Amino Acid Deficiency Research Articles

Recent trends in fish feed ingredients – mini review

Fish farming is commonly described as being extensive, semi-intensive or intensive. The continuous growing of aquaculture sector (around 8 % per year) led systems to be more and more intensive. In intensive farming, the fish are kept at high stocking density, that’s why the fish are dependent on the feed provided. Last decades aquaculture became more and more effective, but it has not been without concern for natural resource use, environmental impact and social judgement. Increasing fishmeal (FM) cost, irregular supply, decreasing availability, and poor quality of FM have put forward highlighting on its partial or complete substitution with other alternative protein sources. The fish feed industry has to be very responsible about to use FM alternatives. Scientists have to share on this burden to achieve sustainable, cost-effective aims. Indicators of sustainability in aquaculture could be energy efficiency, use of water, nutrient utilisation efficiency and production costs. The climate change is also affecting the sector, such as the predicted changes in ocean circulation pattern might also have a negative influence on the reliability of small pelagic stocks that being utilized for FM production or the appearance of mycotoxins on terrestrial plants, what could be possible alternative ingredients. The increase of the amount of non-marine ingredients in feeds is the result of years of investment and research in feed formulation to provide the nutrients the fish need to growth and remain healthy. Significant progress has been made over the past decade in reducing levels of FM in commercial feeds for farmed fish. Common obstacles associated with consumption of plant-based protein diets, because many times we face the facts with amino acid imbalances and deficiencies, high levels of indigestible carbohydrates present in certain grain products, and varying antinutritive factors (ANFs) that negatively affect fish growth and health. In this study the recent trends in alternative fish feed ingredients had been reviewed shortly. In conclusion, complete replacement of fishmeal in fish feeds is more difficult and will require further research efforts to achieve the expected goals, but there are many auspicious alternatives.

2195-P: Identification of the Regulatory Mechanism of mTORC1 Signaling Activity in Pancreatic Beta Cells in GCN2 Knockout Mice

Background and Objective: A single-nucleotide polymorphism (SNP) analysis of Japanese type 2 diabetes (T2DM) patients has revealed a significant correlation between a SNP of GCN2 and T2DM. We have demonstrated that islets isolated from GCN2 knockout (GCN2-/-) mice fed a high-fat diet (HFD) exhibited decreased expression of ATF4, a downstream molecule of GCN2, and pancreatic β-cell failure via hyperactivation of mTORC1 signaling. However, the mechanism of activated mTORC1 signaling has remained unclear. Therefore, our objective is to identify the regulatory mechanism of mTORC1 signaling activity in pancreatic β-cells of GCN2-/- mice fed an HFD. Methods: We analyzed using islets isolated from GCN2-/- mice and wild type mice fed an HFD, and pancreatic β-cell lines. Results: In pancreatic islets isolated from GCN2-/- mice fed an HFD, the phosphorylation of TSC2 was enhanced. Therefore, we speculated that there would be a pathway which regulated mTORC1 signaling via TSC2. It has been reported that TSC2 phosphorylation is regulated by its association with 14-3-3. We started to identify the molecule X downstream of GCN2, which involved in the regulation of mTORC1 signaling activity by competitively inhibiting the binding of TSC2 and 14-3-3. We carried out a proteome analysis to explore the molecules whose binding with 14-3-3 varied depending on the presence or the absence of GCN2. As a result, five molecules were listed, and Asparaginase was identified as the candidate of the molecule X. We revealed that Asparaginase was abundantly expressed in pancreatic islets following heart in mice tissues. In glucose loaded MIN6 cells and amino acid deficient MIN6 cells, the expression of Asparaginase was increased. Furthermore, the increased expression of Asparaginase by amino acid deficiency was decreased by ATF4 knocking down. Conclusion: It was suggested that Asparaginase was involved in the regulation of mTORC1 signaling activity downstream of GCN2-ATF4 pathway. Disclosure M. Kudo: None. S. Asahara: Research Support; Spouse/Partner; AstraZeneca. A. Kanno: None. Y. Kido: Research Support; Self; AstraZeneca.

