The dietary crude protein level could affect ruminal fermentation parameters and the microflora of ruminants. The present study\'s aim was to investigate the effects of different protein level diets on ruminal morphology, fermentation parameters, digestive enzyme activity, microflora and metabolites of Tibetan sheep. Ninety weaned lambs (initial weight of 15.40 ± 0.81 kg, 2 months old) were selected and randomly divided into three groups (six pens/treatment, five rams/pen). Dietary treatments were formulated with 13.03% (high protein, HP), 11.58% (moderate protein, MP) and 10.20% (low protein, LP), respectively. Compared with LP, both papillae length and papillae width were significantly promoted in HP and MP (p < 0.05). The concentrations of ammonia nitrogen, total VFAs, propionic acids and butyric acids in HP were significantly increased compared to those in MP and LP (p < 0.05). The activities of protease and α-amylase in HP were significantly greater than those of LP (p < 0.05). For the ruminal microbial community, higher proportions of phylum Prevotella 1 and Succiniclasticum and genus Rikenellaceae RC9 gut group and Ruminococcus 1 were observed in HP (p < 0.05). A total of 60 differential metabolites (DMs) (28 up, 32 down) between HP and MP; 73 DMs (55 up, 18 down) between HP and LP; and 65 DMs (49 up, 16 down) between MP and LP were identified. Furthermore, four pathways of the biosynthesis of unsaturated fatty acids, tryptophan metabolism, bile secretion and ABC transporters were significantly different (p < 0.05). The abundance of phylum Prevotella 1 was negatively associated with stearic acid and palmitic acid but positively associated with the taurine. The abundance of genus Ruminococcus 1 was negatively associated with stearic acid, oleic acid, erucic acid, Indole-3-acetamide and palmitic acid but positively associated with 6-hydroxymelatonin. In conclusion, a 13.03% CP level improved ruminal morphology, fermentation parameters and digestive enzyme activities through modulating the microbial community and regulating metabolism in Tibetan sheep.

Head and neck squamous cell carcinomas (HNSCCs) are one of the most frequently detected cancers in the world; not all mechanisms related to the expression of keratin in this type of cancer are known. The aim of this study was to evaluate type II cytokeratins (KRT): KRT6A, KRT6B, and KRT6C protein concentrations in 54 tumor and margin samples of head and neck squamous cell carcinoma (HNSCC). Moreover, we examined a possible association between protein concentration and the clinical and demographic variables. Protein concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Significantly higher KRT6A protein concentration was found in HNSCC samples compared to surgical margins. An inverse relationship was observed for KRT6B and KRT6C proteins. We showed an association between the KRT6C protein level and clinical parameters T and N in tumor and margin samples. When analyzing the effect of smoking and drinking on KRT6A, KRT6B, and KRT6C levels, we demonstrated a statistically significant difference between regular or occasional tobacco and alcohol habits and patients who do not have any tobacco and alcohol habits in tumor and margin samples. Moreover, we found an association between KRT6B and KRT6C concentration and proliferative index Ki-67 and HPV status in tumor samples. Our results showed that concentrations of KRT6s were different in the tumor and the margin samples and varied in relation to clinical and demographic parameters. We add information to the current knowledge about the role of KRT6s isoforms in HNSCC. We speculate that variations in the studied isoforms of the KRT6 protein could be due to the presence and development of the tumor and its microenvironment. It is important to note that the analyses were performed in tumor and surgical margins and can provide more accurate information on the function in normal and cancer cells and regulation in response to various factors.

The Chromosome-Centric Human Proteome Project (C-HPP) aims to identify all proteins encoded by the human genome. Currently, the human proteome still contains approximately 2000 PE2-PE5 proteins, referring to annotated coding genes that lack sufficient protein-level evidence. During the past 10 years, it has been increasingly difficult to identify PE2-PE5 proteins in C-HPP approaches due to the limited occurrence. Therefore, we proposed that reanalyzing massive MS data sets in repository with newly developed algorithms may increase the occurrence of the peptides of these proteins. In this study, we downloaded 1000 MS data sets via the ProteomeXchange database. Using pFind software, we identified peptides referring to 1788 PE2-PE5 proteins. Among them, 11 PE2 and 16 PE5 proteins were identified with at least 2 peptides, and 12 of them were identified using 2 peptides in a single data set, following the criteria of the HPP guidelines. We found translation evidence for 16 of the 11 PE2 and 16 PE5 proteins in our RNC-seq data, supporting their existence. The properties of the PE2 and PE5 proteins were similar to those of the PE1 proteins. Our approach demonstrated that mining PE2 and PE5 proteins in massive data repository is still worthy, and multidata set peptide identifications may support the presence of PE2 and PE5 proteins or at least prompt additional studies for validation. Extremely high throughput could be a solution to finding more PE2 and PE5 proteins.

