Supplemental Case Research Articles

The effect of probiotics and casein supplementation on aerobic capacity parameters of male soccer players.

In the realm of sports science, nutrition is a well-established pillar for athletes' training, performance, and post-workout recovery. However, the role of gut microbiota, often overlooked, is a novel and intriguing aspect that can significantly impact athletic performance. With this in mind, our study ventures into uncharted territory, investigating the effect of probiotic and casein supplementation on the aerobic capacity of male soccer players. A double-blinded and placebo-controlled study was conducted with 44 male soccer players (Age: 22.81 ± 2.76 yr, Height: 177.90 ± 6.75 cm, Weight: 67.42 ± 8.44 kg). The participants were subjected to the Bruce test in the beginning; then, they were randomly divided into four groups, each consisting of 11 people: probiotics (PRO), casein (CAS), probiotics with casein (PRO+CAS), and placebo (PLA). PRO group was given one probiotic capsule (containing strains of Lactiplantibacillus plantarum BP06, Lacticaseibacillus casei BP07, Lactobacillus acidophilus BA05, Lactobacillus delbrueckii BD08 bulgaricus, Bifidobacterium infantis BI04, Bifidobacterium longum BL03, Bifidobacterium breve BB02 and Streptococcus salivarius thermophilus BT01, with a total dose of 4.5 × 1011 CFU) during dinner, while the CAS group consumed 20 grams of casein powder 45 minutes before bed. The PRO+CAS group was given one probiotic capsule during dinner and 20 grams of casein powder 45 minutes before bed. The participants in the PLA group were given one red capsule (containing 5 grams of starch) during dinner. All participants were instructed to take the supplements only on training days, three times a week for four weeks. The maximal oxygen consumption (VO2max), Ventilatory Threshold (VT), Time-to-exhaustion (TTE), Respiratory Compensation Point (RCP), Isocapnic area Time (Time-IC), Isocapnic area oxygen consumption (VO2-IC), and Hypocapnic Hyperventilation area Time (Time-HHV), after the Bruce test were Measured. All data were analyzed using SPSS Windows software, mixed repeated measure ANOVA, and Bonferroni post hoc test at p < 0.05 level. The current study's findings illustrated that, after the intervention, TTE (p = 0.01) and RCP (p = 0.01) were significantly improved in PRO+CAS compared to the PLA group. No significant difference was observed between PRO and PLA (p = 0.52), PRO and CAS (p = 0.999), PRO and PRO+CAS (p = 0.9), CAS and PLA (p = 0.65), CAS and PRO+CAS (p = 0.73) in TTE. In addition, no significant difference was observed between PRO and CAS (p = 0.999), PRO and PLA (p = 0.40), PRO and PRO+CAS (p = 0.999), CAS and PLA (p = 0.263), CAS and PRO+CAS (p = 0.999) in RCP. Time-HHV was significantly higher in PRO+CAS (p = 0.000) and CAS (p = 0.047) compared to the PLA group. However, no significant difference was observed in the Time-HHV between PRO and CAS (p = 0.999), PRO and PRO+CAS (p = 0.25), PRO and PLA (p = 0.12), and CAS and PRO+CAS (p = 0.57). Additionally, all the groups had no significant differences in VO2max, VT1, VO2-IC and Time-IC. The findings showed that consuming probiotics and casein could relatively improve the aerobic capacity of male soccer players. Nevertheless, simultaneous consumption of probiotics and casein had a more pronounced effect on aerobic capacity indicators, especially TTE and Time-HHV.

Effects of high-quality protein supplementation on cardiovascular risk factors in individuals with metabolic diseases: A systematic review and meta-analysis of randomized controlled trials

