Isolation and identification of a cellulolytic bacterium from the Tibetan pig's intestine and investigation of its cellulase production

Background: The Tibetan pig is a pig breedwith excellent grazing characteristics indigenous to the Qinghai\u2013Tibet plateau in China. Under conditions of barn feeding, 90% of its diet consists of forage grass, which helps meet its nutritional needs. The present study aimed to isolate and identify a cellulolytic bacterium from the Tibetan pig's intestine and investigate cellulase production by this bacterium. The study purpose is to provide a basic theory for the research and development of herbivore characteristics and to identify a source of probiotics from the Tibetan pig. Results: A cellulolytic bacterium was isolated from a Tibetan pig's intestine and identified based on morphological, physiological, and biochemical characteristics as well as 16S rRNA analysis; it was designated Bacillus subtilis BY-2. Examination of its growth characteristics showed that its growth curve entered the logarithmic phase after 8\u201312 h and the stable growth phase being between 20 and 40 h. The best carbon source for fermentation was 1% corn flour, while 2% peptone and yeast powder compound were the best nitrogen sources. The initial pH during fermentation was 5.5, with 4% inoculum, resulting in a high and stable amount of enzyme in 24\u201348 h. Conclusions: The isolated BY-2 strain rapidly grew and produced cellulase.We believe that BY-2 cellulase can help overcome the shortage of endogenous animal cellulase, improve the utilization rate of roughage, and provide strain sources for research on porcine probiotics

Truncating Mutation in the Autophagy Gene \u3cem\u3eUVRAG\u3c/em\u3e Confers Oncogenic Properties and Chemosensitivity in Colorectal Cancers

Autophagy-related factors are implicated in metabolic adaptation and cancer metastasis. However, the role of autophagy factors in cancer progression and their effect in treatment response remain largely elusive. Recent studies have shown that UVRAG, a key autophagic tumour suppressor, is mutated in common human cancers. Here we demonstrate that the cancer-related UVRAG frameshift (FS), which does not result in a null mutation, is expressed as a truncated UVRAGFS in colorectal cancer (CRC) with microsatellite instability (MSI), and promotes tumorigenesis. UVRAGFS abrogates the normal functions of UVRAG, including autophagy, in a dominant-negative manner. Furthermore, expression of UVRAGFS can trigger CRC metastatic spread through Rac1 activation and epithelial-to-mesenchymal transition, independently of autophagy. Interestingly, UVRAGFS expression renders cells more sensitive to standard chemotherapy regimen due to a DNA repair defect. These results identify UVRAG as a new MSI target gene and provide a mechanism for UVRAG participation in CRC pathogenesis and treatment response

Manufacturing Process Improvement for Automotive Supplier

Tato diplomová práce je zaměřena na zlepšení výrobního procesu v automobilovém průmyslu ve společnosti Brose CZ, spol. s r. o. V teoretické části jsou vysvětleny základní pojmy a metody z oblasti kvality, následně je provedena analýza vybraného procesu. Na základě poznatků jsou navržena řešení sloužící k eliminaci případně k odstranění nedostatků.This thesis is focused on „Manufacturing Process Improvement for Automotive Supplier“ in company Brose CZ s. r. o. Theoretical part covers the basic term, methods from the field of quality. Next part explores the analysis of the selected process. There are proposed corrective and preventive action based on the discovered findings.152 - Katedra podnikohospodářskávýborn

Cellulose Nanocrystals Prepared by Persulfate One-Step Oxidation of Bleached Bagasse Pulp

This article describes a novel one-step method to prepare cellulose nanocrystals (CNCs) from bleached bagasse pulp via ammonium persulfate (APS) oxidation. The obtained persulfate oxidation cellulose nanocrystals (POCNs) were characterized for their microstructure, crystal properties, and chemical composition. The POCNs were successfully prepared with a total yield of 44.6%. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) measurements indicated that the POCNs had an average length of 150 to 300 nm and an average width of 10 to 30 nm, as well as a rod-like morphology. Fourier transform infrared (FTIR) spectroscopy confirmed the introduction of carboxyl groups on the surface of cellulose. The X-ray diffraction (XRD) spectra proved the existence of cellulose type I, with a highly crystalline nature (79.2%), and thermogravimetric analysis (TGA) showed that the thermal stability decreased

Study of Extraction and Enzymatic Properties of Cell-Envelope Proteinases from a Novel Wild Lactobacillus plantarum LP69

