Dietary Whey Research Articles

Comparison of the Faecal Microbiota Composition Following a Dairy By-Product Supplemented Diet in Nero Siciliano and Large White × Landrace Pig Breeds.

The current study compared the faecal microbiota composition of two pig breeds (autochthonous vs. commercial) to understand what happens after the integration of liquid whey in the diet and what the role of the host genetic is. The trial was conducted for 60 days, and the faecal microbiota composition was investigated at three time points, T0, T1 (after 30 days) and T2 (after 60 days) in 30 female pigs (20 commercial crossbred and 10 Nero Siciliano pigs). The animals were divided into four groups (two control and two treatment groups). Generally, in both breeds, Firmicutes (51%) and Bacteroidota (36%) were the most abundant phylum whereas Prevotella, Treponema and Lactobacillus were the most abundant genera. The two breeds have a different reaction to a liquid whey diet. In fact, as shown by PERMANOVA analysis, the liquid whey significantly (p < 0.001) affects the microbiota composition of crossbreeds while not having an effect on the microbiota of the Nero Siciliano. Despite this, in both breeds Bifidobacterium and Ruminococcus have been positively influenced by liquid whey and they promote intestinal health, improve immunity, increase performance, and feed efficiency. In conclusion, the integration of liquid whey had a different effect on the Nero Siciliano and crossbred pig breeds, emphasizing the importance of the host genetic profile in determining the faecal bacterial composition.

Continuous consumption of whey protein affects motor and cognitive functions in a mouse model of cow’s milk hypersensitivity

Abstract Hypersensitization to a food allergen can occur in the absence of immediate allergic reactions, a condition referred to as asymptomatic sensitization. Using mice asymptomatically sensitized to a bovine whey allergen, β-lactoglobulin (BLG, Bos d 5), we have previously shown that chronic exposure to the allergen results in behavior changes with increased neuroinflammation and cortical demyelination. However, whether allergen avoidance can resolve these pathologies is unknown. Thus, we aimed to compare immunological and behavioral changes in sham- or BLG-sensitized mice with or without the removal of dietary whey after the mice had consumed either a control diet (CTL) or a whey-protein-containing diet (WP) for 2 weeks. As previously reported, BLG-sensitized mice did not display overt physical reactions during the 2-week allergen exposure period, while their plasma levels of BLG-specific IgE and IgG1 were elevated compared to the respective sham groups. Following an additional 2 weeks on either the CTL or WP diet, these antibodies remained elevated in some of the sensitized mice in both diet groups. The grip-strength test indicated that sham mice that switched to the CTL diet after the 2-week WP diet increased limb strengths. In contrast, BLG-sensitized mice on either diet, as well as sham mice that stayed on the WP diet, showed a decline in limb strengths, suggesting that WP consumption, whether during sensitization or as part of the daily diet, affected their motor function. Furthermore, removing dietary WP improved spatial memory performance with the cross-maze test in both sham and BLG-sensitized mice. These results suggested that continuous exposure to a dietary allergen may influence motor and cognitive functions. Supported by grants from NIH (R01 AI168563)

Dietary whey protein increases brain leukocytes in mice regardless of their hypersensitivity status

Abstract Cow’s milk allergy (CMA) often manifests as milder reactions and may be linked to neurological problems. Previously, we demonstrated that C57BL/6J mice sensitized to a bovine whey allergen, β-lactoglobulin (BLG, Bos d 5), moderately increased BLG-specific IgE levels and exhibited behavioral changes without severe allergic reactions. When these non-anaphylactic CMA mice were placed on a whey-protein (WP)-containing diet for 2 weeks to simulate continuous dairy consumption, we found neuropathology indicative of neuroinflammation and cortical demyelination. Since immune cells migrate to the central nervous system (CNS) and promote neuroinflammation in demyelinating conditions such as multiple sclerosis, we hypothesized that the number of leukocytes would increase in BLG-sensitized mouse brains to orchestrate neuropathology. To test this hypothesis, we used flow cytometry to determine the number and phenotypes of leukocytes in the brains of naïve, sham, and BLG-sensitized mice after the 2 weeks of the WP diet. The frequencies of cells expressing common leukocyte marker CD45, pan T cell marker CD3, cytotoxic T cell marker CD8, integrin CD11b, myeloid cell marker CD14, and co-stimulatory marker CD86 significantly increased, regardless of the sensitization status. The percentages of these cells were low in mice that never received WP. This result indicated that WP diet consumption alone increased CNS leukocyte populations. Additional immunophenotyping is needed to determine whether the identified cells can be differentiated among the experimental groups. Detailed characterization of CNS leukocyte phenotypes and dynamics will help elucidate the mechanism of CMA-induced neuroinflammation and cortical demyelination.