BackgroundUncertainties still existed about the effect of high-quality protein supplementation on cardiovascular disease (CVD) risk factors, although high-quality proteins such as soy and milk proteins have proposed to be beneficial for cardiometabolic health. MethodsA systematic search in PubMed, Web of Science, Cochrane Library, Scopus, and Embase was conducted to quantify the impact of high-quality protein on CVD risk factors. Results63 RCTs on 4 types of high-quality protein including soy protein, milk protein, whey, and casein were evaluated. Soy protein supplementation decreased systolic blood pressure (SBP, -1.42 [-2.68, -0.17] mmHg), total cholesterol (TC, -0.18 [-0.30, -0.07] mmol/L), and low-density lipoprotein cholesterol (LDL-C, -0.16 [-0.27, -0.05] mmol/L). Milk protein supplementation decreased SBP (-2.30 [-3.45, -1.15] mmHg) and total cholesterol (-0.27 [-0.51, -0.03] mmol/L). Whey supplementation decreased SBP (-2.20 [-3.89, -0.51] mmHg), diastolic blood pressure (DBP, -1.07 [-1.98, -0.16] mmHg), triglycerides (-0.10 [-0.17, -0.03] mmol/L), TC (-0.18 [-0.35, -0.01] mmol/L), LDL-C (-0.09 [-0.16, -0.01] mmol/L) and fasting blood insulin (FBI, -2.02 [-3.75, -0.29] pmol/L). Casein supplementation decreased SBP (-4.10 [-8.05, -0.14] mmHg). In the pooled analysis of four high-quality proteins, differential effects were seen in individuals with different health status. In hypertensive individuals, high-quality proteins decreased both SBP (-2.69 [-3.50, -1.87] mmHg) and DBP (-1.34 [-2.09, -0.60] mmHg). In overweight/obese individuals, high-quality proteins improved SBP (-1.40 [-2.22, -0.59] mmHg), DBP (-2.59 [-3.20, -1.98] mmHg), triglycerides (-0.09 [-0.15, -0.02] mmol/L), TC (-0.14 [-0.22, -0.05] mmol/L), LDL-C (-0.12 [-0.16, -0.07] mmol/L), and HDL-C levels (0.02 [0.01, 0.04] mmol/L). According to the benefits on CVD risks factors, whey ranked top for improving cardiometabolic health in hypertensive or overweight/obese individuals. ConclusionOur study supports a beneficial role of high-quality protein supplementation to reduce CVD risk factors. Further studies are still warranted to investigate the effects of different high-quality proteins on CVD risks in individuals with cardiometabolic disorders.

Campylobacter jejuni and casein hydrolysate addition: Impact on poultry in vitro cecal microbiota and metabolome.

This study investigates the impact of casein hydrolysates on the poultry ceca inoculated with Campylobacter focusing on microbial molecular preferences for different protein sources in the presence of Campylobacter jejuni. Three casein sources (intact casein (IN), casein enzyme hydrolysate (EH), and casein acid hydrolysate (AH)) were introduced to cecal contents in combination with inoculated C. jejuni in an in vitro model system incubated for 48 h at 42°C under microaerophilic conditions. Samples were collected at 0, 24, and 48 h. Genomic DNA was extracted and amplified using custom dual-indexed primers, followed by sequencing on an Illumina MiSeq platform. The obtained sequencing data were then analyzed via QIIME2-2021.11. Metabolite extracts were analyzed with ultra-high-performance liquid orbitrap chromatography-mass spectrometry (UHPLC-MS). Statistical analysis of metabolites was conducted using MetaboAnalyst 5.0, while functional analysis was performed using Mummichog 2.0 with a significance threshold set at P < 0.00001. DNA sequencing and metabolomic analyses revealed that C. jejuni was most abundant in the EH group. Microbial diversity and richness improved in casein supplemented groups, with core microbial differences observed, compared to non-supplemented groups. Vitamin B-associated metabolites significantly increased in the supplemented groups, displaying distinct patterns in vitamin B6 and B9 metabolism between EH and AH groups (P < 0.05). Faecalibacterium and Phascolarctobacterium were associated with AH and EH groups, respectively. These findings suggest microbial interactions in the presence of C. jejuni and casein supplementation are influenced by microbial community preferences for casein hydrolysates impacting B vitamin production and shaping competitive dynamics within the cecal microbial community. These findings underscore the potential of nutritional interventions to modulate the poultry GIT microbiota for improved health outcomes.

Modulation of Swine Gut Microbiota by Phytogenic Blends and High Concentrations of Casein in a Validated Swine Large Intestinal In Vitro Model.

Phytogenic feed additives are gaining popularity in livestock as a replacement for antibiotic growth promotors. Some phytogenic blends (PB) positively affect the production performance, inhibit pathogens within the gut microbiota, and improve the overall health of farm animals. In this study, a swine large intestine in vitro model was used to evaluate the effect of two PBs, alone or in combination with casein, on swine gut microbiota. As a result, the combination of casein with PB1 had the most beneficial effects on swine gut microbiota, as it increased the relative abundance of some commensal bacteria and two genera (Lactobacillus and Oscillospiraceae UCG-002), which are associated with greater production performance in pigs. At the same time, supplementation with PBs did not lead to an increase in opportunistic pathogens, indicating their safety for pigs. Both PBs showed fewer changes in swine gut microbiota compared to interventions with added casein. In contrast, casein supplementation significantly increased beta diversity and the relative abundance of commensal as well as potentially beneficial bacteria. In conclusion, the combination of casein with PBs, in particular PB1, had the most beneficial effects among the studied supplements in vitro, with respect to microbiota modulation and metabolite production, although this data should be proven in further in vivo studies.

