Blood Pressure Elevation of Tubular Specific (P)RR Transgenic Mice and Lethal Tubular Degeneration due to Possible Intracellular Interactions between (P)RR and Alternative Renin Products

The prorenin/renin receptor ((P)RR) is a multifunctional protein that is widely distributed in various organs. Despite intensive research for more than 20 years, this receptor has not been fully characterized. In this study, we generated mice overexpressing the tubular epithelial (P)RR gene ((P)RR-TG mice) to test the previously reported functional role of (P)RR by Ramkumar et al. in 2015 using tubular specific (P)RR KO mice. (P)RR-TG mice were maintained and analyzed in individual metabolic cages and were administered angiotensin II blocker (ARB), direct renin inhibitor (DRI), and bafilomycin, that is, vacuolar ATPase (V-ATPase) antagonist. (P)RR-TG mice were hypertensive and had alkalized urine with lower osmolality and Na⁺ excretion. ARB and DRI, but not bafilomycin, concurrently decreased blood pressure. Bafilomycin acidized urine of (P)RR-TG mice, or equivalently this phenomenon restored the effect of overexpressed transgene, suggesting that (P)RR functioned as a V-ATPase in renal tubules. Afterall, (P)RR-TG mice were mated with alternative renin transgenic mice (ARen2-TG), which we identified as intracellular renin previously, to generate double transgenic mice (DT-TG). Lethal renal tubular damage was observed in DT-TG mice, suggesting that intracellular renin may be a ligand for (P)RR in tubules. In summary, (P)RR did not substantially affect the tissue renin-angiotensin system (RAS) in our model of tubular specific (P)RR gene over-expression, but alternative intracellular renin may be involved in (P)RR signaling in addition to conventional V-ATPase function. Further investigations are warranted.     

Size-dependent separation of graphene oxide by deformation of packed-gel in a chromatographic column

ABSTRACT

A deformable gel-packed chromatographic column was used to separate as-synthesized graphite oxide with different sizes. The synthesized gel (56 µm) was deformed by pressure of the fluid flow and the gaps in the gels showed a range of sizes. A suspension of graphene oxide (0.1 g/L, 10 mL) was injected, and graphene oxide in the elution had a size at 0.56 μm and 0.14 μm, whereas in half upper and bottom domain of the gel layer graphene oxide had a size at 33 µm and 2.9 µm, respectively, demonstrating that graphene oxide suspension was separated by size through gel layer.     

Melatonin-Induced Postconditioning Suppresses NMDA Receptor through Opening of the Mitochondrial Permeability Transition Pore via Melatonin Receptor in Mouse Neurons

Mitochondrial membrane potential regulation through the mitochondrial permeability transition pore (mPTP) is reportedly involved in the ischemic postconditioning (PostC) phenomenon. Melatonin is an endogenous hormone that regulates circadian rhythms. Its neuroprotective effects via mitochondrial melatonin receptors (MTs) have recently attracted attention. However, details of the neuroprotective mechanisms associated with PostC have not been clarified. Using hippocampal CA1 pyramidal cells from C57BL mice, we studied the involvement of MTs and the mPTP in melatonin-induced PostC mechanisms similar to those of ischemic PostC. We measured changes in spontaneous excitatory postsynaptic currents (sEPSCs), intracellular calcium concentration, mitochondrial membrane potential, and N-methyl-D-aspartate receptor (NMDAR) currents after ischemic challenge, using the whole-cell patch-clamp technique. Melatonin significantly suppressed increases in sEPSCs and intracellular calcium concentrations. The NMDAR currents were significantly suppressed by melatonin and the MT agonist, ramelteon. However, this suppressive effect was abolished by the mPTP inhibitor, cyclosporine A, and the MT antagonist, luzindole. Furthermore, both melatonin and ramelteon potentiated depolarization of mitochondrial membrane potentials, and luzindole suppressed depolarization of mitochondrial membrane potentials. This study suggests that melatonin-induced PostC via MTs suppressed the NMDAR that was induced by partial depolarization of mitochondrial membrane potential by opening the mPTP, reducing excessive release of glutamate and inducing neuroprotection against ischemia-reperfusion injury.     

