Kaede is a photoconvertible fluorescence protein that changes from green to red upon exposure violet light. The photoconversion of intracellular has no effect on cellular function. Using transgenic mice expressing the protein, we demonstrated movement cells with photoconverted could be monitored lymphoid organs other tissues as well skin draining lymph node. Analysis kinetics revealed each subset in node, such CD4 + T, CD8 B, and dendritic cells, distinct migration pattern vivo . Thus, mouse...

Tardigrades are able to tolerate almost complete dehydration through transition a metabolically inactive state, called “anhydrobiosis”. Late Embryogenesis Abundant (LEA) proteins heat-soluble involved in the desiccation tolerance of many anhydrobiotic organisms. Tardigrades, Ramazzottius varieornatus, however, express predominantly tardigrade-unique proteins: CAHS (Cytoplasmic Heat Soluble) and SAHS (Secretory proteins, which secreted or localized most intracellular compartments, except...

Alteration of the control DNA replication and mitosis is considered to be a major cause genome instability. To investigate mechanism that controls stability, we used RNA silencing-interference technique (RNAi) eliminate Drosophila geminin homologue from Schneider D2 (SD2) cells. Silencing by RNAi in SD2 cells leads cessation asynchronous overreplication genome, with containing single giant nuclei partial ploidy between 4N 8N content. The effect deficiency completely suppressed cosilencing...

Abstract Methyl esters from crude sunflower oil were produced via methanolysis reaction using sodium hydroxide catalyst. Methanolysis was carried out at different agitation speeds (200–600 rpm), temperatures (25–60 °C), catalyst loadings (0.25–1.00% by weight of oil), and methanol:oil mole ratios (6:1–20:1). Mass‐transfer limitation effectively minimized 400–600 rpm with no apparent lag period. Lowering the temperature resulted in a fall rate prolonging time necessary to achieve maximum...

Cohesive interactions between Porphyromonas gingivalis and plaque-forming bacteria, such as Streptococcus oralis, are considered to play an important role in the colonization of P. periodontal sites. Although fimbriae have been reported mediate coaggregation with S. oralis molecule involved has not identified. We identified coadhesin ATCC 9811 purified it by affinity column chromatography. found that molecular mass protein was approximately 40 kDa. Dot blot Western assays showed binding...

We identified a novel human chondroitin<i>N</i>-acetylgalactosaminyltransferase, designated chondroitin GalNAcT-2 after BLAST analysis of the GenBank^TM data base using sequence previously described chondroitin<i>N</i>-acetylgalactosaminyltransferase (chondroitin GalNAcT-1) as probe. The new cDNA contained an open reading frame encoding protein 542 amino acids with type II transmembrane topology. acid displayed 60% identity to that GalNAcT-1. Like GalNAcT-1, expression soluble form...

We developed a DNA microarray to evaluate the estrogen activity of natural estrogens and industrial chemicals. Using MCF-7 cells, we conducted comprehensive analysis estrogen-responsive genes among approximately 20,000 human genes. On basis reproducible reliable responses estrogen, selected 172 be used for developing customized microarray. this microarray, examined (17beta-estradiol, estriol, estrone, genistein), chemicals (diethylstilbestrol, bisphenol A, nonylphenol, methoxychlor), dioxin....

Interactions of the type 1 phosphatase catalytic subunit (PP1c) and myosin holoenzyme (MBP) were compared using affinity columns. In absence ATP, MBP bound to dephosphorylated myosin, heavy meromyosin (HMM), subfragment 1. contrast, PP1c was not bound. presence binding occurred only with phosphorylated protein. The interaction proteins also demonstrated thiophosphorylated as competitive inhibitors. Kinetics parameters determined. With light chains (P-LC20), major difference between a lower...

Fluorescent proteins with light wavelengths within the optical window are one of improvements in vivo imaging techniques. Near-infrared (NIR) fluorescent protein (iRFP) is a stable, nontoxic that emits fluorescence NIR without addition exogenous substrate. However, studies utilizing an iRFP model have not yet been published. Here, we report generation transgenic mice ubiquitous expression. expression was observed approximately 50% offspring from matings between and WT mice. The serum blood...

Digital PCR (dPCR) is a promising method for performing liquid biopsies that quantifies nucleic acids more sensitively than real-time PCR. However, dPCR shows large fluctuations in the fluorescence intensity of droplets or wells due to insufficient amplification small partitions, limiting multiplexing capability using intensity. In this study, we propose measurement combines with melting curve analysis highly multiplexed genotyping. A sample was digitized into silicon chip up 2 × 104 which...

A highly sensitive and multiplexed quantification technique for nucleic acids is necessary to predict evaluate cancer treatment by liquid biopsy. Digital PCR (dPCR) a technique, but conventional dPCR discriminates multiple targets the color of fluorescent dye probe, which limits multiplexing beyond number colors dyes. We previously developed combined with melting curve analysis. Herein, we improved detection efficiency accuracy analysis detect KRAS mutations in circulating tumor DNA (ctDNA)...