Comparison of methods for estimating basal endogenous losses of amino acids and additivity of digestibility of amino acids in corn and soybean meal for broilers

This study was to compare the estimates of basal endogenous losses (BEL) of amino acids (AA) determined by 3 methods including feeding a nitrogen-free diet (NFD) or a low-casein diet (LCD, containing casein at 30 g/kg diet) or using the regression method. Another objective was to investigate whether the ileal AA digestibility of corn calculated from a casein-supplemented corn diet is additive for a corn-soybean meal (SBM) mixed diet in broilers. On d 31 of age, 168 Ross 308 male broilers were assigned to 8 dietary treatments with 6 replicates in a randomized complete block design. An NFD and 3 diets containing 30, 60, or 90 g/kg of casein were formulated to determine the BEL of AA and ileal AA digestibility of casein. The other 4 diets consisted of a corn diet, SBM diet, casein-supplemented corn diet, and corn-SBM mixed diet. On d 35 of age, digesta from the distal section of the ileum were collected. The BEL of AA in birds fed the LCD were greater (P < 0.05) than those of the NFD and the regression method. There were no differences in the BEL of AA determined between the NFD and the regression method. Apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of AA for corn calculated from the casein-supplemented corn diet were greater (P < 0.05) than those of the corn diet. The predicted AID of Thr in the corn-SBM mixed diet based on the AID of AA for corn in the corn diet was lower (P < 0.05) than the measured AID. However, the predicted AID of AA in the mixed diet based on the AID of AA for corn in the casein-supplemented corn diet did not differ from the measured AID. The predicted SID of AA in the mixed diet did not differ from the measured SID irrespective of casein supplementation. In conclusion, feeding an NFD or using the regression method yields similar BEL of AA, but not feeding an LCD. Casein supplementation in the corn diet increases the ileal AA digestibility for corn, which is additive for the corn-SBM mixed diet.

A Soy Protein Isolate Rich in Genistein and Daidzein and Its Effects on Plasma Isoflavone Concentrations, Platelet Aggregation, Blood Lipids and Fatty Acid Composition of Plasma Phospholipid in Normal Men

The effects of consuming a soy protein isolate beverage powder (60 g/d for 28 d) vs. a casein supplement was evaluated in 20 male subjects who were randomly allocated into the two groups. A dramatic rise in plasma isoflavone concentrations was observed after supplementation in the soy protein group, the levels reaching 907 ± 245 nmol/L for genistein (a 110-fold increase) and 498 ± 102 nmol/L for daidzein (a 150-fold increase) as measured by isotope dilution gas chromatography - mass spectrometry. These concentrations are higher than previously reported for the plasma of Japanese subjects consuming a traditional diet (276 nmol/L and 107 nmol/L, respectively). No significant differences in collagen- or 9,11-dideoxy-11α, 9α-epoxymethanoprostaglandin F2α (U46619)-induced platelet aggregation were observed in platelet-rich plasma from the two groups; the increase in plasma isoflavonoids from soy protein supplementation is not sufficient to significantly inhibit platelet aggregation ex vivo. Similarly, plasma total and HDL-cholesterol were not affected by protein supplementation, possibly because the men were normo-cholesterolemic at entry. Analysis of plasma phospholipid polyunsaturated fatty acid composition showed no differences between soy protein and casein supplementation. Previous investigations reported a significant alteration in fatty acid status in animals fed soy protein relative to those fed casein. The present studies indicate that although soy protein supplementation to a typical Western diet can increase plasma concentrations of isoflavones, this may not necessarily be sufficient to counter heart disease risk factors such as high plasma cholesterol and platelet aggregation.

Whey Versus Casein Supplementation in Physically Active, Healthy Individuals

Purpose: Research shows the benefits of whey isolate protein powder and casein protein powder supplementation in regard to body composition and resting metabolic rate; however, minimal research exists comparing the effectiveness of whey to casein supplementation. Methods: Eighteen (18) physically trained, healthy individuals (23±3.1 yrs.; 171.7cm; 12 males; 6 females) completed a randomized two condition [Combat 100% Isolate (Whey) versus Combat 100% (Casein)] by two time point [Pre-, Post-] intervention. The intervention consisted of subjects supplementing on either whey or casein in conjunction with resistance training over an 8-week period. Pre and post testing included % body fat and metabolic rate. Results: A 2x2, repeated-measures analysis of variance (ANOVA) demonstrated non-significant (p ≥ 0.05) effects on changes in % body fat and resting metabolic rate following the 8-week intervention. Conclusions: The data concludes no differences in the effects of whey versus casein protein supplementation in physically active, healthy individuals

Got Milk?

