Amino Acid Supply Research Articles

SLC38A4 Amino Acid Transporter Expression Is Significantly Lower in Early Preterm Intrauterine Growth Restriction Complicated Placentas.

Intrauterine growth restriction (IUGR), predominantly caused by placental insufficiency, affects partitioning of nutrients to the fetus. The system A sodium-coupled transporters (SNAT or SLC38), of types A1, A2, and A4, control non-essential amino acid uptake and supply. Here, we aimed to investigate the expression of these transporters across different placental disease cohorts and cells. To determine disease impact, transporter expressions at the gene (qPCR) and protein (western blots) level were assessed in gestationally matched placental tissues. Early (<34 weeks), and late (34-36 weeks) onset IUGR cases with/out preeclampsia were compared to preterm controls. We also investigated level of transporter expression in primary trophoblasts under glucose deprivation (n = 6) and hypoxia conditions (n = 7). SLC38A4 protein was significantly downregulated in early preterm pregnancies complicated with IUGR with/out preeclampsia. There were no differences in late preterm IUGR cohorts. Furthermore, we demonstrate for the first time in primary trophoblast cells, that gene expression of the transporters was sensitive to and induced by glucose starvation. SLC38A4 mRNA expression was also significantly upregulated in response to hypoxia. Thus, SLC38A4 expression was persistently low in early preterm IUGR pregnancies, regardless of disease aetiology. This suggests that gestational age at delivery, and consequently IUGR severity, may influence loss of its expression.

Glutamine Produces Ammonium to Tune Lysosomal pH and Regulate Lysosomal Function.

Glutamine is one of the most abundant amino acids in the cell. In mitochondria, glutaminases 1 and 2 (GLS1/2) hydrolyze glutamine to glutamate, which serves as the precursor of multiple metabolites. Here, we show that ammonium generated during GLS1/2-mediated glutaminolysis regulates lysosomal pH and in turn lysosomal degradation. In primary human skin fibroblasts BJ cells and mouse embryonic fibroblasts, deprivation of total amino acids for 1 h increased lysosomal degradation capacity as shown by the increased turnover of lipidated microtubule-associated proteins 1A/1B light chain 3B (LC3-II), several autophagic receptors, and endocytosed DQ-BSA. Removal of glutamine but not any other amino acids from the culture medium enhanced lysosomal degradation similarly as total amino acid starvation. The presence of glutamine in regular culture media increased lysosomal pH by >0.5 pH unit and the removal of glutamine caused lysosomal acidification. GLS1/2 knockdown, GLS1 antagonist, or ammonium scavengers reduced lysosomal pH in the presence of glutamine. The addition of glutamine or NH4Cl prevented the increase in lysosomal degradation and curtailed the extension of mTORC1 function during the early time period of amino acid starvation. Our findings suggest that glutamine tunes lysosomal pH by producing ammonium, which regulates lysosomal degradation to meet the demands of cellular activities. During the early stage of amino acid starvation, the glutamine-dependent mechanism allows more efficient use of internal reserves and endocytosed proteins to extend mTORC1 activation such that the normal anabolism is not easily interrupted by a brief disruption of the amino acid supply.

Characterization of Bacillus velezensis UTB96, Demonstrating Improved Lipopeptide Production Compared to the Strain B. velezensis FZB42.

Bacillus strains can produce various lipopeptides, known for their antifungal properties. This makes them attractive metabolites for applications in agriculture. Therefore, identification of productive wild-type strains is essential for the development of biopesticides. Bacillus velezensis FZB42 is a well-established strain for biocontrol of plant pathogens in agriculture. Here, we characterized an alternative strain, B. velezensis UTB96, that can produce higher amounts of all three major lipopeptide families, namely surfactin, fengycin, and iturin. UTB96 produces iturin A. Furthermore, UTB96 showed superior antifungal activity towards the soybean fungal pathogen Diaporthe longicolla compared to FZB42. Moreover, the additional provision of different amino acids for lipopeptide production in UTB96 was investigated. Lysine and alanine had stimulatory effects on the production of all three lipopeptide families, while supplementation of leucine, valine and isoleucine decreased the lipopeptide bioproduction. Using a 45-litre bioreactor system for upscaling in batch culture, lipopeptide titers of about 140 mg/L surfactin, 620 mg/L iturin A, and 45 mg/L fengycin were achieved. In conclusion, it becomes clear that B. velezensis UTB96 is a promising strain for further research application in the field of agricultural biological controls of fungal diseases.

