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Objective: We used a random approach to analyze amino acid pairs in human Von Hippel-Lindau disease tumor suppressor (G7 protein) in order to determine which amino acid pairs are more sensitive to 109 mutations from human G7 protein. The rationale for this study is based on our hypothesis and findings that a harmful mutation is more likely to occur at randomly unpredictable amino acid pairs and a harmless mutation is more likely to occur at randomly predictable amino acid pairs. We argue that the randomly predictable amino acid pairs should not be deliberately evolved, whereas the randomly unpredictable amino acid pairs should be deliberately evolved with connection of protein function. Design: The amino acid sequence of the human G7 protein and its 109 mutations with point mutant were obtained from the Swiss-Protein data bank. There area total of 212 amino acid pairs in human G7 protein--less than the 400 that are theoretically possible. We calculated the randomly predicted frequency and actual frequency in human G7 protein, the randomly predictable present amino acid pairs, randomly unpredictable present amino acid pairs, randomly predictable absent amino acid pairs, mutations in randomly predictable and unpredictable amino acid pairs, and the difference between actual and randomly predicted frequencies. In this way, we compared the frequency difference in the amino acid pairs affected by mutations. Results: We found that 94.5% of 109 mutations occur at randomly unpredictable amino acid pairs, which account for 78.3% of amino acid pairs in human G7 protein. Conclusion: The randomly unpredictable amino acid pairs ate more sensitive to mutations in human G7 protein. The results also suggest that the human G7 protein has a natural tendency to mutate. [ABSTRACT FROM AUTHOR]

This study is aimed at investigating the effects of dietary supplementation with Artemisia ordosica crude polysaccharides (AOCP) on lactation performance, antioxidant status, and immune status of lactating donkeys and analyzing rectal microbiomes and serum metabolomes. Fourteen lactating Dezhou donkeys with similar age (6.16 ± 0.67 yr of BW ± SD), weight (250.06 ± 25.18 kg), DIM (39.11 ± 7.42 d), and average parity of 3 were randomly allocated into 2 treatments: a control group (CON, basal diet) and an AOCP group (AOCP, basal diet with 1.0 g/kg DM AOCP). Ten weeks were allotted for the experiment, 2 wk for adaptation, and 8 wk for collecting data and samples. The results showed that supplementation of donkey diets with AOCP increased lactation performance, including DMI, milking yield, estimated milk yield, solids-corrected milk, ECM, milk fat yield, milk protein yield, milk lactose yield, milk TS yield, and milk SNF yield. The digestibility of DM, CP, ADF, and NDF was increased in the AOCP group compared with the CON group. The AOCP group increased the concentrations of IgA, IgG, and IgM, the activities of the superoxide dismutase, catalase, and total antioxidant capacity in the serum. Artemisia ordosica crude polysaccharides decreased the concentrations of tumor necrosis factor-α, nitric oxide, reactive oxygen species, and malondialdehyde in the serum. Compared with the CON group, AOCP increased propionate, butyrate, isovalerate, and total VFA concentrations in rectal feces (P < 0.05). The addition of AOCP to increased diversity (Shannon index) and altered structure of the rectal microflora. As a result of AOCP supplementation, there has been a significant improvement in the colonization of beneficial bacteria, including Lactobacillus, Unclassified&#95;f&#95;Prevotellacea, Ruminococcus, and Fibrobacter genera. In contrast, a decrease in the colonization of the Clostridium&#95;sensu&#95;stricto&#95;1 bacterial genus and other pathogenic bacteria was observed. Meanwhile, metabolomics analysis found that AOCP supplementation upregulated metabolites l-tyrosine content while downregulating 9(S)-HODE, choline, sucrose, lysophosphatidylcholine (LysoPC) (18:0), LysoPC (18:1(9Z)), and LysoPC (20:2(11Z,14Z)) concentrations. These altered metabolites were involved in the PPAR signaling pathway, prolactin signaling pathway, glycerophospholipid metabolism, carbohydrate digestion and absorption, and tyrosine metabolism pathways, which were mainly related to antioxidant capacity, immune responses, and protein metabolism in the lactating donkeys. As a consequence of feeding AOCP diets, beneficial bacteria were abundant, and antioxidant and protein metabolism-related pathways were enriched, which may enhance lactation performance in donkeys. Therefore, supplementing AOCP diets is a desirable dietary strategy to improve donkey health and lactation performance., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Artemisia ordosica is one of the main shrubby perennials belonging to Artemisia species of Asteraceae and could be used in folk Chinese/Mongolian medicine to treat symptoms of various inflammatory ailments. The present study was conducted to investigate the protective effects of dietary Artemisia ordosica polysaccharide (AOP) against lipopolysaccharide (LPS) induced oxidative stress in broilers via Nrf2/Keap1 and TLR4/NF-κB pathway. A total of 192 1-day-old Arbor Acres male broilers were randomly allotted to four treatments with 6 replicates (n = 8): (1) CON group, non-challenged broilers fed basal diet; (2) LPS group, LPS-challenged broilers fed basal diet; (3) AOP group, non-challenged broilers fed basal diet supplemented with 750 mg/kg AOP; (4) LPS+AOP group, LPS-challenged broilers fed basal diet supplemented with 750 mg/kg AOP. The trial included starter phase (d 1-14), stress period Ⅰ (d 15-21), convalescence Ⅰ (d 22-28), stress period Ⅱ (d 29-35) and convalescence Ⅱ (d 36-42). During stress period Ⅰ (on d 15, 17, 19 and 21) and stress period Ⅱ (on d 29, 31, 33 and 35), broilers were injected intra-abdominally either with LPS solution or with an equal amount of sterile saline. The results showed that dietary AOP supplementation alleviated LPS-induced reduction in antioxidant enzyme activity and excessive production of ROS, 8-OHdG and PC in serum of broilers challenged with LPS. Moreover, dietary AOP supplementation alleviated the decrease of T-AOC and activities of SOD, CAT and GPx in liver of broilers challenged with LPS by increasing expression of Nrf2, and inhibiting over-expression of Keap1 both at gene and protein level. Additionally, dietary AOP supplementation decreased the over-production of IL-1β and IL-6 in liver of broilers challenged by LPS through decreasing mRNA expression of TLR4, MyD88, NF-κB P65, IL-1β and IL-6, and alleviating the increase of protein expression of TLR4, IKKβ, NF-κB P65, IL-1β, IL-6, and the decrease of protein expression of IkBα. In conclusion, dietary AOP supplementation could alleviate LPS-induced oxidative stress through Nrf2/Keap1 and TLR4/NF-κB pathway., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