Abstract&#x0D; Objective: To test the absorbance of casein protein by comparing two infant feeding methods: formula and breastmilk, in a healthy six-month-old infant. Hypothesis: Casein protein will be better absorbed through the feeding of breastmilk in a healthy six-month-old infant. Methods: Conducted an in vitro experiment testing casein absorbance using pepsin to simulate an infant’s stomach environment and a colorimeter test to get absorbance measurements. Result: There is a statistically insignificant amount of evidence that casein protein is better absorbed through the feeding of breastmilk and therefore, breastmilk can be used interchangeably with infant formula in strict regards to casein supplementation.&#x0D; &#x0D; Key Terms: Casein to whey ratio, colorimeter, pepsin, nutritional tailoring, absorbance, infant formula, breastmilk

Effect of Post-Ruminal Casein Infusion on Milk Yield, Milk Composition, and Efficiency of Nitrogen Use in Dairy Cows

Adequate supply of amino acids can improve the efficiency of nitrogen use. Casein is the predominant milk protein, and its supplementation can improve milk protein synthesis and nitrogen efficiency. We evaluated the effects of post-ruminal supplementation of casein on milk yield and composition and whole-body protein deposition. Two ruminally cannulated Holstein dairy cows (599 kg) were used in a switch-back design, and treatments were an abomasal infusion of 0 or 400 g/day casein. Cows were fed a diet consisting of corn silage, alfalfa hay, wet corn gluten feed, whole cottonseed, and grain mix, and they received 320 g/day dextrose via abomasal infusion to increase energy:metabolizable protein. The experiment used three 8-day periods. Milk, urine, and feces samples were collected to evaluate milk production, milk composition, and nitrogen retention. Abomasal casein infusion increased (p &lt; 0.01) milk protein percentage and milk urea nitrogen. Nitrogen retention (p = 0.03) and urinary N excretion (p &lt; 0.001) were increased and fecal N excretion (p &lt; 0.001) was decreased by casein infusion. Results suggest casein stimulated protein deposition and altered nitrogen use in lactating dairy cattle. Adaptation periods of 4 days were appropriate for evaluating responses to casein supplementation. Our data provide elements that can aid the design of future experiments.

Pre-Sleep Casein Supplementation, Metabolism, and Appetite: A Systematic Review.

Protein intake is an important factor for augmenting the response to resistance training in healthy individuals. Although food intake can help with anabolism during the day, the period of time during sleep is typically characterized by catabolism and other metabolic shifts. Research on the application of nighttime casein protein supplementation has introduced a new research paradigm related to protein timing. Pre-sleep casein supplementation has been attributed to improved adaptive response by skeletal muscle to resistance training through increases in muscle protein synthesis, muscle mass, and strength. However, it remains unclear what the effect of this nutritional strategy is on non-muscular parameters such as metabolism and appetite in both healthy and unhealthy populations. The purpose of this systematic review is to understand the effects of pre-sleep casein protein on energy expenditure, lipolysis, appetite, and food intake in both healthy and overweight or obese individuals. A systematic review following PRISMA guidelines was conducted in CINAHL, Cochrane, and SPORTDiscus during March 2021, and 11 studies met the inclusion criteria. A summary of the main findings shows limited to no effects on metabolism or appetite when ingesting 24–48 g of casein 30 min before sleep, but data are limited, and future research is needed to clarify the relationships observed.

SHORT-TERM EFFECT OF WHEY PROTEIN SUPPLEMENTATION ON THE QUALITY OF LIFE OF PATIENTS WAITING FOR LIVER TRANSPLANTATION: A DOUBLE BLINDED RANDOMIZED CLINICAL TRIAL.