Molecular Markers of Dietary Essential Amino Acid-deficiency (P08-059-19)

ObjectivesThe quality of dietary protein sources became a particularly sensitive issue in the current debates on a rebalancing between animal and vegetable food sources.The ability of a protein to meet the nutritional requirements of essential amino acid (EAA) is the basis for assessing the quality of protein.The objective of this study was to characterize the impact of lysine- and threonine-deficient gluten-based diets on the metabolism of growing rats and to identify molecular markers of these diets. MethodsGrowing rats were fed for 3 weeks with a threonine-supplemented and 70% lysine-deficient gluten diet; a lysine-supplemented and 47% threonine-deficient gluten diet; a gluten diet supplemented in lysine and threonine to meet all the AA requirements, and a control diet with milk protein to meet all the AA requirements.Body weight and food intake were measured daily. At the end of the experiment, tissues and biological fluids were removed. The body composition was analyzed, gene expression measurements involved in protein and lipid metabolism were performed and the urinary metabolome was analyzed by LC-MS. Statistical analysis was done by variance analysis and metabolome analysis by discriminant analysis of independent components. ResultsThese EAA deficiency does not modify the food intake. Lysine deficiency induces a decrease in body weight gain, and lean body mass, associated with an increased in proteolysis and a decreased in proteosynthesis, a decreased in bone mineral density, and no effect on lipid metabolism.Threonine deficiency induces a decrease in body weight gain, and liver and skin weight, without changes in protein metabolism, bone mineral density, and lipid metabolism. After approval of the deficiency model, the metabolomic analysis performed on urine samples revealed the presence of specific discriminating molecules of the diets and types of proteins. ConclusionsEAA deficiency has an impact on the growth, and bone and protein metabolism of growing rats. These deficiency states have resulted in different metabolome profiles that could lead to the identification of specific molecular markers of protein sources and related to EAA deficiencies. Funding SourcesThis study was funded by the UMR Nutrition Physiology and Ingestive Behavior. Supporting Tables, Images and/or Graphs▪

SUN-323 Somatostatin Analogue: An Effective Treatment for Pseudoglucagonoma Syndrome

INTRODUCTION: Pseudoglucagonoma syndrome (PS) refers to the presence of necrolytic migratory erythema (NME), despite normal glucagon levels, and absence of a glucagon-secreting tumor. We present a case of PS successfully treated with octreotide. CASE PRESENTATION: A 42-year-old woman with history of Roux-en-Y bypass surgery, performed 14 years ago, presented to the ER with abdominal pain and rash. Two months before presentation, she took 3 days of furosemide for ankle edema. On the third day of treatment, she developed a painful rash with blisters on the feet that progressed to the arms. She stopped furosemide and consulted our ER. There, her exam showed normal vitals signs and a fatigued-appearing patient. Her skin exam was remarkable for a painful, erythematous, scaling rash with vesicles and bullae in the intergluteal cleft, upper and lower extremities. Initial labs showed decreased albumin levels (1.9 g/dl), and normocytic anemia. Skin biopsy was consistent with zinc deficiency causing acrodermatitis enteropathica vs. NME. An abdominal CT revealed a 2.6 cm mass in the pancreatic head, compatible with pancreatic neoplasm, as well as changes of pancreatitis. Lab results showed deficiency in fat-soluble vitamins, zinc and copper. Infectious and rheumatologic evaluations were negative. Two fasting glucagon levels were normal (185 ng/L and 141 ng/L). A biopsy of the pancreatic mass was unable to be performed because she refused. At this point PS was included as a differential diagnosis and the rash has not improved despite repletion of vitamins and minerals. As a result, empiric immediate-release octreotide trial was started. The starting dose was 50mcg IV twice a day and a rapid titration (1 week) to 200mcg TID over the course of 1 week. Improvement of the skin lesion were observed 24 hours after starting octreotide. DISCUSSION: PS has been associated with malignancies, liver disease, infections, malabsorption, pulmonary or renal glucagonoma and pancreatitis. The pathogenesis of NME is not clear, but is believed to be multifactorial, involving excess of glucagon, deficiency of amino acid, zinc and/or free fatty acids. One hypothesis of glucagon as a cause of NME is that glucagon induces glycogenolysis and gluconeogenesis that may lead to epidermal amino acid depletion, necrolysis, and activation of inflammatory mediators. However, octreotide has been reported to improve NME in patients without glucagonoma, demonstrating that response is independent of glucagon levels. The first step for treatment is to correct the underlying pathology. Octreotide should be considered as second line of treatment. We recommend starting with a low dose of immediate-release octreotide, and increase for a maximum dose of 200 mcg TID. This dose can be maintained until resolution of symptoms and then switched to long-acting release octreotide.

