Amino Acid Analysis Research Articles

Proline inhibits postharvest physiological deterioration of cassava by improving antioxidant capacity.

Cassava (Manihot esculenta Crantz), a crucial global tuber crop, encounters significant economic losses attributed to postharvest physiological deterioration (PPD). The PPD phenomenon in cassava is closely related to the accumulation of reactive oxygen species (ROS), and amino acids play a pivotal role in regulating signaling pathways and eliminating ROS. In this study, the storage performance of eight cassava varieties were conducted. Cassava cultivar SC5 showed the best storage performance among the eight cassava varieties, but the edible cassava cultivar SC9 performed much worse. Comparative analysis of free amino acids was conducted in eight cassava varieties, revealing changes in proline, aspartic acid, histidine, glutamic acid, threonine, and serine. Exogenous supplementation of these six amino acids was performed to inhibit PPD of SC9. Proline was confirmed as the key amino acid for inhibiting PPD. Treatment with optimal exogenous proline of 5g/L resulted in a 17.9% decrease in the deterioration rate compared to untreated cassava. Accompanied by a decrease in H2O2 content and an increase in catalase, superoxide dismutase and ascorbate peroxidase activity. Proline treatment proved to be an effective approach to alleviate cell oxidative damage, inhibit PPD in cassava, and prolong shelf life.

Comparison of δ13C and δ15N of ecologically relevant amino acids among beluga whale tissues

Ecological applications of compound-specific stable isotope analysis (CSIA) of amino acids (AAs) include 1) tracking carbon pathways in food webs using essential AA (AAESS) δ13C values, and 2) estimating consumer trophic position (TP) by comparing relative differences of ‘trophic’ and ‘source’ AA δ15N values. Despite the significance of these applications, few studies have examined AA-specific SI patterns among tissues with different AA compositions and metabolism/turnover rates, which could cause differential drawdown of body AA pools and impart tissue-specific isotopic fractionation. To address this knowledge gap, especially in the absence of controlled diet studies examining this issue in captive marine mammals, we used a paired-sample design to compare δ13C and δ15N values of 11 AAs in commonly sampled tissues (skin, muscle, and dentine) from wild beluga whales (Delphinapterus leucas). δ13C of two AAs, glutamic acid/glutamine (Glx, a non-essential AA) and, notably, threonine (an essential AA), differed between skin and muscle. Furthermore, δ15N of three AAs (alanine, glycine, and proline) differed significantly among the three tissues, with glycine δ15N differences of approximately 10 ‰ among tissues supporting recent findings it is unsuitable as a source AA. Significant δ15N differences in AAs such as proline, a trophic AA used as an alternative to Glx in TP estimation, highlight tissue selection as a potential source of error in ecological applications of CSIA-AA. Amino acids that differed among tissues play key roles in metabolic pathways (e.g., ketogenic and gluconeogenic AAs), pointing to potential physiological applications of CSIA-AA in studies of free-ranging animals. These findings underscore the complexity of isotopic dynamics within tissues and emphasize the need for a nuanced approach when applying CSIA-AA in ecological research.

Investigating the Nutritional and Functional Properties of Protaetia brevitarsis Larvae and Isolated Soy Protein Mixtures as Alternative Protein Sources

The growing demand for sustainable and alternative protein sources has spurred interest in insect-based and plant-based proteins. Protaetia brevitarsis (PB) larvae and isolated soy protein (ISP) are notable in this regard, offering potential health benefits and nutritional enhancements. We assessed the feasibility of PB larvae and ISP mixtures as alternative food ingredients. Methods included the optimized purification and freeze-drying of PB larvae, extraction and refinement of legume proteins, physicochemical and antioxidant capacity evaluations, DPPH radical scavenging activity measurement, total phenolic and flavonoids content quantification, general component analysis, amino acid profiling using HPLC, fatty acid profiling through gas chromatography, and mineral content analysis using inductively coupled plasma spectrometry. The study found that certain PB:ISP ratios, particularly a 7:3 ratio, significantly improved the blend’s antioxidant capacity, as evidenced by DPPH scavenging activity. This ratio also impacted the nutritional profile by altering the mixture’s general components, with a notable increase in moisture, crude protein, and fiber and a decrease in crude fat and ash. Amino acid analysis revealed a balanced presence of essential and non-essential amino acids. The fatty acid profile was rich in unsaturated fatty acids, especially in certain ratios. Mineral analysis showed a complex interplay between PB larvae and ISP, with some minerals decreasing and others increasing in the blend. PB larvae and ISP mixtures have significant potential as alternative protein sources, offering a diversified nutritional profile and enhanced antioxidant properties. The 7:3 ratio of PB larvae to ISP has been shown to be particularly effective, suggesting that this ratio may offer an optimal balance for enhancing the overall nutritional quality of the mixture. This study sets the stage for future research to further explore and optimize the potential of these mixtures for human consumption while considering the challenges of consumer acceptance and long-term safety.