ABSTRACT Background: Chronic liver disease is associated with malnutrition that negatively impacts a patient’s health-related quality of life (HRQoL). Aim: To evaluate the short-term effect of whey protein supplementation on the HRQoL and nutritional and functional status of patients waiting for liver transplantation. Methods: This was a double-blind randomized clinical trial with patients waiting for liver transplantation who were randomized into two groups: WP (whey protein supplementation) and the control (casein supplementation). Both groups received 40 g (20 g in the morning and 20 g in the evening) for 15 days. Nutritional and functional status were evaluated. Energy balance was calculated as the difference between energy intake (24-hour recall) and total energy expenditure (assessed by indirect calorimetry). The chronic liver disease questionnaire was used to assess HRQoL. All measurements were performed before and after the intervention. Results: Fifty-six patients were evaluated. Malnutrition was present in 56.9%, and it was directly associated with a poor HRQoL (p<0.05). No improvement on the nutritional and functional status was observed, in either group after protein supplementation. HRQoL improved after WP and casein supplementation, with no differences between groups (p>0.05). Patients who met protein requirements and had a positive energy balance demonstrated a higher HRQoL score (4.9, p<0.05), without between-group differences. Conclusion: Malnutrition substantially reduces HRQoL. Short-term WP or casein supplementation improved similarly the HRQoL.

Patin (Pangasius hypophthalmus) fish protein concentrate alters insulin-like growth factor (IGF)-1 and igf binding protein (IGFBP)-3 level of sprague dawley neonate rats-induced malnutrition

Malnutrition is caused by inadequate protein intake and affects growth factor. High protein from patin (Pangasius hypophthalmus) fish is a well-known protein source. This study aims to investigate the effect of patin fish protein concentrate (PFPC) in the IGF-1 and IGFBP-3 level of Sprague Dawley (SD) neonate rats-induced malnutrition. Thirty male SD neonate rats were divided randomly into five groups, namely normal control (K1), malnutrition control (K2), malnutrition with PFPC 13.26 mg.g-1 body weight (BW)/day (X1), malnutrition with PFPC 19.89 mg.g-1 BW/d (X2), and malnutrition with casein supplement 13.26 mg.g-1 BW/d (X3). K1 received a standard diet, while the others received a low 8% protein diet (L8PD) since those were born until 21 days. The standard diet was refed for all groups during the intervention (14 days). IGF-1 and IGFBP-3 levels were measured by ELISA. Normal data were analyzed by using One-way ANOVA which then was followed by post-hoc Bonferroni. Meanwhile, the others were analyzed by Kruskal Wallis and followed by Mann-Whitney U-test. Spearman test was used for correlation. PFPC contained 81.07% of protein, 4.08% of fat, 7.24% of moisture, 2.77% of ash, and 4.83% of carbohydrate. These contents had affected the growth factor. As a result, in the PFPC intervention, IGF-1, and IGFBP-3 levels (p &lt;0.05) were decreased, while the controls were increased. The decreased values were shown in IGFBP-3 levels (p &lt;0.05) while the increase was shown in both controls. On the other hand, the increase in body weight was shown in all groups, including control ones. A strong correlation was found between IGF-1 and IGFBP-3. PFPC has additional value on repairing malnutrition that is the best dose in effecting IGF-1 dan IGFBP3 levels is 13.26 mg.g-1 BW/d.

Comparing the effects of whey and casein supplementation on nutritional status and immune parameters in patients with chronic liver disease: a randomised double-blind controlled trial.

Protein supplementation may be beneficial for patients with chronic liver disease (CLD). This study compared the effects of whey protein isolate (WP) and casein (CA) supplementation on nutritional status and immune parameters of CLD patients who were randomly assigned to take 20 g of WP or CA twice per d as a supplement for 15 d. Body composition, muscle functionality and plasmatic immunomarkers were assessed before and after supplementation. Patients were also classified according to the model for end-stage liver disease (MELD) into less (MELD < 15) and more (MELD ≥ 15) severe disease groups. Malnutrition, determined by the Subjective Global Assessment at baseline, was observed in 57·4 % and 54·2 % of patients in the WP and CA groups, respectively (P = 0·649). Protein intake was lower at baseline in the WP group than in the CA group (P = 0·035), with no difference after supplementation (P = 0·410). Both the WP and CA MELD < 15 groups increased protein intake after supplementation according to the intragroup analysis. No differences were observed in body composition, muscle functionality, most plasma cytokines (TNF, IL-6, IL-1β and interferon-γ), immunomodulatory proteins (sTNFR1, sTNFR2, brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor) or immunomodulatory hormones (adiponectin, insulin and leptin) after supplementation in the WP groups at the two assessed moments. WP supplementation increased the levels of interferon-γ-induced protein-10/CXCL10 (P = 0·022), eotaxin-1/CCL11 (P = 0·031) and monocyte chemoattractant protein-1/CCL2 (P = 0·018) and decreased IL-5 (P = 0·027), including among those in the MELD ≥ 15 group, for whom IL-10 was also increased (P = 0·008). Thus, WP consumption by patients with CLD impacted the immunomodulatory responses when compared with CA with no impact on nutritional status.