High‐tryptophan Diet Reduces Weight Gain in Rats via Indole‐3‐propionic Acid, a Gut Microbiota Metabolite

Tryptophan is an essential amino acid. Paradoxically, both tryptophan‐low and tryptophan‐rich diets have been found to decrease weight gain in rats, however, the mechanisms are not clear.Energy and water‐electrolyte balance were evaluated in 12‐week‐old, male, Sprague Dawley rats maintained on either control diet (TC) or tryptophan‐high (TH) or tryptophan‐low (TL) or control diet with neomycin, an antibiotic (TCA), for 10 days. Feces and blood levels of tryptophan and its bacterial metabolites were evaluated using chromatography coupled with mass spectrometry.TL rats showed reduction in body weight. TH showed body weight gain, however, it was significantly lower than in TC and TCA rats. TL rats showed significantly lower blood level of tryptophan and its bacterial derivatives including indoxyl sulfate and indole‐3‐propionic acid (I‐3‐P). TH rats had significantly higher blood tryptophan level comparing to TL rats. I‐3‐P in feces and in portal blood of TH rats was significantly higher comparing to TC, TL and TCA rats. In a separate series of experiments rats treated daily i.p. with I‐3‐P showed significantly smaller weight gain than rats treated with the vehicle.Our study suggests that tryptophan‐rich diet has a negative effect on energy balance via I‐3‐P, a gut bacteria metabolite of tryptophan. In contrast, tryptophan‐deficient diet may reduce weight due to the deficiency of the essential amino acid and decreased anabolism.Support or Funding InformationSupported by the Ministry of Science and Higher Education Republic of Poland, Diamond grant DI2016 007346This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Nutritional and health attributes of milk and milk imitations.

Modern food technology allows designing products aiming to simulate and replace traditional food. In affluent societies there is a rising tendency to consume foods derived from plants including milk imitations or plant drinks based on cereals, nuts, legumes, oil seeds or other plant families. Herein we review production and composition of such drinks, summarize consumers' motivations to change from milk to plant drinks and highlight nutritional and health implications of consuming plant drinks instead of milk, in particular if non-fortified and if consumed by infants, children, adolescents and the elderly. Whereas the macronutrient concentrations of some plant drinks (soy) may approach in some cases (protein) that of cow's milk, the nutritional quality of most plant drinks, e.g., the biological value of protein and the presence and amount of vitamins and essential minerals with high bioavailability does not. If cow's milk is exchanged for non-fortified and non-supplemented plant drinks consumers may risk deficiencies of calcium, zinc, iodine, vitamins B2, B12, D, A, and indispensable amino acids, particularly in infants and toddlers who traditionally consume significant portions of milk. The vegetable nature, appearance and taste of such plant drinks may be appealing to adult consumers and be chosen for adding variety to the menu. However, in young children fed exclusively such plant drinks severe metabolic disturbances may occur. Parents, dietitians, physicians and consumers should be aware of such potential risks, if non-fortified plant drinks are consumed instead of milk.

Activating transcription factor 4‐dependent B cell translocation gene‐1 induction promotes ferroptosis

Ferroptosis is a special type of programmed cell death that is characterized by iron‐dependent lipid peroxidation. The present study investigated the role of B cell translocation gene‐1 (BTG‐1) on sulfur amino acid deficiency (SAAD)‐mediated cell death. SAAD in HepG2 cells depleted the reduced glutathione, accelerated lipid peroxidation, increased cyclooxygenase 2 expression, and promoted cell death. SAAD‐mediated cell death was specifically inhibited by pretreatment with ferrostain or defroxamine, chemical inhibitors of ferroptosis. In parallel with cell death, SAAD increased expression of microtubule‐associated protein light chain 3‐II and facilitated acridine orange positive autophagic flux. Inhibition of autophagy accelerated the SAAD‐mediated cell death. Moreover, SAAD induced BTG‐1 expression in HepG2 cells. By using BTG‐1 promoter haboring reporter gene and siRNA, activating transcription factor 4 (ATF4) was identified as an essential transcription factor for SAAD‐mediated BTG‐1 induction. Knock out of BTG‐1 gene by CRISPR/Cas9 system decreased the cytotoxicity by SAAD. Furthermore, BTG‐1 deficiency increased microtubule‐associated protein light chain 3‐II and autophagic flux. Thus, present study demonstrates that SAAD induces ferroptosis, and the ATF4‐dependent BTG‐1 induction contributes to accelerate ferroptosis by inhibiting autophagy.Support or Funding InformationThis study was supported by the National Research Foundation of Korea (NRF) funded by Korea government (MSIP)(Grant No. 2018R1A5A2025272 and 2018R1A2B6007514).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Decreased plasma l-arginine levels in organic acidurias (MMA and PA) and decreased plasma branched-chain amino acid levels in urea cycle disorders as a potential cause of growth retardation: Options for treatment