Penyembuhan Luka Post Sectio Cesarea (SC) Dengan Ekstrak Ikan Gabus (Channa Striata)

The fulfilment of high protein is one of the determinants of wound healing. Snakehead fish (Channa Striata) is a fish that has high protein and albumin. Protein supplies amino acids needed for tissue repair and regeneration, thereby influencing wound healing. The purpose of this study was to analyse the effect of snakehead fish extract on wound healing in post section-caesarean (SC). The research design used a quasi-experimental with a pre-test and post-test control group design. The population was all postpartum with SC, with as many as 30 mothers at Permata Hati Hospital, Malang. The research sample was 30 respondents divided into two groups (intervention and control groups as many as 15 postpartum, respectively). The intervention group receive a daily dose of 1000 mg of the fish extract and antibiotics, while the control group was given antibiotics only for 14 days. Wound status was assessed with the REEDA scale. The data were analyse using univariate and bivariate with an Independent t-test. Results showed there was a significant difference in wound status between groups (p = 0.0010.05). It can be concluded there was a significant effect of Channa Striata extract on post section-caesarean wounds among participants. Snakehead fish has the highest albumin content and support the wound healing process, especially for post-operative patients. Further research on the benefits of snakehead fish extract in increasing haemoglobin levels among post-partum mothers with anaemia is needed.

Ketone Body Exposure of Cardiomyocytes Impairs Insulin Sensitivity and Contractile Function through Vacuolar-Type H+-ATPase Disassembly-Rescue by Specific Amino Acid Supplementation.

The heart is metabolically flexible. Under physiological conditions, it mainly uses lipids and glucose as energy substrates. In uncontrolled diabetes, the heart switches towards predominant lipid utilization, which over time is detrimental to cardiac function. Additionally, diabetes is accompanied by high plasma ketone levels and increased utilization of energy provision. The administration of exogenous ketones is currently being investigated for the treatment of cardiovascular disease. Yet, it remains unclear whether increased cardiac ketone utilization is beneficial or detrimental to cardiac functioning. The mechanism of lipid-induced cardiac dysfunction includes disassembly of the endosomal proton pump (named vacuolar-type H+-ATPase; v-ATPase) as the main early onset event, followed by endosomal de-acidification/dysfunction. The de-acidified endosomes can no longer serve as a storage compartment for lipid transporter CD36, which then translocates to the sarcolemma to induce lipid accumulation, insulin resistance, and contractile dysfunction. Lipid-induced v-ATPase disassembly is counteracted by the supply of specific amino acids. Here, we tested the effect of ketone bodies on v-ATPase assembly status and regulation of lipid uptake in rodent/human cardiomyocytes. 3-β-hydroxybutyrate (3HB) exposure induced v-ATPase disassembly and the entire cascade of events leading to contractile dysfunction and insulin resistance, similar to conditions of lipid oversupply. Acetoacetate addition did not induce v-ATPase dysfunction. The negative effects of 3HB could be prevented by addition of specific amino acids. Hence, in sedentary/prediabetic subjects ketone bodies should be used with caution because of possible aggravation of cardiac insulin resistance and further loss of cardiac function. When these latter maladaptive conditions would occur, specific amino acids could potentially be a treatment option.