Hairy roots biochemical characteristics of vegetable pea’s morphotype with modified leaf

Hairy roots biochemical characteristics of vegetable pea’s morphotype with modified leaf

Edible Paper Sheets from Alternanthera philoxeroides and Hypophthalmichthys molitrix: Smart Biomass Valorization

Alternanthera philoxeroides and Hypophthalmichthys molitrix offer significant nutritional benefits. This study evaluates the proximate composition, amino acid profile, GC-MS analysis, FT-IR spectroscopy, SEM and EDX, and color values of edible paper sheets (EPSs) derived from Alternanthera philoxeroides incorporating different levels of Hypophthalmichthys molitrix flesh. The protein content in the EPSs varied based on fish flesh incorporation, peaking at 52.66% in Ap100/Hm300 (Non-boil). Protein and carbohydrate contents showed an inverse correlation across EPSs, with the highest carbohydrate content of 60.89% in sample Ap400/Hm0 (Boil). Lipid content was also found to correlate with H. molitrix flesh content in EPSs, ranging from 1.59% to 18.41%. Amino acid analysis identified 11 types, with methionine as the most prevalent, followed by leucine, phenylalanine, and lysine. GC-MS analysis revealed 51 bioactive compounds, including carbonic acid, hentriacontane, and various fatty acids. FT-IR analysis showed characteristic bonds, while color analysis displayed L* values ranging from 24.37 to 30.97. SEM analyses depicted the microstructure, surface view, and elemental composition of the EPSs, and EDX showed an abundance of Ca, N, K, O, C, Mg, Na, P, Cl, Mn, and Fe. Therefore, EPSs prepared from A. philoxeroides and H. molitrix could offer a promising approach for effectively utilizing aquatic biomass and providing both plant and animal nutrients to consumers.

A new green approach to L-histidine and β-alanine analysis in dietary supplements using rapid and simple contactless conductivity detection integrated with high-resolution glass-microchip electrophoresis.

The analysis of dietary supplements is far less regulated than pharmaceuticals, leading to potential quality issues. Considering their positive effect, many athletes consume supplements containing L-histidine and β-alanine. A new microfluidic method for the determination of L-histidine and β-alanine in dietary supplement formulations has been developed. For the first time, capacitively coupled contactless conductivity detection was employed for the microchip electrophoresis of amino acids in real samples. A linear relationship between detector response and concentration was observed in the range of 10-100µmol L-1 for L-histidine (R2 = 0.9968) and β-alanine (R2 = 0.9954), while achieved limits of detection (3 × S/N ratio) were 4.2µmol L-1 and 5.2µmol L-1, respectively. The accuracy of the method was confirmed using recovery experiments as well as CE-UV-VIS and HPLC-UV-VIS techniques. The developed method allows unambiguous identification of amino acids in native form without chemical derivatization and with the possibility of simultaneous analysis of amino acids with metal cations.

Streamlining Protein Fractional Synthesis Rates Using SP3 Beads and Stable Isotope Mass Spectrometry: A Case Study on the Plant Ribosome.