Background and aimPatients with methylmalonic acidemia (MMA) and propionic acidemia (PA) and urea cycle disorders (UCD), treated with a protein restricted diet, are prone to growth failure. To obtain optimal growth and thereby efficacious protein incorporation, a diet containing the essential and functional amino acids for growth is necessary. Optimal growth will result in improved protein tolerance and possibly a decrease in the number of decompensations. It thus needs to be determined if amino acid deficiencies are associated with the growth retardation in these patient groups. We studied the correlations between plasma L-arginine levels, plasma branched chain amino acids (BCAA: L-isoleucine, L-leucine and L-valine) levels (amino acids known to influence growth), and height in MMA/PA and UCD patients. MethodsWe analyzed data from longitudinal visits made in stable metabolic periods by patients registered at the European Registry and Network for Intoxication Type Metabolic Diseases (E-IMD, Chafea no. 2010 12 01). ResultsIn total, 263 MMA/PA and 311 UCD patients were included, all aged below 18 years of age. In patients with MMA and PA, height z-score was positively associated with patients' natural-protein-to-energy prescription ratio and their plasma L-valine and L-arginine levels, while negatively associated with the amount of synthetic protein prescription and their age at visit. In all UCDs combined, height z-score was positively associated with the natural-protein-to-energy prescription ratio. In those with carbamylphosphate synthetase 1 deficiency (CPS1-D), those with male ornithine transcarbamylase deficiency (OTC-D), and those in the hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome subgroup, height z-score was positively associated with patients' plasma L-leucine levels. In those with argininosuccinate synthetase deficiency (ASS-D) and argininosuccinate lyase deficiency (ASL-D), height was positively associated with patients' plasma L-valine levels. ConclusionPlasma L-arginine and L-valine levels in MMA/PA patients and plasma L-leucine and L-valine levels in UCD patients, as well as the protein-to-energy prescription ratio in both groups were positively associated with height. Optimization of these plasma amino acid levels is essential to support normal growth and increase protein tolerance in these disorders. Consequently this could improve the protein-to-energy intake ratio.

Effect of amino acid deficiency on precaecal amino acid digestibility in broiler chickens

In this study, we determined whether deficient dietary amino acid (AA) concentrations influence the precaecal (pc) AA digestibility when determined using the regression approach. We mixed two basal diets. Basal diet 1 was deficient in essential AAs, whereas adequate AA concentrations were ensured in basal diet 2 by adding free AAs. Rapeseed cake and full-fat soya beans as test ingredients were included in the basal diets at levels of 100 and 200, and 150 and 300g/kg, respectively, at the expense of maize starch. Each diet was tested with six replicates of 10 broiler chickens each. The feed intake of the chickens that were fed diets based on basal diet 2 was similar, whereas the feed intake of the chickens that were fed diets based on basal diet 1 differed considerably. The numerical differences in pc AA digestibility determined with basal diet 1 or 2 ranged from 2.6 percentage points to 20.8 percentage points in rapeseed cake and from 0.5 percentage points to 15.2 percentage points in soya beans. Across all measured AAs, the average differences were 10.1 percentage points and 5.4 percentage points in rapeseed cake and soya beans, respectively. The differences in the estimated pc AA digestibility between the basal diets were probably caused by different basal endogenous AA losses in the digesta between treatments as a consequence of different feed intake. Adequate AA concentrations and test ingredient levels that are specifically adjusted to avoid a negative effect on feed intake are recommended for future studies.

Biological evaluation of the influence of protein hydrolysate on the organism of the monks

In this study, performed on the basis of JSC "Plemzverosovkhoz Saltykovsky" on 150 standard minks which were selected according to the principle of analogs, the systemic and local effects of protein hydrolyzate in the composition of the basic diet were evaluated. A complex methodical approach was used, including clinical, hematological, morphological and zootechnical research methods. The digital material was subject o statistical processing. When analyzing the basic diet of minks, it was established that it is balanced in almost all nutritional substances. Nevertheless, based on a comparative study of the amino acid composition of the feed, a deficiency of the essential amino acid of threonine was detected. An evaluation of the dynamics of body weight in experimental animals showed an advantage in the growth of animals receiving hydrolyzate. By the end of the experiment, the animals of the experimental groups outstripped (by 10.1%) the control analogues for weight gain. The results of the analysis of biochemical parameters of blood serum testify to a statistically significant increase in the concentration of the total protein (by 48.6% in test group minks relative to the control group) with a simultaneous decrease (by 27%) of the albumin fraction. The level of ALT in the control group was lower (by 29%) than in the animals of the experimental groups, while increasing (by 35%) the level of AST. Morphometric parameters of the total skin thickness in experimental animals did not have significant differences. At the same time, their ratio in the general cover varies in the direction of diminishing the thickness of the epidermis and the depth of the hair follicles in animals treated with hydrolyzate. Planimetric indicators of skins raw materials in experienced animals increased by 5.5% compared to representatives of the control group. Based on the evaluation of systemic and local effects of protein hydrolyzate, the effectiveness of its use in cellular fur farming has been established.