Different genotype and a liquid whey-supplemented diet influence the resilience of pigs through immune-modulation and anti-inflammatory response

This study evaluated (i) whether weight gain and levels of inflammatory and immune markers including white blood cells (WBC), serum haptoglobin, C-reactive protein, albumin, and globulin fractions change between the Nero Siciliano pig breed and the crossbreed Landrace x Large White (LxLW) reared under the same environmental and farming conditions; and (ii) whether a liquid whey diet supplementation affects the investigated parameters in both genotypes. In this study, 10 crossbreed LxLW and 10 Nero Siciliano pigs were given control feed, representing the control groups (CTRC and CTRNS), whereas 10 crossbreed LxLW and 10 Nero Siciliano pigs were given control feed supplemented with liquid whey for 2 months, representing the experimental groups (WC and WNS). From each pig, body weight and blood were collected before experimental diet supplementation (T0), and one (T1) and two (T2) months after the start of the diet supplemented with whey. The white blood cell count (WBC), serum haptoglobin, C-reactive protein, total proteins, albumin and globulin fraction concentration were assessed. Two-way analysis of variance showed an increasing trend of body weight both in the control and experimental groups of the two pig genotypes throughout the monitoring period (p < 0.01) without a significant effect of genotype and diet (p > 0.05). The concentration of haptoglobin, β1- and β2-globulins was affected by pig genotype, diet supplementation, and time (p < 0.01). The values of WBC, C-reactive protein, albumin, α-globulins, and A/G ratio were affected by diet supplementation (p < 0.01) and time (p < 0.01) without an influence of genotype (p > 0.05). Nero Siciliano pigs showed lower levels of haptoglobin, β1-globulin, and β2-globulin compared to crossbreed LxLW. Nero Siciliano pigs and crossbred LxLW fed with liquid whey showed lower levels of WBC, haptoglobin, C-reactive protein, α-, β1-, and β2-globulins and higher values of albumin compared to control groups. The results reinforced the hypothesis that autochthonous breeds possess higher resilience to farming conditions when compared to allochthonous breeds. Moreover, an immune-modulatory and an anti-inflammatory power of liquid whey dietary supplementation is suggested probably thanks to its content in natural bioactive substances including anti-inflammatory cytokines and anti-oxidative factors.

Lactating sows fed whey protein supplement has eventually increased the blood profile of piglets.

The intension of this study was to examine the effect of dietary whey protein supplementation on the reproduction performance, growth performance and blood profile of sow and their offspring. From Day 114 of lactation to 21days of weaning, a total of 21sows (n=7/ treatment) (Landrace×Yorkshire) were blocked according to average parity (2.4) and allocated to 1 of 3 dietary treatments: (i) CON-corn-soybean meal based basal diet, (ii) WPC-CON+0.047% WPC whey protein concentrate (WPC) and (iii) WPH-CON+0.02% whey protein hydrolysate (WPH). The reproduction performance of sows was not affected by WPC or WPH supplementation. However, piglets that were born to WPC and WPH group sows showed higher body weight at birth (p=0.057) and at weaning (p=0.018). After farrowing, WPC and WPH group sows showed decreased (p=0.043) RBC count and total iron-binding count (TIBC) (p=0.046), whereas at the end of the experiment, the blood profile including red blood cells, iron, haemoglobulin and TIBC was significantly increased (p=0.042, 0.049, 0.051 and 0.052 respectively) in WPC group piglets compared to the CON and WPH groups. Based on the positive impact on the blood profile of piglets, we conclude that whey protein supplement could serve as a potential energy source to suit lactating sows that could eventually benefit the performance of their offspring.