Metastatic Voyage of Ovarian Cancer Cells in Ascites with the Assistance of Various Cellular Components

Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy and has a unique metastatic route using ascites, known as the transcoelomic root. However, studies on ascites and contained cellular components have not yet been sufficiently clarified. In this review, we focus on the significance of accumulating ascites, contained EOC cells in the form of spheroids, and interaction with non-malignant host cells. To become resistant against anoikis, EOC cells form spheroids in ascites, where epithelial-to-mesenchymal transition stimulated by transforming growth factor-β can be a key pathway. As spheroids form, EOC cells are also gaining the ability to attach and invade the peritoneum to induce intraperitoneal metastasis, as well as resistance to conventional chemotherapy. Recently, accumulating evidence suggests that EOC spheroids in ascites are composed of not only cancer cells, but also non-malignant cells existing with higher abundance than EOC cells in ascites, including macrophages, mesothelial cells, and lymphocytes. Moreover, hetero-cellular spheroids are demonstrated to form more aggregated spheroids and have higher adhesion ability for the mesothelial layer. To improve the poor prognosis, we need to elucidate the mechanisms of spheroid formation and interactions with non-malignant cells in ascites that are a unique tumor microenvironment for EOC.     

Observation of atomic steps on single crystal surfaces by a commercial scanning electron microscope.

Atomic steps on (111) and (100) crystal surfaces of Pt were observed using a commercial scanning electron microscope (SEM) in secondary electron mode. By comparing the SEM images and those by reflection electron microscopy (REM), the observed contrast was confirmed to be that from atomic steps on crystal surfaces. The contrast mechanism is briefly discussed. One application of this imaging technique is also shown.     

Applications of microstructures fabricated by proton beam writing to electric-micro filters

Fabrication of high-aspect-ratio microstructures was performed by proton beam writing (PBW) using a microbeam line at Takasaki Ion Accelerators for Advanced Radiation Application (TIARA), JAEA Takasaki, JAPAN. As one of the applications of the high-aspect-ratio structures micro-machined by PBW, we utilized the high-aspect pillars for electric-micro filters of microbes such as Escherichia coli and Yeast based on the dielectrophoretic force. The filter is equipped with high-aspect pillars with a height of ～20 μm and a diameter of ～1 μm on a glass plate. Evaluation of the dielectrophoresis (DEP) device for capturing E. coli and Yeast was made using either observation by optical microscope or photoluminescence (PL) measurements.     

Present Status of the Nd:YAG Thomson Scattering System Development for Time Evolution Measurement of Plasma Profile on Heliotron J

A new high repetition rate Nd:YAG Thomson scattering system is developed for the Heliotron J helical device.A main purpose of installing the new system is the temporal evolution measurement of a plasma profile for improved confinement physics such as the edge transport barrier (H-mode) or the internal transport barrier of the helical plasma.The system has 25 spatial points with ～10mm resolution.Two high repetition Nd:YAG lasers (>550mJ@ 50Hz) realize the measurement of the time evolution of the plasma profile with ～10ms time intervals.Scattered light is collected by a large concave mirror (D=800 mm,f/2.25) with a solid angle of ～100 mstr and transferred to interference filter polychromators by optical fiber bundles in a staircase form.The signal is amplified by newly designed fast preamplifiers with DC and AC output,which reduces the low frequency background noise.The signals are digitized with a multi-event QDC,fast gated integrators.The data acquisition is performed by a VME-based system operated by the CINOS.     

D-Amino Acids in Non-gelling Components of Gelatin

Abstract

As a possible cause of the non-gelling property of the sol fraction of gelatin, we considered that higher D-amino acid content should result in decreased formation of helices. The sol fractions ivere extracted from gelatin gels and hydrolyzed to amino acid mixtures. The rates of reaction with D-amino acid oxidase (D-AOX) were recorded for thehydrolyzates. For a comparison, residual gels were also treated similarly. The sol fractions always gave higher results than the residual gels for the three types of gelatin examined. The result also showed that the rates obtained from the sol fractions were in the order: limed ossein > limed hide > acid pigskin.

To examine the reliability of this procedure the D-form content for each amino acid in the sol from the limed hide gelatin was compared with that of the residual gel. The contents in the sol were higher than those in the residue for most of the amino acids which racemized. The results indicate that the increased racemization of amino acids on the extraction of gelatin from raw materials is a cause of the non-gelling property.     

Mixing speed-controlled gold nanoparticle synthesis with pulsed mixing microfluidic system

Gold nanoparticles with diameters of a few tens of nanometer and a narrow size distribution were synthesized using a pulsed mixing method with a microfluidic system which consists of a Y-shaped mixing microchannel and two piezoelectric valveless micropumps. This mixing method enables control of the mixing speed of gold salts and reducing agent by changing the switching frequency of the micropumps, which was our focus to improve the particle size distribution, which is an essential parameter in gold nanoparticle synthesis. In the proposed method, the mixing time was inversely proportional to the switching frequency and the minimum mixing time was 95 ms at a switching frequency of 200 Hz. During synthesis experiments, the mean diameter of the synthesized gold nanoparticles was found to increase, and the coefficient of variation of particle size was found to decrease with decreasing mixing time. We successfully improved the coefficient of variation to less than 10% for a mean diameter of around 40 nm.