БІЛОКВМІСНА СИРОВИНА РЕГІОНАЛЬНОГО ВИРОБНИЦТВА В ТЕХНОЛОГІЇ М’ЯСОМІСТКОЇ ВАРЕНО-КОПЧЕНОЇ КОВБАСИ

Urgency of the research. The deficiency of the most important human body biologically active substances in modern food products encourages the population to consume significant food in order to ensure a deficiency of amino acids, polyunsaturated fatty acids, minerals and vitamins. Target setting. It is possible to solve this problem by combining different types of raw materials, that is, to create multicomponent food products. Analysis of recent research and publications. Creation of innovative multicomponent products are based on freshwater fish of regional production, mechanically deboned poultry meat and certain types of by-products allows to reduce significantly the cost of sausage and culinary products while improving the quality and nutritional values. Uninvestigated parts of general matters defining. The literary sources analysis indicates the lack of research and development of new innovative recipes and technologies of meatcontaining boiled-smoked sausages with various types of regional aquaculture. The research objective. Investigation of the possibility of using regional aquaculture in the formulation of meatcontaining boiled smoked sausage, analysis of the nutritional value of the developed meat-based boiled-smoked sausage, the study of functional-technological, organoleptic and microbiological indicators of the developed meatcontaining boiled smoked sausage. The statement of basic materials. The formulation and technology of meatcontaining boiled smoked sausage, which includes raw materials of regional origin, namely meat of freshwater regional aquaculture, vegetable proteins as soy isolates and protein of hemp seeds, have been developed. The proposed products have high functional and technological properties of meat systems, indicators of quality and safety of finished products and high protein content with simultaneous decrease of energy value. Conclusions. According to results of investigations, the possibility of increasing the functional and technological properties of the model minced meatcontaining boiled-smoked sausages has been confirmed: moisture content up to 77.20 %, high school - up to 69.66 %, emulsifying capacity - up to 94.32 %, emulsion stability -61, 10 %. The sausages developed are microbiological safe. The high nutritional value on the content of the main nutrients has been confirmed. It is confirmed the possibility of combining meat of freshwater fish with traditional types of meat and vegetable raw materials for increasing the nutritional value and economic efficiency of production of meatcontaining boiled-smoked sausages.

A Pilot Data Analysis of a Metabolomic Liquid Chromatography-Tandem Mass Spectrometry (LC/MS/MS) Based Study of Patients with Sickle Cell/Beta Thalassemia

Background: The complex pathophysiology of Sickle Cell Disease (SCD) makes unlikely that a single therapeutic agent will prevent or reverse all SCD complications. Metabolomic analysis might help in the characterization of the endogenous and exogenous effects of potential new treatments. Metabolites are small molecules that are chemically transformed during metabolism and provide a functional readout of cellular state. Metabolites serve as direct signatures of biochemical activity and are therefore easier to correlate with phenotype. The metabolome is typically defined as the collection of small molecules produced by cells and offers a window for investigating how mechanistic biochemistry relates to cellular phenotype. There are very few reports associated with SCD providing comprehensive measurements of metabolites present in blood. Low arginine bioavailability has been associated with a clinical phenotype of increased hemolytic rate, pulmonary hypertension risk and early mortality. Recently, the FDA approved the use of L-glutamine for the treatment of adults and children with SCD, on the basis of the results of randomized phase 3 clinical trials1, while L-arginine's involvement is under investigation2,3. In this context we aimed to quantify targeted metabolites' abnormalities in patients with Sickle Cell/beta thalassemia (HbS/βThal), to identify pathways that might be of interest to prevent disease complications.Patients and Methods: Thirty adult Caucasian patients with HbS/βThal aged 45.6±10.9y, (43% male), at steady-state were enrolled in the study, while 20 age-matched healthy individuals (45% male) served as controls. Along with measurements of hematologic and blood chemistry parameters, targeted metabolome analyses for 13 aminoacids and 2 aminoacid's derivatives were performed after extraction from dry blood spots (DBSs) on filter paper using LC/MS/MS, with derivatization (AB SCIEX 5500 triple quadrupole QTRAP® LC/MS/MS Systems, Framingham, MA, USA).Results: Multiple metabolite differences are identified in HbS/βThal vs. Controls (Figure1). From metabolites involved in the biosynthesis of glutathione, only L-glutamine's levels were lower in patients with HbS/βThal compared to controls, while 5-oxyproline levels, a catabolic product of glutathione metabolism, were markedly increased in patients with HbS/βThal compared to controls. Urea cycle amino acids, also involved in the production of nitric oxide, L-arginine and L-ornithine concentrations were significantly lower in patients with HbS/βThal compared to controls, with a trend towards lower L-citrulline in patients with HbS/βThal p=0.06). Finally, amino acids involved in catecholamines (dopamine, nor-epinephrine and epinephrine) biosynthesis, such as L-phenylalanine and L-tyrosine and its metabolite succinylacetone levels were significantly lower in patients with HbS/βThal compared to controls. No significant correlations were found between any metabolites and markers of hemolysis, HbF levels or iron burden.Conclusions: This study confirms prior observations concerning aberrations in multiple blood specific amino acid levels in patients with SCD compared to controls, but this is the first report on Caucasian patients with HbS/βThal and on whole blood from DBS. We also identified for the first time in Caucasian patients with HbS/βThal important metabolic abnormalities of glutathione and nitric oxide biosynthesis pathways associated with altered concentrations of the metabolites serving of substrates in these cycles. Importantly, we also demonstrate low levels of L-phenylanine and L-tyrosine, which are essential sources for multiple neurotransmitters biosynthesis and neurobehavioral health. The latter novel observation should be confirmed in larger studies, while measurements of urinary amino acid clearances are necessary to evaluate for potential etiologies of the deficiencies through urinary losses vs. low substrate availability, increased utilization or abnormal metabolism. Patients with SCD are in a precarious state with respect to many amino acid deficiencies, all of which may have clinical consequences warranting further investigation.1N Engl J Med. 2018 Jul 19;379(3):226-235.2Anesth Analg. 2017 Apr;124(4):1369-1370.3Haematologica. 2013 Sep;98(9):1375-82. DisclosuresMorris:Pfizer: Consultancy; Calithera: Consultancy; MAST Therapeutics: Research Funding; UCSF-Benioff Children's Hospital Oakland: Patents & Royalties: For nutritional supplement licensed to Lifetrients; FDA: Research Funding; NIH/NHLBI: Research Funding. Voskaridou:Celgene Corp: Membership on an entity's Board of Directors or advisory committees, Research Funding; Acceleron: Membership on an entity's Board of Directors or advisory committees, Research Funding.

