Biosensing Techniques in Yeast: G-Protein Signaling and Protein-Protein Interaction Assays for Monitoring Ligand Stimulation and Oligomer Formation of Heterologous GPCRs

Guanine nucleotide-binding proteins (G-proteins) act as transducers of external stimuli for intracellular signaling, and control various cellular processes in cooperation with seven transmembrane G-protein-coupled receptors (GPCRs). Because GPCRs constitute the largest family of eukaryotic membrane proteins and enable the selective recognition of a diverse range of molecules (ligands), they are the major molecular targets in pharmaceutical and medicinal fields. In addition, GPCRs have been known to form heteromers as well as homomers, which may result in vast physiological diversity and provide opportunities for drug discovery. G-proteins and their signal transduction machinery are universally conserved in eukaryotes; thereby, the yeast Saccharomyces cerevisiae has been used to construct artificial in vivo GPCR biosensors. In this chapter, we focus on the yeast-based GPCR biosensors that can detect ligand stimulation and oligomer formation, and summarize their techniques using the G-protein signaling and protein-protein interaction assays

Late-onset sick sinus syndrome after carbon monoxide poisoning

Carbon monoxide (CO) is a known, potent poisonous gas that causes hypoxaemia because of its high affinity for haemoglobin. It also induces inflammatory responses that cause tissue injury, particularly to the nervous and cardiovascular systems. Here we present a case of late-onset sick sinus syndrome (SSS) after CO poisoning. Arrhythmia during the acute phase has been recorded in the literature, but to the best of our knowledge, this is the first report of late-onset SSS. Late-onset neuropathy after CO poisoning is well known, and it seems that a similar mechanism develops in cardiac conduction after CO poisoning. This report highlights the importance of follow-up for arrhythmia after CO poisoning

Short-Term Program on Three-Dimensional Printed Self-Help Devices for Occupational Therapy Students: A Pre-Post Intervention Study

Despite the increasing importance of digital fabrication, of which three-dimensional printing is an important aspect, educational programs in this area have not been fully developed. To utilize three-dimensional printing optimally, occupational therapists need to be familiar with this new technology, understand its scope of application, and possess certain levels of skills for producing. The purpose of this study was to examine the effectiveness of a short-term program for occupational therapy students to increase the acceptance of three-dimensional printed devices by acquiring the basic knowledge and skills of making three-dimensional printed self-help devices. The research involved an intervention study with a pre-post design. Participants comprised 112 entry-level occupational therapy students. The program consisted of two 90-minute sessions during 2019 and 2020. It included a three-part lecture series and two types of practice. The conducted pre-post questionnaires were structured into four categories: I. student profile; II. knowledge about digital fabrication technology; III. ideas and attitudes toward three-dimensional printed self-help devices; and IV. impressions and thoughts. After the program, the number of students who acquired basic knowledge of digital fabrication and who felt confident about making three-dimensional printed self-help devices significantly increased (p \u3c 0.05). The study suggested that the program was effective and assisted occupational therapy students to understand the usefulness of this new technology and be comfortable using it

Localization of chondromodulin-I at the feto-maternal interface and its inhibitory actions on trophoblast invasion in vitro

<p>Abstract</p> <p>Background</p> <p>Chondromodulin-I (ChM-I) is an anti-angiogenic glycoprotein that is specifically localized at the extracellular matrix of the avascular mesenchyme including cartilage and cardiac valves. In this study, we characterized the expression pattern of ChM-I during early pregnancy in mice <it>in vivo </it>and its effect on invasion of trophoblastic cells into Matrigel <it>in vitro</it>.</p> <p>Results</p> <p>Northern blot analysis clearly indicated that <it>ChM-I </it>transcripts were expressed in the pregnant mouse uterus at 6.5-9.5 days post coitum. <it>In situ </it>hybridization and immunohistochemistry revealed that ChM-I was localized to the mature decidua surrounding the matrix metalloproteinase-9 (MMP-9)-expressing trophoblasts. Consistent with this observation, the expression of <it>ChM-I </it>mRNA was induced in decidualizing endometrial stromal cells <it>in vitro</it>, in response to estradiol and progesterone. Recombinant human ChM-I (rhChM-I) markedly inhibited the invasion through Matrigel as well as the chemotactic migration of rat Rcho-1 trophoblast cells in a manner independent of MMP activation.</p> <p>Conclusions</p> <p>This study demonstrates the inhibitory action of ChM-I on trophoblast migration and invasion, implying the potential role of the ChM-I expression in decidual cells for the regulated tissue remodeling and angiogenesis at feto-maternal interface.</p

Predominant Magnetic States in Hubbard Model on Anisotropic Triangular Lattices

Using an optimization variational Monte Carlo method, we study the half-filled-band Hubbard model on anisotropic triangular lattices, as a continuation of the preceding study [J. Phys. Soc. Jpn 75, 074707 (2006)]. We introduce two new trial states: (i) A coexisting state of (\pi,\pi)-antiferromagnetic (AF) and a d-wave singlet gaps, in which we allow for a band renormalization effect, and (ii) a state with an AF order of 120^\circ spin structure. In both states, a first-order metal-to-insulator transition occurs at smaller U/t than that of the pure d-wave state. In insulating regimes, magnetic orders always exist; an ordinary (\pi,\pi)-AF order survives up to t'/t\sim 0.9 (U/t=12), and a 120^\circ-AF order becomes dominant for t'/t \gsim 0.9. The regimes of the robust superconductor and of the nonmagnetic insulator the preceding study proposed give way to these magnetic domains.Comment: 11 pages, 14 figure

Targeting cancer cell-specific RNA interference by siRNA delivery using a complex carrier of affibody-displaying bio-nanocapsules and liposomes

BACKGROUND: Small interfering RNA (siRNA) has attracted attention in the field of nucleic acid medicine as a RNA interference (RNAi) application that leads to gene silencing due to specific messenger RNA (mRNA) destruction. However, since siRNA is unstable in blood and unable to cross the cell membrane, encapsulation of siRNA into a carrier is required. RESULTS: In this study, we used a carrier that combined Z(HER2)-displaying bio-nanocapsule (derived from hepatitis B virus surface antigen) and liposomes in a complex in order to investigate the feasibility of effective and target-cell-specific RNAi applications. As a result, by observing RNAi only in HER2-expressing breast cancer cells, using our proposed methodology, we successfully demonstrated target-cell-specific delivery and effective function expression of siRNA. CONCLUSIONS: These findings show that, in the field of nucleic acid medicine, Z(HER2)-BNC/LP can be a useful carrier for siRNA delivery, and could also become a useful tool for gene silencing and to accomplish protein knock-down