The Genetic Variability of Members of the SLC38 Family of Amino Acid Transporters (SLC38A3, SLC38A7 and SLC38A9) Affects Susceptibility to Type 2 Diabetes and Vascular Complications

Type 2 Diabetes (T2D) is a metabolic disease associated with long-term complications, with a multifactorial pathogenesis related to the interplay between genetic and modifiable risk factors, of which nutrition is the most relevant. In particular, the importance of proteins and constitutive amino acids (AAs) in disease susceptibility is emerging. The ability to sense and respond to changes in AA supplies is mediated by complex networks, of which AA transporters (AATs) are crucial components acting also as sensors of AA availability. This study explored the associations between polymorphisms in selected AATs genes and T2D and vascular complications in 433 patients and 506 healthy controls. Analyses revealed significant association of SLC38A3-rs1858828 with disease risk. Stratification of patients based on presence/absence of vascular complications highlighted significant associations of SLC7A8-rs3783436 and SLC38A7-rs9806843 with diabetic retinopathy. Additionally, the SLC38A9-rs4865615 resulted associated with chronic kidney disease. Notably, these genes function as AAs sensors, specifically glutamine, leucine, and arginine, linked to the main nutrient signaling pathway mammalian target of rapamycin complex 1 (mTORC1). Thus, their genetic variability may contribute to T2D by influencing the ability to properly transduce a signal activating mTORC1 in response to AA availability. In this scenario, the contribution of dietary AAs supply to disease risk may be relevant.

Different dietary ratios of arginine, methionine and lysine for turkeys: effects on whole-body composition and nutrient utilization efficiency in the early growth stage

Abstract The growth rate, tissue development and health status of turkeys are affected by the dietary supply of essential amino acids (AAs) such as lysine (Lys), methionine (Met) and arginine (Arg). According to various guidelines, the diets fed to turkeys in the first 4 weeks of the rearing period should contain 1.60–1.76% Lys, 1.60–1.80% Arg and 0.55–0.70% Met. This study investigated the effects of different ratios of Arg and Met in starter diets with 1.60% of Lys content, i.e. a low level, on the whole-body composition of turkeys and nutrient retention efficiency. The experiment lasted for 28 days and it had a two-factorial randomized design with three levels of Arg (90%, 100% and 110%) and two levels of Met (30% or 45%), relative to the content of dietary Lys, with six groups of eight replicates per group and 18 turkeys per replicate. Different dietary Arg and Met levels, relative to Lys, did not affect AA concentrations in the body protein or the proximate whole-body composition (crude protein, crude fat, gross energy and ash) of turkeys. The higher Met level positively influenced energy, protein and Arg utilization and their retention efficiency as well as the growth performance of turkeys. An increase in the dietary level of Arg to 100% and 110% of Lys content had no influence on the growth performance of turkeys and it reduced the retention efficiency of Arg. Differences in the body weight gain, feed intake and Lys utilization of turkeys fed diets with different Arg levels resulted also from the strong interaction between Met and Arg levels. At the lower level of Met in the diet, a decrease in Arg relative to Lys from 100% to 90% resulted in decreased (P = 0.003) feed intake (from 50.34 g to 46.72 g) and growth rate (from 31.84 g to 29.79 g), which was not noted for the higher Met content (45% relative to Lys). At the lowest level of Arg in the diet (90% of Lys content), a decrease in Met relative to Lys from 45% to 30% caused a decrease in Lys retention efficiency (from 61.51% to 56.29%). The results of this study suggest that the higher content of Met in the diet (45% to Lys) and the Arg level corresponding to 90% of Lys content contribute to optimizing nutrient retention efficiency in turkeys during the first 28 days of their life.

Bacterial Metalo-Aminopeptidases as Targets in Human Infectious Diseases.

Human infectious diseases caused by bacteria are a worldwide health problem due to the increased resistance of these microorganisms to conventional antibiotics. For this reason, the identification of novel molecular targets and the discovery of new antibacterial compounds are urgently required. Metalo-aminopeptidases are promising targets in bacterial infections. They participate in crucial processes for bacterial growth and pathogenesis, such as protein and peptide degradation to supply amino acids, protein processing, access to host tissues, cysteine supply for redox control, transcriptional regulation, site-specific DNA recombination, and hydrogen sulfide production. Although several of these enzymes are not essential, they are required for virulence and maximal growth in conditions of nutrient limitation and high temperatures. In this review, we describe the structural, functional, and kinetic properties of some examples of bacterial metalo-aminopeptidases, in the context of their use as antibacterial targets. In addition, we present some inhibitors reported for these enzymes. It is necessary to conduct a meticulous work to validate these peptidases as good/bad targets and to identify inhibitors with potential therapeutic use.

