Unambiguous discrimination of all 20 proteinogenic amino acids and their modifications by nanopore.

Natural proteins are composed of 20 proteinogenic amino acids and their post-translational modifications (PTMs). However, due to the lack of a suitable nanopore sensor that can simultaneously discriminate between all 20 amino acids and their PTMs, direct sequencing of protein with nanopores has not yet been realized. Here, we present an engineered hetero-octameric Mycobacterium smegmatis porin A (MspA) nanopore containing a sole Ni2+ modification. It enables full discrimination of all 20 proteinogenic amino acids and 4 representative modified amino acids, Nω,N'ω-dimethyl-arginine (Me-R), O-acetyl-threonine (Ac-T), N4-(ß-N-acetyl-D-glucosaminyl)-asparagine (GlcNAc-N) and O-phosphoserine (P-S). Assisted by machine learning, an accuracy of 98.6% was achieved. Amino acid supplement tablets and peptidase-digested amino acids from peptides were also analyzed using this strategy. This capacity for simultaneous discrimination of all 20 proteinogenic amino acids and their PTMs suggests the potential to achieve protein sequencing using this nanopore-based strategy.

Supplemental dietary genistein improves the laying performance and antioxidant capacity of Hy-Line brown hens during the late laying period.

The present study evaluated the effects of 3 supplemental levels of dietary genistein ingested during the late laying period (66-73 wk) of laying hens. A total of 384 Hy-Line brown hens (66 wk old) were randomly divided into 4 groups (6 replicates of 16 hens in each group), the basal diet group (CON), and groups for the basal diet supplemented with 80, 120, and 400 mg/kg of genistein, G1, G2, and G3, respectively. The results of the present study showed an increased laying rate in groups G2 and G3 (linear, P < 0.01), and decreased feed-egg ratios (linear, P < 0.05) and broken egg rate (P < 0.01) in all genistein-treated groups compared with the CON group. Moreover, the G2 group showed an increase in eggshell strength and ratio (linear, P < 0.05), whereas all genistein-treated groups saw a decrease in the L* value (linear, P < 0.01) and an increase in the a* value (linear, P < 0.05) compared with the CON group. Additionally, all genistein-treated groups had an increase in the total antioxidant capacity of plasma (linear, P < 0.05), along with reduced plasma, ovarian, and yolk malondialdehyde levels (linear, P < 0.05), compared with the CON group. The G2 group had an increase in both the superoxide dismutase activity of plasma (P < 0.01) and the total antioxidant capacity of the ovaries (linear, P < 0.05), compared with the CON group. The G3 group had an increase in both the glutathione peroxidase concentration of plasma (linear, P < 0.05) and the total antioxidant capacity of the ovaries (linear, P < 0.01), compared with the CON group. The transcript levels of nuclear factor erythroid 2-related factor 2, superoxide dismutase 1, and catalase were increased in all of the genistein-treated groups (P < 0.05) compared with the CON group, whereas heme oxygenase 1 and glutamate-cysteine ligase modifier subunit were increased only in the G2 group (P < 0.05). In conclusion, supplementation with 120 mg/kg dietary genistein had the best effect on improving the laying rate, eggshell quality, and antioxidant capacity in Hy-Line brown hens during the late laying period.

Effects of dietary N-carbamylglutamate supplementation on growth performance, tissue development and blood parameters of yellow-feather broilers.

The effects of N-carbamylglutamate (NCG) on the growth performance, tissue development, and blood parameters of broilers are unknown. In this study, 2 linked experiments were conducted to investigate the effects of 4 graded dietary levels and 3 dietary stages of NCG in a Chinese indigenous yellow-feather broiler breed during 2 growth phases: 1 to 18 d and 19 to 36 d. The dietary levels of NCG were 0.05%, 0.10%, 0.15%, and 0.20%, and dietary stages were designed to add NCG during the starter stage or grower stage or throughout the experimental period. At the age of 18 d, graded doses of NCG from 0.05 to 0.20% in the diet produced quadratic (P < 0.05) positive responses in body weight, width of intermuscular fat cingulum, liver weight, serum blood urea nitrogen, and serum low-density lipoprotein as well as linear (P < 0.05) positive responses in albumin serum concentration. The average feed per gain and mortality were unaffected by dietary NCG levels. Among 3 dietary treatments, only NCG dietary treatments throughout the experimental period improved the body weight and daily weight gain linearly (P < 0.05). The daily weight gain under the 3 dietary treatments used indicated that the most fitting dose is 0.1% NCG among the 4 dietary levels of NCG (P < 0.05). At this dose, muscle weight increased, whereas subcutaneous adipose as well as the serum contents of uric acid, triglyceride, and albumin decreased. Considering the growth performance and tissue development under the conditions used in this study, the best-fit model for NCG requirements of Chinese yellow-feather broilers was estimated from regression analysis to be 0.09 to 0.12% dietary NCG treatments during the grower stage. The modified blood parameters indicated that NCG dietary effects on broiler growth may be accompanied by modified homeostasis of arginine metabolism, lipid deposition, protein synthesis, and immune response.

Comprehensive evaluation of the role of soy and isoflavone supplementation in humans and animals over the past two decades.

Soy and soy-based foods are considered healthy, particularly in many Asia-Pacific countries, where soy products have long been consumed. Soy and soy-related products have been found to help prevent the occurrence of cardiovascular diseases and certain types of cancer, such as breast and prostate cancer. These products can also have antioxidative effects that alleviate hot flashes during menopause and bone loss. These biological and therapeutic functions are primarily due to the isoflavones derived from soy, whose structure is similar to the structure of 17-ß-oestradiol. Despite the many health benefits for humans and animals, the application of isoflavones remains controversial because of their anti-oestrogenic properties. We focused on general information regarding isoflavones, as well as their structure, function, and application. We summarized evidence showing that dietary or supplemental isoflavones exert protective effects on the health of humans and animals. Based on the literature, we conclude that soy foods and isoflavones may be effective and safe; however, more high-quality trials are needed to fully substantiate their potential use.