Search

The protein complex of recombinant human insulin-like growth factor-1 and insulin‑like growth factor binding protein‑3 (rhIGF-1/rhIGFBP-3; mecasermin rinfabate), is an investigational product for the prevention of complications of prematurity. Delivery of rhIGF-1/rhIGFBP-3 is by continuous central line intravenous infusion in preterm infants until endogenous IGF-1 production begins. Protein-specific analytical methodologies were developed to evaluate the compatibility of rhIGF- 1/rhIGFBP-3 at low protein concentrations (∼2.5-10 μg/mL) expected when co-administered with other required medications in the NICU. Highly sensitive detection of the biologic potential degradants (fragments) and/or molecular modifications (oxidized species, aggregates) required the use of reversed-phase high-performance liquid chromatography and size-exclusion ultra-performance liquid chromatography coupled with mass spectrometric detection. We report on the quantification of rhIGF-1/rhIGFBP-3, its components and degradants, to a limit of quantitation of 3.1 μg/mL upon mixing with 24 commonly administered neonatal medications. Methods developed for the rhIGF-1/rhIGFBP-3 admixtures, optimized in studies with furosemide, caffeine citrate and ampicillin, demonstrated good reproducibility, linearity, and limit of detection/quantitation. Using these methods, no increase in degradation of rhIGF-1/rhIGFBP-3 components and no increase in oxidation or aggregation level was observed with caffeine citrate, while admixtures of rhIGF-1/rhIGFBP-3 with ampicillin yielded lower mass recovery of rhIGF-1/rhIGFBP-3 components, which likely resulted from adduct formation. Furosemide was found to be physically incompatible with rhIGF-1/rhIGFBP-3. Our findings support the use of these methodologies for detection of protein modifications under various clinical administration conditions, and additionally supplement physical compatibility data studies of ultra-low concentrations of rhIGF-1/rhIGFBP-3 post co-administration to preterm infants with other medications (manuscript in-preparation).

Amyloid fibrils were initially known for the implications in neurodegenerative diseases, such as Alzheimer’s and Parkinson’s diseases. In this context, the interaction between polyphenols and amyloid fibrils has attracted intensive interest in the journey of pursuing therapies to slow down the progress of amyloidosis-related diseases. It is generally concluded that polyphenols inhibit the formation of amyloid fibrils and remodel or cut the mature ones, which are observed in low protein concentrations. Unfolding and self-assembly of food-grade proteins result in amyloid fibrils. Our very recent studies demonstrated that polyphenols absorb and deposit on the surface of amyloid fibrils forming the supramolecular nanofilaments which self-assemble in multi-length scales to form hierarchical structures. This finding has created the food hybrid hydrogels with unique properties of incomparable high loading content and stability of polyphenols, target delivery to colon and re-construction of gut microbiota for health promotion. It paves ways for food molecular processing.

The cellular membrane has been identified to play a critical role in various biological processes including the assembly of biological systems. Membranes are complex, primarily two-dimensional assemblies with varied lipid compositions depending on the particular region of the cell. Supported lipid bilayers are considered as appropriate models for physio-chemical studies of membranes including numerous single molecule techniques. Atomic force microscopy (AFM) as a topographic technique is a fully appropriate single molecule technique capable of direct observation of molecular processes on membranes. However, reliable experimental AFM studies require the preparation of the bilayer with a sub-nanometer smooth morphology, which remains stable over long-time observation. Here we present the methodology, which allows one to prepare a smooth, stable, structurally homogeneous lipid bilayer without the presence of any trapped vesicles. We described the application of such lipid bilayers to probe time-dependent early stages of aggregation of monomeric amyloid proteins. Importantly, the proposed methodology can be extended to bilayers with various compositions, by incorporating different lipids for on-membrane aggregation study including cholesterol. Furthermore, this methodology development allowed us to monitor the aggregation of amyloid protein at its physiologically relevant low protein concentration. The flexibility of altering the membrane composition allows to identify the specific role of a particular lipid towards the aggregation kinetics, revealing the plausible mechanism of disease development.

High-solid anaerobic digestion of the very small particle fraction of mechanically-sorted organic fraction of municipal solid waste (OFMSW) was examined in mesophilic digestion tests in a conventional laboratory (0.013 m3) and a pilot (0.300 m3) reactor. The non-biodegradable and recalcitrant molecules together with the low protein and starch contents of the small-particles of OFMSW limited the methane generation potential of substrate. In the conventional AD system, methane yields remained low at 0.139 m3kgVS-1 due to formation of a non-reacting layer on digestate surface, which restricted utilization of the available in OFMSW digestible organics. The absence of surface solid crust in the pilot unit favoured consumption of a greater proportion of volatile solids of the OFMSW. Dry AD was remarkably stable over the entire period and negligibly effected by the toxic H2S yields. Methane generation (0.167 m3kgVS-1) was increased 1.2-fold compared to the conventional system due to a better mixing of substrate and microorganisms achieved inside the pilot reactor, which led to an increase of the digested volatile organics. Digestate presented low stability and high heavy metal content, both of which restrain its implementation as soil conditioner or fertilizer in agriculture. A secondary co-digestion treatment may be required for the neutralization of digestate.

Purpose Phenylalanine hydroxylase deficiency, or phenylketonuria (PKU), is a rare autosomal recessive metabolic disorder. Early diagnosis via newborn screening (NBS) and initiation of treatment prevent the development of cognitive impairment and other co-morbidities. The purpose of this study is to describe the natural history of PKU in the United States, including prevalence of co-morbidities and predictors of outcomes. Methods We analyzed data from a self-report survey in the NBS-PKU Connect online registry. We describe the participants' nutrition management strategies, barriers to management, outcomes of bone disorders, skin, and psychological co-morbidities, and the use of special education or other special services. Predictors of outcomes were identified and assessed, including the impact of sex, age, age at diagnosis, blood phenylalanine concentration, use of sapropterin, use of medical food, adherence to prescribed diet, use of low protein modified foods, whether they had ever been off-diet, and use of tyrosine supplementation. Results The 219 respondents included individuals with PKU or hyperphenylalanemia (n = 78), or their caregivers (n = 141). Most (84.3%) started treatment before the age of two weeks. About one-third indicated that they had been off-diet at some point in their lives, and 81.4% reported that they currently adhered to their prescribed diet, with adherence to prescribed diet decreasing with age. Blood phenylalanine concentration was under the recommended threshold of 360 μmol/L for 68.5% of participants. One-quarter of respondents reported psychological co-morbidities, with anxiety and ADD/ADHD being the most common. The incidence of psychological co-morbidities increased with age and with ever having been off diet. Special education or other special services were more likely to be reported by individuals who were diagnosed after one week of age. Skin disorders such as acne and eczema were more common in females than males, and a minority of participants reported bone disorders. Conclusions Despite recommendations to maintain blood phenylalanine concentrations in the therapeutic range throughout life, it is not uncommon for adults with PKU to discontinue dietary management of their disorder. Early diagnosis was associated with reduced need for special education or other special services, and continuous treatment was associated with decreased psychological co-morbidities.

More food waste (FW) is desired to be treated in a certain processing period, while the degradation pattern of biochemical fractions during FW bioevaporation was significantly influenced by the organic loading (OL). Lower OL facilitated the lipids degradation, while higher OL favored the protein degradation. It was the more porous structure and abundant oxygen accelerated the lipids degradation, and the rapid proliferation of aerobic microorganisms compensated for the low protein degradation in lower OL. Detailly, 76.8% of the lipids was degraded in the trial with OL of 1.04 kg VSFW/kg TSBS (Trial A), but in the trial with OL of 3.16 kg VSFW/kg TSBS (Trial C) it was only 0.5%. For protein, the degradation was different that 17.5% of the protein was degraded in Trial A, whereas 69.1% was degraded in Trial C. Lipids degradation contributed 63.0% to the metabolic heat in Trial A, but its contribution in Trial C was only 0.5%. For protein, it contributed 4.1% to the metabolic heat in Trial A, but in Trial C it accounted for 53.6%. In addition, the degradation of carbohydrates (71.6-80.8%) and their contribution to metabolic heat (32.8-45.9%) were comparable in all trials, thus OL had little effect on carbohydrates degradation. Results from this study could provide important guideline for FW practical disposal during their biological treatment.