SLC38A2 provides proline and alanine to regulate postnatal bone mass accrual in mice.

Amino acids have recently emerged as important regulators of osteoblast differentiation and bone formation. Osteoblasts require a continuous supply of amino acids to sustain biomass production to fuel cell proliferation, osteoblast differentiation and bone matrix production. We recently identified proline as an essential amino acid for bone development by fulfilling unique synthetic demands that are associated with osteoblast differentiation. Osteoblasts rely on the amino acid transporter SLC38A2 to provide proline to fuel endochondral ossification. Despite this, very little is known about the function or substrates of SLC38A2 during bone homeostasis. Here we demonstrate that the neutral amino acid transporter SLC38A2 is expressed in osteoblast lineage cells and provides proline and alanine to osteoblast lineage cells. Genetic ablation of SLC38A2 using Prrx1Cre results in decreased bone mass in both male and female mice due to a reduction in osteoblast numbers and bone forming activity. Decreased osteoblast numbers are attributed to impaired proliferation and osteogenic differentiation of skeletal stem and progenitor cells. Collectively, these data highlight the necessity of SLC38A2-mediated proline and alanine uptake during postnatal bone formation and bone homeostasis.

The Environmental Impact of Lowering Dietary Crude Protein in Finishing Pig Diets—The Effect on Ammonia, Odour and Slurry Production

Excess nitrogen excretion, ammonia and odour are environmental pollutants associated with pig production. Reducing dietary crude protein (CP) will lower the amount of nitrogen excreted, reducing the potential for ammonia emissions, if diets are adequately formulated to supply amino acids and production performance is maintained. Crude protein content in diets for finishing pigs has been lowered recently, but the quantitative effect of this reduction on ammonia, odour, and slurry output is not well-established. The relationship between ammonia and odour is equivocal, and the effect on slurry production is unclear. The objective of this study was to investigate the effect of lowering dietary CP on ammonia emission, odour emission and slurry output of finishing pigs. Thirty entire boar pigs were individually housed in groups of six, from 10 weeks of age (30 ± 3.0 kg) and offered standard diets. At 75 kg (±1.5 kg) pigs were assigned to one of three treatment diets; (1) 180 g/kg CP, 11.0 g/kg total lysine (High CP), (2) 150 g/kg CP, 11.1 g/kg total lysine (Medium CP), and (3) 130 g/kg CP, 9.0 g/kg total lysine (Low CP). After three weeks on the experimental diets pigs were moved, six each week, to individual calorimetry chambers to measure ammonia and odour emissions. Pigs were offered treatment diets ad libitum. At the end of the recording period, the pigs were removed from the chamber, weighed and feed disappearance recorded to calculate intake. The slurry in each chamber was collected and analysed. The reduction in CP in the diet from 180 g/kg to 150 g/kg resulted in a 22% reduction in ammonia emissions, and from 180 g/kg to 130 g/kg resulted in a 47% reduction (p < 0.001). Slurry output from pigs offered the 130 g/kg CP diet was reduced by 39% (p < 0.001) and dry matter increased by 35% compared to slurry from pigs offered the 180 g/kg CP diet (p < 0.05). Water usage (p = 0.017), slurry output and nitrogen and phosphate in the slurry (p < 0.05, respectively) were found to decrease linearly with decreasing dietary CP content. There was no significant effect of reducing CP on performance or odour emission but hydrogen sulphide emissions decreased linearly (p < 0.010) with decreasing dietary CP. There was a weak positive relationship between odour emission and ammonia (linear: R2 = 0.25, p = 0.005) with odour emission reduced as ammonia emission reduced. Reducing dietary CP in finishing pig diets could reduce ammonia emissions, water usage and slurry and nutrient output from pig production.