Dietary whey and egg proteins interact with inulin fiber to modulate energy balance and gut microbiota in obese rats

Diets supplemented with protein and fiber are well known to reduce food intake and weight gain; however, less is known about the combined effects of protein and prebiotic fiber on energy balance and gut microbiota composition. We compared effects of diets containing high egg or whey protein with cellulose or prebiotic (inulin) fiber on energy balance, gut microbiota, hormones, and metabolites. Male obese rats (n=8/group) were allocated to four diets: Egg albumen+Cellulose (EC), Egg albumen+Inulin (EI), Whey protein+Cellulose (WC), and Whey protein+Inulin (WI). Results revealed that diet-induced hypophagia was transient with EC and prolonged with EI and WI, compared to WC. Importantly, CCK-1 receptor antagonist (Devazepide) attenuated the hypophagic effects of EC, EI, and WI. Further, EC, EI and WI decreased respiratory quotient, energy expenditure, weight and adiposity gains, and improved glycemia, relative to WC. Propranolol (β1-β2-receptor blocker) attenuated diet-induced changes in energy expenditure. Transcript abundance of thermogenic markers in brown adipose tissue, plasma hormones, and metabolites especially acyl-carnitines and glycerophospholipids, were differentially altered by diets. Diet explained 25% of compositional differences in cecal microbiomes, but diets with same fiber type did not differ. Microbiota differing between groups also strongly correlated with gut hormones and metabolites. Species most strongly correlated to a marker for butyrate production were in highest abundance in inulin diets. Together, these findings indicate that inulin enriched diets containing egg or whey protein improved energy balance, decreased adiposity, and modulated gut microbiota and metabolites, with CCK signaling partly mediating the satiety effects of diets.

Dietary Whey Protein Supplementation Increases Immunoglobulin G Production by Affecting Helper T Cell Populations after Antigen Exposure.

Whey protein is a by-product of cheese and casein manufacturing processes. It contains highly bioactive molecules, such as epidermal growth factor, colony-stimulating factor, transforming growth factor-α and -β, insulin-like growth factor, and fibroblast growth factor. Effects of whey protein on immune responses after antigen (hemagglutinin peptide) injection were evaluated in rats. Experimental diets were formulated based on NIH-31M and supplemented with 1% amino acids mixture (CON) or 1% whey protein concentrate (WPC) to generate isocaloric and isonitrogenous diets. Rats were fed the experimental diets for two weeks and then exposed to antigen two times (Days 0 and 14). Blood was collected on Days 0, 7, 14, and 21 for hematological analysis. The WPC group showed decreased IgA and cytotoxic T cells before the antigen injection (Day 0) but increased IgG, IL-2, and IL-4 after antigen injection due to increased B cells and T cells. Helper T cells were increased at Days 14 and 21, but cytotoxic T cells were not affected by WPC. WPC may activate adaptive immunity (IgG) against antigen by modulating helper T cells. Bioactive molecules might contribute to the immune-enhancing effects of whey protein concentrate.

Whey peptides exacerbate body weight gain and perturb systemic glucose and tissue lipid metabolism in male high-fat fed mice

Consumption of milk-derived whey proteins has been demonstrated to have insulin-sensitizing effects in mice and humans, in part through the generation of bioactive whey peptides. While whey peptides can prevent insulin resistance in vitro, it is unclear whether consumption of whey peptides can prevent obesity-induced metabolic dysfunction in vivo. We sought to determine whether whey peptides consumption can protect from high fat (HF) diet-induced obesity and dysregulation of glucose homeostasis. Male C57BL/6J mice were fed either a low or HF diet for 13 weeks. HF diet fed mice were provided drinking water with no addition (control), undigested whey protein isolate (WPI, 1 mg ml-1) or whey protein hydrolysate (WPH, 1 mg ml-1) throughout the diet regimen. Mice consuming WPH gained more body weight and were more glucose intolerant compared to those consuming WPI or water only. Despite increased body weight gain, perigonadal adipose tissue weight and lipid accumulation were unchanged. However, excess lipids accumulated ectopically in the liver and skeletal muscle in mice consuming WPH, which was associated with elevated inflammatory markers systemically and in adipose tissue, liver, and skeletal muscle. In skeletal muscle, mitochondrial fat oxidation and electron transport chain proteins were decreased with WPH consumption, indicative of mitochondrial dysfunction. Taken together, our results demonstrate that WPH, but not WPI, exacerbates HF-induced body weight gain and impairs glucose homeostasis, which is accompanied by increased inflammation, ectopic fat accumulation and mitochondrial dysfunction. Thus, our results argue against the use of dietary whey peptide supplementation as a preventative option against HF diet-induced metabolic dysfunction.

