Additional files 1: of N-carbamylglutamate restores nitric oxide synthesis and attenuates high altitude-induced pulmonary hypertension in Holstein heifers ascended to high altitude

Table S1. Ingredients and chemical composition of the total mixed ration. (DOCX 14 kb

Immunization and antibody detection in the serum of SF and WL.

<p>Vaccines, injection routes, doses, and sample times are listed (a). Lower antibody levels against H5N1 Re4 were detected in the serum of SF, and obvious differences were calculated between SF and WL in females at 6 and 10 weeks of age (b). Antibody level against infectious bursal disease virus (IBDV) was low in the serum of SF, but no difference was detected between the SF and WL (c). *<i>P</i> < 0.05.</p

Data_Sheet_1_Transcriptomic and metabolomic analysis reveals the influence of carbohydrates on lignin degradation mediated by Bacillus amyloliquefaciens.zip

IntroductionLigninolytic bacteria can secrete extracellular enzymes to depolymerize lignin into small-molecular aromatics that are subsequently metabolized and funneled into the TCA cycle. Carbohydrates, which are the preferred carbon sources of bacteria, influence the metabolism of lignin-derived aromatics through bacteria.MethodsIn this study, untargeted metabolomics and transcriptomics analyses were performed to investigate the effect of carbohydrates on lignin degradation mediated by Bacillus amyloliquefaciens MN-13, a strain with lignin-degrading activity that was isolated in our previous work.ResultsThe results demonstrated that the cell growth of the MN-13 strain and lignin removal were promoted when carbohydrates such as glucose and sodium carboxymethyl cellulose were added to an alkaline lignin-minimal salt medium (AL-MSM) culture. Metabolomics analysis showed that lignin depolymerization took place outside the cells, and the addition of glucose regulated the uptake and metabolism of lignin-derived monomers and activated the downstream metabolism process in cells. In the transcriptomics analysis, 299 DEGs were screened after 24 h of inoculation in AL-MSM with free glucose and 2 g/L glucose, respectively, accounting for 8.3% of the total amount of annotated genes. These DEGs were primarily assigned to 30 subcategories, including flagellar assembly, the PTS system, RNA degradation, glycolysis/gluconeogenesis, the TCA cycle, pyruvate metabolism, and tryptophan metabolism. These subcategories were closely associated with the cell structure, generation of cellular energy, and precursors for biosynthetic pathways, based on a − log 10 (P adjust) value in the KEGG pathway analysis.ConclusionIn summary, the addition of glucose increased lignin degradation mediated by the MN-13 strain through regulating glycolysis, TCA cycle, and central carbon metabolism.</p

CD3⁺ cells in the spleen and thymus of SF and WL.

<p>(A) CD3⁺ cells were observed in the spleen (a, b) and thymus (e, f) of SF and in the spleen (c, d) and thymus (g, h) of WL. Scale bar = 100 μm. (B) There were significant differences during early development between the numbers of CD3⁺ cells in the spleen and thymus of SF and WL. *<i>P</i> < 0.05, **<i>P</i> < 0.01.</p

TUNEL analysis of apoptosis in SF and WL.

<p>(A) Apoptotic cells were observed in the bursa of Fabricius (a) and thymus (b) of SF, and the bursa of Fabricius (c) and thymus (d) of WL. Scale bar = 100 μm. (B) There were significant differences between the numbers of apoptotic cells in the spleen, thymus, and bursa of Fabricius in SF, compared with those in WL. **<i>P</i><0.01.</p

Melanocyte morphology.

<p>Numerous melanocytes (indicated by a blue arrow), with the appearance of dendritic cells and with round nuclei and abundant melanin, were observed in the skin (a), testis (b), thymus (c), and ovary (d). Macrophages in the thymus were stained by 3, 4-dihydroxy-l-phenylalanine (DOPA; c, green arrow). Scale bar = 100 μm.</p

Organ indices and immune gene expression in the F2 population.

<p>(a) Lower indices of spleen, thymus, and bursa of Fabricius were detected in Black-boned chickens, with a significance difference (<i>P</i> < 0.05) in the thymus. (b) Lower expression levels of <i>IFN-γ</i> and <i>IL-4</i> were detected in Black-boned chickens, compared with those in non-Black-boned chickens (<i>P</i> < 0.05). Higher expression level of <i>GAL-7</i> was detected in Black-boned chickens, compared with that in non-Black-boned chickens, but the difference was not statistically significant. B, Black-boned chicken. Non B, non-Black-boned chicken.</p

Expression of <i>IFN-γ</i>, <i>IL-4</i> and <i>GAL-7</i> in the spleen of SF and WL.

<p>Lower expression levels of <i>IFN-γ</i> (a) and <i>IL-4</i> (b) were detected in SF at 10 and 6 weeks of age, respectively, but higher expression levels were detected in SF at 23 weeks of age. High expression level of <i>GAL</i>-7 was detected in SF before 6 weeks of age, but low expression levels were detected at 10 and 23 weeks of age. (c). *<i>P</i> < 0.05, **<i>P</i> < 0.01.</p

Bu-1⁺ cells in the spleen, thymus, and bursa of Fabricius of SF and WL.

<p>(A) Bu-1⁺ cells were observed in the spleen (a, b), thymus (e, f), and bursa of Fabricius (i, j) of SF and in the spleen (c, d), thymus (g, h), and bursa of Fabricius (k, l) of WL. Scale bar = 100 μm. (B) There were significant differences in the numbers of Bu-1⁺ cells in the spleen, thymus, and bursa of Fabricius of SF, compared with those in WL, during early development. *<i>P</i> < 0.05, **<i>P</i><0.01.</p