Effect of Cannabis Seeds on Some Biomedical Parameters in Male Rats


 
 
 
 Cannabis, sometimes known as hemp, is a plant that originated in Central Asia. Cannabis seeds nutritional profile is astounding. Cannabis seed, most digestible, balanced, natural, and complete supply of protein, amino acids, and necessary fats found anyplace in nature when it is in its pristine organic natural condition. Cannabis seeds can be used in a variety of medical fields. Each 100 gm of the seeds used in the study contains 26, 37 and 20 gm of protein, fat and carbohydrate respectively. The present study used two groups of male rats (control and treatment groups). Rats were 11 weeks of average age. 10 mg/kg of body weight of powdered cannabis seeds were administrated orally to treatment group daily for 15 days. Alkaline Phosphatase, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase (ALP, GPT, and GOT), creatinine and uric acid was done for estimating the effect of cannabis seeds on renal and liver function. Enzymes Alkaline phosphatase, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase (ALP, GPT, and GOT) as well as creatinine, uric acid and very low-density lipoprotein (VLDL) were not significantly (p<0.05) different in both groups. Level of Cholesterol, triglyceride and c-reactive protein were significantly (p<0.05) reduced. Serum total protein, albumin and globulin were significantly (p<0.05) elevated in treatment group comparing to control group. The aim of this study is to examine physiological impact of cannabis whole seeds. Identifying the relationship(s) between liver and renal functions and cannabis seeds, as well as the anti-inflammatory effect of cannabis seeds and possible of using cannabis seeds as herbal medicine for health care in male rats.
 
 
 

PSVI-17 Effect of Post-Ruminal Amino Acid Supply on Organ Mass and Small Intestinal Length in Holstein Bull Calves fed a Starch-Based Milk Replacer

Abstract Forty Holstein bull calves were fed 1 of 3 starch-based milk replacers for either 10 or 35 days to determine responses in organ mass and small intestinal length. Calves (87.2 ± 7.7 kg) were bottle-fed a cornstarch-based milk replacer (727 g of cornstarch daily) alone, milk replacer with added casein (154.7 g/d) or milk replacer with added glutamate (45.5 g/d) for 10 or 35 d. Prior to receiving treatments calves were fed a commercial milk replacer for either 12 or 15 weeks so that measures of organ mass and small intestinal length were collected at the same day of age. Measures of organ mass were achieved after calves were euthanized and digesta was removed. Duodenum was defined as the first 1-m caudal to the pyloric sphincter. Jejunum and ileum were defined as the cranial and caudal halves of the remaining small intestines. Data were analyzed using the MIXED procedure of SAS. Average daily gain was 68% greater (P = 0.02) for calves fed casein compared to control, but glutamate did not differ from casein or control. Mass of the omasum, abomasum, jejunum, ileum, heart, or kidney and length of the jejunum or ileum were not affected by treatment (P ≥ 0.18); however, additions of casein increased (P ≤ 0.05) mass of the rumen, duodenum, large intestine, liver, and spleen. Greater duration of treatment increased rumen, abomasum, duodenum, jejunum, large intestine, heart, liver, and spleen mass (P ≤ 0.05), but there was treatment by duration interaction (P ≥ 0.18). Interestingly, there was an interaction (P < 0.01) for jejunal and ileal length in response to treatment and duration. Length of each segment increased with duration of treatment for calves fed casein or glutamate but lengths decreased in calves fed control for greater amounts of time.

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production.