Characterization of casein-derived peptide bioactivity: Differential effects on angiotensin-converting enzyme inhibition and cytokine and nitric oxide production

Lactic acid bacteria (LAB) are used as starter cultures in the production of fermented dairy products and have the potential to confer bioactivity relevant to cardiovascular health, as they possess extensive proteolytic systems that liberate small bioactive peptides from larger milk proteins. Certain casein-derived peptides released by various LAB strains during fermentation have been shown to reduce hypertension and to modulate the immune system. We investigated the growth and peptide production of 2 LAB strains, Lactobacillus helveticus R0389 and Lactocaseibacillus rhamnosus R0011, their immunomodulatory activities, as well as their abilities to inhibit the angiotensin-converting enzyme (ACE). Peptide fractions collected from the cell-free supernatant of both medium-grown and milk fermentation cultures were assessed for ACE-inhibitory activity and their effects on the production of proinflammatory and regulatory cytokines by human THP-1 monocytes. Cultures were grown in medium, with or without supplementation with 0.1% casein, or in 3.25% milk fermented with each LAB strain. Casein supplementation increased the growth rate of both LAB strains, and significantly increased ACE-inhibitory activity of peptide fractions collected from both L. helveticus R0389 and L. rhamnosus R0011 cultures grown for 12 h. Fermentation peptide fractions of L. rhamnosus R0011 showed comparable ACE-inhibitory activity to known ACE inhibiting peptides Val-Pro-Pro and Ile-Pro-Pro (up to 79% inhibition) with a significant difference between culture peptide fractions and acidified and nonacidified control fractions collected after 6 d of fermentation. Many milk and casein-derived peptides reported in previous studies have been identified as part of a larger bioactive fraction. We synthesized a group of these peptides to individually assess both ACE-inhibitory and immunomodulatory activity. The known ACE inhibitors Val-Pro-Pro and Ile-Pro-Pro showed similar ACE inhibition to previously published results, while also inducing the production of the regulatory cytokine IL-10 by monocytes in the presence and absence of a proinflammatory stimulant. These synthesized peptides could also induce the production of nitric oxide (NO), a potent vasodilator, in human endothelial cell cultures. Investigating the relationships among these bioactive properties could improve the use of probiotic organisms and their secreted products in the food industry.

Pre-sleep protein in casein supplement or whole-food form has no impact on resting energy expenditure or hunger in women.

The purpose of this study was to determine the effect of a whole-food protein (cottage cheese, CC) consumed before sleep on next-morning resting energy expenditure (REE), RER and appetite compared with an isoenergetic/isonitrogenous casein protein (CP) supplement and placebo (PL) in active women. In a beverage-blinded, randomised, cross-over design, ten active women (age, 23·1 (sd 1·9) years; body fat, 22·0 (sd 4·6) %) consumed pre-sleep CC (30 g of protein, 10 g of carbohydrate and 0 g of fat) or energy- and protein-matched liquid CP or PL (0 kJ). Participants arrived at 18.00 hours for an overnight stay in the laboratory. At 30-60 min before normal bed time (2 h post standard meal), participants consumed CC, CP or PL before measurement of REE. Upon waking (05.00-08.00 hours), REE was repeated and subjective appetite was recorded. Statistical analyses were conducted using repeated-measures ANOVA (SPSS). Significance was accepted at P≤0·05. There were no significant differences in acute REE (CC, 7217 (sd 1368); CP, 7188 (SD 895); PL, 7075 (sd 1108) kJ/d, P=0·95), acute RER (0·79 (sd 0·05), P=0·56), morning REE (CC, 5840 (sd 1225); CP, 5694 (sd 732); PL, 5991 (sd 903) kJ/d, P=0·79) or morning RER (0·77 (sd 0·03), P=0·52). Subjective measures of appetite were not different between groups. In active women, pre-sleep consumption of CC does not alter REE or RER more than a CP or PL beverage. These data suggest that the metabolic response from whole-food protein do not differ from the metabolic response of liquid protein.