Lactobacilli cell-envelope proteinases (CEPs) have been widely used in the development of new streams of blockbuster nutraceuticals because of numerous biopharmaceutical potentials; thus, the development of viable methods for CEP extraction and the improvement of extraction efficiency will promote their full-scale application. In this study, CEP from a novel wild Lactobacillus plantarum LP69 was released from cells by incubating in calcium-free buffer. The extraction conditions of CEP were optimized by response surface methodology with the enzyme activity and specific activity as the detective marker. The optimal extraction conditions were: time of 80 min, temperature of 39 °C and buffer pH of 6.5. Under these conditions, enzyme activity and specific activity were (23.94 ± 0.86) U/mL and (1.37 ± 0.03) U/mg, respectively, which were well matched with the predicted values (22.12 U/mL and 1.36 U/mg). Optimal activity of the crude CEP occurred at pH 8.0 and 40 °C. It is a metallopeptidase, activated by Ca2+, inhibited by Zn2+ and ethylene-diamine-tetra-acetic acid, and a serine proteinase which is inhibited by phenylmethylsulfonyl fluoride. Kinetic studies showed that CEP from LP69 could hydrolyze whey protein, lactoglobulin and casein. Our study improves the extraction efficiency of CEPs from LP69, providing the reference for their industrial development

LncRNA Neat1 targets NonO and miR-128-3p to promote antigen-specific Th17 cell responses and autoimmune inflammation

Abstract Long non-coding RNAs (lncRNAs) interaction with RNA-Binding proteins (RBPs) plays an important role in immunological processes. The generation of antigen-specific Th17 cells is closely associated with autoimmune pathogenesis. However, the function of lncRNA-RBP interactions in the regulation of pathogenic Th17 cell responses during autoimmunity remains poorly understood. Here, we found that lncRNA Neat1, highly expressed in Th17 cells, promoted antigen-specific Th17 cell responses. Both global and CD4+ T cell-specific knockdown of Neat1 protected mice against the development of experimental autoimmune uveitis (EAU). Mechanistically, Neat1 regulated RNA-Binding protein NonO, thus relieving IL-17 and IL-23R from NonO-mediated transcriptional repression and supporting antigen-specific Th17 cell responses. In addition, Neat1 also modulated miR-128-3p/NFAT5 axis to increase the expression of IL-17 and IL-23R, leading to augmented Th17 cell responses. Our findings elucidate a previously unrecognized mechanistic insight into the action of Neat1 in promoting antigen-specific Th17 responses and autoimmunity, and may facilitate the development of therapeutic targets for T cell-mediated autoimmune diseases

Isolation and identification of a cellulolytic bacterium from the Tibetan pig's intestine and investigation of its cellulase production

Background: The Tibetan pig is a pig breed with excellent grazing characteristics indigenous to the Qinghai–Tibet plateau in China. Under conditions of barn feeding, 90% of its diet consists of forage grass, which helps meet its nutritional needs. The present study aimed to isolate and identify a cellulolytic bacterium from the Tibetan pig's intestine and investigate cellulase production by this bacterium. The study purpose is to provide a basic theory for the research and development of herbivore characteristics and to identify a source of probiotics from the Tibetan pig. Results: A cellulolytic bacterium was isolated from a Tibetan pig's intestine and identified based on morphological, physiological, and biochemical characteristics as well as 16S rRNA analysis; it was designated Bacillus subtilis BY-2. Examination of its growth characteristics showed that its growth curve entered the logarithmic phase after 8–12 h and the stable growth phase being between 20 and 40 h. The best carbon source for fermentation was 1% corn flour, while 2% peptone and yeast powder compound were the best nitrogen sources. The initial pH during fermentation was 5.5, with 4% inoculum, resulting in a high and stable amount of enzyme in 24–48 h. Conclusions: The isolated BY-2 strain rapidly grew and produced cellulase. We believe that BY-2 cellulase can help overcome the shortage of endogenous animal cellulase, improve the utilization rate of roughage, and provide strain sources for research on porcine probiotics

Detection and comparison of microRNAs in the caprine mammary gland tissues of colostrum and common milk stages

Abstract Background MicroRNAs (miRNAs) have a great influence on various physiological functions. A lot of high-throughput sequencing (HTS) research on miRNAs has been executed in the caprine mammary gland at different lactation periods (common milk lactation and dry period), but little is known about differentially expressed miRNAs in the caprine mammary gland of colostrum and peak lactation periods. Result This study identified 131 differentially expressed miRNAs (P  1 or log2 colostrum NE/peak lactation NE  1 or log2 colostrum NE/peak lactation NE < −1). The expressions of 10 randomly selected miRNAs was analyzed through stem-loop real-time quantitative PCR (RT-qPCR). Their expression patterns were the same with Solexa sequencing results. Pathway analysis suggested that oestrogen, endocrine, adipocytokine, oxytocin and MAPK signalling pathways act on the development of mammary gland and milk secretion importantly. In addition, the miRNA-target-network showed that the bta-miR-574 could influence the development of mammary gland and lactation by leptin receptor (LEPR), which was in the adipocytokine signalling pathway. Chr5_3880_mature regulated mammary gland development and lactation through Serine/threonine-protein phosphatase (PPP1CA), which was in the oxytocin signalling pathway. Conclusions Our finding suggested that the profiles of miRNAs were related to the physiological functions of mammary gland in the colostrum and peak lactation periods. The biological features of these miRNAs may help to clarify the molecular mechanisms of lactation and the development of caprine mammary gland