Development of CRISPR Interference (CRISPRi) Platform for Metabolic Engineering of and Its Application for Engineering Riboflavin Biosynthesis.

, a hetero-fermentative type of lactic acid bacteria, is a crucial probiotic candidate because of its ability to promote human health. However, inefficient gene manipulation tools limit its utilization in bioindustries. We report, for the first time, the development of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) interference (CRISPRi) system for engineering .

Carrier Depletion near the Grain Boundary of a SiC Bicrystal.

Silicon carbide (SiC) bicrystals were prepared by diffusion bonding, and their grain boundary was observed using scanning transmission electron microscopy. The n-type electrical conductivity of a SiC single crystal was confirmed by scanning nonlinear dielectric microscopy (SNDM). Dopant profiling of the sample by SNDM showed that the interface acted as an electrical insulator with a ~2-μm-thick carrier depletion layer.

Development of bicistronic expression system for the enhanced and reliable production of recombinant proteins in Leuconostoc citreum.

The lactic acid bacteria (LAB) Leuconostoc citreum are non-sporulating hetero-fermentative bacteria that play an important role in the fermented food industry. In this study, for the enhanced and reliable production of recombinant proteins in L. citreum, we developed a bicistronic design (BCD) expression system which includes a short leader peptide (1 cistron) followed by target genes (2 cistron) under the control of a single promoter.

Clinical characteristics of non-small cell lung cancer patients who experienced acquired resistance during gefitinib treatment.

Background: The NSCLC patients who experienced good clinical responses to an EGFR-TKI will inevitably develop acquired resistance. A great deal of research is being carried out to discover the molecular mechanisms underlying this resistance. In comparison, few studies have been conducted to find out about the clinical characteristics of acquired resistance in the patients who had responded to an EGFR-TKI.

Overproduction of a C5a receptor antagonist (C5aRA) in Escherichia coli.

The C5aR antagonist (C5aRA)(1), which blocks the interaction of C5a anaphylatoxin and its receptor C5aR, is one of the most potent therapeutic agents for the treatment of various autoimmune diseases and acute inflammatory conditions. Here we developed an efficient C5aRA production system using Escherichia coli. To produce functional C5aRA, which contains three disulfide bonds, we used E.

Dysfunction of endothelial progenitor cells under diabetic conditions and its underlying mechanisms.

Cardiovascular complications have been major concerns in the treatment of diabetes, and up to 80% of all deaths in diabetic patients are linked to cardiovascular problems. Impaired angiogenesis is one of the most serious symptoms associated with diabetes, resulting in delayed wound healing and lower limb amputation. Endothelial progenitor cells (EPCs), a subpopulation of adult stem cells, are recruited from bone marrow to the injured vessel to promote endothelial regeneration and neovascularization, playing an important role in angiogenesis.

High-level production of a kringle domain variant by high-cell-density cultivation of Escherichia coli.

Human kringle domains (KDs) are ubiquitously expressed binding modulators that fold into seven flexible loops and it has been previously demonstrated that KDs can be engineered toward target-specific binding proteins as a non-antibody protein scaffold. Here, we report a method for efficient expression of a KD derivative (KD548)-a promising anti-cancer agent-by high-cell-density culture of Escherichia coli at a preparative scale production. The correct folding of KD548 requires three disulfide bonds.

Recombinant antibodies: engineering and production in yeast and bacterial hosts.

After the appearance of the first FDA-approved antibody 25 years ago, antibodies have become major therapeutic agents in the treatment of many human diseases, including cancer and infectious diseases, and the use of antibodies as therapeutic/diagnostic agents is expected to increase in the future. So far, a variety of strategies have been devised for engineering of these fascinating molecules to develop superior properties and functions. Recent progress in systems biology has provided more information about the structures and cellular networks of antibodies, and, in addition, recent development of biotechnology tools, particularly in regard to high-throughput screening, has made it possible to perform more intensive engineering on these substances.

The PI3K-Akt mediates oncogenic Met-induced centrosome amplification and chromosome instability.

The oncogenic ability of aberrant hepatocyte growth factor receptor (Met) signaling is thought to mainly rely on its mitogenic and anti-apoptotic effects. Recently, however, cumulating evidences suggest that genomic instability may be a crucial factor in tumorigenesis. Here, we address whether oncogenic Met receptor is linked to the centrosome abnormality and genomic instability.

[A case of undifferentiated (embryonal) liver sarcoma mimicking klatskin tumor in an adult].

Undifferentiated sarcoma is an uncommon primary malignant tumor of the liver typically occurring in older children. It is also referred to as malignant mesenchymoma, fibromyxosarcoma, or mesenchymal sarcoma. We experienced a case of undifferentiated sarcoma in 72-year-old male.