Daytime and nighttime casein supplements similarly increase muscle size and strength in response to resistance training earlier in the day: a preliminary investigation

BackgroundCasein protein consumed before sleep has been suggested to offer an overnight supply of exogenous amino acids for anabolic processes. The purpose of this study was to compare supplemental casein consumed earlier in the day (DayTime, DT) versus shortly before bed (NightTime, NT) on body composition, strength, and muscle hypertrophy in response to supervised resistance training.MethodsThirteen males participated in a 10-week exercise and dietary intervention while receiving 35 g casein daily. Isocaloric diets provided 1.8 g protein/kg body weight.ResultsBoth groups increased (p < 0.05) in lean soft tissue (DT Pre: 58.3 ± 10.3 kg; DT Post: 61.1 ± 11.1 kg; NT Pre: 58.3 ± 8.6 kg; NT Post: 60.3 ± 8.2 kg), cross-sectional area (CSA, DT Pre: 3.4 ± 1.5 cm2; DT Post: 4.1 ± 1.7 cm2; NT Pre: 3.3 ± 1.6 cm2; NT Post: 3.7 ± 1.6 cm2) and strength in the leg press (DT Pre: 341 ± 87.3 kg; DT Post: 421.1 ± 94.0 kg; NT Pre: 450.0 ± 180.3 kg; NT Post: 533.9 ± 155.4 kg) and bench press (DT Pre: 89.0 ± 27.0 kg; DT Post: 101.0 ± 24.0 kg; NT Pre 100.8 ± 32.4 kg; NT Post: 109.1 ± 30.4 kg) with no difference between groups in any variable (p > 0.05).ConclusionsBoth NT and DT protein consumption as part of a 24-h nutrition approach are effective for increasing strength and hypertrophy. The results support the strategy of achieving specific daily protein levels versus specific timing of protein ingestion for increasing muscle mass and performance.Trial registrationClinicalTrials.gov Identifier: NCT03352583.

Comparison between pre-exercise casein peptide and intact casein supplementation on glucose tolerance in mice fed a high-fat diet

We hypothesized that along with exercise, casein peptide supplementation would have a higher impact on improving glucose tolerance than intact casein. Male 6-week-old ICR mice were provided a high-fat diet to induce obesity and glucose intolerance. The mice were randomly divided into 4 treatment groups: control (Con), endurance training (Tr), endurance training with intact casein supplementation (Cas+Tr), and endurance training with casein peptide supplementation (CP+Tr). The mice in each group were orally administrated water, intact casein, or casein peptide (1.0 mg/g body weight, every day), and then subjected to endurance training (15-25 m/min, 60 min, 5 times/week for 4 weeks) on a motor-driven treadmill 30 min after ingestion. Our results revealed that total intra-abdominal fat was significantly lower in CP+Tr than in Con (p < 0.05). Following an oral glucose tolerance test, the blood glucose area under the curve (AUC) was found to be significantly smaller for CP+Tr than for Con (p < 0.05). Moreover, in the soleus muscle, glucose transporter 4 (GLUT4) protein levels were significantly higher in CP+Tr than in Con (p < 0.01). However, intra-abdominal fat, blood glucose AUC, and GLUT4 protein content in the soleus muscle did not alter in Tr and Cas+Tr when compared with Con. These observations suggest that pre-exercise casein peptide supplementation has a higher effect on improving glucose tolerance than intact casein does in mice fed a high-fat diet.

Metabolic acidosis stimulates the production of the antimicrobial peptide cathelicidin in rabbit urine.

Intercalated cells of the collecting duct (CD) are critical for acid-base homeostasis and innate immune defense of the kidney. Little is known about the impact of acidosis on innate immune defense in the distal nephron. Urinary tract infections are mainly due to Escherichia coli and are an important risk factor for development of chronic kidney disease. While the effect of urinary pH on growth of E. coli is well established, in this study, we demonstrate that acidosis increases urine antimicrobial activity due, at least in part, to induction of cathelicidin expression within the CD. Acidosis was induced in rabbits by adding NH4Cl to the drinking water and reducing food intake over 3 days or by casein supplementation. Microdissected CDs were examined for cathelicidin mRNA expression and antimicrobial activity, and cathelicidin protein levels in rabbit urine were measured. Cathelicidin expression in CD cells was detected in kidney sections. CDs from acidotic rabbits expressed three times more cathelicidin mRNA than those isolated from normal rabbits. Urine from acidotic rabbits had significantly more antimicrobial activity (vs. E. coli) than normal urine, and most of this increased activity was blocked by cathelicidin antibody. The antibody had little effect on antimicrobial activity of normal urine. Urine from acidotic rabbits had at least twice the amount of cathelicidin protein as did normal urine. We conclude that metabolic acidosis not only stimulates CD acid secretion but also induces expression of cathelicidin and, thereby, enhances innate immune defense against urinary tract infections via induction of antimicrobial peptide expression.

