The subionospheric VLF/LF propagation is extensively used to investigate the lower ionospheric perturbation in possible association with earthquakes. An extensive period of data over 7 yr from January 2001 to December 2007 and a combination of different propagation paths in and around Japan are used to examine the statistical correlation between the VLF/LF propagation anomaly (average nighttime amplitude, dispersion, and nighttime fluctuation) and earthquakes with magnitude >6.0. It is then found that the propagation anomaly exceeding the 2σ (standard deviation) criterion indicating the presence of ionospheric perturbation is significantly correlated with earthquakes with shallow depth (<40 km). Finally, the mechanism of seismoionospheric perturbations is discussed.

Abstract. Relatively long-period (4 years) data on different propagation paths by means of Japanese-Pacific VLF/LF network observation, are used to obtain further statistical significance on the correlation of ionospheric perturbations as revealed by VLF/LF propagation anomalies with earthquakes. Earthquakes with magnitude greater than 6.0, taken place only within the fifth Fresnel zone of each great-circle path are selected for the correlation study. It is finally found based on the superimposed epoch analysis that the nighttime trend (average amplitude) exhibits a significant decrease exceeding 2σ (σ : standard deviation) several days before the earthquake and the nighttime fluctuation exceeds the corresponding 2σ again several days before the earthquake when the earthquake depth is smaller than 30 km (shallow earthquakes). However, when we treat all earthquakes including deep earthquakes, the trend shows a significant decrease (just approaching 2σ line), and the nighttime fluctuation shows a less significant broad enhancement before the EQ.

The appearance of ionospheric perturbations in possible association with earthquakes (EQs) seems to be confirmed on the basis of both event and statistical studies, but the generation mechanism of those seismo-ionospheric perturbations is not well understood, though a few possible hypotheses have been proposed. Two hypotheses are promising candidates; one is so-called "chemical channel", in which radon emanation results in the change in atmospheric conductivity, then in the atmospheric electric field, leading to the redistribution of ionospheric plasma, and the other is the excitation of atmospheric oscillations due to some precursory changes of ground surface, which propagate upward and induce the ionospheric perturbation. We support the second "atmospheric oscillation channel", so that in this paper we present several evidences (or facts) in support of this hypothesis. First, we present some case studies of the enhancement of AGW (atmospheric gravity wave) modulation in subionospheric VLF/LF propagation data, and then a recent statistical treatment of this AGW modulation in VLF/LF data is reported. Further studies on the satellite observation and also on the satellite-ground coordinated measurements are presented in order to indicate the AGW as the agent of lithosphere-ionosphere coupling. Finally, our equipment recently developed to measure the Doppler-shift of a short-distance subionospheric LF signal is used to indicate the direct evidence of the AGW hypothesis.

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This simple, accurate, and reproducible colorimetric method for determining the activity of angiotensin-I converting enzyme is based upon colorimetry of the quinoneimine dye produced from the substrate p-hydroxyhippuryl-L-histidyl-L-leucine by action of this enzyme through the following series of reactions. The enzyme acts on the substrate to yield p-hydroxyhippuric acid and L-histidyl-L-leucine. The former is then hydrolyzed in the presence of hippuricase to produce p-hydroxybenzoic acid and glycine. Finally, oxidative coupling of p-hydroxybenzoic acid with 4-aminoantipyrine is catalyzed by peroxidase in the presence of hydrogen peroxide, producing a quinoneimine dye, the concentration of which is measured at its absorbance maximum at 505 nm to evaluate the activity of ACE. The Km value for the above-mentioned substrate is 0.32 mmol/L, the optimum pH is 8.3. Results by the present method and Cushman and Cheung's method (Biochem. Pharmacol. 20: 1637, 1971) correlate closely (r = 0.986). The within-run CV is 2.93%.

Peptidylarginine deiminases (PADs) are posttranslational modification enzymes that convert protein arginine to citrulline residues in a calcium ion-dependent manner. Previously, we reported the abnormal accumulation of citrullinated proteins and the increase in the amount of PAD2 in hippocampi from Alzheimer's disease (AD) patients. Moreover, glial fibrillary acidic protein (GFAP), an astrocyte-specific marker protein, and vimentin were identified as citrullinated proteins by using two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. To clarify the substrate specificity of PADs against GFAP, we prepared recombinant human (rh)PAD1, rhPAD2, rhPAD3, rhPAD4, and rhGFAP. After incubation of rhGFAP with rhPAD1, rhPAD2, rhPAD3, and rhPAD4, citrullinated (cit-)rhGFAP was detected by Western blotting. The citrullination of rhGFAP by rhPAD2 was unique, specific, and time dependent; additionally, rhPAD1 slightly citrullinated rhGFAP. We then generated eight anti-cit-rhGFAP monoclonal antibodies, CTGF-125, -128, -129, -1212, -1213, -1221, -122R, and -1224R, which reacted specifically with cit-rhGFAP. Two of those eight monoclonal antibodies, CTGF-122R and -1224R, reacted with both cit-rhGFAP and rhGFAP in Western blots. By using the CTGF-1221 antibody and a tandem mass spectrometer, we identified the two independent citrullination sites (R270Cit and R416Cit) of cit-rhGFAP. Immunohistochemical analysis with CTGF-1221 antibody revealed cit-GFAP staining in the hippocampus of AD brain, and the cit-GFAP-positive cells appeared to be astrocyte-like cells. These collective results strongly suggest that PAD2 is responsible for the citrullination of GFAP in the progression of AD and that the monoclonal antibody CTGF-1221, reacting with cit-GFAP at R270Cit and R416Cit, is useful for immunohistochemical investigation of AD brains.

Abstract.As the target earthquake we have taken a huge earthquake (EQ) named Miyagi-oki earthquake on 16 August 2005 (with magnitude of 7.2) and we have analyzed the 4 month period including the date of this EQ. In addition to our previous analysis on the nighttime average amplitude (trend) and nighttime fluctuation, we have proposed the use of fluctuation power spectra in the frequency rage of atmospheric gravity waves (period=10 min to 100 min) as a third parameter of subionospheric VLF/LF propagation characteristics. Then it is found that this third parameter would be of additional importance in confirming the presence of seismoionospheric perturbations. Finally, we have discovered an important role of lunar tidal effect in the VLF/LF data, which appears one and two months before this large EQ.

Abstract. Huge five earthquakes with magnitude greater than 6.0 took place in Asia (include Philippines, Indonesia, etc.) during the period from the beginning of August 2008 to the end of Junuary 2009, and the corresponding data of subionospheric VLF propagation between the NWC transmitter (Australia, 19.8 kHz) and a few Japanese stations (distance 6∼8 Mm) are examined. As the result of our analysis by means of (1) trend (average nighttime amplitude), (2) dispersion, (3) nighttime fluctuation, and (4) atmospheric gravity wave enhancement, three earthquakes from the five taking place within the fifth Frenel zone are found to have accompanied a precursory signature in VLF propagation. On the other hand, there were observed no such precursory signatures for the remaing two earthquakes. One is too deep (>400 km) and another is too distant from the great-circle path. These characteristics of seismo-ionospheric perturbations would be of essential importance in studying the spatial/temporal properties of seismo-ionospheric perturbations for medium-distance propagation.