Semi-modified okara whey diet increased insulin secretion in diabetic rats fed a basal or high fat diet.

Lifestyle and diet preferences are primarily responsible for developing type 2 diabetes. In this study, okara was manufactured into okara whey crackers (OWC) to investigate its dietary role in controlling diabetes in streptozotocin-diabetic rats with and without a high-fat diet. Forty-eight rats were divided into eight groups. G1-G4 were nondiabetic and fed a basal diet, a basal diet with 30% crackers, high fat diet, and a high-fat diet with 30% crackers, respectively. G5-G8 were diabetic groups that received similar diets as previous groups. Blood glucose, liver function, lipid pattern, pancreas and liver histopathology, and insulin immunohistochemistry were performed. OWC improved measured parameters and histopathology of the liver and pancreas in diabetic rats. The area % of positive insulin cells was increased in G6 (5.20%) and G8 rats (2.83%) fed OWC compared to diabetic rats (1.17%). In conclusion, the use of 30% OWC in a semi-modified diet has controlled the hyperglycemia and hyperlipidemia associated with diabetes.

The Effect of Whey on Performance, Gut Health and Bone Morphology Parameters in Broiler Chicks.

Whey is a highly nutritious byproduct of the cheese industry that can be used effectively in the animal feed industry. However, the use of whey in poultry diets is limited by its high lactose and mineral contents. The objective of the present study was to evaluate the effect of different concentrations of whey in poultry diets on the performance, intestinal microbiota and physico-chemical parameters of the intestinal ecosystem, as well as on the bone morphology and its strength in broiler chicks. One hundred and twenty-eight, day-old, male broiler chicks were randomly allocated into four treatment groups of 32 chicks each. The treatment groups were: group A, which served as negative control and groups B, C and D, supplemented with 1, 2 and 5% of dietary whey, respectively. Performance of the groups was evaluated throughout the experiment. Following necropsies, the gastrointestinal tract from each bird was removed, divided into its anatomical parts and intestinal samples were taken for microbiological analysis and for pH and viscosity measurement as well. Tibiotarsus was also collected for morphometric analysis and strength evaluation. The statistical analysis of the experimental data revealed that the dietary supplementation of 1 and 2% of whey improved significantly (p ≤ 0.05) the body weight, while the addition of 5% of whey reduced significantly (p ≤ 0.05) the body weight. Furthermore, the addition of 1, 2 and 5% of dietary whey increased significantly (p ≤ 0.05) the pH of jejunum digesta and reduced significantly (p ≤ 0.05) the pH of caecum digesta compared to the control group. The addition of 1 and 2% of whey reduced significantly (p ≤ 0.05) the viscosity in the jejunum and ileum digesta, compared to the addition of 5% of whey which reduced significantly (p ≤ 0.05) the viscosity in jejunum digesta but increased significantly (p ≤ 0.05) the viscosity in ileum digesta. Moreover, the addition of 1, 2 and 5% of dietary whey increased significantly (p ≤ 0.05) the caecal counts of Lactobacillus spp. and Lactococcus lactis, while the addition of 5% of whey reduced significantly (p ≤ 0.05) the tibiotarsus length. It can be concluded that the addition of low quantities of whey up to 2% promoted the performance and gut health of birds, while the addition of higher quantities of whey at the level of 5% had a detrimental effect on the performance and tibiotarsus length.