Remodeling mTORC1 Responsiveness to Amino Acids by the Herpes Simplex Virus UL46 and Us3 Gene Products Supports Replication during Nutrient Insufficiency.

By sensing fundamental parameters, including nutrient availability, activated mechanistic target of rapamycin complex 1 (mTORC1) suppresses catabolic outcomes and promotes anabolic processes needed for herpes simplex virus 1 (HSV-1) productive growth. While the virus-encoded Us3 Ser/Thr kinase is required to activate mTORC1, whether stress associated with amino acid insufficiency impacts mTORC1 activation in infected cells and virus reproduction was unknown. In contrast to uninfected cells, where amino acid withdrawal inhibits mTORC1 activation, we demonstrate that mTORC1 activity is sustained in HSV-1-infected cells during amino acid insufficiency. We show that in the absence of Us3, the insensitivity of mTORC1 to amino acid withdrawal in infected cells was dependent on the host kinase Akt and establish a role for the HSV-1 UL46 gene product, which stimulates phosphatidylinositol (PI) 3-kinase signaling. Significantly, virus reproduction during amino acid insufficiency was stimulated by the viral UL46 gene product. By synergizing with Us3, UL46 reprograms mTORC1 such that it is insensitive to amino acid withdrawal and supports sustained mTORC1 activation and virus reproduction during amino acid insufficiency. This identifies an unexpected function for UL46 in supporting virus reproduction during physiological stress and identifies a new class of virus-encoded mTORC1 regulators that selectively uncouple mTORC1 activation from amino acid sufficiency.IMPORTANCE Mechanistic target of rapamycin complex 1 (mTORC1) is a multisubunit cellular kinase that coordinates protein synthesis with changing amino acid levels. During amino acid insufficiency, mTORC1 is repressed in uninfected cells, dampening protein synthesis and potentially restricting virus reproduction. Here, we establish that HSV-1 alters the responsiveness of mTORC1 to metabolic stress resulting from amino acid insufficiency. Unlike in uninfected cells, mTORC1 remains activated in HSV-1-infected cells deprived of amino acids. Synergistic action of the HSV-1 UL46 gene product, which stimulates PI 3-kinase, and the Us3 kinase supports virus reproduction during amino acid withdrawal. These results define how HSV-1, a medically important human pathogen associated with a range of diseases, uncouples mTORC1 activation from amino acid availability. Furthermore, they help explain how the virus reproduces during physiological stress. Reproduction triggered by physiological stress is characteristic of herpesvirus infections, where lifelong latency is punctuated by episodic reactivation events.