Ribosomes are an archetypal ribonucleoprotein assembly. Due to ribosomal evolution and function, r-proteins share specific physicochemical similarities, making the riboproteome particularly suited for tailored proteome profiling methods. Moreover, the structural proteome of ribonucleoprotein assemblies reflects context-dependent functional features. Thus, characterizing the state of riboproteomes provides insights to uncover the context-dependent functionality of r-protein rearrangements, as they relate to what has been termed the ribosomal code, a concept that parallels that of the histone code, in which chromatin rearrangements influence gene expression. Compared to high-resolution ribosomal structures, omics methods lag when it comes to offering customized solutions to close the knowledge gap between structure and function that currently exists in riboproteomes. Purifying the riboproteome and subsequent shot-gun proteomics typically involves protein denaturation and digestion with proteases. The results are relative abundances of r-proteins at the ribosome population level. We have previously shown that, to gain insight into the stoichiometry of individual proteins, it is necessary to measure by proteomics bound r-proteins and normalize their intensities by the sum of r-protein abundances per ribosomal complex, i.e., 40S or 60S subunits. These calculations ensure that individual r-protein stoichiometries represent the fraction of each family/paralog relative to the complex, effectively revealing which r-proteins become substoichiometric in specific physiological scenarios. Here, we present an optimized method to profile the riboproteome of any organism as well as the synthesis rates of r-proteins determined by stable isotope-assisted mass spectrometry. Our method purifies the r-proteins in a reversibly denatured state, which offers the possibility for combined top-down and bottom-up proteomics. Our method offers a milder native denaturation of the r-proteome via a chaotropic GuHCl solution as compared with previous studies that use irreversible denaturation under highly acidic conditions to dissociate rRNA and r-proteins. As such, our method is better suited to conserve post-translational modifications (PTMs). Subsequently, our method carefully considers the amino acid composition of r-proteins to select an appropriate protease for digestion. We avoid non-specific protease cleavage by increasing the pH of our standardized r-proteome dilutions that enter the digestion pipeline and by using a digestion buffer that ensures an optimal pH for a reliable protease digestion process. Finally, we provide the R package ProtSynthesis to study the fractional synthesis rates of r-proteins. The package uses physiological parameters as input to determine peptide or protein fractional synthesis rates. Once the physiological parameters are measured, our equations allow a fair comparison between treatments that alter the biological equilibrium state of the system under study. Our equations correct peptide enrichment using enrichments in soluble amino acids, growth rates, and total protein accumulation. As a means of validation, our pipeline fails to find "false" enrichments in non-labeled samples while also filtering out proteins with multiple unique peptides that have different enrichment values, which are rare in our datasets. These two aspects reflect the accuracy of our tool. Our method offers the possibility of elucidating individual r-protein family/paralog abundances, PTM status, fractional synthesis rates, and dynamic assembly into ribosomal complexes if top-down and bottom-up proteomic approaches are used concomitantly, taking one step further into mapping the native and dynamic status of the r-proteome onto high-resolution ribosome structures. In addition, our method can be used to study the proteomes of all macromolecular assemblies that can be purified, although purification is the limiting step, and the efficacy and accuracy of the proteases may be limited depending on the digestion requirements. Key features • Efficient purification of the ribosomal proteome: streamlined procedure for the specific purification of the ribosomal proteome or complex Ome. • Accurate calculation of fractional synthesis rates: robust method for calculating fractional protein synthesis rates in macromolecular complexes under different physiological steady states. • Holistic ribosome methodology focused on plants: comprehensive approach that provides insights into the ribosomes and translational control of plants, demonstrated using cold acclimation [1]. • Tailored strategies for stable isotope labeling in plants: methodology focusing on materials and labeling considerations specific to free and proteinogenic amino acid analysis [2].

Nutritional composition and phenolic contents of Gracilariopsis longissima, Padina tetrastromatica and Ulva intestinalis from the Bay of Bengal, Bangladesh coast

Seaweeds have become the focus of experts in recent years due to their biological characteristics and the variety of uses they have for both humans and animals. Biochemical composition, amino acids, fatty acids, and phenolic components were analyzed to explore the nutritive value of Gracilariopsis longissima, Padina tetrastromatica, and Ulva intestinalis from the Bay of Bengal, Bangladesh coast. Proximate composition and mineral content were determined according to the AOAC method. The high-performance liquid chromatography amino acid analysis system was used for the amino acid analysis and the fatty acid profile of the extracted oils was assessed as their methyl esters. The Folin–Ciocalteu technique was used to estimate the phenolic content and the aluminum chloride colorimetric technique was used to calculate the total flavonoid content. The three different species of seaweed had significantly different proximate compositions (P < 0.05), with G. longissima having the highest protein content. Except for sulfur, the mineral contents were likewise considerably higher (P < 0.05) in G. longissima. Although the amounts of the essential amino acids were greater than 50 % of the total amino acids in the three studied seaweed species, the total amino acid composition of these three species differed significantly (P < 0.05). The findings indicated that lipid levels were low in all the assessed species, but unsaturated fatty acid levels were high, with G. longissima exhibiting the highest amounts. The results showed that, compared to the other species, G. longissima had a substantially higher (P < 0.05) level of total phenolic and flavonoid content. The three studied seaweed appear to be excellent for nutrition based on their overall nutritional profiles. However, due to high protein, unsaturated fatty acid, essential amino acid, and total phenolic and flavonoid content, G. longissima is the most promising seaweed that will be helpful for pharmaceutical and multifunctional food applications.