Use of Whey Protein Concentrates in Broiler Diets

In this study, we investigated the response of broiler chickens to dietary whey protein concentrate (WPC) as a prebiotic supplement (0, 0.15, 0.20, and 0.25%) on performance, carcass criteria, blood metabolites, oxidative status of meat and liver, and meat quality traits. A total of 200, one-wk-old, unsexed chicks were randomly allotted to 4 treatment groups in a completely randomized design experiment. Results revealed that supplementing broiler diets with WPC did not affect live body weight and body weight gain values in comparison with control during the overall period (1–6 wk of age). A reduction in daily feed intake was observed in WPC-treated groups compared to that in the control. However, improvements (P < 0.05) in the feed conversion ratio in all WPC groups were recorded compared to that in the control, throughout the experimental period. The WPC group had the best values of dressing and breast percentages. A gradual reduction in serum creatinine, triglycerides, total cholesterol, and malondialdehyde as well as increasing levels of the antioxidative enzymes glutathione peroxidase, glutathione-S-transferase, catalase, and superoxide dismutase were recorded with increasing WPC levels. Dietary WPC supplementation improved (P < 0.05) most of the meat quality traits. Thus, WPC supplementation can enhance broiler performance and meat quality without biohazards. In addition, WPC has several health benefits, which may reflect positively on human health as consumers of broiler meat.

Metabolic Response to Short-term Protein Supplementation in Reproductive-Aged Women with Polycystic Ovary Syndrome (PCOS) (P08-070-19)

ObjectivesWomen with polycystic ovary syndrome (PCOS) are commonly insulin resistant, hyperinsulinemic and hyperglycemic, increasing their risk for development of other metabolic, cardiovascular and reproductive disturbances. Dietary whey protein may attenuate postprandial glycemic excursions due to its effect on the gut-brain axis. The aim of the study is to evaluate 7-day whey protein supplementation on metabolic parameters in young women with PCOS. MethodsA repeated measures (RM) design was used. Three women with and six without PCOS (PCO and CON respectively) underwent three 150-min oral glucose tolerance tests (OGTT): (i) OGTT without protein preload, (ii) OGTT on first and (iii) seventh day of protein preload. Participants consumed 35 g protein daily with protein intake 30 min before carbohydrate load on the test days. Blood samples collected at −30 (fasting/pre-preload), 0 (pre-glucose), 15, 30, 60, 90, 120 and 150 min of OGTTs were analyzed for glucose, insulin, and incretins. (Participant recruitment, data collection and analyses in progress.) ResultsMixed design RM - MANOVA indicates that day of test and time did not have a significant effect on glucose levels within and between groups (P ≥ .05). However, day (P = .027) and time (P = .047) had a significant impact on insulin concentrations in both, PCO and CON groups. Insulin was significantly higher at 30 min compared to −30 and 0 min timepoints (P = .016 and P = .032 respectively) on the first and seventh days of preload. ConclusionsAn insulinogenic response elicited by 35 g protein preload may blunt glycemic responses, moderating them throughout the test period in women with PCOS and age-matched healthy controls. Timing of supplementation may play a vital role in achieving long term glucose homeostasis in PCOS. Funding SourcesTexas Woman’s University Research Enhancement Program & Glanbia Nutritionals, Inc.

Performance and development of gastric compartments of calves fed with cheese whey and transition milk

ABSTRACT: This study aimed to evaluate whether cheese whey associated with transitional milk or not, could serve as a suitable alternative to whole milk for calf rearing. For this, twenty-four male calves of crossbred Holstein-Zebu, were distributed in a completely randomized design with three treatments (8 calves per treatment) and fed for 60 days with one of the following liquid diets: i) 100% whole milk, ii) 50% whole milk and 50% cheese whey, and iii) 30% transitional milk and 70% cheese whey. There was no difference in food intake between treatments. The analysis showed that whole milk replacement did not alter the stomach weight or the ruminal cell proliferation when compared to treatment using 100% whole milk; however, the transitional milk/cheese whey group showed larger ruminal papillae. These results suggested that a transitional milk-cheese whey diet associated with solid food and proper nutrient complements might enhance the post-natal development of rumen papillae in calves, which might contribute to prepare them for weaning.

Regulation of intestinal growth in response to variations in energy supply and demand.

SummaryThe growth of the intestine requires energy, which is known to be met by catabolism of ingested nutrients. Paradoxically, during whole body energy deficit including calorie restriction, the intestine grows in size. To understand how and why this happens, we reviewed data from several animal models of energetic challenge. These were bariatric surgery, cold exposure, lactation, dietary whey protein intake and calorie restriction. Notably, these challenges all reduced the adipose tissue mass, altered hypothalamic neuropeptide expression and increased intestinal size. Based on these data, we propose that the loss of energy in the adipose tissue promotes the growth of the intestine via a signalling mechanism involving the hypothalamus. We discuss possible candidates in this pathway including data showing a correlative change in intestinal (ileal) expression of the cyclin D1 gene with adipose tissue mass, adipose derived‐hormone leptin and hypothalamic expression of leptin receptor and the pro‐opiomelanocortin gene. The ability of the intestine to grow in size during depletion of energy stores provides a mechanism to maximize assimilation of ingested energy and in turn sustain critical functions of tissues important for survival.