The biosynthesis of antifungal lipopeptides iturin and fengycin has attracted broad interest; however, there is a bottleneck in its low yield in wild strains. Because the key metabolic mechanisms in the lipopeptides synthesis pathway remain unclear, genetic engineering approaches are all ending up with a single or a few gene modifications. The aim of this study is to develop a systematic engineering approach to improve the antifungal activity and biosynthesis of iturin and fengycin in Bacillus amyloliquefaciens. First, blocking the carbon overflow metabolic pathway to increase precursor supply of the branched-chain amino acids by knockout of bdh, disrupting sporulation to extend the stage for producing antifungal lipopeptides by deletion of kinA, blocking of siderophore synthesis to enhance the availability of amino acids and fatty acids by deletion of dhbF, and increasing Spo0A∼P by deletion of rapA, could improve the antifungal activity by 24%, 10%, 13% and 18%, respectively. Second, the double knockout strain ΔbdhΔkinA, triple knockout strain ΔbdhΔkinAΔdhbF and quadruple knockout strain ΔkinAΔbdhΔdhbFΔrapA could improve the antifungal activity by 38%, 44% and 53%, respectively. Finally, overexpression of sfp in ΔkinAΔbdhΔdhbFΔrapA further increased the antifungal activity by 65%. After purifying iturin and fengycin as standards for quantitative analysis of lipopeptides, we found the iturin titer was 17.0 mg/L in the final engineered strain, which was 3.2-fold of the original strain. After fermentation optimization, the titer of iturin and fengycin reached 31.1 mg/L and 175.3 mg/L in flask, and 123.5 mg/L and 1200.8 mg/L in bioreactor. Compared to the original strain, the iturin and fengycin titer in bioreactor increased by 22.8-fold and 15.9-fold in the final engineered strain, respectively. This study may pave the way for the commercial production of green antifungal lipopeptides, and is also favorable for understanding the regulatory and biosynthetic mechanism of iturin and fengycin.

Phenotypic response of yeast metabolic network to availability of proteinogenic amino acids.

Genome-scale metabolism can best be described as a highly interconnected network of biochemical reactions and metabolites. The flow of metabolites, i.e., flux, throughout these networks can be predicted and analyzed using approaches such as flux balance analysis (FBA). By knowing the network topology and employing only a few simple assumptions, FBA can efficiently predict metabolic functions at the genome scale as well as microbial phenotypes. The network topology is represented in the form of genome-scale metabolic models (GEMs), which provide a direct mapping between network structure and function via the enzyme-coding genes and corresponding metabolic capacity. Recently, the role of protein limitations in shaping metabolic phenotypes have been extensively studied following the reconstruction of enzyme-constrained GEMs. This framework has been shown to significantly improve the accuracy of predicting microbial phenotypes, and it has demonstrated that a global limitation in protein availability can prompt the ubiquitous metabolic strategy of overflow metabolism. Being one of the most abundant and differentially expressed proteome sectors, metabolic proteins constitute a major cellular demand on proteinogenic amino acids. However, little is known about the impact and sensitivity of amino acid availability with regards to genome-scale metabolism. Here, we explore these aspects by extending on the enzyme-constrained GEM framework by also accounting for the usage of amino acids in expressing the metabolic proteome. Including amino acids in an enzyme-constrained GEM of Saccharomyces cerevisiae, we demonstrate that the expanded model is capable of accurately reproducing experimental amino acid levels. We further show that the metabolic proteome exerts variable demands on amino acid supplies in a condition-dependent manner, suggesting that S. cerevisiae must have evolved to efficiently fine-tune the synthesis of amino acids for expressing its metabolic proteins in response to changes in the external environment. Finally, our results demonstrate how the metabolic network of S. cerevisiae is robust towards perturbations of individual amino acids, while simultaneously being highly sensitive when the relative amino acid availability is set to mimic a priori distributions of both yeast and non-yeast origins.

Amino Acids Supplied through the Autophagy/Endocytosis Pathway Promote Starch Synthesis in Physcomitrella Protonemal Cells.