Two experiments were conducted to determine the effects of different protein levels on the growth performance, feed efficiency and nutritional values, and phase feeding of the 2-spotted cricket (Gryllus bimaculatus de Geer). In experiment 1, 4 crude protein (CP) diets were formulated to contain 18%, 20%, 22%, or 24% CP, respectively. A sample of 7-day-old 3,600 crickets was equally divided into 24 plastic boxes (150 crickets each) in a completely randomized design with 4 diets and 6 replications. In experiment 2, 2-phase feedings were used. For starting period (days 7-18), crickets in all treatments were fed a diet containing 22% CP. During the growing period (days 19-35), 3 groups of crickets were fed diets containing 18%, 20%, and 22% CP. In the overall period of experiment 1, the crickets fed with 22% CP diet had greater body weight compared to those fed with 18% CP diet. In addition, the crickets fed with 22% CP diet had the lowest feed conversion ratio (FCR). The broken-line model indicated the growth pattern changed on day 18. In experiment 2, the crickets fed with 20% CP diet from days 19 to 35 had greater growth performance and lower FCR than those fed with 18% CP, but not different from those fed with 22% CP. In conclusion, 22% CP can increase growth performance by improving the feed efficiency of crickets. The implementation of 2-phase feedings using 20% CP, during the growing period, could be considered as a cost-effective strategy for sustainable cricket production.

We investigated the effects of crude protein (CP) levels and exogenous enzymes on growth performance, meat quality, toxic gas emissions, and colonic microbiota community in 200 finishing pigs. Four groups corresponded to 4 diets: 16.74% CP (high-protein level, HP) and 14.73% CP (medium protein level, MP) diet supplemented with or without 1-g/kg multi-enzymes (ENZs, including 1000-U/kg protease, 2500-U/kg α-amylase, and 10,000-U/kg β-glucanase), using a 2 × 2 factorial arrangement. After 7 weeks of trial, ENZs supplementation increased (P < 0.05) the average daily gain (ADG) of finishing pigs during weeks 4 to 7 and in the overall period and improved gross energy utilization. Dietary HP improved (P < 0.05) ADG during the overall period. The MP diet-treated pigs had higher intramuscular fat (IMF) content in the longissimus dorsi muscle (P < 0.01). ENZs supplementation to the MP diets lowered muscle IMF content (P < 0.01). Additionally, pigs fed the HP diet released (P < 0.05) more NH3 and H2S in excrement. The HP diet enhanced (P < 0.05) intestinal microbial richness, represented by higher observed_ amplicon sequence variants and Chao1. Administration of ENZs to the HP diet increased (P < 0.05) the Shannon and Pielou\'s evenness. Dietary MP promoted Firmicutes proliferation. Supplementary HP diet increased the relative abundances of Spirochaetota, Verrucomicrobiota, Desulfobacterota, and Fibrobacterota (P < 0.05). Supplemental ENZ elevated (P < 0.05) Actinobacteriota and Desulfobacterota abundances. ENZ supplementation to the HP diet increased the abundances of Bacteroidota, Desulfobacterota, and Proteobacteria but lowered their abundances in the MP diet. Taken together, the HP diet or ENZs\' supplements improved growth performance. Although the interaction between CP levels and ENZs had no effect on growth performance, it modulated colonic flora and muscle IMF content.

Protein is an essential macronutrient. The nutritional needs of dietary proteins are met by digestion and absorption in the small intestine. Indigestible proteins are further metabolized in the gut and produce metabolites via protein fermentation. Thus, protein indigestibility exerts a wide range of effects on gut microbiota composition and function. This review aims to discuss protein digestibility, the effects of food factors, such as protein sources, intake level, and amino acid composition, and making meat analogues. Besides, it provides an inventory of antinutritional factors and processing techniques that influence protein digestibility and, consequently, the diversity and composition of intestinal microbiota. Future studies are warranted to understand the implication of plant-based analogues on protein digestibility and gut microbiota and to elucidate the mechanisms concerning protein digestibility to host gut microbiota using various omics techniques.

UNASSIGNED: The crude protein level in the diet will affect the fermentation parameters, microflora, and metabolites in the rumen of ruminants. It is of great significance to study the effect of crude protein levels in supplementary diet on microbial community and metabolites for improving animal growth performance. At present, the effects of crude protein level in supplementary diet on rumen fermentation parameters, microbial community, and metabolites of Jersey-Yak (JY) are still unclear.
UNASSIGNED: The purpose of this experiment was to study the appropriate crude protein level in the diet of JY. The rumen fermentation indexes (volatile fatty acids and pH) were determined by supplementary diets with crude protein levels of 15.16 and 17.90%, respectively, and the microbial community and metabolites of JYs were analyzed by non-target metabonomics and metagenome sequencing technology, and the changes of rumen fermentation parameters, microbial flora, and metabolites in the three groups and their interactions were studied.
UNASSIGNED: The crude protein level in the supplementary diet had significant effects on pH, valeric acid, and the ratio of acetic acid to propionic acid (p < 0.05). The protein level had no significant effect on the dominant microflora at the phylum level (p > 0.05), and all three groups were Bacteroides and Firmicutes. The results of metabolite analysis showed that the crude protein level of supplementary diet significantly affected the metabolic pathways such as Bile secretion and styrene degradation (p < 0.05), and there were different metabolites between the LP group and HP group, and these different metabolites were related to the dominant microbial to some extent. To sum up, in this experiment, the effects of crude protein level in supplementary diet on rumen microorganisms and metabolites of JY and their relationship were studied, which provided the theoretical basis for formulating a more scientific and reasonable supplementary diet in the future.