The Effects of Dietary Whey Lactose and Lactobacillus Agilis Bacteria on the Growth Performance, Physicochemical Conditions of the Digestive Tract and the Caecal Microbial Ecology of Broiler Chickens

Abstract The principal goal of this study was to assess the responses of broiler chickens raised on floor litter to the 2% dietary level of lactose (LAC) originating from dried whey fed in combination with live culture of Lactobacillus agilis bacteria (90 million cells/kg diet) in terms of the performance and basic postslaughter parameters, the lumen pH in some alimentary tract segments, concentration of volatile fatty acids (VFA ) in the total (T) and undissociated (UD) forms and count of selected microbial populations in the caeca determined by fluorescent in-situ hybridisation (FISH). A parallel aim was to evaluate the outcomes from the combined supplementation in comparison with feeding of LAC and the L. agilis bacteria as separate dietary supplements. Six hundred and forty Ross 308 chickens were placed in 16 floor pens (40 birds per pen having equal sex ratio) and were provided with free access to the feed (a mash maize-wheat-soybean meal-based diet) and water. Dietary treatments provided from day 8 to 42 of age were: LAC-free basal diet (CON), LAC-containing diet (CON + LAC), LAC-free diet with the addition of L. agilis (CON + BAC) and LAC-containing diet with addition of L. agilis (CON + LAC + BAC). The LAC supplementation caused significant decreases in the luminal pH of the crop, ileum and caeca, and the addition of the L. agilis bacteria reduced the pH in the crop and caeca. The overall concentration of total (T) volatile fatty acids was higher in the caeca of broilers receiving the LAC-containing diets. Both the LAC and the BAC supplements, independently from one another, resulted in significantly greater caecal levels of UD acetate, propionate and butyrate. The FISH analysis revealed that counts of Bacteroides sp./Prevotella sp. group were higher after the inclusion of LAC in the feed. All three dietary supplementations significantly reduced the total counts of the family Enterobacteriaceae and decreased the number of naturally occurring C. perfringens bacteria compared with the basal control diet (CON). Neither LAC inclusion nor BAC addition to the diet affected the counts of the Clostridium coccoides/Eubacterium rectale group. The synergistic effects of the simultaneous supplementation of LAC and L. agilis were found on the T butyrate concentration and on C. perfringens and the Enterobacteriaceae counts. No improvements in the body weight gains and post-slaughter traits were observed due to uncombined and combined supplementation with 2% LAC and L. agilis, indicating that the shifts in composition of the caecal microbiota toward a healthier composition by using these additives were not large enough to create the positive growth rate and processing yields responses in broilers maintained in a litter-floor environment.

Immune Response and Disease Resistance of Barramundi, Lates calcarifer (Bloch), Offered Diets Supplemented with Replete Levels of Tryptophan or Whey

AbstractA basal diet containing 45% protein, 10% lipid, and 1.95% tryptophan (Trp) was formulated, and then supplemented with crystalline Trp (0, 0.1, 0.5, and 1% of the diet) or whey (16.9%) to evaluate the physiological effects of juvenile (initial weight: 0.57 ± 0.15 g) barramundi over a 58‐d rearing period. Significant reductions in growth and feed intake were observed in fish offered diets supplemented with Trp as compared to those offered the basal diet or the whey diet. Brain serotonin increased in fish fed the 1% Trp and whey diets albeit a decrease in cannibalism was not observed. Phagocytic activity, and Mx gene expression in spleens were not significantly changed among all groups. There were significant increases in lysozyme activity in the 1% Trp and whey groups, respiratory burst in the whey group, and Mx gene expression in head kidneys in the 1% Trp group 12 h after nerve necrosis virus (NNV) injection and in the whey 12 group and 24 h after NNV injection. Cumulative mortalities of fish fed the whey diet were significantly lower than those of fish fed the basal diet after Aeromonas hydrophila infection, but not Trp groups. Based on these results, diets supplemented with whey had reduced mortality when challenged with A. hydrophila and consequently function as an immunostimulant.