The physiological implications of autophagy in plant cells have not been fully elucidated. Therefore, we investigated the consequences of autophagy in the moss Physcomitrella by measuring biochemical parameters (fresh and dry weights; starch, amino acid, carbohydrate, and NH3 content) in wild-type (WT) and autophagy-deficient atg5 Physcomitrella cells. We found higher starch levels and a higher net starch synthesis rate in WT cells than in atg5 cells cultured in a glucose-containing culture medium, whereas net starch degradation was similar in the two strains cultured in a glucose-deficient culture medium. Additionally, the treatment of cells with the autophagy inhibitor 3-methyladenine suppressed starch synthesis. Loading bovine serum albumin into atg5 cells through endocytosis, i.e., supplying proteins to vacuoles in the same way as through autophagy, accelerated starch synthesis, whereas loading glutamine through the plasma membrane had no such effect, suggesting that Physcomitrella cells distinguish between different amino acid supply pathways. After net starch synthesis, NH3 levels increased in WT cells, although the change in total amino acid content did not differ between WT and atg5 cells, indicating that autophagy-produced amino acids are oxidized rapidly. We conclude that autophagy promotes starch synthesis in Physcomitrella by supplying the energy obtained by oxidizing autophagy-produced amino acids.

Cryptochrome 1 as a state variable of the circadian clockwork of the suprachiasmatic nucleus: Evidence from translational switching

The suprachiasmatic nucleus (SCN) of the hypothalamus is the principal clock driving circadian rhythms of physiology and behavior that adapt mammals to environmental cycles. Disruption of SCN-dependent rhythms compromises health, and so understanding SCN time keeping will inform management of diseases associated with modern lifestyles. SCN time keeping is a self-sustaining transcriptional/translational delayed feedback loop (TTFL), whereby negative regulators inhibit their own transcription. Formally, the SCN clock is viewed as a limit-cycle oscillator, the simplest being a trajectory of successive phases that progresses through two-dimensional space defined by two state variables mapped along their respective axes. The TTFL motif is readily compatible with limit-cycle models, and in Neurospora and Drosophila the negative regulators Frequency (FRQ) and Period (Per) have been identified as state variables of their respective TTFLs. The identity of state variables of the SCN oscillator is, however, less clear. Experimental identification of state variables requires reversible and temporally specific control over their abundance. Translational switching (ts) provides this, the expression of a protein of interest relying on the provision of a noncanonical amino acid. We show that the negative regulator Cryptochrome 1 (CRY1) fulfills criteria defining a state variable: ts-CRY1 dose-dependently and reversibly suppresses the baseline, amplitude, and period of SCN rhythms, and its acute withdrawal releases the TTFL to oscillate from a defined phase. Its effect also depends on its temporal pattern of expression, although constitutive ts-CRY1 sustained (albeit less stable) oscillations. We conclude that CRY1 has properties of a state variable, but may operate among several state variables within a multidimensional limit cycle.

Research of a Dry Extruded Mixture of Protein Rich Plant Composition

Abstract Since plant protein is generally accepted as sustainable and environmentally friendly, production of new plant protein-based products is expanding and a raw material is diversified to ensure a high-value and complete meal. The objective of the study was to compare chemical composition and amino acid provision of pea-buckwheat, pea-barley and pea-oat extruded mixtures with the aim to look for new products to diversify diet using the best plant protein resources. Within the research, amino acid scores of hull-less barley, buckwheat, hull-less oat and grey pea as an indicator of raw material protein quality were evaluated. The highest scores of essential amino acids were found in oat and pea protein. Amino acid scoring patterns of raw material – oat ‘Stendes Emilija’ and grey pea ‘Bruno’ fully meet the requirements for essential amino acid supply. The protein of buckwheat is the richest in lysine (4.3 g 100g−1 protein) among studied cereals, although the difference in the essential amino acids was not significant. The results of the research showed that pea-barley and pea-buckwheat composite mixtures meet human needs better if pea protein is used in the proportion 65:35 with barley or buckwheat flour than in the proportion 35:65. The high protein content – 43.33% and 43.96%, the sum and composition of essential amino acids, and high level of total dietary fibre – 11.9% and 13.7% are the main reasons to consider the mixtures (including as a base for smoothies) as a complete meal.