Recently, homozygous variants in PTPA were identified as the disease cause for two pedigrees with early-onset parkinsonism and intellectual disability. Although the initial link between PTPA and parkinsonism has been established, further replication was still necessary.
To evaluate the genetic role of PTPA in Parkinson\'s disease (PD).
We analyzed rare variants of PTPA in cohorts of Asian and European ancestries (Ncase  = 2743, Ncontrol  = 8177) with whole-exome sequencing, and further explored the functional effect of the target variant.
One patient with early-onset PD from a consanguineous family carried the homozygous variant p.Met329Val, while her parents and elder sister with heterozygous p.Met329Val were healthy. This patient developed minor cognitive decline within 1 year, with a Montreal Cognitive Assessment (MoCA) score dropping from 28 to 25. Functional exploration with overexpression studies suggested that this variant was associated with decreased protein phosphatase 2A (PTPA) protein level by affecting protein stability, but not mRNA expression.
These results have broadened the mutation spectrum of PTPA, and paved the way for further research into the role of PTPA in PD. © 2023 International Parkinson and Movement Disorder Society.

The MAPT gene, encoding the microtubule-associated protein tau on chromosome 17q21.31, is result of an inversion polymorphism, leading to two allelic variants (H1 and H2). Homozygosity for the more common haplotype H1 is associated with an increased risk for several tauopathies, but also for the synucleinopathy Parkinson\'s disease (PD). In the present study, we aimed to clarify whether the MAPT haplotype influences expression of MAPT and SNCA, encoding the protein α-synuclein (α-syn), on mRNA and protein levels in postmortem brains of PD patients and controls. We also investigated mRNA expression of several other MAPT haplotype-encoded genes. Postmortem tissues from cortex of fusiform gyrus (ctx-fg) and of the cerebellar hemisphere (ctx-cbl) of neuropathologically confirmed PD patients (n = 95) and age- and sex-matched controls (n = 81) were MAPT haplotype genotyped to identify cases homozygous for either H1 or H2. Relative expression of genes was quantified using real-time qPCR; soluble and insoluble protein levels of tau and α-syn were determined by Western blotting. Homozygosity for H1 versus H2 was associated with increased total MAPT mRNA expression in ctx-fg regardless of disease state. Inversely, H2 homozygosity was associated with markedly increased expression of the corresponding antisense MAPT-AS1 in ctx-cbl. PD patients had higher levels of insoluble 0N3R and 1N4R tau isoforms regardless of the MAPT genotype. The increased presence of insoluble α-syn in PD patients in ctx-fg validated the selected postmortem brain tissue. Our findings in this small, but well controlled cohort of PD and controls support a putative biological relevance of tau in PD. However, we did not identify any link between the disease-predisposing H1/H1 associated overexpression of MAPT with PD status. Further studies are required to gain a deeper understanding of the potential regulatory role of MAPT-AS1 and its association to the disease-protective H2/H2 condition in the context of PD.

BACKGROUND: Appropriate levels of cholesterol are necessary for the mother and developing fetus, but theirexcess may cause preeclampsia. The ABCA1 transporter mediates the secretion of cholesterol and is highly regulated at the transcriptional level via the nuclear liver X receptors (LXRs).
METHODS: Sixteen preeclamptic and 39 normotensives healthy women with uncomplicated pregnancies were involved in the case-control study. The placental levels of ABCA1, LXRA and LXRB mRNA were quantified by real-time quantitative PCR. The concentrations of ABCA1, LXRA and LXRB proteins from the placenta were determined using an enzyme-linked immunosorbent assay Results: We found in the logistic regression model significantly lower placental expression of LXRB mRNA (crude OR = 0.26, 95% CI: 0.07-0.94, p = 0.040) and LXRA protein level (crude OR = 0.19, 95% CI: 0.05-0.69, p = 0.012) in late-onset preeclamptic women compared to healthy pregnant women. The values remained statistically significant after adjustment for possible confounders.
CONCLUSIONS: Our results suggest that high placenta LXRA mRNA and LXRA protein expression levels decrease the risk of late-onset preeclampsia. These nuclear receptors could play a role in the development of preeclampsia through disturbances of lipid metabolism.