help in preventing malnutrition and nosocomial infections during critical illness. With all these benefits of whey protein, its use can potentially lead to shorter duration of mechanical ventilation and hospital stay which can also mean less cost of care delivery.
Whey Proteins ; Weaning ; Respiration, Artificial ; Intensive Care Units ; Randomized Controlled Trial

Athletes ; Flavones/administration & dosage* ; Humans ; Hydrocortisone/blood* ; Seasons ; Sports Nutritional Physiological Phenomena ; Testosterone/blood* ; Whey Proteins/administration & dosage*

Postprandial hyperglycemia is associated with the risk of diabetes mellitus, cardiovascular disease, and mortality. Nutrition therapy is an important component of the management of postprandial hyperglycemia. Postprandial glucose levels are determined by several factors, such as the quantity and composition of nutrients, gastric emptying rates, secretion of incretin hormones, insulin secretion, glucose uptake by peripheral tissues, and endogenous glucose production. Nutrient preload and food order (or meal sequence) are dietary approaches targeting these factors. Nutrient preload reduces postprandial glucose excursion by enhancing insulin secretion, augmenting the secretion of glucagonlike peptide-1, and delaying gastric emptying. Carbohydrates-last food order improves glycemic control, increases the secretion of glucagon-like peptide-1, and decreases insulin requirements. Therefore, both nutrient preload and manipulation of food order can be an effective, safe, and feasible strategy for treating hyperglycemia in individuals with diabetes mellitus.
Carbohydrates ; Cardiovascular Diseases ; Diabetes Mellitus ; Gastric Emptying ; Gastrointestinal Hormones ; Glucagon-Like Peptide 1 ; Glucose* ; Hyperglycemia ; Incretins ; Insulin* ; Meals ; Mortality ; Nutrition Therapy ; Whey Proteins

Whey protein concentrate (WPC 80) and sodium caseinate were hydrolyzed by Protamex to 5%, 10%, 15%, and 20% degree of hydrolysis (DH). WPC 80, sodium caseinate and their hydrolysates were then analyzed, compared and evaluated for their nutritional qualities. Their chemical composition, protein solubility, amino acid composition, essential amino acid index (EAA index), biological value (BV), nutritional index (NI), chemical score, enzymic protein efficiency ratio (E-PER) and in vitro protein digestibility (IVPD) were determined. The results indicated that the enzymatic hydrolysis of WPC 80 and sodium caseinate by Protamex improved the solubility and IVPD of their hydrolysates. WPC 80, sodium caseinate and their hydrolysates were high-quality proteins and had a surplus of essential amino acids compared with the FAO/WHO/UNU (1985) reference standard. The nutritive value of WPC 80 and its hydrolysates was superior to that of sodium caseinate and its hydrolysates as indicated by some nutritional parameters such as the amino acid composition, chemical score, EAA index and predicted BV. However, the E-PER was lower for the WPC hydrolysates as compared to unhydrolyzed WPC 80 but sodium caseinate and its hydrolysates did not differ significantly. The nutritional qualities of WPC 80, sodium caseinate and their hydrolysates were good and make them appropriate for food formulations or as nutritional supplements.
Amino Acids ; chemistry ; Caseins ; chemistry ; metabolism ; Dietary Proteins ; analysis ; Evaluation Studies as Topic ; Hydrolysis ; Milk Proteins ; chemistry ; metabolism ; Models, Statistical ; Nutritive Value ; Protein Hydrolysates ; chemistry ; Solubility ; Temperature ; Time Factors ; Tryptophan ; chemistry ; Whey Proteins

PURPOSE: Cow's milk protein is one of the most common and strongest food allergen. We investigated the effects of heat treatment on the distribution and antigenicities of major allergens from cow's milk. We also compared the protein distribution and antigenicities among cow's milk formula and its substitutes. METHODS: We heated alpha-casen, beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), and crude extract of cow's milk in 100degrees C boiling water for 1 hour. We prepared crude extracts from cow's milk formula, partially hydrolyzed milk formula (pHF) and extensively hydrolyzed milk formula (eHF). The protein compositions of all the samples were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigenicities were determined by IgE immunoblotting with pooled serum collected from 11 patients with milk allergy. RESULTS: After heating, no significant alteration was found in casein, and the aggregates of ALA and BLG were detected with molecular weights of about 30 and 45 kDa, respectively. The antigenicities of newly detected aggregates were increased. The new aggregates of BLG with increased antigenicities were also found in heated milk total protein. Major milk allergens were not found in pHF, and residual components with a molecular weight below 10 KDa did not show IgE-binding activity. We failed to observe the residual components and antigenicities of eHF. CONCLUSION: Changes in protein distribution and antigenicity of milk total protein induced by heat treatment may not be significantly different from those of each major allergen. The residual components of pHF could have little IgE-binding capacity, and there may be few or no antigenic components in eHF.
Allergens ; Caseins ; Complex Mixtures ; Electrophoresis ; Heating ; Hot Temperature* ; Humans ; Hydrolysis* ; Immunoblotting ; Immunoglobulin E ; Lactalbumin ; Lactoglobulins ; Milk ; Milk Hypersensitivity ; Milk Proteins* ; Molecular Weight ; Sodium ; Water

Protein self-assemblies at the micro- and nano-scale are of great interest because of their morphological diversity and good biocompatibility. High-throughput screening of protein self-assembly at different scales and morphologies using protein crystallization screening conditions is an emerging method. When using this method to screen protein self-assembly conditions, some apparently transparent droplets are often observed, in which it is not clear whether self-assembly occurs. We explored the interaction between β-lactoglobulin and the protein crystallization kit Index™ C10 and observed the presence of micro- and nano-scale protein self-assemblies in the transparent droplets. The diverse morphology of the micro- and nano-scale self-assemblies in the transparent droplets formed by mixing different initial concentrations of β-lactoglobulin and Index™ C10 was further investigated by scanning electron microscope. Self-assembly process of fluorescence-labelled β-lactoglobulin was monitored continuously by laser confocal microscope, allowing real-time observation of the liquid-liquid phase separation phenomenon and the morphology of the final self-assemblies. The internal structure of the self-assemblies was gradually ordered over time by in-situ X-ray diffraction. This indicates that the self-assembly phenomenon within transparent droplets, observed in protein self-assembly condition screening experiments, is worthy of further in-depth exploration.
Crystallization ; Lactoglobulins

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection