Korea Research Institute of Bioscience and Biotechnology (KRIBB) is a government research institute in Daejeon, South Korea. It is dedicated to biotechnology research across a broad span of expertise, from basic studies for the fundamental understanding of life phenomena to applied studies such as drug discovery, novel biomaterials, integrated biotechnology and bioinformation.KRIBB was established in 1985. Its accomplishments include the advancement of welfare and medical technology, an increase in food production, a cleaner environment and new bio-materials and energy sources.It has identified reasons for the failure of animal cloning, conducted a comparative study of chimpanzee genes and successfully analyzed the structure of the reactive oxygen species switch protein, which became the first study by Korean scholars to be published in Cell, an international scientific journal. The institute was ranked first in the discovery of new microorganisms, including the indigenous microorganisms of the Dokdo Islets, for four consecutive years.Its recent accomplishments include the development of a genome capable of controlling cancer cell proliferation and the identification of a neuropeptide Y-based growth control mechanism, with possibilities for new treatments for cancer, diabetes, obesity and ageing. Nano–bio sensor research led to the development of the world's smallest surface plasmon resonance biochip.
A thorough understanding of living systems necessitates detailed mapping of the high‐dimensional molecular landscapes within intact, thick biological tissues. Recent advances in hydrogel‐based tissue processing and clearing techniques have facilitated 3D visualization of biological systems while preserving their native spatial context. Although various polymer hydrogels have been developed for tissue processing, systematic elucidation of their workflow‐dependent properties from a materials science perspective remains lacking, hindering the establishment of design principles for multifunctional performance. Here, a physicochemical design framework for polymer hydrogels in tissue processing is presented, derived from a comprehensive analysis of materials employed in four representative platforms. This analysis encompasses chemo‐rheological parameters, swelling behavior, micro/nanopore morphology, diffusion kinetics, mechanical performance, and thermochemical stability, all of which influence the functional and structural outcomes of the platform. Furthermore, mock tissue‐hydrogel hybrids are engineered using chemically‐fixed protein chunks to evaluate changes in mechanical, morphological, and thermochemical properties in a context relevant to their application. By linking hydrogel design parameters to physicochemical properties and downstream performance in tissue processing, labeling, and imaging, this study establishes a structure‐function blueprint that directly informs rational polymer engineering for high‐throughput, multiscale, and multiplexed molecular tissue imaging applications and beyond.
IntroductionBoehmeria nivea (L.) Gaud. has traditionally been regarded as a medicinal food with applications in various inflammatory disorders. However, its role in chronic obstructive pulmonary disease (COPD) has not yet been clarified.MethodsIn this study, the preventive efficacy of the ethyl acetate fraction of B. nivea (L.) Gaud. leaves (EA-BN) was evaluated in a COPD model established by intratracheal instillation of lipopolysaccharide (LPS; 0.5 mg/kg body weight) and cigarette smoke condensate (CSC; 12.5 mg/kg body weight) in male C57BL/6N mice. The experimental groups received dexamethasone (3 mg/kg) as a positive control or EA-BN at doses of 100 and 200 mg/kg.ResultsEA-BN administration significantly reduced T helper 1 cytokine levels and decreased macrophage and neutrophil counts in bronchoalveolar lavage fluid. Histological analyses revealed that EA-BN mitigated alveolar destruction and inflammatory infiltration, whereas pulmonary function tests demonstrated improvements in the FEV0.1/FVC ratio and lung elastance in the LPS/CSC-induced COPD. Additionally, EA-BN alleviated oxidative stress by promoting the nuclear translocation of Nrf2 and enhancing the expression of its downstream targets, HO-1 and NQO1, leading to a reduction in reactive oxygen species and nitric oxide production. EA-BN downregulated thioredoxin-interacting protein and NLRP3 inflammasome activation, thereby suppressing caspase-1 and IL-1β expression, whereas also attenuating apoptosis by modulating the Bax/Bcl-2/caspase-3 pathway.DiscussionCollectively, these findings suggest that EA-BN possesses antioxidant, anti-inflammatory, and anti-apoptotic properties, supporting its potential as a preventive agent against COPD.
