Isolation and Characterization of the Genes Involved in the Berberine Synthesis Pathway in Asian Blue Cohosh, Caulophyllum robustum.

Caulophyllum robustum, commonly named Asian blue cohosh, is a perennial herb in the family Berberidaceae. It has traditionally been used for folk medicine in China. We isolated berberine from the leaves, stem, roots, and fruits of C. robustum, and this is the first report on berberine in this species. Transcriptome analysis was conducted for the characterization of berberine biosynthesis genes in C. robustum, in which, all the genes for berberine biosynthesis were identified. From 40,094 transcripts, using gene ontology (GO) analysis, 26,750 transcripts were assigned their functions in the categories of biological process, molecular function, and cellular component. In the analysis of genes expressed in different tissues, the numbers of genes in the categories of intrinsic component of membrane and transferase activity were up-regulated in leaves versus stem. The berberine synthesis genes in C. robustum were characterized by phylogenetic analysis with corresponding genes from other berberine-producing species. The co-existence of genes from different plant families in the deepest branch subclade implies that the differentiation of berberine synthesis genes occurred early in the evolution of berberine-producing plants. Furthermore, the copy number increment of the berberine synthesis genes was detected at the species level.

Comparison of bacterial community profiles from large intestine specimens, rectal swabs, and stool samples.

The human gastrointestinal tract contains a complex and dynamic population of microorganisms, known as the gut microbiota. Although interest in the role of the gut microbiota in human health has increased in recent years, there remains no standard sampling protocol for analyzing these organisms. Here, we aimed to characterize the microbial composition of distinct segments of the large intestine and to determine whether rectal swabs are suitable for identifying colon microbiota. A total of 100 participants who underwent screening colonoscopy from October 2019 to October 2020 were included in this study. Large intestinal samples (ascending colon, descending colon, sigmoid colon, and rectum) were aspirated by colonoscopy. Rectal swabs were collected before colonoscopy, and stool samples were collected before patients began colonoscopy preparation. All samples were subjected to 16S ribosomal RNA gene sequencing. We identified differences in the number of phylum-level operational taxonomic units among large intestinal samples, rectal swabs, and stool. Five major phyla were detected in all samples (Firmicutes, Bacteroides, Proteobacteria, Actinobacteria, Fusobacteria), although their relative abundances varied. Notably, we found that the microbial compositions of rectal swabs were most similar to those of the sigmoid colon and rectum, whereas the microbiota in stool were relatively different than those from the large intestine and rectal swabs. Our results reveal the existence of microbial heterogeneity within different large intestinal compartments and further suggest that rectal swabs are an acceptable and practical tool for gut microbiota analysis. KEY POINTS: • Our findings highlight local microbiome variations within different regions of the large intestine. • Stool samples do not appear to fully recapitulate the gut microbiome. • Our data from a large population-based cohort indicate that rectal swabs can be used to study the gut microbiome.

Human act and AR1 sequences differentially regulate murine and human D1A dopamine receptor promoters.

The D1A dopamine receptor gene underlies complex transcriptional regulation in order to achieve the tissue-specific expression. Transcription in the D1A genes proceeds from two distinct promoters utilized for the tissue-specific regulation of these genes. Furthermore, analysis of the human D1A dopamine receptor gene has revealed that the region between nucleotides -1173 and -1136 (ActAR1) of the gene might be important for its neural cell-specific expression. To investigate the function of D1A dopamine receptor promoters in the brain cell-specific expression of transgenes, we analyzed the regulatory patterns of two distinct protein-binding regions of ActAR1, i.e., an Act sequence (-1174/-1154) and an AR1 sequence (-1154/-1136), toward murine and human D1A promoters. Transient expression analyses using various chloramphenicol acetyltransferase constructs revealed that Act could not activate murine or human D1A promoters, and that AR1 could effectively stimulate these promoters in a cell type-non-specific manner. Only ActAR1, a combination of Act and AR1, could activate murine and human D1A promoters in a prominent cell type-specific manner. Abundant protein binding to Act was detected by gel mobility shift assay using nuclear extracts from SK-N-MC, NS20Y, OK, and C6 but faint protein binding using nuclear extracts from HepG2. Furthermore, strong protein binding to AR1 was detected using nuclear extracts from SK-N-MC, NS20Y, HepG2 but faint protein binding from C6 extracts and no detectable protein binding from OK extracts. These observations suggest that the tissue-specific expression of the D1A gene is due, at least in part, to the differential expression of these activator proteins that bind to Act and AR1.