Whey protein effects on energy balance link the intestinal mechanisms of energy absorption with adiposity and hypothalamic neuropeptide gene expression

We tested the hypothesis that dietary whey protein isolate (WPI) affects the intestinal mechanisms related to energy absorption and that the resulting energy deficit is compensated by changes in energy balance to support growth. C57BL/6 mice were provided a diet enriched with WPI with varied sucrose content, and the impact on energy balance-related parameters was investigated. As part of a high-sucrose diet, WPI reduced the hypothalamic expression of pro-opiomelanocortin gene expression and increased energy intake. The energy expenditure was unaffected, but epididymal weight was reduced, indicating an energy loss. Notably, there was a reduction in the ileum gene expression for amino acid transporter SLC6a19, glucose transporter 2, and fatty acid transporter 4. The composition of the gut microbiota also changed, where Firmicutes were reduced. The above changes indicated reduced energy absorption through the intestine. We propose that this mobilized energy in the adipose tissue and caused hypothalamic changes that increased energy intake, acting to counteract the energy deficit arising in the intestine. Lowering the sucrose content in the WPI diet increased energy expenditure. This further reduced epididymal weight and plasma leptin, whereupon hypothalamic ghrelin gene expression and the intestinal weight were both increased. These data suggest that when the intestine-adipose-hypothalamic pathway is subjected to an additional energy loss (now in the adipose tissue), compensatory changes attempt to assimilate more energy. Notably, WPI and sucrose content interact to enable the component mechanisms of this pathway.

Pulmonary Pseudomonas aeruginosa infection induces autophagy and proteasome proteolytic pathways in skeletal muscles: effects of a pressurized whey protein-based diet in mice

ABSTRACTBackground: Pulmonary Pseudomonas aeruginosa infection in cystic fibrosis patients is associated with skeletal muscle atrophy. In this study, we investigated the effects of P. aeurginosa infection and a whey protein-rich diet on skeletal muscle proteolytic pathways.Design: An agar bead model of pulmonary P. aeurginosa infection was established in adult C57/Bl6 mice. Protein ubiquitinaiton, lipidation of LC3B protein and expressions of autophagy-related genes and ubiquitin E3 ligases were quantified using immunoblotting and qPCR. The effects of pressure-treated whey protein diet on muscle proteolysis were also evaluated.Results: Pulmonary P. aeurginosa infection reduced diaphragm, tibialis anterior, and soleus muscle weights and increased protein ubiquitination, LC3B protein lipidation, and the expressions of Lc3b, Gabarapl1, Bnip3, Parkin, Atrogin-1, and MuRF1 genes in each muscle. These changes were greater in the tibialis as compared to soleus and diaphragm. Proteolysis indicators increased within one day of infection but were not evident after seven days of infection. A pressurized whey diet attenuated LC3B protein lipidation, expressions of autophagy-related genes (BNIP3), pro-inflammatory cytokines, and protein ubiquitination.Conclusions: We conclude that pulmonary P. aeruginosa infection activates the autophagy, and the proteasome pathways in skeletal muscles and that a pressurized whey protein diet attenuates muscle proteolysis in this model.

Dietary whey reduces energy intake and alters hypothalamic gene expression in obese phyto-oestrogen-deprived male rats.