Flavour characterization of twelve species of edible algae

Integration of algae as ingredients in food products may contribute to the flavour characteristics, on top of providing nutritional requirements. In this study, characterization of the volatile and non-volatile flavour compounds was performed on twelve species of macroalgae and microalgae representing various groups of algae. Nine macroalgae species from three divisions were selected, namely Phaeophyta (brown alga): Laminaria japonica, Undaria pinnatifida, Sargassum fusiforme and Fucus vesiculosus; Rhodophyta (red alga): Porphyra haitanensis, Porphyra yezoensis, Palmaria palmata and Gelidium amansii, and Chlorophyta (green alga): Ulva lactuca. In addition, three microalgae species include Arthrospira platensis, Chlorella vulgaris and Dunaliella salina. Volatile compounds were analysed by solid-phase microextraction (SPME) and gas chromatography–mass spectrometry-flame ionization detector (GC–MS/FID). Amino acids and nucleotides were analysed using amino acid analyser and high-performance liquid chromatography (HPLC) respectively. Based on the identified volatile and non-volatile compounds, no obvious trend was found within each group of algae. However, some algae variants have been found to contain a unique aroma profile of selected volatile and non-volatile compounds. D. salina was found to be high in volatile aldehydes, ketones, and alcohols, primarily products of carotenoid degradation. U. lactuca contains the highest volatile sulfur content, while F. vesiculosus was found to be high in aldehyde and alcohol compounds, which are potentially fishy odours. C. vulgaris was found to have the highest value of equivalent umami concentration (EUC) followed by roasted P. haitanensis, indicating that both algae have the highest umami potential. Lastly, microalgae species A. platensis and C. vulgaris contained the highest amounts of sulfurous tasting amino acids cysteine and methionine respectively.

Sensory quality of emulsions prepared with the seaweed Ulva spp. or a derived protein ingredient

The successful commercialization of seaweed, whether as a whole biomass or a protein ingredient, partly relies on its sensory quality. This study explored the effect of different Ulva species, biomass washing, and pH-shift-based protein extraction on the sensory quality of emulsions with 0, 5, and 10% oil. A trained panel assessed the sensory profile, complemented by analyses of volatile compounds, total ash, and amino acids. Saltiness emerged as the primary distinction between emulsions with unwashed U. linza and U. fenestrata, due to higher ash in the former. Washing U. fenestrata retained sensory qualities despite reduced ash and increased content of the lipid oxidation-marker pentanal. Protein extraction up-concentrated total amino acids 2.9-fold, and yielded emulsions with reduced particle sensation and grassy flavor, while bitterness, sourness, dark color, pentanal, hexanal, and 2-ethylfuran increased. Increased oil content of emulsions lowered their grassy odor which correlated with reduced hexanal content. Overall, these findings can contribute to the development of food products containing seaweed or its protein ingredients that match consumer preferences.

Hemolymph metabolism of black soldier fly (Diptera: Stratiomyidae), response to different supplemental fungi.

The black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae), is commonly used for organic waste recycling and animal feed production. However, the often inadequate nutrients in organic waste necessitate nutritional enhancement of black soldier fly larvae, e.g., by fungal supplementation of its diet. We investigated the amino acid composition of two fungi, Candida tropicalis (Castell.) Berkhout (Saccharomycetales: Saccharomycetaceae) and Pichia kudriavzevii Boidin, Pignal & Besson (Saccharomycetales: Pichiaceae), from the black soldier fly gut, and commercial baker's yeast, Saccharomyces cerevisiae Meyen ex E.C. Hansen (Saccharomycetales: Saccharomycetaceae), and their effects on larval growth and hemolymph metabolites in fifth-instar black soldier fly larvae. Liquid chromatography-mass spectrometry was used to study the effect of fungal metabolites on black soldier fly larval metabolism. Amino acid analysis revealed significant variation among the fungi. Fungal supplementation led to increased larval body mass and differential metabolite accumulation. The three fungal species caused distinct metabolic changes, with each over-accumulating and down-accumulating various metabolites. We identified significant alteration of histidine metabolism, aminoacyl-tRNA biosynthesis, and glycerophospholipid metabolism in BSF larvae treated with C. tropicalis. Treatment with P. kudriavzevii affected histidine metabolism and citrate cycle metabolites, while both P. kudriavzevii and S. cerevisiae treatments impacted tyrosine metabolism. Treatment with S. cerevisiae resulted in down-accumulation of metabolites related to glycine, serine, and threonine metabolism. This study suggests that adding fungi to the larval diet significantly affects black soldier fly larval metabolomics. Further research is needed to understand how individual amino acids and their metabolites contributed by fungi affect black soldier fly larval physiology, growth, and development, to elucidate the interaction between fungal nutrients and black soldier fly physiology.