Glutamic acid (Glu) and aspartic acid (Asp) are acidic amino acids with regulatory roles in nutrition, energy metabolism, and oxidative stress. This study aimed to evaluate the effects of low-protein diets supplemented with Glu and Asp on the intestinal barrier function and energy metabolism in weaned piglets challenged with hydrogen peroxide (H2O2). Forty piglets were randomly divided into 5 groups: NC, PC, PGA, PG, and PA (n = 8 for each group). Pigs in the NC and PC groups were fed a low-protein diet, while pigs in the PGA, PG, or PA groups were fed the low-protein diet supplemented with 2.0% Glu +1.0% Asp, 2.0% Glu, or 1.0% Asp, respectively. On day 8 and 11, pigs in the NC group were intraperitoneally injected with saline (1 mL/kg BW), while pigs in the other groups were intraperitoneally administered 10% H2O2 (1 mL/kg BW). On day 14, all pigs were sacrificed to collect jejunum and ileum following the blood sample collection in the morning. Notably, low-protein diets supplemented with Glu or Asp ameliorated the intestinal oxidative stress response in H2O2-challenged piglets by decreasing intestinal expression of genes (P < 0.05) (e.g., manganese superoxide dismutase [MnSOD], glutathione peroxidase [Gpx]-1, and Gpx-4) encoding oxidative stress-associated proteins, reducing the serum concentration of diamine oxidase (P

INTRODUCTION: Protein energy wasting (PEW) is the most important risk factor for morbidity and mortality in hemodialysis patients. Inadequate dietary protein intake is a frequent cause of PEW. Recent studies have identified fibroblast growth factor 21 (FGF21) as an endocrine protein sensor. This study aims to investigate the potential of FGF21 as a biomarker for protein intake and PEW and to investigate intradialytic FGF21 changes.METHODS: Plasma FGF21 was measured using an enzyme-linked immunoassay. Complete intradialytic dialysate and interdialytic urinary collections were used to calculate 24-h urea excretion and protein intake. Muscle mass was assessed using the creatinine excretion rate and fatigue was assessed using the Short Form 36 and the Checklist Individual Strength.RESULTS: Out of 59 hemodialysis patients (65 ± 15 years, 63% male), 39 patients had a low protein intake, defined as a protein intake less than 0.9 g/kg/24-h. Patients with a low protein intake had nearly twofold higher plasma FGF21 compared to those with an adequate protein intake (FGF21 1370 [795-4034] pg/mL versus 709 [405-1077] pg/mL;P < 0.001). Higher plasma FGF21 was associated with higher odds of low protein intake (Odds Ratio: 3.18 [1.62-7.95] per doubling of FGF21; P = 0.004), independent of potential confounders. Higher plasma FGF21 was also associated with lower muscle mass (std β: -0.34 [-0.59;-0.09];P = 0.009), lower vitality (std β: -0.30 [-0.55;-0.05];P = 0.02), and more fatigue (std β: 0.32 [0.07;0.57];P = 0.01). During hemodialysis plasma FGF21 increased by 354 [71-570] pg/mL, corresponding to a 29% increase.CONCLUSION: Higher plasma FGF21 is associated with higher odds of low protein intake in hemodialysis patients. Secondarily, plasma FGF21 is also associated with lower muscle mass, less vitality, and more fatigue. Lastly, there is an intradialytic increase in plasma FGF21. FGF21 could be a valuable marker allowing for objective assessment of PEW.

Background and aims It has been demonstrated that maternal low protein during development induces mitochondrial dysfunction and oxidative stress in the heart. Moderate-intensity exercise in early life, conversely, increases the overall cardiac health. Thus, we hypothesize that moderate-intensity exercise performed during young age could ameliorate the deleterious effect of maternal protein deprivation on cardiac bioenergetics. Methods and results We used a rat model of maternal protein restriction during gestational and lactation period followed by an offspring treadmill moderate physical training. Pregnant rats were divided into two groups: normal nutrition receiving 17% of casein in the diet and undernutrition receiving a low-protein diet (8% casein). At 30 days of age, the male offspring were further subdivided into sedentary (NS and LS) or exercised (NT and LT) groups. Treadmill exercise was performed as follows: 4 weeks, 5 days/week, 60 min/day at 50% of maximal running capacity. Our results showed that a low-protein diet decreases oxidative metabolism and mitochondrial function associated with higher oxidative stress. In contrast, exercise rescues mitochondrial capacity and promotes a cellular resilience to oxidative stress. Up-regulation of cardiac sirtuin 1 and 3 decreased acetylation levels, redeeming from the deleterious effect of protein restriction. Conclusion Our findings show that moderate daily exercise during a young age acts as a therapeutical intervention opposing the harmful effects of a maternal diet restricted in protein.

Sterile filtration is an integral step in the manufacturing process of biological therapeutics. Protein adsorption to the surface of the filter is an unfortunate, common occurrence that can result in manufacturing difficulties, such as filter fouling or product loss. Although many filters have surface modifications to minimize adsorption, under certain conditions binding can still occur. We observed the loss of high molecular weight species (HMWS) during sterile filtration of eight different therapeutic monoclonal antibodies formulated at low protein concentrations across a commonly used hydrophilic polyvinylidene fluoride or polyvinylidene difluoride (PVDF) filter membrane. The protein absorption was specific to HMWS, and each antibody exhibited different degrees of filter adsorption. Debye screening length parameters of the solution (e.g. ionic strength) were adjusted, and influenced the amount of HMWS lost during filtration. Additionally, HMWS of a representative antibody (mAb1) were observed to be more positively charged than other size variants by ion-exchange chromatography. From these results, it is concluded that this HMWS loss is due to electrostatic interactions between HMWS and the filter surface. This adsorption can be reduced by increasing the ionic strength of the buffer matrix, demonstrating the influence of the Debye screening length in the filtration of low concentration proteins.

Developing ultra-high concentration biotherapeutics drug products can be challenging due to increased viscosity, processing, and stability issues. Excipients used to alleviate these concerns are traditionally evaluated at lower protein concentrations. This study investigates whether classically known modulators of stability and viscosity at low (50 mg/mL) to high (50 - 150 mg/mL) protein concentrations are beneficial in ultra-high (150 mg/mL) concentration protein formulations and drug products. This study evaluates the effect of arginine monohydrochloride, proline, and lysine monohydrochloride on viscosity and concentratability at different high and ultra-high protein concentrations using a monoclonal antibody, mAbN, formulation as a candidate protein system. The effect of excipients on the viscosity and concentratability (rate and extent) was different at high versus ultra-high protein concentrations. These results highlight that classical excipients in literature known to modulate protein interactions at low protein concentrations and reduce viscosity at high protein concentrations may need to be evaluated at target protein concentrations in a product-specific manner while developing ultra-high concentration biologics drug products.

Amino acids (AA) are the second most expensive component of broiler diets after energy, and therefore their excessive inclusion in diets can result in significant increase in broiler feed cost. In addition to the increased feed cost, excessive intake of AA by broilers results in increased N excretion, which has negative impact on environment, and welfare of birds and caretakers. Also, excessive intake of AA by broilers results in increased excretion of water and hence increased litter moisture, which has negative impact on environment and welfare of birds. Finally, excessive intake of AA can result in increased production of toxic compounds in hindgut from AA post small intestinal digestion. Thus, methods to reduce dietary CP level without compromising performance of broilers will reduce feed costs and improve the environment, and may improve poultry health and welfare. The dietary level of CP can be reduced without compromising performance of broilers if the resulting low-CP diets are supplemented with crystalline AA to meet the AA requirements of broilers. By supplementing the diets with crystalline AA, the CP values in broiler starter, in this paper any period between hatch and approximately days 21 of age and finisher, in this paper any period between day 21 and day 42 of age, diets can be reduced from approximately 220 and 200 g/kg to 190 and 170 g/kg, respectively, without compromising growth performance of the broilers provided the digestible Lys, Met, Thr, and Val contents in low-CP diets are similar to those in the high-CP diets. The level of CP in broiler starter and finisher diets can be further reduced to 180 g/kg and 160 g/kg, respectively, without compromising growth performance of the broilers if the low-CP diets in addition are supplemented by Gly and the digestible content of Lys, Met, Thr and Val, and Gly are equivalent of the low-CP diets to those in the high-CP diets. Publications delineated in this text indicate several strategies to consider for reducing CP in broiler diets including: feed enzymes, soybean isoflavones, fatty acid type, and fiber type. All these strategies are discussed and gaps in knowledge are identified.