A study was conducted to evaluate the effects of forage-to-concentrate (F:C) ratio and forage particle length (FPL) on intake, duodenal flow, and digestibility of individual AA in the intestine of lactating dairy cows. The experiment was designed as a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments using 4 lactating dairy cows (parity 2) with ruminal and duodenal cannulas. Low (35:65) and high (60:40) F:C ratios (dry matter basis) were combined with 2 FPL of alfalfa silage (short vs. long; 7.9 vs. 19.1 mm). Few interactions between F:C and FPL for duodenal flow and intestinal digestibility of AA occurred, but interactions were detected for intakes of several AA. Intake of essential AA and nonessential AA decreased with increasing F:C, and the intake of several individual AA increased or decreased with increasing FPL. Increasing F:C decreased duodenal flows of essential AA, nonessential AA, and microbial AA due to consistent decreased flows of most individual AA (except Glu). Degradability of most individual AA in the rumen was not affected by F:C ratio or FPL except that the degradability of His was greater with high than low F:C diets, and the degradability of Ser was greater with long versus short FPL diets. However, the degradability of individual AA within diet varied considerably. Overall, F:C ratio and FPL did not affect intestinal digestibility of AA and rumen undegradable protein AA, whereas the digestibility of individual AA in the intestine varied considerably regardless of dietary treatment. These results indicate that increasing F:C ratio decreased AA supply due to decreased flow of AA to the duodenum but altering FPL did not affect AA supply. The results also revealed the necessity to consider both the flows and digestibility of individual AA when optimizing ration formulation to meet AA requirements of dairy cows., (Crown Copyright © 2020. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

The repeated amino-acid sequences in Citrobacter Freundii beta-lactamase may be indispensable for its function, because such repetitions cannot be simply attributed to a chance. In order to fully explore the functional units in Citrobacter Freundii beta-lactamase, it may need to analyse all the amino acid pairs, triplets, etc. along Citrobacter Freundii beta-lactamase from one terminal to the other terminal, to count their frequencies and calculate their probabilities. The amino-acid sequence of Citrobacter Freundii beta-lactamase was counted according to two-, three- and four-amino-acid sequences. The counted frequency and probability were compared with the predicted frequency and probability. The amino acid sequences, which appear in Citrobacter Freundii beta-lactamase and can be predicted from its amino acid composition according to a purely random mechanism, should not be deliberately evolved and conserved. By contrast, the amino acid sequences, which appear in Citrobacter Freundii beta-lactamase but cannot be predicted from its amino acid composition according to a purely random mechanism, should be deliberately evolved and conversed. Accordingly 99 (26.053%) and 33 (8.684%) of 380 two-amino-acid sequences can be predicted by the frequency and probability according to a purely random mechanism. Some kinds of amino acid sequences, which absent in Citrobacter Freundii beta-lactamase and can be predicted from its amino acid composition according to a purely random mechanism, should not be deliberately excluded from Citrobacter Freundii beta-lactamase. By contrast, some kinds of amino acid sequences, which absent in Citrobacter Freundii beta-lactamase and cannot be predicted from its amino acid composition according to a purely random mechanism, should be deliberately excluded from Citrobacter Freundii beta-lactamase. Accordingly 89 (48.370%) and 41 (22.283%) of 184 kinds of absent two-amino-acid sequences can be predicted by the frequency and probability according to a purely random mechanism, and 7236 (99.848%) of 7247 kinds of absent three-amino-acid sequences can be predicted by the frequency according to a purely random mechanism. The amino acids, whose probabilities in following certain preceding amino acids can be predicted from Citrobacter Freundii beta-lactamase amino acid composition according to a purely random mechanism, should not be deliberately evolved and conversed, accordingly 2 (0.526%) of 380 counted first order Markov transition probabilities for the second amino acid in two-amino-acid sequences match the predicted conditional probabilities.