Precision feeding approach affecting growth, nutrient utilization, feed conversion efficiency and economics of feeding weaned Murrah buffalo calves

The biological value of protein affects the growth of pre-ruminant calves and most deficient (critical) amino acids are methionine and lysine, which have an important role to play in balancing of amino acids for efficient utilization of feed protein (CP). Planning was made to develop economical calf starter for weaned buffalo (Bubalus bubalis) calves. Equal-energy calf starters (CS) were prepared with variable protein and amino acid levels, CS1 for control group (26% CP) was made with natural feed ingredients only, while designer CSs for groups CS2 and CS3, had 2 levels of critical amino acids with reduced CP content (21 and 20%, respectively) and fed to 3 groups of 6 calves (age 30 to 45 days and 58.40 kg mean body weight) each for 3 months. Higher growth rate and savings on the cost of feed per kg weight gain was achieved without affecting the digestibility of organic nutrients.

A cupin domain is involved in α-amylase inhibitory activity

Proteinaceous α-amylase inhibitors have specialized activities that make some strong inhibition of α-amylases. New α-amylase inhibitors continue to be discovered so far. A proteinaceous α-amylase inhibitor CL-AI was isolated and identified from chickpea seeds. CL-AI, encoded by Q9SMJ4, was a storage legumin precursor containing one α-chain and one β-chain, and each chain possessed a same conserved cupin domain. Amino acid mutation and deficiency of cupin domain would lead to loss of α-amylase inhibitory activity, indicating that it was essential for inhibitory activity. CL-AI(α + β) in its single stranded state in vivo had inhibitory activity. After it was processed into one α-chain and one β-chain, the two chains were connected to each other via disulfide bond, which would cover the cupin domains and lead to the loss of inhibitory activity. The CL-AI(α + β), α-chain and β-chain could inhibit various α-amylases and delay the seed germination of wheat, rice and maize as well as the growth and development of potato beetle larva. Two cupin proteins, Glycinin G1 in soybean and Glutelinin in rice were also found to have inhibitory activity. Our results indicated that the cupin domain is involved in α-amylase inhibitory activity and the proteins with a cupin domain may be a new kind of proteinaceous α-amylase inhibitor.

Integrated Remodeling of Gut-Liver Metabolism Induced by Moderate Protein Restriction Contributes to Improvement of Insulin Sensitivity.

ScopeProtein restriction (PR) is beneficial for relieving metabolic disorders and aging‐related diseases. However, extreme PR could result in malnutrition due to severe deficiency of essential amino acids. Therefore, the effect of moderate PR on insulin sensitivity is investigated.Methods and resultsThe growing and adult pigs are subjected to moderate PR by 15–30%. Plasma insulin concentration and insulin resistance index HOMA‐IR are significantly decreased upon moderate PR. Furthermore, IRS1/PI3K/AKT pathway in the basal state is enhanced in both liver and skeletal muscle. The adapted metabolism in the liver upon moderate PR is in support of improving insulin sensitivity. The liver shares a coordinated metabolic adaption in terms of energy metabolism and amino acid metabolism with the small intestine. Particularly, alteration of the metabolic footprint appeared in the portal venous blood, representing metabolites to be absorbed into liver after intestinal metabolism, is also in favor of improvement of insulin sensitivity.ConclusionIn summary, the study proves that moderate PR could improve insulin sensitivity from childhood to adulthood in a pig model, and sheds a new light on the role of integrated remodeling of gut and liver metabolism in the improved insulin sensitivity induced by moderate PR.

Adequacy of Plant-Based Proteins in Chronic Kidney Disease

Concerns regarding protein and amino acid deficiencies with plant-based proteins have precluded their use in chronic kidney disease (CKD) patients. Many of these concerns were debunked years ago, but recommendations persist regarding the use of "high-biological value" (animal-based) proteins in CKD patients, which may contribute to worsening of other parameters such as blood pressure, metabolic acidosis, and hyperphosphatemia. Plant-based proteins are sufficient in meeting both quantity and quality requirements. Those eating primarily plant-based diets have been observed to consume approximately 1.0g/kg/day of protein, or more. CKD patients have been seen to consume 0.7-0.9g/kg/day of mostly plant-based protein without any negative effects. Furthermore, those substituting animal-based proteins for plant-based proteins have shown reductions in severity of hypertension, hyperphosphatemia, and metabolic acidosis. Plant-based proteins, when consumed in a varied diet, are not only nutritionally adequate but have pleiotropic effects which may favor their use in CKD patients.

The Global Response of Cronobacter sakazakii Cells to Amino Acid Deficiency.