BACKGROUND: Recently, we showed that there are higher protein, lysine, and phenylalanine requirements in late stages of pregnancy compared with early stages. Animal studies have suggested an increased dietary need for specific dispensable amino acids in pregnancy; whether such a need exists in human pregnancies is unknown. OBJECTIVE: The objective of the current study was to examine whether healthy pregnant women at midgestation (20–29 wk) and late gestation (30–40 wk) have a dietary demand for glycine, a dispensable amino acid, using the indicator amino acid oxidation method and measurement of plasma 5-oxoproline concentrations. METHODS: Seventeen healthy women (aged 26–36 y) randomly received different test glycine intakes (range: 5–100 mg·kg(−1)·d(−1)) during each study day in midgestation (∼26 wk, n = 17 observations in 9 women) and late gestation (∼35 wk, n = 19 observations in 8 women). Diets were isocaloric with energy at 1.7 × resting energy expenditure. Protein was given as a crystalline amino acid mixture based on egg protein composition at current estimated average requirement (EAR; 0.88 g·kg(−1)·d(−1)). Breath samples were collected at baseline and isotopic steady state to measure oxidation of L-[1–(13)C]phenylalanine to (13)CO(2) (F(13)CO(2)). Plasma was collected at the sixth hour of the study day. Linear regression crossover analysis and simple linear regression were used to assess responses in F(13)CO(2) and plasma 5-oxoproline concentrations to different glycine intakes. RESULTS: No statistically significant responses were observed in midgestation. However, in late gestation, lower glycine intakes resulted in higher rates of F(13)CO(2) (suggesting low protein synthesis) with a breakpoint for phenylalanine oxidation at >37 mg glycine·kg(−1)·d(−1) and higher plasma 5-oxoproline (suggesting low glycine availability) with a breakpoint >27 mg glycine·kg(−1)·d(−1). CONCLUSIONS: The findings suggest that glycine should be considered a “conditionally” indispensable amino acid during late gestation, especially when protein intakes are at 0.88 g·kg(−1)·d(−1), the current EAR. This trial was registered at clinicaltrials.gov as NCT02149953.

A total of 945 male Ross 308 broiler chicks were used in a growth study to explore the interaction between dietary crude protein concentration and available phosphorus. Nine experimental treatments were constructed factorially by offering low, medium, or standard protein concentrations without or with low, standard, or high available phosphorus. Diets were based on corn, wheat, and soybean meal and all nutrients other than protein/amino acids and available phosphorus were maintained at or above breeder guidelines. Additional synthetic amino acids were used in the diets with low protein concentration in attempt to maintain digestible amino acid supply. Diets were offered to 7 replicate pens of 15 chicks per pen from day 8 to 35. Growth performance was measured during the grower (day 8–24) and finisher (day 25–35) periods. On day 35 carcass composition was determined, blood was drawn for various biochemical measurements and the tibia was excised for mechanical and compositional analyses. Birds that received the low-protein diet had lower terminal body weight and higher feed conversion ratio compared with those that received diets with adequate crude protein content. However, addition of available phosphorus to the low-protein diet resulted in significant reductions in weight-corrected feed conversion that were not evident in the diet with adequate protein content. Bone architecture was only moderately influenced by dietary treatment but birds that ingested the diets containing low and medium protein concentrations had relatively heavier abdominal fat pad weight. Blood biochemistry, especially ammonia, uric acid, and phosphorus, was influenced by both dietary protein and available phosphorus and trends suggested that both axes are involved in protein accretion and catabolism. It can be concluded that performance losses associated with feeding low protein diets to broiler chickens may be partially restored by additional available phosphorus. The implications for use of exogenous enzymes such as protease and phytase and protein nutrition per se warrants further examination.

Four experiments were conducted to estimate the optimal standardized ileal digestible (SID) level of branched-chain amino acids in low-protein diets during the starter, grower, and finisher periods, using the response surface methodology, and to study their effects on performance and mRNA expression of genes involved in the mechanistic target of rapamycin (mTOR) pathway of broiler chickens from 8 to 21 D of age. In experiments 1, 2, and 3, a total of 1,500 Cobb male broiler chickens were assigned to 15 diets of a central composite rotatable design (CCD) of response surface methodology containing 5 levels of SID Leu, Val, and Ile with 5 replicate pens of 20 birds each. A 3-factor, 5-level CCD platform was used to fit the second-order polynomial equation of broiler performance. In experiment 4, a total of 540 8-day-old Cobb male broiler chickens were distributed in a completely randomized 2 x 3 x 3 factorial arrangement with 2 SID Leu levels (1.28 or 1.83%), 3 SID Val levels (0.65, 0.90, or 1.20%), and 3 SID Ile levels (0.54, 0.79, or 1.09%) for a total of 18 treatments with 5 replicate cages of 6 birds each. High Leu levels impaired (P

Summary Background & aims Sarcopenia, or age-related muscle loss, is an enormous health problem in an aging world because of its many clinical and societal adverse effects. The uncovering of healthy dietary patterns is an important strategy to prevent or delay sarcopenia. We used K-means clustering to identify subgroups of men and women based on nutritional and health-related factors and investigated risk factors for low muscle mass in the subgroups and in the study population as a whole. Methods We analyzed a total 10,863 participants over 40 years of age who participated in the Korea National Health and Nutrition Survey from 2008 to 2011. Dual energy X-ray absorptiometry was used to determine the appendicular lean mass (ALM) of the participants. Participants with low ALM adjusted BMI (ALM/BMI) were then identified using the criteria of the Foundation for the National Institutes of Health sarcopenia project. K-means clustering and multivariate logistic regression were used to analyze associations between nutritional and health-related variables and low ALM/BMI in the population as a whole and in the individual clusters. Results A total of 712 (15.8%) men and 869 (13.7%) women had low ALM/BMI. Five clusters were identified in men and women, respectively. Two clusters of men and one cluster of women exhibited an increased risk of low ALM/BMI. Old age, low total energy intake, low levels of physical activity, and a high number of chronic diseases were consistent risk factors for low ALM/BMI in all Korean men and women. Low protein was a common risk factor for low ALM/BMI in men. After dividing all subjects by the K-means clustering algorithm, two risk factors (high fat intake and smoking) and four factors (low intakes of carbohydrate, protein and fat, and high alcohol consumption) were additionally proposed in Korean men and women, respectively. Conclusions Age, low total energy intake, low level of physical activity, and an increased number of chronic diseases were consistent risk factors for low ALM/BMI in men and women. Cluster-specific risk factors were also noted in men and women.

A 56 days feeding trial was conducted to investigate the effects of enzyme-treated soy protein (ETSP) supplementation in low protein diets on immune function of immune organs (head kidney, spleen and skin) in on-growing grass carp. A total of 540 on-growing grass carp (initial average weight: 325.72 ± 0.60 g) were fed six diets, which included a normal protein diet (28% crude protein) and five low protein diets (26% crude protein) supplemented with graded levels of ETSP (0.0, 0.8, 1.2, 1.6 and 2.0%). At the end of feeding period, a challenge test was performed by infection with Aeromonas hydrophila for two weeks. The results indicated that (1) reducing dietary protein content from 28 to 26% decreased antibacterial substances and aggravated inflammatory responses of above three immune organs; (2) under the condition of reducing protein level in diet, 0.8–1.2% ETSP supplementation reversed these above adverse effects on immune function of above three immune organs; (3) suitable ETSP supplementation-decreased inflammatory responses were partly associated with [IκB kinase β (IKKβ)/inhibitor of κBα (IκBα)/nuclear factor kappa B (NF-κB) p65 and p52 or NF-κB p65] signaling and [target of rapamycin (TOR)/(S6K1, 4E-BP)] signaling in above three immune organs. (4) On the basis of C3 content (head kidney), C4 content (spleen) and skin hemorrhage and lesion, the optimal ETSP supplementation levels in low protein diets were estimated to be 1.48%, 1.61% and 1.03%, respectively. In summary, ETSP supplementation in low protein diets improved immune function of head kidney, spleen and skin in on-growing grass carp.

Originally, the kwashiorkor is a pathology justified by the low consumption of proteins and high carbohydrates in weaned children. However, today, it can appear due to multifactorial causes, one of the hypotheses being the presence of aflatoxins in foods consumed by the child population and detected in biological fluids. The objective of this work is to scoping review the presence of aflatoxins in kwashiorkor, marasmus and marasmic-kwashiorkor from organs and biological samples in children. Results reflected that the presence of aflatoxins in kwashiorkor is greater compared to marasmic-kwashiorkor and marasmus in the organs and biological samples analyzed. The relationship of this mycotoxin with the pathology shows that it can affect both genders, even up to 12 years, in addition they are detected in eight biological samples and organs, except in the spleen, and in ten African countries and in the Philippines. The appearance of this pathology has been associated in children when after weaning they consume foods with low protein content and rich in carbohydrates, but coincidentally coincides with foods where the growth of aflatoxigenic fungi is more prevalent, and even the presence of other fungi that can generate other mycotoxins, such as ochratoxin A and fumonisin B1.