Effects of dietary total phosphorus concentration and casein supplementation on the determination of true phosphorus digestibility for broiler chickens

A study was conducted to compare the regression-driven estimates of true precaecal digestibility (TPD) of phosphorus (P) in soybean meal (SBM) for broiler chickens fed diets with or without casein supplementation at moderate or low total P concentration. A total of 768 male Ross 308 broilers were allocated to 12 diets in a 2 × 2 × 3 factorial arrangement of two total P concentration (moderate-P or low-P), two diet types (with or without casein) and three SBM levels (low, medium or high). There were 8 chicks per cage and 8 replicate cages per treatment group in a randomised complete block design. The birds were fed experimental diets form d 14 to 21 post-hatching. Chromic dioxide was used as an indigestible marker. The results showed that dietary casein supplementation improved body weight (BW) gain, feed intake and feed efficiency of broilers (p < .01). Broilers fed the moderate-P diets had greater dietary total P intake, precaecal flow and total tract output of P, precaecal digested P, and total tract retained P compared with birds fed the low-P diets (p < .01). Dietary casein supplementation increased precaecal digested P, total tract retained P, apparent precaecal digestibility (APD) and total tract retention (TTR) of P (p < .01). Broilers fed the moderate-P diets had a linear decrease in the APD and TTR of P with increasing SBM levels (p < .01). Precaecal digested P and total tract retained P increased with graded inclusion of SBM levels (linear, p < .01). Regression of precaecal digested P against dietary P intake showed that chicks fed the moderate-P diets had lower estimates of TPD of P in SBM than that for chicks fed the low-P diets (p < .05), while dietary casein supplementation had no effects on the determined values of TPD of P. In conclusion, the regression-driven true phosphorus digestibility in assay feed ingredients for broiler chickens was affected by dietary total P concentration.

L‐carnitine supplement and running training improve skeletal muscle recovery after hindlimb suspension induced muscle atrophy

Several studies demonstrated that endurance exercise has an effect on protein synthesis through the activation of Akt1/mTOR pathway. Along with Akt1/mTOR pathway, PGC‐1α effects on protein synthesis through the inhibition of muscle specific E3 ligases expression such as atrogin‐1 and MuRF1. PGC‐1α expression in skeletal muscle can be induced by a stimulus of endurance exercise. Recently, L‐carnitine down‐regulates atrogin‐1 and MuRF1 mRNA expression and reduces muscle wasting in a rat model of cancer cachexia. Also, L‐carnitine supplement can increase the level of IGF‐1, which in turn activates the downstream Akt1/mTOR signaling pathway. Altogether, these findings suggest that L –carnitine and combined with running exercise could promote muscle recovery after disuse induced muscle atrophy through the inhibition of E3 ligases expression and Akt1/mTOR pathway activation. However, the effect of LC supplement in growing rats on muscle atrophy and recovery of muscle mass combined with running exercise after muscle atrophy induced by disuse has not yet been reported. Therefore, the purpose of this study was to clarify whether L‐carnitine supplement and combined with running exercise improve muscle mass recovery after disuse induced muscle atrophy through the inhibition of UPP and activation of Ak1/mTOR pathway. Male Wistar rats were randomly assigned to 4 groups; sedentary after hindlimb suspension with 1,250 mg/kg casein supplement group (Sed‐Con), sedentary after hindlimb suspension with 1,250 mg/kg l‐carnitine supplement group (Sed‐Car), running exercise after hindlimb suspension with 1,250 mg/kg casein supplement group(Ex‐Con), and running exercise after hindlimb suspension with 1,250 mg/kg l‐carnitine supplement group (Ex‐Car) for 4wk. Soleus muscle significantly greater in exercised groups compared to sedentary groups. MuRF1 mRNA expression is significantly decreased by L‐carnitine supplement in sedentary and exercise groups. PGC‐1α and Akt1 mRNAs expression in muscle is significantly increased by the exercise and L‐carnitine supplement. However, PGC‐1α protein expression is significantly increased in Ex‐Car group compared to Sed‐Con group. mTOR mRNA expression is only increased in Ex‐Car group groups compared to the Sed‐Con group. In conclusion, the main findings of this study is that L‐carnitine combined running exercise enhanced the muscle recovery after hindlimb suspension induced muscle atrophy in soleus muscle possibly through of the increased expression of PGC‐1α by inhibiting the muscle specific E3 ligases expression and activation of Akt1/mTOR pathway. These results provide important information that understanding of the mechanisms underlying muscle recovery after hindlimb suspension induced muscle atrophy by the L‐carnitine combined with running exercise.