Optimizing amino acid analysis: Enhancing throughput and precision with advanced fluorescence detection on high-performance liquid chromatography (HPLC)

Optimizing amino acid analysis: Enhancing throughput and precision with advanced fluorescence detection on high-performance liquid chromatography (HPLC)

Analytical data of Acacia nilotica var. Nilotica gum

This study aimed to characterize the exudate gum from Acacia nilotica var. nilotica in Sudan and compare its physicochemical properties to Acacia seyal var. seyal and Acacia senegal var. senegal (gum Arabic). Samples were collected from six different states in Sudan over three seasons. The gum had a moisture content of 10.50%, ash content of 1.86%, pH value of 5.19, specific optical rotation of +94.70, intrinsic viscosity of 10.44 cm3 g-1, nitrogen content of 0.024%, protein content of 0.16%, acid equivalent weight of 1907.82, and total uronic acid content of 10.18%. Sugar content analysis revealed arabinose (41.20%), galactose (17.43%), and rhamnose (10.68%). Potassium was the predominant cation, followed by calcium, magnesium, sodium, lead, and iron. Acacia nilotica was classified as part of the Gummeferae series and exhibited a positive specific optical rotation. The Number average molecular weight (Mn) was estimated using osmometric measurements and gel permeation chromatography. The gum had a higher molecular weight and lower intrinsic viscosity compared to gum Arabic, suggesting a spheroidal shape of molecule. Amino acid analysis showed similarities with gum Arabic, with hydroxyproline and serine as principal amino acids. Variations in cationic composition were attributed to differences in soil type among collection locations. KEY WORDS: Physicochemical characterization, Acacia senegal var. senegal gum, Acacia seyal var. seyal gum, Acacia nilotica var. nilotica gum, gammiferae Bull. Chem. Soc. Ethiop. 2024, 38(4), 839-852. DOI: https://dx.doi.org/10.4314/bcse.v38i4.2

Impact of non‐thermal processing on the nutritional quality and bioactive properties of Industrial hempseed flours and protein isolate

AbstractThere is an unmet demand for plant‐based ingredients with desirable nutritional, techno‐functional properties, and health benefits. In this study, the composition, nutritional quality, and bioactives in industrial hempseed flours and protein isolate generated by milling, germination, isoelectric precipitation (IEP), and enzyme‐assisted extraction (EAE) were evaluated. Moisture, ash, fat, protein, phytic acid, tannin, and trypsin inhibitor content of the hempseed flours and protein isolate were 2.80%–6.46%, 5.07%–28.89%, 0.00%–31.44%, 22.71%–89.94%, 0.55%–1.05%, 274.24–1300.76 μg/g, and 0.00–42.66 U/g, respectively. IEP resulted in the highest protein content (89.94%), indicative of its effective to isolate hempseed protein. Germination and IEP significantly reduced phytic acid and tannin contents by 1.53‐ and 3.63‐fold, respectively. All processing methods improved in vitro protein digestibility (IVPD). SDS‐PAGE analysis revealed comparable band patterns in milled and protein isolate, with a strong 50 kDa band attributed as edestin. Amino acid analysis showed that EAE augmented total essential amino acids, particularly protease. Milling and pronase treatment yielded the highest and lowest IVPD‐corrected amino acid score (IVPDCAAS) of 76% and 47%, respectively. Milled and germinated flours contained varying amounts of γ‐tocopherol, lutein, zeaxanthin, α‐carotene, and β‐carotene. Germinated flour exhibited elevated levels of total phenolic (14.36 mg/g), and flavonoid (1.76 mg/g) contents, FRAP, TEAC, and DPPH compared to the other flours suggesting superior antioxidant capacity. Strong positive correlations (r &gt;0.70) were found between IVPD and phytic acid for the protein isolate, total phenolic content, and FRAP for IEP and germinated flours. Overall, hempseed flours with diverse properties could be produced using non‐thermal processing.

