Identification of genes related to growth and amino acid metabolism from the transcriptome profile of the liver of growing laying hens.

The growing period is a critical period for the growth and development of hens and affects their production performance during the laying period. During the early stage of growing, bone and muscle growth is rapid, making it necessary to provide sufficient amino acids (AA) to support the growth and development of laying hens. In this experiment, RNA-Sequencing (RNA-Seq) was applied to compare the liver tissues from 6- to 12-wk-old growing laying hens to identify candidate genes related to growth and AA transport and metabolism. In the liver tissues, 596 differentially expressed genes (DEG) were identified, of which 424 genes were up-regulated and 172 were down-regulated. Through enrichment analysis and DEGs analysis, some DEGs and pathways related to AA transport and metabolism were identified. Additionally, there were significantly increased activities in the liver of glutamate dehydrogenase (GDH), glutamic oxaloacetic transaminase (GOT), and glutamate pyruvate transaminase (GPT). Meanwhile, the level of serum insulin-like growth factor binding protein-5 (IGFBP-5) significantly elevated, and insulin-like growth factor-1 (IGF-1) levels significantly reduced at 12 wk compared to 6 wk. The AA contents in the breast muscle were not significantly altered, while the levels of the free AA in the serum underwent significant changes. This study discovered that the transport and metabolism of AAs in growing laying hens at different ages changed, which influenced the growth and development of growing laying hens. This contributes to future research on the mechanisms of growth and AA metabolism during the growing period of laying hens.

Nanosensor Chemical Cytometry: Advances and Opportunities in Cellular Therapy and Precision Medicine.

With the definition of therapeutics now encompassing transplanted or engineered cells and their molecular products, there is a growing scientific necessity for analytics to understand this new category of drugs. This Perspective highlights the recent development of new measurement science on label-free single cell analysis, nanosensor chemical cytometry (NCC), and their potential for cellular therapeutics and precision medicine. NCC is based on microfluidics integrated with fluorescent nanosensor arrays utilizing the optical lensing effect of a single cell to real-time extract molecular properties and correlate them with physical attributes of single cells. This new class of cytometry can quantify the heterogeneity of the multivariate physicochemical attributes of the cell populations in a completely label-free and nondestructive way and, thus, suggest the vein-to-vein conditions for the safe therapeutic applications. After the introduction of the NCC technology, we suggest the technological development roadmap for the maturation of the new field: from the sensor/chip design perspective to the system/software development level based on hardware automation and deep learning data analytics. The advancement of this new single cell sensing technology is anticipated to aid rich and multivariate single cell data setting and support safe and reliable cellular therapeutics. This new measurement science can lead to data-driven personalized precision medicine.

Effects of Methyl Sulfonyl Methane and Selenium Yeast on Fatty Liver Syndrome in Laying Hens and Their Biological Mechanisms.

The purpose of this study was to explore the effects of MSM and Se-Y on FLS in laying hens during the late peak laying period and the underlying biological mechanisms. Therefore 240 55-week-old Jing-fen No. 6 laying hens were randomly divided into five groups, with eight replicates in each group and six laying hens in each replicate. The hens were fed a basal diet (Control) and diets supplemented with 350 and 700 mg/kg MSM and 25 and 50 mg/kg Se-Y, respectively, for four weeks. The results showed that MSM and Se-Y had no significant effects on the performance of laying hens. With the increasing dosage of MSM and Se-Y, the symptoms of liver steatosis in laying hens were reduced, and MSM and Se-Y could significantly reduce the content of malondialdehyde (MDA) in serum and liver (p < 0.05) and increase the contents of total superoxide dismutase (T-SOD) and glutathione peroxidase (GPX) in serum and liver (p < 0.05). The RNA-seq results showed that 700 mg/kg MSM significantly downregulated the expression levels of the ATP5I, ATP5G1, CYCS, and UQCRQ genes in the liver, and 50 mg/kg Se-Y significantly downregulated the expression levels of MAPK10, SRC, BMP2, and FGF9 genes in the liver. In conclusion, dietary supplementation with MSM and Se-Y can effectively reduce the FLS of laying hens in the late peak laying period and increase their antioxidant capacity. The underlying biological mechanism may be related to the downregulation of genes involved in liver oxidative phosphorylation and inflammation-related pathways.