Background: A high protein intake in early life is associated with a risk of obesity later in life. The essential amino acid requirements of formula-fed infants have been reassessed recently, enabling a reduction in total protein content and thus in protein intake. Objectives: We aimed to assess the safety of an infant formula with a modified amino acid profile and a modified low-protein (mLP) content in healthy term-born infants. Outcomes were compared with a specifically designed control (CTRL) infant formula. Methods: In this double-blind, randomized controlled equivalence trial, infants received either mLP (1.7 g protein/100 kcal; n = 90) or CTRL formula (2.1 g protein/100 kcal; n = 88) from enrollment (age ≤ 45 d) to 6 mo of age. A breastfed group served as a reference (n = 67). Anthropometry and body composition were determined at baseline, 17 wk (including safety blood parameters), and 6 mo of age. The primary outcome was daily weight gain from enrollment up until the age of 17 wk (at an equivalence margin of ±3.0 g/d). Results: Weight gain from baseline (mean ± SD age: 31 ± 9 d) up to the age of 17 wk was equivalent between the mLP and CTRL formula groups (27.9 and 28.8 g/d, respectively; difference:-0.86 g/d; 90% CI:-2.36, 0.63 g/d). No differences in other growth parameters, body composition, or in adverse events were observed. Urea was significantly lower in the mLP formula group than in the CTRL formula group (-0.74 mmol/L; 95% CI:-0.97,-0.51 mmol/L; P < 0.001). Growth rates, fat mass, fat-free mass, and several essential amino acids were significantly higher in both formula groups than in the breastfed reference group. Conclusions: Feeding an infant formula with a modified amino acid profile and a lower protein content from an average age of 1 mo until the age of 6 mo is safe and supports an adequate growth, similar to that of infants consuming CTRL formula. This trial was registered at www.trialregister.nl as Trial NL4677.

Early life adversities (stress, infection and mal/undernutrition) can affect neurocognitive, hippocampal and immunological functioning of the brain throughout life. Substantial evidence suggests that maternal protein malnutrition contributes to the progression of neurocognitive abnormalities and psychopathologies in adolescence and adulthood in offspring. Maternal malnutrition is prevalent in low and middle resource populations. The present study was therefore undertaken to evaluate the effects of dietary Spirulina supplementation of protein malnourished mothers during pregnancy and lactation on their offspring’s reflex, neurobehavioral and cognitive development. Spirulina is a Cyanobacterium and a major source of protein and is being used extensively as a dynamic nutraceutical against aging and neurodegeneration. Sprague Dawley rats were switched to low protein (8% protein) or normal protein (20% protein) diet for 15 days before conception. Spirulina was orally administered (400 mg/kg/b.wt.) to subgroups of pregnant females from the day of conception throughout the lactational period. We examined several parameters including reproductive performance of dams, physical development, postnatal reflex ontogeny, locomotor behavior, neuromuscular strength, anxiety, anhedonic behavior, cognitive abilities and microglia populations in the F1 progeny. The study showed improved reproductive performance of Spirulina supplemented protein malnourished dams, accelerated acquisition of neurological reflexes, better physical appearance, enhanced neuromuscular strength, improved spatial learning and memory and partly normalized PMN induced hyperactivity, anxiolytic and anhedonic behavior in offspring. These beneficial effects of Spirulina consumption were also accompanied by reduced microglial activation which might assist in restoring the behavioral and cognitive skills in protein malnourished F1 rats. Maternal Spirulina supplementation is therefore proposed as an economical nutraceutical/supplement to combat malnutrition associated behavioral and cognitive deficits.

BACKGROUND Rare coding mutations underlying deficiencies of antithrombin and proteins C and S contribute to familial venous thromboembolism (VTE). It is uncertain whether rare variants play a role in the etiology of VTE in the general population. OBJECTIVES We conducted a deep whole-exome sequencing (WES) study to investigate the associations between rare coding variants and the risk of VTE in two population-based prospective cohorts. PATIENTS/METHODS Whole-exome sequencing was performed in the Longitudinal Investigation of Thromboembolism Etiology (LITE), which combines the Atherosclerosis Risk in Communities (ARIC) study (316 incident VTE events among 3159 African Americans [AAs] and 458 incident VTEs among 7772 European Americans [EAs]) and the Cardiovascular Healthy Study (CHS; 60 incident VTEs among 1751 EAs). We performed gene-based tests of rare variants (allele frequency

The present study investigated the effects of supplementing a low protein (LP) diet supplemented with key essential amino acids (AA) to broilers on growth performance, intestinal tract function, blood metabolites, and nitrogen excretion when the animals were maintained under various sanitary conditions for 35 D after hatching. Three hundred eighty-four one-day-old male broilers (Ross 308) were randomly allotted to groups that received one of 6 dietary treatments in a 2 × 3 factorial arrangement (i.e., 2 environmental conditions and 3 dietary treatments) to give 8 replicates per treatment. Broilers were challenged with 2 environmental conditions (sanitary vs. poor sanitary). The dietary treatments were (1) high protein (HP) diet, (2) LP diet, and (3) LP diet with synthetic key essential AA (LPA): the LP diet was supplemented with synthetic AA up to the required levels for broilers. On day 14, birds consumed the LP diet impaired growth performance compared with those fed the HP diet, while the average daily weight gain-to-feed conversion ratio of birds fed the LPA diet improved to the level of birds fed the HP diet under poor sanitary conditions (P < 0.05). Broilers raised under poor sanitary conditions and fed the LP diet displayed higher (P < 0.05) zonula occludens (ZO-1) expression on day 14 than broilers fed either the HP or LPA diet. Under sanitary conditions, birds fed HP and LPA diets showed higher villus height and crypt depth compared with those of broilers fed the LP diet on day 35. Moreover, broilers raised in the poor sanitary environment had higher (P < 0.05) serum endotoxins than those raised in the sanitary environment. Broilers fed the LPA diet showed reduced (P < 0.05) nitrogen excretion on days 14 and 35 compared with those fed the LP and HP diets independent of the environment. In conclusion, the LPA diet did not impair growth performance under poor sanitary conditions for 14 D after hatch while resulting in lower nitrogen excretion in any environment conditions throughout the experiment.

The native reversible self-association of monoclonal antibodies has been associated with high viscosity, liquid-liquid, and liquid-solid phase separation. We investigated the native reversible self-association of an IgG1, which exerts this association even at low protein concentrations, in detail to gain further understanding of this phenomenon by extensive characterization of the association as a function of multiple factors, namely pH, temperature, salt concentration, and protein concentration. The nature of the self-association of the full-length IgG1 as well as the corresponding Fab and Fc fragment was studied by viz. size exclusion chromatography combined with multiangle light scattering, batch dynamic and static light scattering, analytical ultracentrifugation, small angle X-ray scattering, asymmetric flow field flow fractionation coupled with multiangle light scattering, and intrinsic fluorescence. We rationalized the self-association as a combination of hydrophobic and electrostatic interactions driven by the Fab fragments. Finally, we investigated the long-term stability of the IgG1 molecule.

Reducing dietary protein has been of interest to the global poultry industry to improve bird health, welfare, and industry sustainability. Low protein (LP) diets are typically glycine (Gly) deficient and produce poor performance. Supplementing the diet with Gly or precursors of Gly can overcome this deficiency. A feeding experiment was conducted with 330 Ross 308 off-sex males across 5 treatments in a randomized design using 11 birds per pen replicated 6 times. Grower and finisher treatments were fed from day 7 to 21 and day 21 to 35, respectively. The objective was to test the efficacy of supplementation with Gly and Gly equivalents (Glyequiv), serine (Ser) and threonine (Thr), in plant-based LP diets on bird performance against a standard protein (SP) diet containing meat and bone meal. Glycine, Ser, or Thr were supplemented on Glyequiv basis to an approximately 3% lower CP diet to achieve the same digestible Gly and Ser level as the SP diet. Nitrogen efficiency, serum uric acid, blood plasma amino acids (AA) and AA digestibility were also investigated to monitor potential metabolic effects. Birds fed the LP diet were only 3.3% lower in final body weight than the SP treatment (2,556 vs. 2,641 g) while the supplementation of Gly or Ser had no effect. Supplementation of Thr reduced final body weight by 9.5% (P < 0.05). Reducing CP increased N efficiency by 9.6% (P < 0.05) and decreased blood serum uric acid by 26.9% (P < 0.001) in the finisher treatments. Glycine and Ser supplementation in LP diets had no effect on these parameters. The LP diet reduced AA digestibility and blood plasma AA while the supplementation with either Gly, Ser, or Thr increased overall AA digestibility (P < 0.05) but had no overall effect on blood plasma AA. Further research is required into Gly metabolism; however, Thr supplementation depressed growth and therefore is not feasible to cover Gly deficiency in LP diets on a Glyequiv basis.