Cronobacter species can cause necrotizing enterocolitis and meningitis in neonates and infants, their infection is closely relevant to their responses to extreme growth conditions. In this study, the response of Cronobacter species to amino acid deficiency has been investigated. Four Cronobacter species formed smooth colonies when grown on the solid LB medium, but formed mucoid colonies when grown on the amino acid deficient M9 medium. When the mucoid colonies were stained with tannin mordant, exopolysaccharide around the cells could be discerned. The exopolysaccharide was isolated, analyzed, and identified as colanic acid. When genes wcaD and wcaE relevant to colanic acid biosynthesis were deleted in Cronobacter sakazakii BAA-894, no exopolysaccharide could be produced, confirming the exopolysaccharide formed in C. sakazakii grown in M9 is colanic acid. On the other hand, when genes rcsA, rcsB, rcsC, rcsD, or rcsF relevant to Rcs phosphorelay system was deleted in C. sakazakii BAA-894, colanic acid could not be produced, suggesting that the production of colanic acid in C. sakazakii is regulated by Rcs phosphorelay system. Furthermore, C. sakazakii BAA-894 grown in M9 supplemented with amino acids could not produce exopolysaccharide. Transcriptomes of C. sakazakii BAA-894 grown in M9 or LB were analyzed. A total of 3956 genes were differentially expressed in M9, of which 2339 were up-regulated and 1617 were down-regulated. When C. sakazakii BAA-894 was grown in M9, the genes relevant to the biosynthesis of exopolysaccharide were significantly up-regulated; on the other hand, the genes relevant to the flagellum formation and chemotaxis were significantly down-regulated; in addition, most genes relevant to various amino acid biosynthesis were also significantly regulated. The results demonstrate that amino acid deficiency has a global impact on C. sakazakii cells.

Acquired urea cycle amino acid deficiency and hyperammonaemic encephalopathy in a cat with inflammatory bowel disease and chronic kidney disease.

Case summaryA 5-year-old male neutered Persian cat was referred for investigation of a 4 week history of weight loss, inappetence and intermittent vomiting. Chronic kidney disease (CKD) and inflammatory bowel disease were diagnosed, and despite immunosuppressive therapy and assisted enteral nutrition, the cat experienced persistent anorexia, vomiting and severe weight loss. After 2 additional weeks of treatment, the cat developed acute-onset neurological signs associated with severe hyperammonaemia and was euthanased. Plasma amino acid assessment revealed deficiency of several amino acids involved in the urea cycle, including arginine.Relevance and novel informationTo our knowledge, this is the first reported case of an acquired urea cycle amino acid deficiency without nutritional deprivation in a cat. Several contributing factors were suspected, including intestinal malabsorption and CKD. This case demonstrates the importance of urea cycle amino acids in feline metabolism and possible necessity for parenteral supplementation, particularly in the context of persistent weight loss despite adequate enteral nutrition.

Effect of Homeopathic Medicine (Alfalfa), Methionine and Lysine Supplementation in Low Protein Based Diets on the Performance of Broiler Chicken

An experiment was conducted, with objective to determine the feed consumption, final live weight, weight gain, feed efficiency and economy in broiler chicken fed with locally available low protein based diets (20 percent CP in starter and 18 percent CP in finisher) supplemented with methionine, lysine and homeopathic medicine alfalfa. For this hundred fifty day-old birds of commercial broiler (Vencobb) were randomly assigned to five dietary treatments(10 chicks/treatment) and replicated three times in CRD with treatments composition of low protein based diets (LPB) (T0), LPB+ 100g/100 met + 300g/100 lysine (T1), LPB + 200g/100 met +300g/100lysine (T2), LPB + Alfalfa 5ml/100 chicks 2 day in week with drinking water (T3) and LPB + Alfalfa 5ml/100 chicks 3 day in week with drinking water (T4). Synthetic methionine and lysine was used for supplementation of deficient amino acid. Same condition was applied in finisher diets of broiler chicken. The result showed that chicks fed with LPB in both starter and finisher diet had significantly (P<0.05) lower feed intake, final live weight , total weight gain, feed efficiency and income over feed and chicks cost (IOFCC) with compromising chicks fed with LPB with addition of methionine and lysine each and also homeopathic medicine alfalfa in starter and finisher period. Highest feed intake, final live weight, weight gain, feed efficiency was obtained with treatment (T1) followed by treatment (T2). However, positive growth response and better health status of chicks was also observed from supplementation of homeopathic medicine alfalfa with LPB and non-significant difference on Income Over Feed and Chicks Cost was obtained with treatment (T1). So, it was concluded that with supplemental methionine, lysine and alfalfa, the CP level of broiler chicks could be reduced up to 20 percent in starter and 18 percent in finisher diets without adverse effect on feed intake, final live weight, weight gain, feed efficiency and Income overfeed and chicks cost, provided under locally based feed ingredients.Int. J. Appl. Sci. Biotechnol. Vol 6(2): 174-180