Isotopic evidence of high reliance on plant food among Later Stone Age hunter-gatherers at Taforalt, Morocco

The transition from hunting-gathering to agriculture stands as one of the most important dietary revolutions in human history. Yet, due to a scarcity of well-preserved human remains from Pleistocene sites, little is known about the dietary practices of pre-agricultural human groups. Here we present the isotopic evidence of pronounced plant reliance among Late Stone Age hunter-gatherers from North Africa (15,000–13,000 cal BP), predating the advent of agriculture by several millennia. Employing a comprehensive multi-isotopic approach, we conducted zinc (δ66Zn) and strontium (87Sr/86Sr) analysis on dental enamel, bulk carbon (δ13C) and nitrogen (δ15N) and sulfur (δ34S) isotope analysis on dentin and bone collagen, and single amino acid analysis on human and faunal remains from Taforalt (Morocco). Our results unequivocally demonstrate a substantial plant-based component in the diets of these hunter-gatherers. This distinct dietary pattern challenges the prevailing notion of high reliance on animal proteins among pre-agricultural human groups. It also raises intriguing questions surrounding the absence of agricultural development in North Africa during the early Holocene. This study underscores the importance of investigating dietary practices during the transition to agriculture and provides insights into the complexities of human subsistence strategies across different regions.

Metabolomic analysis of rumen-protected branched-chain amino acids in primiparous dairy cows.

Peripartal cows are susceptible to a negative energy balance due to inadequate nutrient intake and high energy requirements for lactation. Improving the energy metabolism of perinatal dairy cows is crucial in increasing production in dairy cows. In this study, we investigated the impact of rumen-protected branched-chain amino acid (RPBCAA) on the production performance, energy and lipid metabolism, oxidative stress, and immune function of primiparous dairy cows using metabolomics through a single-factor experiment. Twenty healthy primiparous Holstein cows were selected based on body condition scores and expected calving date, and were randomly divided into RPBCAA (n = 10) and control (n = 10) groups. The control group received a basal diet from calving until 21d in milk, and the RPBCAA group received the basal diet and 44.6 g/d RPLeu, 25.14 g/d RPIle, and 25.43 g/d RPVal. In comparison to the control group, the supplementation of RPBCAA had no significant effect on milk yield and milk composition of the dairy cows. Supplementation with RPBCAA significantly increased the concentrations of insulin, insulin growth factor 1, glucagon, and growth hormones, which are indicators of energy metabolism in postpartum cows. The very low density lipoprotein, fatty acid synthase, acetyl coenzyme A carboxylase, and hormone-sensitive lipase contents of the RPBCAA group were significantly greater than that of the control group; these metrics are related to lipid metabolism. In addition, RPBCAA supplementation significantly increased serum glutathione peroxidase and immunoglobulin G concentrations and decreased malondialdehyde concentrations. Liquid chromatography-mass spectrometry (LC-MS) analysis revealed 414 serum and 430 milk metabolic features. Supplementation with RPBCAA primarily increased concentrations of amino acid and lipid metabolism pathways and upregulated the abundance of serotonin, glutamine, and phosphatidylcholines. In summary, adding RPBCAA to the daily ration can influence endocrine function and improve energy metabolism, regulate amino acid and lipid metabolism, mitigate oxidative stress and maintain immune function on primiparous cows in early lactation.

The TRAF3-DYRK1A-RAD54L2 complex maintains ACE2 expression to promote SARS-CoV-2 infection.

The expression of ACE2 is essential for the entry of SARS-CoV-2 into host cells. We identify a new complex-the TDR complex-that acts to maintain the abundance of ACE2 in host cells. The identification and characterization of the TDR complex provide new targets for the development of therapeutics against SARS-CoV-2 infection. By analysis of SARS-CoV-2 virus replicating in cells expressing low levels of ACE2, we identified mutations in spike (P1079T) and nucleocapsid (S194L) that overcome the restriction of limited ACE2. Functional analysis of these key amino acids in S and N extends our knowledge of the impact of SARS-CoV-2 variants on virus infection and transmission.