The current trial was conducted to study effects of dietary valine (Val) supplementation in low crude protein (CP) diets on performance, gut development and immune responses in broiler chickens. A total of 540 day-old broiler chicks (Ross 308) were assigned to 6 dietary treatments, comprising 6 replicates of 15 birds based on a factorial arrangement of treatments (2 × 3) in a completely randomized design during 1–12, 12–24 and 24–42 days of experiment. Dietary treatments included 3 diets with 100% (CP100; starter:217 g/kg, grower: 200 g/kg, finisher: 180 g/kg), 90% (CP90; starter: 195.3 g/kg, grower: 180 g/kg, finisher: 162 g/kg) and 85% (CP85; starter: 184.5 g/kg, grower: 170 g/kg, finisher: 153 g/kg) of dietary crude protein requirement of Ross 308 broiler chickens each supplemented with or without Val. Valine was supplemented to CP85 and CP90 diets in order to fulfill Val requirement of Ross 308 broiler chickens in which CP85 contained 9 g/kg digestible Val in starter, 7.9 g/kg digestible Val in grower and 7.1 g/kg digestible Val in finisher periods and CP90 contained 9 g/kg digestible Val in starter, 7.9 g/kg digestible Val in grower and 7.1 g/kg digestible Val in finisher periods. However, Val supplemented to CP100 with equal magnitude supplemented to CP90 in order to offer chickens with Val beyond their requirement. (starter: 9.9 g/kg digestible Val; grower: 8.7 g/kg digestible Val; finisher: 7.9 g/kg digestible Val). Supplemental Val could restore the compromising effect of CP85 and CP90 on growth of broilers during starter and growing phases equal to broilers received CP100 (P

Cyclic pentapeptides containing the amino acid sequence arginine-glycine-aspartic (RGD) have been widely applied to target αvβ3 integrin, which is upregulated in various tumors during tumor-induced angiogenesis. Multimeric cyclic RGD peptides have been reported to be advantageous over monomeric counterparts for angiogenesis imaging. Here, we prepared mono-, di-, and trimeric cyclic arginine-glycine-aspartic-D-phenylalanine-lysine (c (RGDfK)) derivatives by conjugation with the natural chelator fusarinine C (FSC) using click chemistry based on copper (I)-catalyzed azide-alkyne cycloaddition (CuAAC). The αvβ3 binding properties of 68Ga-labeled mono-, di-, and trimeric c(RGDfK) peptides were evaluated in vitro as well as in vivo and compared with the references monomeric [68Ga]GaNODAGA-c(RGDfK) and trimeric [68Ga]GaFSC(suc-c(RGDfK))3. All 68Ga-labeled c(RGDfK) peptides displayed hydrophilicity (logD = −2.96 to −3.80), low protein binding and were stable in phosphate buffered-saline (PBS) and serum up to 2 h. In vitro internalization assays with human melanoma M21 (αvβ3-positive) and M21-L (αvβ3-negative) cell lines showed specific uptake of all derivatives and increased in the series: mono- In conclusion, we developed a novel strategy for direct, straight forward preparation of mono-, di-, and trimeric c(RGDfK) conjugates based on the FSC scaffold. Interestingly, the best αvβ3 imaging properties were found for the dimeric [68Ga]GaMAFC(c(RGDfK)aza)2.

Soybean (Glycine max [L.] Merr.) is an important source of human dietary protein and vegetable oil. A strong negative correlation between protein and oil contents has hindered efforts to improve soybean seed quality. The metabolic and genetic bases of soybean seed composition remain elusive. We evaluated metabolic diversity in a soybean near-isogenic line (NIL) population derived from parents (JD12 and CMSD) with contrasting seed oil contents. Using GC-TOF/MS, we compared seed primary metabolites of high protein/low oil lines, low protein/high oil lines, and their parents. Principal-components analysis showed that metabolic profiles of all progeny lines could be discriminated based on protein and oil contents. Univariate analysis revealed wide variation and transgressive segregation of metabolites in the population. Twenty-eight annotated metabolites, in particular free asparagine, free 3-cyanoalanine, and L-malic acid, were correlated with seed protein content or seed oil content or seed protein and oil content. These results shed light on the metabolic and genetic basis of soybean seed composition. Keywords: Metabolomics, Seed composition, Protein, Oil, Near-isogenic population

The objective of this study is to explore the effect of high-resistant starch (RS), low-protein flour as a source of RS on patients with early type 2 diabetic nephropathy (DN) through the clinical intervention trial.This was a single center, randomized, comparative, open-label trial. Seventy-five patients with early DN, aged 18 to 80 y, were recruited and randomly assigned to two groups. During the 12-week intervention, the control group patients (38 cases) followed protein-restriction diet daily with a common staple. The intervention group (37 cases) received 50 g of high-RS, low-protein flour instead of a common staple of equal quality at lunch and dinner each day. The blood glucose, blood lipids, nutritional parameters, indicators of renal function, oxidative stress, and inflammatory markers were measured.Compared with the control group, high-RS, low-protein flour intake led to a significant reduction in fasting blood glucose, HbA1c, total cholesterol, and triglycerides levels (P .05 for all). The changes in serum uric acid (UA) and β2-microglobulin (β2-MG) level were observed after high-RS, low-protein flour intervention (uric acid [mean ± standard deviation]: -24.7 ± 38.5 μmol/L, P = .001; β2-MG: 0.5 ± 0.9 mg/L, P = 0.018). In addition, high-RS, low-protein flour intake increased serum superoxide dismutase level by 10.1 ± 27.7 U/mL (P .05); however, it did not change the interleukin-6 and Tumor Necrosis Factor α (TNF-α) concentration.Twelve-week intervention with high-RS, low-protein flour improved the blood glucose and blood lipid levels, decreased the serum uric acid (UA) and urine β2-MG, and enhanced the ability to prevent antioxidative stress in patients with early DN.

The rice industry requires new technologies to increase production in response to world population growth. These include technologies that ensure rice quality in postharvest processing. Protein content defines rice quality and palatability. Differences in kernel thickness and maturity affect the protein level of harvested rice. We therefore analysed the relationship between kernel thickness and maturity on protein content of Japonica brown rice collected during postharvest processing. We fractionated the rice into six thickness fractions and classified each fraction into mature, immature and chalky levels of maturity. Results showed that thickness fraction and maturity level affected protein content. Each increase in thickness produced an increase in the percentage of sound whole kernel and a decrease in protein content. The mature sample had the lowest protein content among maturity levels within each thickness fraction. Based on sound whole kernel and protein content values, we classified the mature sample in higher thickness fractions as having high-quality appearance and palatability. There were significant differences in protein content between different thickness fractions of the mature sample suggested that we can sort rice protein by thickness fraction. Moreover, differences in protein content suggested that we can use the thickness grader and colour sorter machines to sort rice into high or low protein content fractions. This information can contribute to more accurate quality constituent determination through the development of a rice protein sorter. This would support the production of high-quality rice and help address the demand for more efficient technology for rice postharvest processing.

Autophagy is a highly regulated, biological process that provides energy during periods of stress and starvation. This conserved process also acts as a defense mechanism and clears microbes from the host cell. Autophagy is impaired in Cystic Fibrosis (CF) patients and CF mice, as their cells exhibit low expression levels of essential autophagy molecules. The genetic disorder in CF is due to mutations in the cystic fibrosis transmembrane conductance regulator (cftr) gene that encodes for a chloride channel. CF patients are particularly prone to infection by pathogens that are otherwise cleared by autophagy in healthy immune cells including Burkholderia cenocepacia (B. cenocepacia). The objective of this study is to determine the mechanism underlying weak autophagic activity in CF macrophages and find therapeutic targets to correct it. Using reduced representation bisulfite sequencing (RRBS) to determine DNA methylation profile, we found that the promoter regions of Atg12 in CF macrophages are significantly more methylated than in the wild-type (WT) immune cells, accompanied by low protein expression. The natural product epigallocatechin-3-gallate (EGCG) significantly reduced the methylation of Atg12 promoter improving its expression. Accordingly, EGCG restricted B. cenocepacia replication within CF mice and their derived macrophages by improving autophagy and preventing dissemination. In addition, EGCG improved the function of CFTR protein. Altogether, utilizing RRBS for the first time in the CF field revealed a previously unrecognized mechanism for reduced autophagic activity in CF. Our data also offers a mechanism by which EGCG exerts its positive effects in CF.

The rheological properties of egg white adsorbed layer at pH 9 and 7 at air/water interface is investigated in a wide range of dilute to reconstructed egg white solution using pendant drop tensiometer. The aggregation of proteins in the solution bulk influences the adsorption behavior as well as the dilational visco-elasticity (measured at various frequencies) of adsorbed layer. The comparison of the dilational visco-elasticity as measured at equilibrium and under dynamic conditions shows pH effects of the egg white molecules. Furthermore, the formation of dynamic adsorption layers of egg white proteins at the surface of rising bubbles was investigated by measuring the local rising velocity at low protein concentrations at pH 9 and 7. The velocity profiles of the rising bubble were in agreement with the dynamic surface tension and dilational visco-elasticity results. The time required to establish the adsorption layers of egg white at pH 7 is shorter than at pH 9 due to the conformation of the protein molecules close to the isoelectric point.