Fatty Liver Disease Caused by High-Alcohol-Producing Klebsiella pneumoniae.

The underlying etiology of nonalcoholic fatty liver disease (NAFLD) is believed to be quite varied. Changes in the gut microbiota have been investigated and are believed to contribute to at least some cases of the disease, though a causal relationship remains unclear. Here, we show that high-alcohol-producing Klebsiella pneumoniae (HiAlc Kpn) is associated with up to 60% of individuals with NAFLD in a Chinese cohort. Transfer of clinical isolates of HiAlc Kpn by oral gavage into mice induced NAFLD. Likewise, fecal microbiota transplant (FMT) into mice using a HiAlc-Kpn-strain-containing microbiota isolated from an individual with NASH induced NAFLD. However, selective elimination of the HiAlc Kpn strain before FMT prevented NAFLD in the recipient mice. These results suggest that at least in some cases of NAFLD an alteration in the gut microbiome drives the condition due to excess endogenous alcohol production.

Effects of a Lactulose-Rich Diet on Fecal Microbiome and Metabolome in Pregnant Mice.

Lactulose, a safe and beneficial molecule, can be used in food as a prebiotic and as an osmotic laxative during pregnancy. This work evaluated the effects of dietary lactulose on the gut microenvironment of pregnant mice using the fecal microbiota and metabolomic profiling. After 2 weeks of feeding, the Bifidobacterium and Bacteroides abundances in the mouse feces were significantly increased in the LAC-high (the diet supplemented with 15% lactulose) group. A total of 15 metabolites, including 1-monoolein, glucose-6-phosphate, and short-chain fatty acids, were increased significantly in the LAC-high group. The serum glucose and total cholesterol concentrations were significantly decreased, while the progesterone level was significantly increased in the lactulose-fed mice. In the LAC-high group, the colonic pH and intestinal permeability were decreased, while the immunoglobulins in the colonic epithelial cells and the small intestinal absorption capacity were significantly increased. These findings indicated that lactulose supplementation benefitted pregnancy performance in mice.

Characterizing the Biology of Lytic Bacteriophage vB_EaeM_φEap-3 Infecting Multidrug-Resistant Enterobacter aerogenes.

Carbapenem-resistant Enterobacter aerogenes strains are a major clinical problem because of the lack of effective alternative antibiotics. However, viruses that lyze bacteria, called bacteriophages, have potential therapeutic applications in the control of antibiotic-resistant bacteria. In the present study, a lytic bacteriophage specific for E. aerogenes isolates, designated vB_EaeM_φEap-3, was characterized. Based on transmission electron microscopy analysis, phage vB_EaeM_φEap-3 was classified as a member of the family Myoviridae (order, Caudovirales). Host range determination revealed that vB_EaeM_φEap-3 lyzed 18 of the 28 E. aerogenes strains tested, while a one-step growth curve showed a short latent period and a moderate burst size. The stability of vB_EaeM_φEap-3 at various temperatures and pH levels was also examined. Genomic sequencing and bioinformatics analysis revealed that vB_EaeM_φEap-3 has a 175,814-bp double-stranded DNA genome that does not contain any genes considered undesirable for the development of therapeutics (e.g., antibiotic resistance genes, toxin-encoding genes, integrase). The phage genome contained 278 putative protein-coding genes and one tRNA gene, tRNA-Met (AUG). Phylogenetic analysis based on large terminase subunit and major capsid protein sequences suggested that vB_EaeM_φEap-3 belongs to novel genus "Kp15 virus" within the T4-like virus subfamily. Based on host range, genomic, and physiological parameters, we propose that phage vB_EaeM_φEap-3 is a suitable candidate for phage therapy applications.

Targeting the Gut Microbiota to Investigate the Mechanism of Lactulose in Negating the Effects of a High-Salt Diet on Hypertension.