For in vitro DNA assembly, enzymes with exonuclease activities have been utilized to generate relatively long recessed ends on DNA fragments, which can anneal to other DNA fragments if they have complementary nucleotide sequences. The combined construct can be directly delivered to competent cells, where the gaps and nicks between the fragments are completely rectified. We introduce a versatile sequence- and ligation-independent cloning (SLIC) method called 'DNA Assembly with Phosphorothioate (PT) and T5 Exonuclease' (DAPE), which generates precise lengths of 3' overhangs at both ends of linearized DNA. In contrast to conventional SLIC techniques, which are not suitable for cloning DNA fragments smaller than 50 base pairs (bp) due to overzealous exonuclease activity, such as with gRNA and epitope tags, DAPE can efficiently and precisely assemble several fragments in a single reaction regardless of the size of the DNA. Thus, DAPE, as an advanced toolkit for DNA cloning and synthetic biology, may further expedite the construction of more elaborate multi-gene circuitry.
l-theanine (γ-glutamylethylamide) is a bioactive amino acid widely valued for its functional and nutraceutical applications. While enzymatic synthesis using γ-glutamylmethylamide synthetase (GMAS) has been extensively studied, the potential of γ-glutamylcysteine synthetase (GCS) as an alternative biocatalyst remains underexplored. In this study, a high-substrate, buffer-free whole-cell conversion system was established for l-theanine production using Escherichia coli expressing either GMAS from Methylovorus mays (MmGMAS) or the GCS from E. coli (ecGCS), with an integrated ATP regeneration mechanism driven by polyphosphate kinase 2 (PPK2). In silico predictions using CatPred, a machine learning-based catalyst prediction tool, indicated that ecGCS exhibits catalytic efficiency comparable to or exceeding that of MmGMAS, which was consistent with the experimental results. Among six PPK2 variants tested, the enzyme from Rhodobacter sphaeroides (PPK2-6) was identified as the most suitable ATP regeneration module, enabling approximately 90% reduction in ATP input while maintaining high l-theanine yields. Both MmGMAS-PPK2-6 and ecGCS-PPK2-6 systems produced 47.9 g/L (34.4%) and 44.5 g/L (31.9%) of l-theanine, respectively, starting from 800 mM substrates. Furthermore, it was confirmed that l-theanine production was not compromised in the absence of exogenous buffers, which may also facilitate downstream processing. This work represents the first demonstration of efficient l-theanine production using native ecGCS under process-relevant conditions, and highlights its potential as a complementary or alternative platform to GMAS-based biosynthesis.
Alcoholic beverages have been concerned not only for gastronomic delight but also for certain impacts on health, such as obesity, diabetes, and cardiovascular diseases. In this study, we assessed the bioactive functions of 1,1-Diethoxyethane (1,1-DEE), a flavoring compound formed during the aging process of wine by flor yeast, using both cultured cell lines and a high-fat diet (HFD) mouse model. 1,1-DEE was identified in the batches of ethanol that induced oxidation of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) using gel mobility shift assay and gas chromatography-mass spectrometry. PTEN was reversibly oxidized when exposed to 1,1-DEE, but 1,2-DEE did not induce PTEN oxidation. Mechanistically, 1,1-DEE treatment enhanced the production of mitochondrial reactive oxygen species, accompanying by oxidation of PTEN and subsequent activation of Akt signaling. 1,1-DEE treatment elevated Akt activation when combined with insulin, compared with insulin alone, and alleviated palmitate-induced insulin resistance in C2C12 myoblasts. Moreover, the oral administration of 1,1-DEE alleviated glucose intolerance and insulin resistance in HFD-fed mice. 1,1-DEE also mitigated HFD-induced body weight gain and hepatic dyslipidemia without reduction of food intake. Transcriptome analysis revealed significant genes involved in the improvement of insulin sensitivity and dyslipidemia. Thus, 1,1-DEE may serve as a promising therapeutic agent for the intervention of obesity, diabetes, and dyslipidemia.