High-protein intake in early childhood may cause negative health effects in later life. However, low protein powdered follow-on milks are challenging to manufacture due to stickiness caused by increased free fat and lower glass transition points. Skim milk powder and palm oil emulsions, ranging from 0.25 to 5% w/w protein were assessed for their stability and subsequently spray dried to produce powders ranging from 1 to 20% w/w protein. Powders containing 2–8% w/w protein had high free fat levels despite the initial emulsion being stable, suggesting small adjustments in protein concentration can have a significant impact on emulsion stability during drying and atomisation. Reducing the protein content via substitution with lactose reduced the powders glass transition point (Tg) from 65 to 49 °C (powder contained 4%w/w protein, 67%w/w carbohydrate and 28%w/w fat). Substituting lactose with maltodextrin (DE6) at a 45%w/w level of inclusion increased the Tg of the powders from 50 to 113 °C, however, viscosity increased significantly. Interestingly, using glucose syrup (DE39) to substitute lactose at a 15%w/w level of inclusion, increased the powder Tg by 21 °C with no significant change in emulsion viscosity compared to a control with lactose as the sole carbohydrate. XPS analysis showed fat type (milk, palm, sunflower) influenced powder surface fat coverage. The inner fat contained increased levels of saturated fatty acids (palmitic and stearic) compared to the surface and encapsulated fat. In conclusion the Tg of low protein spray dried emulsions can be increased and the surface fat decreased, helping to reduce susceptibility to stickiness.

ATP-dependent phospholipid flippase activity crucial for generating lipid asymmetry was first detected in red blood cell (RBC) membranes, but the P4-ATPases responsible have not been directly determined. Using affinity-based MS, we show that ATP11C is the only abundant P4-ATPase phospholipid flippase in human RBCs, whereas ATP11C and ATP8A1 are the major P4-ATPasesinmouse RBCs. Wealso found that ATP11A and ATP11B are present at low levels. Mutations in the gene encoding ATP11C are responsible for blood and liver disorders, but the disease mechanisms are not known. Using heterologous expression, we show that the T415N substitution in the phosphorylation motif of ATP11C, responsible for congenital hemolytic anemia, reduces ATP11C expression, increases retention in the endoplasmic reticulum, and decreases ATPase activity by 61% relative to WT ATP11C. The I355K substitution in the transmembrane domain associated with cholestasis and anemia in mice was expressed at WT levels and trafficked to the plasma membrane but was devoid of activity. We conclude that the T415N variant causes significant protein misfolding, resulting in low protein expression, cellular mislocalization, and reduced functional activity. In contrast, the I355K variant folds and traffics normally but lacks key contacts required for activity. We propose that the loss in ATP11C phospholipid flippase activity coupled with phospholipid scramblase activity results in the exposure of phosphatidylserine on the surface of RBCs, decreasing RBC survival and resulting in anemia.

Optimal protein intake during critical illness is unknown. Conflicting results on nutritional support during the first week of ICU stay have been published. We addressed timing of protein intake and outcomes in ICU patients requiring prolonged mechanical ventilation.We retrospectively collected nutritional and clinical data on the first 7 days of ICU admission of adult critically ill patients, who were mechanically ventilated in our ICU for at least 7 days and admitted between January 1st 2011 and December 31st 2015. Based on recent literature, patients were divided into 3 protein intake categories,0.8 g/kg/day, 0.8-1.2 g/kg/day and1.2 g/kg/day. Our primary aim was to identify the optimum protein dose and timing related to the lowest 6 month mortality. Secondary endpoints were ventilation duration, need for renal replacement therapy (RRT), ICU length of stay (LOS) and mortality and hospital LOS and mortality.In total 455 patients met the inclusion criteria. We found a time-dependent association of protein intake and mortality; low protein intake (0.8 g/kg/day) before day 3 and high protein intake (0.8 g/kg/day) after day 3 was associated with lower 6-month mortality, adjusted HR 0.609; 95% CI 0.480-0.772, p 0.001) compared to patients with overall high protein intake. Lowest 6-month mortality was found when increasing protein intake from0.8 g/kg/day on day 1-2 to 0.8-1.2 g/kg/day on day 3-5 and1.2 g/kg/day after day 5. Moreover, overall low protein intake was associated with the highest ICU, in-hospital and 6-month mortality. No differences in ICU LOS, need for RRT or ventilation duration were found.Our data suggest that although overall low protein intake is associated with the highest mortality risk, high protein intake during the first 3-5 days of ICU stay is also associated with increased long-term mortality. Therefore, timing of high protein intake may be relevant for optimizing ICU, in-hospital and long-term mortality outcomes.

The Maillard conjugation of proteins and reducing saccharides is used to modify the technological functionality of whey proteins. In this study, whey protein isolate (WPI) was conjugated with maltodextrin (at 1:1 ratio and two total solid contents of 100 and 200 mg mL−1) through the Maillard reaction and used to form cold-set gels. The glycation reaction increased the strength of hydrogen bonding of whey proteins and preferentially modified α-lactalbumin, in comparison with β-lactoglobulin. It also increased the reducing power of binary protein-saccharide solution and allowed formation of self-standing cold-set WPI gel at a low protein content (i.e., ≈50 mg mL−1). Microscopic imaging showed micro-phase separated maltodextrin domains, interrupting the protein network, in gels made of protein-maltodextrin physical mixtures, whereas Maillard conjugation resulted in more homogenous microstructures at both total solid contents. The Maillard reaction increased gel firmness and water-holding capacity and caused a reduction in the extent of gel swelling.

The effects of corn processing and dietary protein level and their interaction on intake, digestibility, and feeding behavior in dairy cow was examined using eight multiparous Holstein dairy cows in mid-lactation (105 ± 9 days in milk and 47.2 ± 3 kg/day milk production at the start of the experiment) that were placed in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments. Experimental diets contained either finely ground corn (FGC) or steam-flaked corn (SFC) based on either low protein (LP, 14.8%) or high protein (HP, 16.2%) content. Diets contained 40% forage (corn silage and alfalfa hay) and 40% corn grain. Dry matter intake did not differ between HP and LP diets. In contrast, cows fed LP had greater total tract digestibility compared with that for HP-fed cows. Cows fed LP had greater chewing time and lower chewing rate (g of DM/min) compared to those for cows fed HP. Cows receiving SFC had a lower intake of nutrients and tended to have greater ADF and starch digestibility than cows fed FGC. There was an interaction between corn processing and level of protein with respect to the apparent total tract digestibility with cows fed FGC with HP resulted in the lowest digestibility. Steam-flaked corn had lower density and greater moisture and mean particle size than the FGC. In comparison with FGC, SFC did not pass through the 8-mm sieve and caused significant increase in physical effectiveness factor (pef) and physically effective fiber (peNDF) in SFC diets compared to that for FGC diets. Cows fed SFC sorted against longer particles (P = 0.04) to a greater extent than cows fed FGC. Dietary protein level had no detectable effect on sorting index. Chewing time did not differ for cows fed SFC or FGC; however, cows fed SFC had lower chewing and rumination rate (g of DM/min). Chewing time per kg of DM and CP were greater and per kg of peNDF were lower in cows fed SFC rather than cows fed FGC. Results indicated that corn processing and dietary protein level may interact to affect nutrient digestibility, but did not affect the feeding and chewing behavior of lactating cows under our experimental conditions. Results also showed that dietary peNDF content and chewing activity can be effectively manipulated by steam flaking of corn, and cows fed SFC had greater peNDF and energy intake despite lower DMI.

Dietary protein plays a major role in determining the rate of fish growth and overall health. Given that the liver is an important organ for metabolism and detoxification, we hypothesized that optimal dietary protein levels may benefit liver function. Herein, we investigated the effects of dietary protein level on serum biochemistry, liver histology and transcriptome profiling of juvenile bighead carp Aristichthys nobilis fed for 8 weeks on a diet supplemented with high protein (HP, 40%), low protein (LP, 24%) or optimal protein (OP, 32%; controls). The results revealed a significant change in liver morphology in LP and HP groups compared with the OP group, coupled with increased serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity. RNA sequencing (RNA-Seq) analysis of the liver transcriptome yielded 47 million high-quality reads using an Illumina platform, which were de novo assembled into 80,777 unique transcript fragments (unigenes) with an average length of 1021 bp. Subsequent bioinformatics analysis identified 878 and 733 differentially expressed unigenes (DEGs) in liver in response to LP and HP diets, respectively. KEGG enrichment analysis of DEGs identified immune and metabolism-related pathways, including Toll-like receptor signaling, PI3K-Akt signaling, NF-κB signaling, complement and coagulation, peroxisome, nitrogen metabolism, PPAR signaling, and glycolysis and gluconeogenesis pathways. Transcriptome profiling results were validated by quantitative real-time PCR for 16 selected DEGs. The findings expand our understanding of the molecular mechanisms underlying the effects of dietary protein level on liver function in bighead carp.