SCOPE: High-salt diets (HSDs) are widely considered to cause health problems such as gut microecological imbalances, constipation, and hypertension. This study explores how lactulose as a safe molecule can stimulate bodily responses to alleviate salt-sensitive hypertension by regulating the gut microbiotas of HSD-fed mice. METHODS AND RESULTS: After 4 weeks, the blood pressures of mice fed a high-salt plus lactulose diet (HSLD) are significantly lower than those of the HSD-fed mice. The HSD increases the abundances of Alistipes and Ruminococcaceae_UCG_009 and reduced the abundance of Lactobacillus in the gut, while lactulose supplementation increases the abundances of Bifidobacterium, Alloprevotella, and Subdoligranulum. Fecal metabolic profiling shows significant increases in metabolites involved in ATP-binding cassette transporter pathways, and tryptophan metabolism is significantly reduced in the HSLD group compared with the HSD group. Lactulose maintains the intestinal microenvironmental health in the HSD-fed mice by improving glycolipid metabolism, decreasing the small intestinal interleukin-17a (IL-17a) and interleukin-22 (IL-22) mRNA levels and serum IL-17a and IL-22 levels, relieving constipation, increasing fecal sodium, and reducing intestinal permeability. CONCLUSION: Lactulose negates salt-sensitive hypertension. Regulating the gut microbiota is a potential treatment for salt-sensitive hypertension.

Identification and molecular characterization of Serratia marcescens phages vB_SmaA_2050H1 and vB_SmaM_2050HW.

Serratia marcescens is a rod-shaped, Gram-negative bacterium causing nosocomially acquired infections. Bacteriophages are natural opponents of their pathogenic bacterial hosts and could be an alternative to traditional antibiotic treatments. In this study, two S. marcescens-specific bacteriophages, vB_SmaA_2050H1 and vB_SmaM_2050HW, were isolated from two different waste samples in China. Phage plaque assays, transmission electron microscopy, host-range determination, and one-step growth curve analyses were performed for both phages. vB_SmaA_2050H1 was classified as belonging to the family Ackermannviridae, and vB_SmaM_2050HW was classified as belonging to the family Myoviridae. One-step growth curve analysis showed that the latent and rise period of vB_SmaA_2050H1 were 80 min and 50 min, respectively, with a burst size of approximately 103 phage particles per infected cell. For vB_SmaM_2050HW, latent and rise periods of 40 min and 60 min, respectively, were determined, with a burst size of approximately 110 phage particles per infected cell. vB_SmaA_2050H1 infected 10 of the 15 (66.67%) S. marcescens strains tested, while vB_SmaM_2050HW infected 12 (80%) of the strains. Whole-genome sequencing and annotation of each of the phage genomes revealed genome sizes of 159,631 bp and 276,025 bp for vB_SmaA_2050H1 and vB_SmaM_2050HW, respectively, with the respective genomes containing 213 and 363 putative open reading frames. Sequence analysis of the genomes revealed that vB_SmaA_2050H1 is a member of the ViI-like family, while vB_SmaM_2050HW is a novel virulent bacteriophage. These findings provide further insights into the genomic structures of S. marcescens bacteriophages.

Characterization of Tail Sheath Protein of N4-Like Phage phiAxp-3.

Achromobacter phage phiAxp-3, an N4-like bacteriophage, specifically recognize Achromobacter xylosoxidans lipopolysaccharide (LPS) as its receptor. PhiAxp-3 tail sheath protein (TSP, ORF69) shares 54% amino acid sequence identity with the TSP of phage N4 (gp65); the latter functions as a receptor binding protein and interacts with the outer membrane receptor NfrA of its host bacterium. Thus, we hypothesized that ORF69 is the receptor-binding protein of phiAxp-3. In the present study, a series of ORF69 truncation variants was constructed to identify the part(s) of this protein essential for binding to A. xylosoxidans LPS. Phage adsorption and enzyme-linked immunosorbent assay showed that amino acids 795-1195 of the TSP, i.e., ORF69(795-1195), are sufficient and essential for receptor and binding. The optimum temperature and pH for the functions of ORF69 and ORF69(795-1195) are 4/25°C and 7, respectively. In vitro cytotoxicity assays showed that ORF69 and ORF69(795-1195) were respectively toxic and non-toxic to a human immortalized normal hepatocyte cell line (LO2; doses: 0.375-12 µg). The potential of this non-toxic truncated version of phiASP-3 TSP for clinical applications is discussed.