Analysis of carcass traits, meat quality, amino acid and fatty acid profiles between different duck lines

To investigate the effect of genetic selection on meat quality in ducks, twenty of each fast growth ducks (LCA) and slow growth ducks (LCC) selected from F6 generation of Cherry Valley ducks (♂) x Liancheng white ducks (♀) were analyzed for carcass characteristics, meat quality (physicochemical and textural characteristics), amino acid and fatty acid profiles at 7 wk. Results showed that live body weight, slaughter weight, eviscerated yield and abdominal fat percentage of LCA were significantly higher than those in LCC ducks (P < 0.01). Moreover, the average area and diameter of myofiber were larger in LCA than LCC ducks (P < 0.01). The breast and thigh muscles of LCA exhibited significantly lower water holding capacity and thermal loss compared with LCC ducks (P < 0.01). In addition, the content of nonessential amino acids (Glu, Asp, and Arg) in breast muscles and Asp, Ser, Thr, and Met in thigh muscles was higher in LCC than LCA ducks (P < 0.05). The proportion of polyunsaturated fatty acids (PUFA) in breast muscles of LCC was higher than LCA ducks (P < 0.05). However, the content of saturated fatty acids (SFA) in breast and thigh muscles of LCA was higher compared with LCC ducks (P < 0.05). The proportion of monounsaturated fatty acids (MUFA) in thigh muscles was significantly higher in LCC compared with LCA ducks (P < 0.01). Finally, multiple traits were evaluated by applying principal component analysis (PCA) and the results indicated that PUFA and SFA in breast muscles of LCA played important roles in meat quality, followed by Warner-Bratzler shear force (WBSF) and MUFA. However, water holding capacity (WHC) had a dominant effect in meat quality of thigh muscles in both LCA and LCC ducks.

Luteolin-incorporated fish collagen hydrogel scaffold: An effective drug delivery strategy for wound healing

In clinical practice, wound care has always been challenging. Hydrogels play a key role in facilitating active wound recovery by absorbing exudates, maintaining moisture, and alleviating pain through cooling. In this study, type I collagen was isolated from the skin of crucian carp (Carassius carassius) and verified by amino acid analysis, FTIR, and SDS-PAGE. By adopting a new approach, luteolin was added to collagen hydrogels in situ after being dissolved in an alkaline solution. XRD and SEM confirmed the luteolin was incorporated and entirely distributed throughout the hydrogel. The plastic compression improved the young's modulus of hydrogel to 15.24 ± 0.59 kPa, which is adequate for wound protection. The drug loading efficiency was 98 ± 1.47 % in the selected formulation. The luteolin-incorporated hydrogel enabled regulated drug release. We assessed the cytotoxicity using MTT and live-dead assays, as well as examined the hemocompatibility to determine the biocompatibility of the hydrogel. In vivo experiments showed that the hydrogel with luteolin had the highest wound closure rate (94.01 ± 2.1 %) and improved wound healing with granular tissue formation, collagen deposition, and re-epithelialization. These findings indicate that this efficient drug delivery technology can accelerate the process of wound healing.

Physiological high temperatures alter the amino acid metabolism of bovine early antral follicles.

Heat stress reduces the developmental competence of bovine oocytes during the growth phase; however, the detailed mechanisms remain unclear. Amino acids play various critical roles in follicular development, including protein synthesis and as energy sources. We performed in vitro growth (IVG) culture of oocyte-cumulus-granulosa complexes (OCGCs) to assess the amino acid metabolism of small follicles at high temperatures. We isolated OCGCs from early antral follicles (0.5-1.0 mm) and subjected them to IVG culture for 12 days. OCGCs in the heat shock group were cultured under a temperature cycle of (38.5°C: 5 h, 39.5°C: 5 h, 40.5°C: 5 h, and 39.5°C: 9 h) to reproduce the body temperature of lactating cows under a hot environment. OCGCs in the control group were cultured at a constant temperature of 38.5°C for 24 h. Of the surviving OCGCs, those showing similar morphology and size between the groups were selected for amino acid analysis. We analyzed the free amino acids and their metabolites in the culture medium and calculated the depletion or appearance of molecular species. The depletion of three essential amino acids (isoleucine, leucine, and valine), two non-essential amino acids (aspartic acid and glycine), and ornithine was higher in the heat shock group (P < 0.05). Alanine depletion was lower in the heat shock group (P < 0.05). We concluded that heat exposure alters the amino acid metabolism of OCGCs isolated from early antral follicles, which might be involved with the diminished developmental potential of oocytes during summer.