The objective of this experiment was to evaluate the interactive effects of corn processing and dietary protein level on milk production and composition, as well as rumen and plasma metabolites in lactation dairy cows. Eight multiparous mid lactating Holstein cows, averaging 105 ± 9 d in milk and producing 47.2 ± 3 kg milk/d were randomly assigned in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments. Experimental diets contained either finely ground corn (FGC) or steam flaked corn (SFC) each with either low protein (LP, 14.8%) or high protein (HP, 16.2%) content. Data were analyzed using PROC MIXED of SAS with significance declared at P ≤ 0.05. Cows fed HP diets had greater milk production and lower milk fat percentage than those for cows fed LP diets. Cows on LP diets however had greater medium-chain fatty acid (FA) and saturated FA but lower monounsaturated FA, polyunsaturated FA and unsaturated FA compared with cows on HP diets. Reduction of milk fat for cows on HP was associated with lower amount of short and medium chain FA and higher unsaturated FA content in milk fat. Blood glucose (74.7 vs 69.3 mg/dL) and cholesterol (275.2 vs 262.9 mg/dL) were greater for cows fed SFC than cows fed FGC. Corn processing had no detectable effect on milk production; however, cows on FGC had greater milk fat percentage and production and lower milk: feed ratio than cows fed SFC. Cows fed SFC had greater amount of short chain FA and polyunsaturated FA and lower C16:0 (30.5 vs 29.3) and C18:0 (9.1 vs 8.2) in milk fat than cows fed FGC. Treatments had no effect on ruminal VFA concentration and pH. Milk and blood urea, and rumen nitrogen (N) were lower for cows when fed SFC or LP diets. Cows fed SFC or LP had greater N efficiency than cows fed FGC or HP diets. These findings showed that cows supplemented with SFC or fed LP diets had a more efficient use of dietary N. Results indicate that employing a 14.8% CP diet is insufficient for maximum milk production even with corn flaking. Cows fed LP diets had similar milk: feed ratio and greater N efficiency. Compared with FGC diets, SFC diets improved feed and N efficiency while reducing N loss and increasing the glucose concentration.

Heparin, a highly sulfated linear polysaccharide, with anticoagulation function and blood compatibility is widely used as a biomaterials in medical application, but the most importance of heparin is its structure function as the macromolecular space arm. In this study, heparin as a spacer was covalently immobilized on the chloromethylated polystyrene microspheres (Ps) and then connected with l -phenylalanine forming the Ps-Hep-Phe structure, which was developed for endotoxin adsorption in hemoperfusion. The grafting density of heparin reach the maximum when the initial concentration of heparin solution was 5 mg/mL. The adsorbents with the heparin as a spacer showed the prolonged clotting times, low protein adsorption, and reduced the hemolysis rate, indicating that heparin-modified adsorbents have great blood compatibility. The adsorption capacity of Ps-Hep-Phe for endotoxin was 25.15 EU/g in dynamic adsorption, higher than that of Ps. Therefore, this study imply that heparin would be promising for modification of adsorbents in hemoperfusion.

Tibetan sheep are indigenous to the Qinghai-Tibetan Plateau and graze natural pasture all year round; small-tailed Han sheep were introduced to the plateau, are raised mainly in feedlots and graze natural pasture only in summer. A low energy intake could have a negative effect on N utilization and N balance in ruminants. We hypothesized that Tibetan sheep require less energy for N balance and utilize N more efficiently than Han sheep when offered a low protein and low energy diet. To test this hypothesis, we examined the effect of dietary energy on whole-body N retention and urea kinetics in Tibetan (47.7 ± 2.46 kg BW) and Han (46.2 ± 3.42 kg BW) sheep. Diets were formulated to be low in crude protein (˜7%) with different metabolizable energy levels (6.73, 7.65, 8.57 and 9.49 MJ/kg). Urea kinetics were determined using continuous intrajugular infusion of 15N15N-urea for 72 h. N retention was greater (P

To explore the effects of reducing the Cp levels on intestinal barrier function, low Cp (LP) and NRC standard Cp (NP) diets were fed to pigs from 45 to 160 days, and in vitro experiments were performed using monolayers of IPEC-J2 cells. The number of goblet cells, expression of proteins related to cell junction, amino acid transport, glucose transport, transepithelial electrical resistance (TEER), dextran permeability, and IL-6 secretion level were detected in pigs. The results demonstrated that a moderate reduction of Cp levels did not affect intestinal morphology, as demonstrated by a normal villi height, crypt depth and normal numbers of goblet cells. The maintenance of the intestinal structure obtained with LP was also confirmed by stable mRNA expression levels of muc2 and E-cadherin in the jejunum. We also found that LP did not affect the protein expression of cationic amino acid transporter 1 (CAT-1) and alanine serine cysteine transporter 1 (ASCT1) from 45 to 160 days. Moreover, the excitatory amino acid transporter 3 (EAAT3), sodium-glucose cotransporter 1 (SGLT1) and glucose transporter (GLUT2) protein expression levels in the jejunum were significantly increased at a certain age during the rearing period. Furthermore, we also demonstrated that a reduction in protein concentration up to 15% in the cultural medium of IPEC-J2 cells did not impact the mucosal barrier function. This study demonstrated that a moderate reduction of the protein level did not affect intestinal mucosal barrier function and morphology in the jejunum.

Animal welfare is an important aspect of organic broiler (OB) production which involves e.g. up to twice the rearing period compared with conventional production and feedstuffs without prophylactic additives or synthetic amino acids. As market demand for organic broiler meat increases, the formerly modest OB production in Sweden is now rapidly increasing and research is needed to support this development. It is essential to use genotypes that adapt well to the rearing environment, in order to achieve satisfactory animal welfare. Health problems related to fast growth rate are a common animal welfare issue in modern broilers and this applies to systems with long rearing periods. In this study, the Welfare Quality® assessment protocol for poultry was used for assessment of lameness, contact dermatitis, cleanliness, thermal comfort, litter quality and the human-animal relationship in two modern broiler genotypes; the fast-growing Ross 308 (R) and the slower-growing Rowan Ranger (RR). In total, 645 day-old chicks (328 R and 317 RR) were reared until 10 weeks of age, which is the rearing period formerly used in organic production in Sweden, and fed a low-protein (L), high-protein (H) or mussel-meal (M) diet containing 14.5%, 17.0% or 15.6% crude protein, respectively. Broiler welfare was assessed on three occasions (at weeks 2, 6 and 9). The results showed rapid deterioration in welfare for fast-growing broilers when kept beyond 6 weeks. Mortality rate and incidence of lameness and contact dermatitis increased and litter quality, thermal comfort and plumage cleanliness decreased. Indications of poor welfare were also observed in the slower-growing hybrid, but to a lesser extent and later during rearing. Diet type only had minor effects on bird welfare, although R birds grew faster on the M diet. Thus the slower-growing RR hybrid is preferable to the fast-growing broiler type in production systems with a long rearing period. However, the RR growth rate can be regarded as moderate and, to avoid health problems related to fast growth rate, hybrids that grow even more slowly should be considered for OB production systems with a long rearing period.

Summary: Background: With the increase in the prevalence of overweight, percentage of overweight patients with gastric cancer has also increased. This 5-year retrospective cohort study was performed to investigate long-term changes in the metabolic and nutritional parameters of early gastric cancer (EGC) patients with overweight after gastrectomy. Methods: EGC patients who underwent gastrectomy were followed up over a 5-year period. We included 393 patients (261 men, 132 women) who had an initial body mass index (BMI) of ≥23 kg/m2, and analyzed the longitudinal changes in the metabolic and nutritional parameters. Results: Body weight and random glucose, alanine aminotransferase (ALT), hemoglobin, and serum calcium levels significantly decreased, while serum protein and albumin levels increased in both men and women after gastrectomy. The odds ratios (ORs) for BMI ≥ 25 kg/m2 (P 40 IU/dL (men; P

Spray-dried powders were produced from milk serum protein concentrate and lactose in varying ratios, and the rehydration characteristics of the powders were evaluated. The dissolution rate was estimated with a flow-cell based technique, and the external and internal distribution of the powder components were evaluated with X-ray photoelectron spectroscopy and confocal Raman microscopy, respectively. The surface of the powder particles is more or less covered by a thin protein layer. A phase segregation between protein and lactose is observed in the interior of the particle resulting in a protein rich layer in the vicinity of the surface. However, the protein layer in the vicinity of the particle surface tends to become thinner as the bulk protein concentration increases in the powders (from 10 to 60% w/w). The time for the spontaneous imbibition to occur show a linear correlation with the protein surface coverage. The dissolution rate of powders containing 0.1% w/w protein is around 60 times faster than for a powder containing 1% w/w protein but the dissolution rate of powders containing 1% and 100% w/w differ only by a factor of 2. Thus, it is suggested that the outer protein layer becomes denser at the interface as the protein content increases in the powders, thereby causing poorer rehydration characteristics of the powders (especially for low protein concentrations 0.1–1% w/w). This insight has relevance for the formulation of whey protein powders with improved rehydration characteristics.