Removing dietary phyto-oestrogens in adult male rats causes obesity and diabetes. As whey proteins have been reported to reduce food intake and improve glucose homoeostasis, we investigated whether they could attenuate susceptibility to obesity and diabetes due to phyto-oestrogen deprivation. To this end, thirty male Wistar rats were fed a high-phyto-oestrogen (HP) or a phyto-oestrogen-free (PF) diet for 10 weeks; six rats from each group were killed. The remaining HP animals (six animals) continued receiving the HP diet for 6 weeks. The remaining PF rats (twelve rats) were divided in two groups: one was given the PF diet and the other a variation of the PF diet plus whey protein (PF-W). Body weight, food intake and adipose tissue weights were recorded. Hypothalamic mRNA expressions of orexigenic (neuropeptide Y, agouti-related protein (AgRP)) and anorexigenic (pro-opiomelanocortin (POMC), cocaine-amphetamine-related transcript (CART)) neuropeptides were quantified by real-time PCR. Serum glucose, insulin and total thyroxine (T4), thyroid-stimulating hormone, testosterone and oestradiol were assessed. After 10 weeks of PF diet, increased body weight, adiposity and energy intake, with up-regulation of AgRP and down-regulation of POMC', were observed. Longer treatment exacerbated these results, increased total T4 levels, reduced oestradiol levels and impaired glucose homoeostasis. PF-W reduced energy intake and increased POMC expression; however, body weight and adiposity remained unchanged. PF-W could not prevent the hormonal changes or the high circulating glucose levels induced by phyto-oestrogen deprivation, but reduced fasting insulin. These data demonstrate that, although 6 weeks of whey administration could not prevent obesity in phyto-oestrogen-deprived rats, the reduction in energy intake and circulating insulin could be beneficial with longer treatments.

Abstract 4324: Undenatured whey protein isolate exhibit chemopreventive activity against HPV-16 induced carcinogenesis of CD271+ oral mucosa stem cells

Abstract Oral cancer, and associated HPV-16 infection, and tobacco smoking is a major public health concern in the Kamrup district of India. To facilitate research on oral cancer chemoprevention, we have set up a collaborative research team between Forsyth Institute, an oral medicine specialized center affiliated to Harvard School of Dental Medicine, and KaviKrishna Laboratory, a non-for profit private laboratory set up located in Guwahati, the major city of Kamrup district, India. To study the cellular and molecular mechanism of HPV-16 mediated carcinogenesis, we have developed an in vitro model of HPV-16 mediated carcinogenesis of oral squamous stem cells. In this model (Bhuyan et al. The potential role of oral mucosa stem cell altruistic behavior as the initiating event of malignant transformation. AACR abstract Control # 16-A-6618-AACR), the treatment of oral mucosa cells derived from healthy volunteer with HPV-16 derived E6 protein led to expansion of a p53 deficient CD271+ expressing oral mucosa stem cells (OMSC). Noted that CD271 cell surface marker was recently identified as a putative marker for OMSCs. Using this in vitro carcinogenesis model, we evaluated the potential chemopreventive role of dietary whey protein. The E6 treated p53 deficient CD271+ cells were treated with DMEM/F12 media containing 0.2% of Immunocal, an undenatured whey protein (Tsai WY et al. Nutr Cancer 2000. PMID: 11525598). The p53 status as well as in vitro self-renewal activity of the CD271+ cells were examined by ELISA, transcriptional activity assay, and methylcellulose based clonogenic assay. We also fed Immunocal 20 gm/daily for 6 months to 5 individual with oral leukoplakia lesion with HPV-16 positivity. The CD271+ cells obtained from the leukoplakia lesion of clinical subjects were subjected to p53 status, and clonogenic assay. The data was compared with CD271+ cells obtained from leukoplakia lesion of subjects taking regular diet without whey protein supplements. We found that addition of Immunocal whey protein led to 8-fold decrease in the expansion of CD271+ cells following E6 treatment in the in vitro model of OMSC culture. Importantly, Immunocal treatment prevented the suppression of p53 in OMSCs. In the preliminary clinical study, the dietary intake of Immunocal led to complete loss of leukoplakia lesion in the 4/5 individual. In contrast, the control subjects exhibited the presence of CD271+ cells in the leukoplakia lesion having low p53 status, and high clonogenic activity. We are incorporating a larger number of subjects to study the potential chemopreventive activity of whey protein, using the locally available affordable whey protein extracts. Conclusion: This study indicate that whey protein extract, which is available in the local villages of Kamrup district could serve as a chemopreventive agent against oral cancer. Citation Format: Sora Sandhya, Rika Tsuchida, Sukanya Gayan, Rashmi Bhuyan, Joyeeta Talukdar, Bidisha Pal, Seema Bhuyan, Jugal Ch Das, Amal Ch Kataki, Debabrata Baishya, Bikul Das. Undenatured whey protein isolate exhibit chemopreventive activity against HPV-16 induced carcinogenesis of CD271+ oral mucosa stem cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4324.