Whole-body imaging with single-cell resolution by tissue decolorization

The development of whole-body imaging at single-cell resolution enables system-level approaches to studying cellular circuits in organisms. Previous clearing methods focused on homogenizing mismatched refractive indices of individual tissues, enabling reductions in opacity but falling short of achieving transparency. Here, we show that an aminoalcohol decolorizes blood by efficiently eluting the heme chromophore from hemoglobin. Direct transcardial perfusion of an aminoalcohol-containing cocktail that we previously termed CUBIC coupled with a 10 day to 2 week clearing protocol decolorized and rendered nearly transparent almost all organs of adult mice as well as the entire body of infant and adult mice. This CUBIC-perfusion protocol enables rapid whole-body and whole-organ imaging at single-cell resolution by using light-sheet fluorescent microscopy. The CUBIC protocol is also applicable to 3D pathology, anatomy, and immunohistochemistry of various organs. These results suggest that whole-body imaging of colorless tissues at high resolution will contribute to organism-level systems biology

Energy Performance Evaluation of a Desiccant Air Handling System to Maximize Solar Thermal Energy Use in a Hot and Humid Climate

A desiccant air handling unit is one of the major types of dehumidification handling systems and requires hot water or hot air to regenerate sorption materials. If solar thermal energy is used as the heat source for regeneration, in general, a backup electrical heater, backup boiler, or combined heat and power (CHP) is installed in order to maintain a stable hot water supply. In this study, effective control is proposed for a desiccant air handling system that uses solar thermal energy (flexible control), and its energy performance is compared to that of a traditional control (the fixed control) through a system simulation. The diurnal behavior shows that the system with a fixed control without a backup boiler cannot process the latent load properly (28 GJ of unprocessed latent load for July and August). On the other hand, the system with a flexible control without a backup boiler is able to process required latent heat load. Based on the fact that the fixed control needs a backup boiler to process the latent load, the system with a fixed control with a backup boiler is considered for the energy performance comparison. The simulation results show that the primary energy-based coefficient of performance (hereafter, COP) of the system with a flexible control without a backup boiler reaches 1.56. On the other hand, the primary energy-based COP of the system with a fixed control with a backup boiler reaches only 1.43. This proves that the flexible control contributes to the higher energy performance of the system and maximizes the use of solar thermal energy more than the fixed control

Formation of an Active Form of the Interleukin-2/15 Receptor beta-Chain by Insertion of the Intracisternal A Particle in a Radiation-Induced Mouse Thymic Lymphoma and Its Role in Tumorigenesis

Although many reports suggest that aberrant regulation of cytokine signaling pathways via the interleukin-2 receptor (IL-2R) induces tumorigenic transformation, constitutively active IL-2R in tumors has not been reported. We searched for genomic alteration of the IL-2/15R beta-subunit gene (IL-2/15R beta) in cytokine-independent cell lines established from radiation-induced mouse thymic lymphomas. In the TL34 cell line and its primary tumor, one of the IL-2/15R beta alleles was rearranged by the insertion of an intracisternal A particle (IAP) retrotransposon. The IAP-IL2/15R beta chimeric gene expressed chimeric mRNA in which IAP-coding Gag-Pol mRNA was fused to IL-2/15R beta mRNA and coded for Gag-Pol-IL-2/15R beta chimeric protein. Forced expression of the Gag-Pol-IL-2/15R beta chimeric cDNA in a mouse cytotoxic T-cell line (CTLL-2) converted IL-2-dependent cell growth to IL-2-independent growth, suggesting that the chimeric protein activates some of the IL-2 signaling pathways necessary for cell proliferation. Downregulation of the expression of the Gag-Pol-IL-2/15R beta chimeric protein in TL34 by antisense RNA inhibited cell growth, and concomitantly reduced the level of c-myc protein. These results suggest that the Gag-Pol-IL-2/15R beta is a constitutively active form that transmits proliferative signals by expressing downstream target genes, including c-myc. Thus, we demonstrated that the chimeric receptor gene produced by the insertion of an IAP functions as an oncogene by providing IL-2-independent autonomous growth potential

Radiation-induced deletions in the 5\u27 end region of Notch1 lead to the formation of truncated proteins and are involved in the development of mouse thymic lymphomas

Notch1 protein is a transmembrane receptor that directs various cell fate decisions. Active forms of Notch1 consisting of a transmembrane domain and an intracellular domain (Notch1TM) or only an intracellular domain (Notch1IC) function as oncoproteins. To elucidate the effect of Notch1 abnormalities in radiation-induced lymphomagenesis, we determined the structure of the Notch1 gene and examined the frequency and the sites of Notch1 rearrangements in radiation-induced mouse thymic lymphomas. The Notch1 gene consists of 37 exons, including three exons upstream of the previously reported exon 1. The transcript starting from exon 1 was the major transcript whereas the transcripts read upstream from exon 1a, in which amino acid sequences in the N-terminal region were changed, were minor. More than 50% of radiation-induced thymic lymphomas exhibited Notch1 rearrangements, suggesting that Notch1 acts as a major oncogene in radiation-induced lymphomagenesis. We identified three rearranged sites: novel sites in the 5\u27 end region encompassing exons 1 and 2, the previously identified juxtamembrane extracellular region, and the 3\u27 end region. The 5\u27 deletion and the insertion of murine leukemia virus in the juxtamembrane region led to the production of abnormal transcripts starting from cryptic transcription start sites located halfway through the Notch1 gene and resulted in transcripts lacking most of the extracellular domain. As a result of these rearrangements, truncated Notch1 polypeptides resembling Notch1TM or Notch1IC were formed. In contrast, the 3\u27 deletion led to the production of a C-terminal PEST motif-deleted transcript. The downstream target gene Hes1 was transcribed in a lymphoma with insertion of murine leukemia virus, but not in a lymphoma with a 5\u27 deletion. These results indicate that in addition to Hes1 expression, other Notch1 pathway(s) have a role in thymic lymphomagenesis and suggest the presence of a novel mechanism for oncogenic activation of Notch1 by 5\u27 deletion