Heterologous and High Production of Ergothioneine in Escherichia coli

Ergothioneine (ERG) is a histidine-derived thiol compound suggested to function as an antioxidant and cytoprotectant in humans. Therefore, experimental trials have been conducted applying ERG from mushrooms in dietary supplements and as a cosmetic additive. However, this method of producing ERG is expensive; therefore, alternative methods for ERG supply are required. Five Mycobacterium smegmatis genes, egtABCDE, have been confirmed to be responsible for ERG biosynthesis. This enabled us to develop practical fermentative ERG production by microorganisms. In this study, we carried out heterologous and high-level production of ERG in Escherichia coli using the egt genes from M. smegmatis. By high production of each of the Egt enzymes and elimination of bottlenecks in the substrate supply, we succeeded in constructing a production system that yielded 24 mg/L (104 μM) secreted ERG

Adsorption of Urinary Proteins on the Conventionally Used Urine Collection Tubes: Possible Effects on Urinary Proteome Analysis and Prevention of the Adsorption by Polymer Coating

One possible factor determining recovery of trace amount of protein biomarker candidates during proteome analyses could be adsorption on urine tubes. This issue, however, has not been well addressed so far. Recently, a new technical device of surface coating by poly(2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)) (poly(MPC-co-BMA)) has been developed mainly to prevent the adsorption of plasma proteins. We assessed whether conventionally used urine tubes adsorb trace amount of urinary proteins and, if any, whether the surface coating by poly(MPC-co-BMA) can minimize the adsorption. Proteinuric urine samples were kept in poly(MPC-co-BMA)-coated and noncoated urine tubes for 15 min and possibly adsorbed proteins and/or peptides onto urine tubes were analyzed by SDS-PAGE, 2-DE, and the MALDI-TOF MS. It was found that a number of proteins and/or peptides adsorb on the conventionally used urine tubes and that surface coating by poly(MPC-co-BMA) can minimize the adsorption without any significant effects on routine urinalysis test results. Although it remains to be clarified to what extent the protein adsorption can modify the results of urinary proteome analyses, one has to consider this possible adsorption of urinary proteins when searching for trace amounts of protein biomarkers in urine

Review of Schismatogobius (Gobiidae) from Japan, with the description of a new species

Three species of Schismatogobius de Beaufort 1912, distinguished by their morphology and mitochondrial DNA sequences, were found in freshwater streams in the Ryukyu Archipelago, Japan. Although two species were previously known from Japan (S. roxasi Herre 1936 and S. ampluvinculus Chen, Shao, and Fang 1995), the taxonomy needs to be revised. To identify these species, the holotype morphology of S. marmoratus (Peters 1868), S. bruynisi de Beaufort 1912, and S. roxasi, originally described from the Philippines and Indonesia, were examined and re-described here, because relatively little information about their diagnostic characters was provided in the original descriptions. The three Japanese species were identified as S. ampluvinculus, S. marmoratus, and a new species. They were distinguished from each other and from their congeners by the banding pattern of the body, markings on the pectoral fins, pigment patterns on the ventral surface of the head and pelvic fin, body depth at the pelvic-fin origin, pre-anal length, and pectoral-fin ray counts. Although the new species had been regarded as S. roxasi in previous publications, we show that it is actually not S. roxasi and that it also differs from all other nominal species of Schismatogobius. This is described as a new species, S. ninja. Additionally, this is the first record of S. marmoratus from Japan

A Possible Trifunctional β-Carotene Synthase Gene Identified in the Draft Genome of Aurantiochytrium sp. Strain KH105

Labyrinthulomycetes have been regarded as a promising industrial source of xanthophylls, including astaxanthin and canthaxanthin, polyunsaturated fatty acids such as docosahexaenoic acid and docosapentaenoic acid, ω-3 oils, and terpenic hydrocarbons, such as sterols and squalene. A Thraustochytrid, Aurantiochytrium sp. KH105 produces carotenoids, including astaxanthin, with strong antioxidant activity. To gain genomic insights into this capacity, we decoded its 97-Mbp genome and characterized genes for enzymes involved in carotenoid biosynthesis. Interestingly, all carotenogenic genes, as well as other eukaryotic genes, appeared duplicated, suggesting that this strain is diploid. In addition, among the five genes involved in the pathway from geranylgeranyl pyrophosphate to astaxanthin, geranylgeranyl phytoene synthase (crtB), phytoene desaturase (crtI) and lycopene cyclase (crtY) were fused into single gene (crtIBY) with no internal stop codons. Functionality of the trifunctional enzyme, CrtIBY, to catalyze the reaction from geranylgeranyl diphosphate to β-carotene was confirmed using a yeast assay system and mass spectrometry. Furthermore, analyses of differential gene expression showed characteristic up-regulation of carotenoid biosynthetic genes during stationary and starvation phases under these culture conditions. This suggests genetic engineering events to promote more efficient production of carotenoids. We also showed an occurrence of crtIBY in other Thraustochytrid species

Mild inflammation persists in the glenohumeral joint of patients with shoulder instability:Cross-sectional study

