Table_2_Influence on the fermentation quality, microbial diversity, and metabolomics in the ensiling of sunflower stalks and alfalfa.xlsx

With the rapid development of the livestock industry, finding new sources of feed has become a critical issue that needs to be addressed urgently. China is one of the top five sunflower producers in the world and generates a massive amount of sunflower stalks annually, yet this resource has not been effectively utilized. Therefore, in order to tap into the potential of sunflower stalks for animal feed, it is essential to explore and develop efficient methods for their utilization.In this study, various proportions of alfalfa and sunflower straw were co-ensiled with the following mixing ratios: 0:10, 2:8, 4:6, 5:5, 6:4, and 8:2, denoted as A0S10, A2S8, A4S6, A5S5, A6S4, and A8S2, respectively. The nutrient composition, fermentation quality, microbial quantity, microbial diversity, and broad-spectrum metabolomics on the 60th day were assessed. The results showed that the treatment groups with more sunflower straw added (A2S8, A4S6) could start fermentation earlier. On the first day of fermentation, Weissella spp.dominated overwhelmingly in these two groups. At the same time, in the early stage of fermentation, the pH in these two groups dropped rapidly, which could effectively reduce the loss of nutrients in the early stage of fermentation.In the later fermentation period, a declining trend in acetic acid levels was observed in A0S10, A2S8, and A4S6, while no butyric acid production was detected in A0S10 and A2S8 throughout the process. In A4S6, butyric acid production was observed only after 30 days of fermentation. From the perspective of metabolites, compared with sunflower ensiling alone, many bioactive substances such as flavonoids, alkaloids, and terpenes are upregulated in mixed ensiling.</p

Table_3_Influence on the fermentation quality, microbial diversity, and metabolomics in the ensiling of sunflower stalks and alfalfa.xlsx

With the rapid development of the livestock industry, finding new sources of feed has become a critical issue that needs to be addressed urgently. China is one of the top five sunflower producers in the world and generates a massive amount of sunflower stalks annually, yet this resource has not been effectively utilized. Therefore, in order to tap into the potential of sunflower stalks for animal feed, it is essential to explore and develop efficient methods for their utilization.In this study, various proportions of alfalfa and sunflower straw were co-ensiled with the following mixing ratios: 0:10, 2:8, 4:6, 5:5, 6:4, and 8:2, denoted as A0S10, A2S8, A4S6, A5S5, A6S4, and A8S2, respectively. The nutrient composition, fermentation quality, microbial quantity, microbial diversity, and broad-spectrum metabolomics on the 60th day were assessed. The results showed that the treatment groups with more sunflower straw added (A2S8, A4S6) could start fermentation earlier. On the first day of fermentation, Weissella spp.dominated overwhelmingly in these two groups. At the same time, in the early stage of fermentation, the pH in these two groups dropped rapidly, which could effectively reduce the loss of nutrients in the early stage of fermentation.In the later fermentation period, a declining trend in acetic acid levels was observed in A0S10, A2S8, and A4S6, while no butyric acid production was detected in A0S10 and A2S8 throughout the process. In A4S6, butyric acid production was observed only after 30 days of fermentation. From the perspective of metabolites, compared with sunflower ensiling alone, many bioactive substances such as flavonoids, alkaloids, and terpenes are upregulated in mixed ensiling.</p

Table_4_Influence on the fermentation quality, microbial diversity, and metabolomics in the ensiling of sunflower stalks and alfalfa.xlsx

With the rapid development of the livestock industry, finding new sources of feed has become a critical issue that needs to be addressed urgently. China is one of the top five sunflower producers in the world and generates a massive amount of sunflower stalks annually, yet this resource has not been effectively utilized. Therefore, in order to tap into the potential of sunflower stalks for animal feed, it is essential to explore and develop efficient methods for their utilization.In this study, various proportions of alfalfa and sunflower straw were co-ensiled with the following mixing ratios: 0:10, 2:8, 4:6, 5:5, 6:4, and 8:2, denoted as A0S10, A2S8, A4S6, A5S5, A6S4, and A8S2, respectively. The nutrient composition, fermentation quality, microbial quantity, microbial diversity, and broad-spectrum metabolomics on the 60th day were assessed. The results showed that the treatment groups with more sunflower straw added (A2S8, A4S6) could start fermentation earlier. On the first day of fermentation, Weissella spp.dominated overwhelmingly in these two groups. At the same time, in the early stage of fermentation, the pH in these two groups dropped rapidly, which could effectively reduce the loss of nutrients in the early stage of fermentation.In the later fermentation period, a declining trend in acetic acid levels was observed in A0S10, A2S8, and A4S6, while no butyric acid production was detected in A0S10 and A2S8 throughout the process. In A4S6, butyric acid production was observed only after 30 days of fermentation. From the perspective of metabolites, compared with sunflower ensiling alone, many bioactive substances such as flavonoids, alkaloids, and terpenes are upregulated in mixed ensiling.</p

Additional file 1: of Acupuncture as an early treatment for idiopathic sudden sensorineural hearing loss (ISSNHL) patients with flat or high-frequency drop audiograms: study protocol for a randomized controlled trial

SPIRIT 2013 Checklist: Recommended items to address in a clinical trial protocol and related documents*. (DOC 136 kb