High-rate activated sludge (HRAS) is an essential cornerstone of the pursuit towards energy positive sewage treatment through maximizing capture of organics. The capture efficiency heavily relies on the degree of solid separation achieved in the clarifiers. Limitations in the floc formation process commonly emerge in HRAS systems, with detrimental consequences for the capture of organics. This study pinpointed and overcame floc formation limitations present in full-scale HRAS reactors. Orthokinetic flocculation tests were performed with varying shear, sludge concentration, and coagulant or flocculant addition. These were analyzed with traditional and novel settling parameters and extracellular polymeric substances (EPS) measurements. HRAS was limited by insufficient collision efficiency and occurred because the solids retention time (SRT) was short and colloid loading was high. The limitation was predominantly caused by impaired flocculation rather than coagulation. In addition, the collision efficiency limitation was driven by EPS composition (low protein over polysaccharide ratio) instead of total EPS amount. Collision efficiency limitation was successfully overcome by bio-augmenting sludge from a biological nutrient removal reactor operating at long SRT which did not show any floc formation limitations. However, this action brought up a floc strength limitation. The latter was not correlated with EPS composition, but rather EPS amount and hindered settling parameters, which determined floc morphology. With this, an analysis toolkit was proposed that will enable design engineers and operators to tackle activated solid separation challenges found in HRAS systems and maximize the recovery potential of the process. (C) 2018 Elsevier Ltd. All rights reserved.

Made available in DSpace on 2019-10-06T16:52:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The effect of extruded foods intake with different contents of starch, protein and hydroxyproline were evaluated on water balance, oxalate renal excretion, urine chemical composition and the relative urine supersaturation with calcium oxalate (RSSCaOx) in cats. Three complete and balanced diets were formulated with different proportions of starch, protein and hydroxyproline: STARCH, high starch and low protein and hydroxyproline content diet; SOY, high protein and low hydroxyproline and starch content diet; HYDROX, high hydroxyproline and protein, and low starch content diet. Twenty-four cats were used, with 8 replicate cats per diet. Animals were submitted to an adaptation period of 10 days, followed by 7 days for total feces and urine collection and 1 day for blood collection. Data were submitted to analysis of variance and means compared by Tukey's test (P < 0.05). Non-normal data were analyzed using Kruskal-Wallis and Dunn tests (P < 0.05). According to each treatment, cats had significantly different intake of starch, crude protein and hydroxyproline (P < 0.05). For diets higher in protein and lower in starch, coefficient of total tract apparent digestibility (CTTAD) of organic matter (OM), crude protein (CP), and gross energy (GE) were higher (P < 0.05). The daily urinary volume and water intake via drinking bowl (P < 0.001) were higher for cats fed with high protein foods (SOY and HYDROX) (P < 0.05). Renal excretion of Cl, S, uric acid and urea increased for animals receiving SOY and HYDROX diets compared with cats on the STARCH diet (P < 0.05), following the increase in intake of protein and these minerals. Although high protein consumption is generally associated with low urinary pH, on the present study cats fed the SOY diet produced urine with higher pH values than those fed the STARCH diet, although the former is higher in protein (P = 0.004). Oxalate urine concentration and RSSCaOx were higher (P < 0.001) and oxalate renal excretion tended to be higher (P = 0.109) for cats fed STARCH and HYDROX diets than those fed with the SOY food, suggesting higher hepatocyte production of oxalate after higher starch and hydroxyproline consumption by cats. In conclusion, although the limitation of the use of practical diets with differences on chemical composition, elevated starch intake with low protein consumption reduced urine production, and the high starch and hydroxyproline increased oxalate urine concentration and RSSCaOx. However, none of these diets induced the production of supersaturated urine with calcium oxalate, because all cats produced urine in the undersaturated zone. Sao Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences Sao Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences CNPq: 131979/2014-0

Distinct CD4+T cell epitopes have been associated with spontaneous control of HIV-1 replication, but analysis of antigen-dependent factors that influence epitope selection is lacking. To examine these factors, we used a cell-free antigen processing system that incorporates soluble HLA-DR (DR1), HLA-DM (DM), and cathepsins along with full-length protein antigens for epitope identification by LC-MS/MS. HIV-1 Gag, Pol, Env, Vif, Nef, Tat, and Rev were examined using this system. We identified 35 novel epitopes, including glycopeptides. Epitopes from smaller HIV-1 proteins with crystal structures mapped to regions of low protein stability and higher solvent accessibility. HIV-1 antigens associated with limited CD4+T cell responses were processed efficiently, while some protective epitopes were inefficiently processed. 66% of epitopes obtained from cell-free processing induced memory CD4+T cell responses in HIV-1+ donors, including 10 of 19 novel epitopes. Thus, an in vitro processing system can identify novel HIV-1 epitopes and reveal factors influencing epitope dominance.

The effect of different calcium levels (10–40 mM) during protein denaturation on the characteristics of cowpea protein gels obtained by heating or high hydrostatic pressure treatment (HHPT) at low protein content (7.5%w/w) was studied. Calcium allowed the formation of self-standing gels. For the lowest levels of treatments (70 °C and 400 MPa) calcium avoided a complete protein denaturation, which in turn resulted in weak matrices. For the highest levels (90 °C and 600 MPa) the complete protein denaturation in the presence of calcium allowed that this ion strengthened the matrices by establishing associations between exposed reactive sites. For HHPT, hardness and elastic module increased for each increase in calcium concentration, while for heating a plateau was reached from 15 mM. Differences in interactions that stabilized aggregates (more hydrophobic ones for heating and more calcium bridges for pressurizing) could explain the differences between heat- and HHPT-induced gels. Most of the gels belonged to the aggregate type, which entails opacity and low water holding capacity.

Defatted rice bran (DFRB) is an inexpensive and easily available agricultural byproduct. Existence of anti-nutritional factors (ANFs), high fiber and low protein content, susceptible to oxidation and rancidity make DFRB currently underutilized. In this study, Bacillus subtilis with high enzyme activities, Saccharomyces cerevisiae with high single-cell proteins concentration and Lactiplantibacillus plantarum with excellent acid secreting capacity were screened to co-fermented DFRB with phytase, and multiple physicochemical analyses combined with high-throughput sequencing were applied to provide insights into the dynamics of the physicochemical characteristics and the complex microbiome during the two-stage co-fermentation of DFRB. The results showed that co-fermentation effectively improved the nutritional value by degrading ANFs (trypsin inhibitors and phytic acid), fiber (acid detergent fiber and neutral detergent fiber) and allergenic protein, and increasing the trichloroacetic acid soluble protein, amino acids and organic acid. In addition, co-fermentation prevented lipid oxidation by enhancing antioxidant activity and reducing the activity of lipase and lipoxygenase. High-throughput sequencing results suggested that co-fermentation optimized microbial community of DFRB by increasing desirable Lactobacillus, Pediococcus, Saccharomyces and Talaromyces and reducing undesirable bacteria (Enterobacter and Pseudomonas) and animal and plant-pathogenic fungi (Blumeria, Alternaria, Fusarium, etc.). Furthermore, high-throughput sequencing and gas chromatography-mass spectrometry (GC-MS) were adopted to predict microbial metabolic functions and metabolic pathways during whole DFRB co-fermentation.

In this study, heat-denatured and alcalase-hydrolyzed whey and soy protein films/coatings containing marjoram essential oil (MEO) and thyme extract (TE) were investigated comparatively in terms of physicochemical and functional properties. Heat-denaturated high protein solutions produced self-standing, smooth, transparent and viscoelastic films in the presence of glycerol. Enzymatic hydrolysis provided transparency, elasticity and extensibility to the films of low protein solutions. The addition of MEO and TE triggered hardness and brittleness especially in hydrolyzed high protein films and remarkably changed film color, thickness and viscosity possibly affecting surface adherence of films. Heat denatured films exhibited higher water vapor permeability (WVP) than alcalase-hydrolyzed ones. The lowest hardness was obtained in hydrolyzed low protein films with considerable adhesiveness. Hydrolyzed protein solutions coated to cheese provided better protection against moisture loss and MEO added films caused elevated hardening of cheese. MEO and TE incorporated in film matrices were seemed to suppress mycelial growth and sporulation of Aspergillus niger. This study presented valuable new findings on functional protein films produced by enzymatic hydrolysis as an alternative to heat treatment.