Heparan, the common unsulfated precursor of heparan sulfate (HS) and heparin, is synthesized on glycosaminoglycan-protein linkage region tetrasaccharide GlcUA-Gal-Gal-Xyl attached to respective core proteins presumably by HS co-polymerases encoded EXT1 EXT2, genetic defects which result in hereditary multiple exostoses humans. Although both EXT2 exhibit GlcNAc transferase GlcUA activities required for synthesis, no chain polymerization has been demonstrated vitro using recombinant enzymes....

Cdt1 is a licensing factor for DNA replication, the function of which tightly controlled to maintain genome integrity. Previous studies have indicated that cell cycle-dependent degradation triggered at S phase prevent re-replication. In this study, we found degraded upon damage induced by either UV treatment or γ-irradiation (IR). Although IR-triggered was caffeine-insensitive, UV-triggered caffeine-sensitive. This indicates cells treated with utilize checkpoint pathway, differs from IR. A...

In this report, we investigate the C. elegans CBFbeta homologue, BRO-1. bro-1 mutants have a similar male-specific sensory ray loss phenotype to rnt-1 (the homologue of mammalian CBFbeta-interacting Runx factors), caused by failed cell divisions in seam lineages. Our studies indicate that BRO-1 and RNT-1 form proliferation-promoting complex, increases both affinity specificity RNT-1-DNA interactions. Overexpression bro-1, like rnt-1, leads an expansion number co-overexpression results...

Blood vessel development and network patterning are controlled by several signaling molecules, including VEGF, FGF, TGF-ß, Ang-1,2. Among these, the role of VEGF-A in morphogenesis is best understood. The biological activity depends on its reaction with specific receptors Flt1 Flk1. Roles endothelial cell proliferation, migration, survival, vascular permeability, induction tip filopodia have been reported. In this study, we generated Flt1-tdsRed BAC transgenic (Tg) mice to monitor gene...

The peroxidation of lipids and changes in the activities related enzymes gastric mucosa were studied a rat model mucosal injury induced by nonsteroidal anti-inflammatory drug indomethacin. area erosion amount thiobarbituric acid reactive substances (TBARS) significantly increased beginning 4h after administration Xanthine oxidase (XOD) activity also immediately drug. Although XOD was suppressed allopurinol treatment, induction increase TBARS not. Myeloperoxidase (MPO), marker enzyme...

The 130 kDa myosin-binding subunit (MBS) of smooth muscle myosin phosphatase was detected in cytoskeletal, cytosolic, and membrane fractions T24 cells. Also, MBS distributed between cytoplasm plasmalemma mitotic REF52 These observations prompted this study the interaction(s) phospholipids with phosphatase. Using a sedimentation assay, gizzard bound to vesicles acidic phospholipids, i.e. phosphatidylserine (PS), phosphatidylinositol, phosphatidic acid (PA). Neutral did not bind. Binding PS...

alpha1-Acid glycoprotein (AGP) showed multiple peaks on separation using capillary electrophoresis in a chemically modified with dimethylpolysiloxane at slightly acidic conditions. We analyzed glycoforms of AGP species after by ion-exchange chromatography, Con A affinity and Cu(II)-chelating chromatography. The thus obtained were digested N-glycosidase F, the released carbohydrate chains high-performance liquid chromatography labeling 3-aminobenzoic acid. results afforded basic information...

MOSFETs in the sub-0.1- mu m regime were investigated using a nonplanar device simulator CADDETH-NP. It was found that even this regime, short-channel effect can be suppressed grooved gate because of concave corner insulator. with length 0.05 or less no threshold voltage lowering made by optimizing radius, junction depths, and channel doping.< <ETX xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">&gt;</ETX>

An extended-gate CMOS sensor array with enzyme-immobilized microbeads for redox-potential glucose detection is demonstrated the first time. Redox-potential has possibility to achieve high accuracy because it not affected by buffer conditions. Despite this high-accuracy property, requires a sufficient amount of enzyme, which leads increased cost. In order reduce enzyme consumption while maintaining capability, we have introduced microbeads. By using microbeads, can be efficiently positioned...

Pancreatic intraepithelial neoplasia (PanIN) and intraductal papillary mucinous neoplasms (IPMNs) are pancreatic ductal adenocarcinoma (PDAC) precursor lesions. Detecting these precursors monitoring their progression crucial for early PDAC diagnosis. Digital PCR (dPCR) is a highly sensitive nucleic acid quantification technique offers cost‐effective option patient follow‐up. However, the clinical utility of conventional dPCR restricted by multiplexing constraints, particularly due to...

The insulin response to the sulfonylurea glibenclamide was markedly impaired in pancreatic beta-cell line MIN6 cells with chronic treatment (MIN6-Glib). intracellular calcium concentration increased only slightly MIN6-Glib. While properties of voltage-dependent channels were not altered, conductance K(ATP) channels, primary target glibenclamide, significantly reduced ATP-sensitive K+ (K(ATP)) comprise inwardly rectifying channel member Kir6.2 subunits and receptor (SUR) 1 subunits. have both...