Anaerobic plasmalogen production in recombinant Escherichia coli carrying plasmalogen synthase gene from Selenomonas ruminantium.

Escherichia coli expressing SrPlsAR from Selenomonas ruminantium produces plasmalogen, comprising almost 60% of the total phospholipid content under anaerobic conditions. Both plasmenylethanolamine and plasmenylglycerol were detected, and the major acyl aldehyde derived from sn-1 vinyl ether was C16:1. Plasmalogen synthesis is affected by mutations in ATP-binding sites and Cys expected to be involved in the formation of the [4Fe-4S] cluster.

Evaluation of psychological stress in confined environments using salivary, skin, and facial image parameters.

Detecting the influence of psychological stress is particularly important in prolonged space missions. In this study, we determined potential markers of psychological stress in a confined environment. We examined 23 Japanese subjects staying for 2 weeks in a confined facility at Tsukuba Space Center, measuring salivary, skin, and facial image parameters.

A semi high-throughput method for screening small bispecific antibodies with high cytotoxicity.

Small bispecific antibodies that induce T-cell-mediated cytotoxicity have the potential to damage late-stage tumor masses to a clinically relevant degree, but their cytotoxicity is critically dependent on their structural and functional properties. Here, we constructed an optimized procedure for identifying highly cytotoxic antibodies from a variety of the T-cell-recruiting antibodies engineered from a series of antibodies against cancer antigens of epidermal growth factor receptor family and T-cell receptors. By developing and applying a set of rapid operations for expression vector construction and protein preparation, we screened the cytotoxicity of 104 small antibodies with diabody format and identified some with 10-times higher cytotoxicity than that of previously reported active diabody.

Complete genome sequence of Selenomonas ruminantium subsp. lactilytica will accelerate further understanding of the nature of the class Negativicutes.

Selenomonas ruminantium subsp. lactilytica, a strictly anaerobic ruminal bacterium, possesses typical Gram-negative cell surface structure comprising cytoplasmic membrane, peptidoglycan layer and outer membrane, whereas its 16S rRNA-based taxonomy shows that the bacteria belongs to Gram-positive Firmicutes. Complete genome analysis showed that genes or gene clusters involved in Gram-negative cell structure were scattered in the S.

Phosphatidylethanolamine plasmalogen enhances the inhibiting effect of phosphatidylethanolamine on γ-secretase activity.

Plasmalogens (Pls) are widely distributed in the biological membrane of animals and certain anaerobic bacteria, but their functions in the cell membrane are still poorly understood. Decrease of phosphatidylethanolamine plasmalogen (PEPls) in the brain tissue of patients with Alzheimer's disease prompted us to investigate the effect of the membrane phosphorus lipid composition on the activity of γ-secretase that produces amyloid-beta protein (Aβ). To clarify the effect of phospholipids, including PEPls, on Aβ production, γ-secretase activity was measured in an in vitro assay using yeast microsomes and reconstituted liposomes.

Directed evolution of endoglucanase III (Cel12A) from Trichoderma reesei.

The stability and specific activity of endo-beta-1,4-glucanase III from Trichoderma reesei QM9414 was enhanced, and the expression efficiency of its encoding gene, egl3, was optimized by directed evolution using error-prone PCR and activity screening in Escherichia coli RosettaBlue (DE3) pLacI as a host. Relationship between increase in yield of active enzyme in the clones and improvement in its stability was observed among the mutants obtained in the present study. The clone harboring the best mutant 2R4 (G41E/T110P/K173M/Y195F/P201S/N218I) selected in via second-round mutagenesis after optimal recombinating of first-round mutations produced 130-fold higher amount of mutant enzyme than the transformant with wild-type EG III.

Functional characteristics of the skin surface of children approaching puberty: age and seasonal influences.

The aim of this study was to determine differences in the functional properties of the stratum corneum of children and adults, focusing on the influence of approaching puberty. Biophysical measurements were made of the stratum corneum of 32 healthy Japanese children aged 10-14 years and their mothers in summer and the following winter. The children showed significantly lower skin surface hydration.

Characterization of the catalytic domains of Trichoderma reesei endoglucanase I, II, and III, expressed in Escherichia coli.

The genes encoding the catalytic domains (CD) of the three endoglucanases (EG I; Cel7B, EG II; Cel5A, and EG III; Cel12A) from Trichoderma reesei QM9414 were expressed in Escherichia coli strains Rosetta-gami B (DE3) pLacI or Origami B (DE3) pLacI and were found to produce functional intracellular proteins. Protein production by the three endoglucanase transformants was evaluated as a function of growth temperature. Maximal productivity of EG I-CD at 15 degrees C, EG II-CD at 20 degrees C and EG III at 37 degrees C resulted in yields of 6.

Glucocorticoids enhance Toll-like receptor 2 expression in human keratinocytes stimulated with Propionibacterium acnes or proinflammatory cytokines.

Toll-like receptors (TLRs) on keratinocytes are important cell surface receptors involved in the innate and acquired immune response to invading microorganisms. In acne vulgaris, TLR2 activation by Propionibacterium acnes (P. acnes) may induce skin inflammation via induction of various proinflammatory molecules that stimulate the invasion of inflammatory cells.