Table_1_Influence on the fermentation quality, microbial diversity, and metabolomics in the ensiling of sunflower stalks and alfalfa.xlsx

With the rapid development of the livestock industry, finding new sources of feed has become a critical issue that needs to be addressed urgently. China is one of the top five sunflower producers in the world and generates a massive amount of sunflower stalks annually, yet this resource has not been effectively utilized. Therefore, in order to tap into the potential of sunflower stalks for animal feed, it is essential to explore and develop efficient methods for their utilization.In this study, various proportions of alfalfa and sunflower straw were co-ensiled with the following mixing ratios: 0:10, 2:8, 4:6, 5:5, 6:4, and 8:2, denoted as A0S10, A2S8, A4S6, A5S5, A6S4, and A8S2, respectively. The nutrient composition, fermentation quality, microbial quantity, microbial diversity, and broad-spectrum metabolomics on the 60th day were assessed. The results showed that the treatment groups with more sunflower straw added (A2S8, A4S6) could start fermentation earlier. On the first day of fermentation, Weissella spp.dominated overwhelmingly in these two groups. At the same time, in the early stage of fermentation, the pH in these two groups dropped rapidly, which could effectively reduce the loss of nutrients in the early stage of fermentation.In the later fermentation period, a declining trend in acetic acid levels was observed in A0S10, A2S8, and A4S6, while no butyric acid production was detected in A0S10 and A2S8 throughout the process. In A4S6, butyric acid production was observed only after 30 days of fermentation. From the perspective of metabolites, compared with sunflower ensiling alone, many bioactive substances such as flavonoids, alkaloids, and terpenes are upregulated in mixed ensiling.</p

Analyses of cell viabilities receiving various plasmids expressing PrPs and/or TPPP.

<p>The cell viabilities of HeLa (A) and SHSY5Y (B) cells transfected with various plasmids were measured by a commercially Cell Counting Kit 24 and 48 h after transfection. Cells treated with colchicines (10 µM) were used as controls. The average data of each preparation was calculated based on three independent experiments and represented as mean ± S.D.</p

Morphological assays of the effects of various recombinant TPPP proteins on the fibril formation of synthetic PrP106–126 <i>in vitro</i> with a transmission electronic microscopy.

<p>0.5 mg/ml synthetic peptide PrP106–126 was incubated in the absence (A) or presence of TPPP1–219 (B), TPPP50–219 (C) and TPPP100–219 (D) at 37°C for 72 h, respectively. The molar ratio of PrP106–126 to TPPP was 2∶1. Magnification was 97,000×. Scale bar represented 100 nm.</p

Protein purification of various recombinant TPPP and PrP proteins.

<p>A. 15% SDS-PAGE assay for various purified recombinant TPPP proteins. M: standard molecular weights of protein. B. Western blot assay for various recombinant TPPP proteins with anti-TPPP mAb. C. Western blot assay for various recombinant PrP proteins with different antibodies. His-PrP(23–231) and His-PrP(90–231) were blotted with anti-PrP pAb. GST, GST-PrP(106–126) and GST-PrP(23–90) were blotted with anti-GST mAb. Protein molecular markers are shown on the left.</p

Molecular interactions between various recombinant TPPP and PrP.

<p>A. GST pull-down assay of various recombinant TPPP with His-PrP23–231. The TPPP-PrP complexes were precipitated with glutathione agarose beads. The bound PrP was detected by PrP specific 3F4 mAb. B. Co-immunoprecipitation assay of various TPPPs with His-PrP23–231. The TPPP-PrP complexes were precipitated by anti-TPPP pAb and the bound PrP was detected with PrP specific 3F4 mAb. C. His pull-down assay of various His-tagged PrPs with GST-TPPP1–219. The TPPP-PrP complexes were precipitated with Ni-NTA agarose beads. The bound TPPP was detected by anti-TPPP pAb. D. Co-immunoprecipitation assay of various His-tagged PrPs with GST-TPPP1–219. The TPPP-PrP complexes were precipitated by anti-TPPP pAb and the bound PrP was detected with PrP specific 3F4 mAb. E. Co-immunoprecipitation assay of various GST-tagged PrP with GST-TPPP1–219. The TPPP-PrP complexes were precipitated by anti-TPPP mAb and the bound PrP was detected with anti-PrP pAb. F. Co-immunoprecipitation assay of various PrPs in the context of full-length with TPPP1–219. The TPPP-PrP complexes were captured by anti-PrP pAb and the bound TPPP was detected with anti-TPPP mAb. PrP-input represents the purified PrP23–231 and TPPP-input represents the purified GST-TPPP1–219, directly loaded as controls of Western blots. Protein molecular markers are shown on the left.</p

Morphological analyses of the influences of TPPP and PrP on the structures of microtubule in HeLa cells by a confocal microscopy.

<p>The structures of microtubules of the cells treated with various agents were monitored 48 h post-transfection. Panel A: colchicines (10 µM). Panel B: pcDNA3.1-CytoPrP. Panel C: pcDNA3.1-CytoPrP and pcDNA3.1-TPPP. Panel D: pcDNA3.1. Panel E: pcDNA3.1-PG5. Panel F: pcDNA3.1-TPPP.</p