Mulberry Leaf Supplements Effecting Anti-Inflammatory Genes and Improving Obesity in Elderly Overweight Dogs.

Overweight and obesity, associated with various health complications, refer to abnormal or excessive fat accumulation conditions that harm health. Like humans, obesity is a growing problem in dogs, which may increase the risk of serious diseases such as diabetes and cancer. Mulberry leaf has shown potential anti-obesity and anti-diabetes effects in several studies. Our research studied the impact of mulberry leaf supplements in healthy old overweight dogs for 12 weeks. Blood and fecal samples were collected from the dogs before and after treatment for different analyses, including whole transcriptome and gut microbiome analysis. The Body Condition Score (BCS) and blood glucose levels were significantly decreased in all mulberry treatment groups, which justifies the anti-obesity effect of mulberry leaf in dogs. Throughout the whole transcriptome study, the downregulation of PTX3 and upregulation of PDCD-1, TNFRSF1B, RUNX3, and TICAM1 genes in the high mulberry group were found, which have been associated with anti-inflammatory effects in the literature. It may be an essential gene expression mechanism responsible for the anti-inflammatory and, subsequently, anti-obesity effects associated with mulberry leaf treatment, as confirmed by real-time polymerase chain reaction analysis. In microbiome analysis, Papillibacter cinnamivorans, related to the Mediterranean diet, which may cause anti-inflammatory effects, were abundant in the same treatment group. Further studies may be required to establish the gene expression mechanism and role of abundant bacteria in the anti-obesity effect of mulberry supplements in dogs. Overall, we propose mulberry leaves as a portion of food supplements for improving blood glucose levels and the anti-inflammation of blood in companion dogs.

Effect of Organic or Inorganic Mineral Premix in the Diet on Laying Performance of Aged Laying Hens and Eggshell Quality.

In this study, we examined the effect of diets supplemented with organic and inorganic mineral premixes on the laying performance and eggshell quality of aged laying hens. A total of 600 68-week-old Hy-Line Brown laying hens were randomly assigned to 1 of 3 dietary treatments, repeated 5 times: Mash type basal diet, basal diet supplemented with an inorganic mineral premix (1.0 g/kg), and basal diet supplemented with an organic mineral premix (1.8 g/kg). The results showed that eggshell strength was higher (p < 0.01) in the inorganic mineral diet group than in the organic mineral and basal diet groups. Further, the levels of Fe and Mn in the liver were higher (p < 0.05) in the inorganic and organic mineral diet groups than in the basal diet group. The concentrations of Fe and Mg in the spleen were different (p < 0.05) among the treatment groups, with the highest levels reported in the organic mineral premix group. The concentrations of Cu, Zn, and Mn in the eggshell were different (p < 0.05) among the groups, with the highest levels reported in the inorganic and organic mineral premix diet groups. In conclusion, a diet containing organic mineral premix improved eggshell strength and had no detrimental effect on the laying performance of aged laying hens.

Evaluation of Fermented Oat and Black Soldier Fly Larva as Food Ingredients in Senior Dog Diets.

The aim of this study was to evaluate the suitability of fermented oat (FO) and black soldier fly larva (BSFL) as food ingredients for dogs. A total of 20 spayed female dogs were divided into four treatment groups, with 5 dogs per group. The four treatment groups consisted of a control group, a diet with 10% FO, one with 5% BSFL, and one with 10% FO and 5% BSFL, and each experimental food was fed for 12 weeks. The feeding of FO and/or BSFL did not affect the daily food intake, body weight, body condition score, fecal score, or skin condition of the dogs. In all the experimental groups, no significant differences in serum IgG, IL-10, or TNF-α levels were observed upon the feeding of FO and/or BSFL. Some hematological (white blood cell and basophils) and serum biochemical parameters (phosphorous, globulin, and alkaline phosphatase) showed significant differences with FO and/or BSFL feeding compared to the control group, but they were within the normal reference range. No adverse clinical signs related to these parameters being affected by FO and BSFL were observed. The feeding of BSFL for 12 weeks reduced the serum cholesterol level (p < 0.05) at the end of the experiment. Our findings suggest the suitability of FO and BSFL as food materials for dogs.

Molecular cloning and characterization of porcine ribosomal protein L21.

Ribosomal protein L21 (RPL21) is a structural component of the 60S subunit of the eukaryotic ribosome. This protein has an important role in protein synthesis and the occurrence of hereditary diseases. Pig is a common laboratory model, however, to the best of our knowledge, its RPL21 gene has not been cloned to date. In this study, we cloned and identified the full-length sequence of the pig RPL21 gene for the first time. In addition, we examined its expression pattern and function by using overexpression or knockdown approaches. As a result, we obtained a 604 bp segment that contains a 483 bp open reading frame encoding 160 amino acids. The pig RPL21 gene is located in the "+" strand of chromosome 11, which spans 2167 bp from 4199792 to 4201958. Pig RPL21 protein has nine strands and two helices in its secondary structure. Pig RPL21 is predominantly expressed in ovary and lung, at lower levels in kidney, small intestine, and skin, and at the lowest levels in heart and liver. Furthermore, RPL21 expression is closely connected with cell proliferation and cell cycle arrest. The results are intended to provide useful information for the further study of pig RPL21.

Construction of a Dual-Fluorescence Reporter System to Monitor the Dynamic Progression of Pluripotent Cell Differentiation.

Oct4 is a crucial germ line-specific transcription factor expressed in different pluripotent cells and downregulated in the process of differentiation. There are two conserved enhancers, called the distal enhancer (DE) and proximal enhancer (PE), in the 5' upstream regulatory sequences (URSs) of the mouse Oct4 gene, which were demonstrated to control Oct4 expression independently in embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs). We analyzed the URSs of the pig Oct4 and identified two similar enhancers that were highly consistent with the mouse DE and PE. A dual-fluorescence reporter was later constructed by combining a DE-free-Oct4-promoter-driven EGFP reporter cassette with a PE-free-Oct4-promoter-driven mCherry reporter cassette. Then, it was tested in a mouse ESC-like cell line (F9) and a mouse EpiSC-like cell line (P19) before it is formally used for pig. As a result, a higher red fluorescence was observed in F9 cells, while green fluorescence was primarily detected in P19 cells. This fluorescence expression pattern in the two cell lines was consistent with that in the early naïve pluripotent state and late primed pluripotent state during differentiation of mouse ESCs. Hence, this reporter system will be a convenient tool for screening out ESC-like naïve pluripotent stem cells from other metastable state cells in a heterogenous population.