K6 Physiological impact of amino acid supply during heat stress in ruminants

K6 Physiological impact of amino acid supply during heat stress in ruminants

Effect of shrimp waste fermentation feed supplement use on the balance of protein efficiency and color of native chicken egg yolk

Background: Supplement feed with an optimal composition will increase livestock productivity through increased feed digestibility and feed consumption that will provide a balance between amino acid supply and energy to grow, and produce. One of the fishery wastes that can be used as feed supplements is shrimp waste. The purpose of the study was to determine and obtain the level of use of feed supplements of fermented shrimp waste in the ration which resulted in a balance value of protein efficiency and the best egg yolk colour in the native chickens. Materials and Methods: The experiment used 20 native chickens aged 40 weeks in 20-unit cages. The study used the Complete Randomized Design method with 5 kinds of treatment, namely R0 (ration without the use of feed supplement fermentation of shrimp waste), R1 (ration with the use of feed supplement 0.5% fermentation of shrimp waste), R2 (ration with the use of feed supplement 1.0% fermentation of shrimp waste), R3 (ration with the use of feed supplement 1.5% fermentation of shrimp waste), and R4 (ration with the use of feed supplement 2.0% fermentation of shrimp waste), each treatment is repeated four times. The changes observed were ration consumption, protein consumption, egg weight, protein efficiency balance, and yolk colour. Results: Shrimp waste fermentation products as feed supplements have a significant influence (P<0.05) on feed consumption, protein consumption, egg weight, the balance of protein efficiency, and the colour of the yolk. The use of shrimp waste fermentation products gives optimal results at the rate of 1.5% (R3) in the ration against feed consumption and protein consumption, the weight of eggs optimal results at the level of 2.0 % (R4), and the balance of protein efficiency and against the colour of the yolk optimal results at the level of 1.0% (R2). Conclusions: The use of shrimp waste fermentation products of 1.0 levels in the ration results in an optimal balance of protein efficiency and yolk colour. The pattern of the relationship between treatments to the balance of protein efficiency is obtained in linear form with the equation y = 0.1494x + 3.7593 (R2 =93.71%). and in the colour of the yolk in linear form with the equation y = 0.85x + 7.4883 (R2=96.33%). Shrimp waste fermentation products can be used at a rate of 1.0-2.0% in the feed formula of the local poultry layer phase and livestock other than local poultry.

Comparative amino acid composition and quality parameters of Moringa oleifera testa and cotyledon

Introduction: Moringa oleifera is a drought-resistant plant, widely used in the tropical region. The leaves and stems have been extensively utilised in foods and neutraceuticals preparation, with less attention to the seeds. In this study, amino acid (AA) compositions of M. oleifera testa and cotyledon were examined comparatively. Methods: Samples were separately defatted, hydrolyed, and neutralised. The AA solution was purified by cation-exchange solid-phase extraction, derivatised and analysed by gas chromatography. Results: Glutamic (acidic amino acid) and phenylalanine (essential amino acid, EAA) were the most concentrated in both samples. Total EAA (g/100g crude protein, cp) was higher in cotyledon (51.0) than testa (41.9). Predicted protein efficiency ratios (P-PERs) were higher in testa (0.605-1.530) than cotyledon 0.286-1.460). EAA index ranged between 0.951-1.13 (soybean comparison) and 83.0-96.9 (egg comparison) with corresponding biological value of 78.7-93.9. The following AA had scores >1.0 in comparison to whole hen’s egg, testa: glycine (Gly), glutamic acid (Glu), phenylalanine (Phe), histidine (His), and cysteine (Cys); cotyledon (Gly), proline (Pro), Glu, Phe, His, arginine (Arg) and Cys. In comparison with requirements of pre-school children, six AA (6/9 or 66.7%) had scores >1.0 in each sample. In provisional AA scoring pattern, isoleucine (Ile) (1.25) and Phe + tyrosine (Tyr) (1.68) had scores >1.0 in testa while methionine (Met) + Cys, Phe+Tyr, and tryptophan (Trp) in cotyledon. However, tryptophan and lysine were the limiting AAs in testa and cotyledon, respectively. Conclusion: The study showed that both anatomical parts would complement each other in terms of amino acid supply.