OBJECTIVE: Approximately two-thirds of patients with history of shoulder dislocation may develop osteoarthritis (OA) of the glenohumeral joint. However, the biochemical mechanisms underlying the association between dislocation and OA are largely unknown. This study aimed to investigate macrophage markers and inflammatory cytokine expression associated with shoulder instability (SI) in comparison to rotator cuff tears (RCTs). DESIGN: This study included 30 patients with SI and 30 patients with RCTs. Synovial membrane samples were harvested from the rotator interval during the arthroscopic anatomical repair for both groups. The localization of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and cluster of differentiation (CD) 68 in synovial membranes was determined by immunohistochemistry. Transcript-level expressions of the inflammatory cytokines (TNFA and IL1B) and macrophage markers pan-CD68 and -M1 (CD80 and CD86) were quantified. CD80 and CD86 expression in macrophages from the SI group was confirmed using flow cytometry. RESULTS: TNF-α, IL-1β, and CD68 were expressed in the synovial lining layer of the synovial tissue in both groups. In addition, the mRNA expressions of TNFA, IL1B, CD68, and CD80 were significantly higher in the SI group compared to the RCT group (P ​= ​0.012, 0.014, 0.022, 0.003, respectively). In samples from the SI group, 96.3% of CD68+/CD14+ macrophages were CD86-positive, whereas 2.5% of CD68+/CD14+/CD86+ cells were CD80-positive. CONCLUSIONS: Patients with SI had higher mRNA levels of TNFA, IL1B, CD68, and CD80 than those with RCTs. These findings may partially explain the biochemical mechanism underlying the frequent development and progression of osteoarthritis in patients with SI

A new species of Xenoturbella from the western Pacific Ocean and the evolution of Xenoturbella

BackgroundXenoturbella is a group of marine benthic animals lacking an anus and a centralized nervous system. Molecular phylogenetic analyses group the animal together with the Acoelomorpha, forming the Xenacoelomorpha. This group has been suggested to be either a sister group to the Nephrozoa or a deuterostome, and therefore it may provide important insights into origins of bilaterian traits such as an anus, the nephron, feeding larvae and centralized nervous systems. However, only five Xenoturbella species have been reported and the evolutionary history of xenoturbellids and Xenacoelomorpha remains obscure.ResultsHere we describe a new Xenoturbella species from the western Pacific Ocean, and report a new xenoturbellid structure - the frontal pore. Non-destructive microCT was used to investigate the internal morphology of this soft-bodied animal. This revealed the presence of a frontal pore that is continuous with the ventral glandular network and which exhibits similarities with the frontal organ in acoelomorphs.ConclusionsOur results suggest that large size, oval mouth, frontal pore and ventral glandular network may be ancestral features for Xenoturbella. Further studies will clarify the evolutionary relationship of the frontal pore and ventral glandular network of xenoturbellids and the acoelomorph frontal organ. One of the habitats of the newly identified species is easily accessible from a marine station and so this species promises to be valuable for research on bilaterian and deuterostome evolution

The Pharmacological Effects of Herbs on Catecholamine Signaling

Herbs have many biologically and pharmacologically active compounds such as flavonoids and stilbenes. They have been used in remedies for various disorders. Here we review the effects of herbs on catecholamine synthesis and secretion in cultured bovine adrenal medullary cells. Ikarisoside A (1.0–100 μM), a flavonol glycoside, inhibited the catecholamine secretion induced by acetylcholine (0.3 mM). This inhibition was associated with the suppression of 22Na+ and 45Ca2+ influx induced by acetylcholine. The ethanol extract (0.0003–0.005%) of matsufushi (extract of pine nodules) inhibited the catecholamine secretion induced by acetylcholine. SJ-2, one of the stilbene compounds isolated from matsufushi, inhibited acetylcholine-induced catecholamine secretion. Matsufushi extract and SJ-2 reversibly inhibited acetylcholine-induced Na+ currents in Xenopus oocytes expressed with α3β4nicotinic acetylcholine receptors. Sweet tea is the processed leaves of Hydrangea macrophylla. The extract of sweet tea (0.3–1.0 mg/ml) suppressed catecholamine secretion induced by acetylcholine (0.3 mM). Moreover, sweet tea (0.1–1.0 mg/ml), ikarisoside A (1.0–100 μM), and matsufushi (0.001–0.003%) or SJ-2 (10–30 μM) inhibited acetylcholine-induced 14C-catecholamine synthesis from 14C-tyrosine. These findings indicate that ikarisoside A, matsufushi (or SJ-2), and sweet tea inhibit the catecholamine secretion and synthesis induced by acetylcholine in cultured bovine adrenal medullary cells and probably in sympathetic neurons

NBRP, National Bioresource Project of Japan and plant bioresource management

The National BioResource Project has been organized and established to promote research activities using valuable bioresources. A total of twenty-eight bioresources for ten animals, nine plants and nine microorganisms/cell lines developed or collected in Japan were selected for the project. Resources are categorized into several different groups in the project; genetic resources, germplasm, genome resources and their information. Choices of how many resources must be preserved and maintained and in which categories are dependent on the status of the research community of each organism. These resources, if utilized systematically and intelligently, are powerful means for leading new scientific discoveries. Some examples can be seen in this paper. This paper reviews plant bioresources with the main focus on rice resource activities within the project

Two new species of Rhinogobius (Gobiiformes: Oxudercidae) from Palawan, Philippines, with their phylogenetic placement

Two new species of Rhinogobius found in streams on central part of Palawan Island, Philippines are described. The two new species, Rhinogobius estrellae and Rhinogobius tandikan share unique transverse rows of sensory papillae on the cheek with Rhinogobius similis Gill, 1859, but differ from the latter in fin ray counts, arrangement of the scales, etc. The two new species are distinguished from each other by the pectoral-fin ray count, the longitudinal- and predorsal-scale counts, and colouration of the body. Rhinogobius estrellae new species and R. tandikan new species have been found allopatrically in a stream within Malatgao River system flowing into the Sulu Sea and in the Cayulo River flowing into the South China Sea, respectively. The Malatgao River system is the southernmost habitat of the genus Rhinogobius. Rhinogobius similis had been considered as the only member of the most basal lineage of this genus, but our mitochondrial genome analysis suggested that the two new species are additional members of this lineage. They are considered to be relicts of their common ancestor with R. similis, which probably had a wider distribution.journal articl