Chronic irradiation with low-dose-rate ¹³⁷Cs-γ rays inhibits NGF-induced neurite extension of PC12 cells via Ca²⁺/calmodulin-dependent kinase II activation

Chronic irradiation with low-dose-rate ¹³⁷Cs-γ rays inhibits the differentiation of human neural progenitor cells and influences the expression of proteins associated with several cellular functions. We aimed to determine whether such chronic irradiation influences the expression of proteins associated with PC12 cells. Chronic irradiation at 0.027 mGy/min resulted in inhibition of NGF-induced neurite extension. Furthermore, irradiation enhanced the nerve growth factor (NGF)-induced increase in the phosphorylation of extracellular signal–regulated kinase (ERK), but did not affect the phosphorylation of NGF receptors, suggesting that irradiation influences pathways unassociated with the activation of ERK. We then examined whether irradiation influenced the Akt−Rac1 pathway, which is unaffected by ERK activation. Chronic irradiation also enhanced the NGF-induced increase in Akt phosphorylation, but markedly inhibited the NGF-induced increase in Rac1 activity that is associated with neurite extension. These results suggest that the inhibitory effect of irradiation on neurite extension influences pathways unassociated with Akt activation. As Ca²⁺ /calmodulin-dependent kinase II (CaMKII) is known to inhibit the NGF-induced neurite extension in PC12 cells, independent of ERK and Akt activation, we next examined the effects of irradiation on CaMKII activation. Chronic irradiation induced CaMKII activation, while application of KN-62 (a specific inhibitor of CaMKII), attenuated increases in CaMKII activation and recovered neurite extension and NGF-induced increases in Rac1 activity that was inhibited by irradiation. Our results suggest that chronic irradiation with low-dose-rate γ-rays inhibits Rac1 activity via CaMKII activation, thereby inhibiting NGF-induced neurite extension

Effects of Microalloying on the Impact Toughness of Ultrahigh-Strength TRIP-Aided Martensitic Steels

The effects of the addition of Cr, Mo, and/or Ni on the Charpy impact toughness of a 0.2 pct C-1.5 pct Si-1.5 pct Mn-0.05 pct Nb transformation-induced plasticity (TRIP)-aided steel with a lath-martensite structure matrix (i.e., a TRIP-aided martensitic steel or TM steel) were investigated with the aim of using the steel in automotive applications. In addition, the relationship between the toughness of the various alloyed steels and their metallurgical characteristics was determined. When Cr, Cr-Mo, or Cr-Mo-Ni was added to the base steel, the TM steel exhibited a high upper-shelf Charpy impact absorbed value that ranged from 100 to 120 J/cm2 and a low ductile–brittle fracture appearance transition temperature that ranged from 123 K to 143 K (−150 °C to −130 °C), while also exhibiting a tensile strength of about 1.5 GPa. This impact toughness of the alloyed steels was far superior to that of conventional martensitic steel and was caused by the presence of (i) a softened wide lath-martensite matrix, which contained only a small amount of carbide and hence had a lower carbon concentration, (ii) a large amount of finely dispersed martensite-retained austenite complex phase, and (iii) a metastable retained austenite phase of 2 to 4 vol pct in the complex phase, which led to plastic relaxation via strain-induced transformation and played an important role in the suppression of the initiation and propagation of voids and/or cleavage cracks.ArticleMETALLURGICAL AND MATERIALS TRANSACTIONS A-PHYSICAL METALLURGY AND MATERIALS SCIENCE. 44A(11):5006-5017 (2013)journal articl

Bisbenzamidine derivative, pentamidine represses DNA damage response through inhibition of histone H2A acetylation

<p>Abstract</p> <p>Background</p> <p>MRE11 is an important nuclease which functions in the end-resection step of homologous recombination (HR) repair of DNA double-strand breaks (DSBs). As MRE11-deficient ATLD cells exhibit hyper radio-sensitivity and impaired DSB repair, MRE11 inhibitors could possibly function as potent radio-sensitizers. Therefore, we investigated whether a bisbenzamidine derivative, pentamidine, which can inhibit endoexonuclease activity, might influence DSB-induced damage responses <it>via </it>inhibition of MRE11.</p> <p>Results</p> <p>We first clarified that pentamidine inhibited MRE11 nuclease activity and also reduced ATM kinase activity in vitro. Pentamidine increased the radio-sensitivity of HeLa cells, suggesting that this compound could possibly influence DNA damage response factors in vivo. Indeed, we found that pentamidine reduced the accumulation of γ-H2AX, NBS1 and phospho-ATM at the sites of DSBs. Furthermore, pentamidine decreased HR activity <it>in vivo</it>. Pentamidine was found to inhibit the acetylation of histone H2A which could contribute both to inhibition of IR-induced focus formation and HR repair. These results suggest that pentamidine might exert its effects by inhibiting histone acetyltransferases. We found that pentamidine repressed the activity of Tip60 acetyltransferase which is known to acetylate histone H2A and that knockdown of Tip60 by siRNA reduced HR activity.</p> <p>Conclusion</p> <p>These results indicate that inhibition of Tip60 as well as hMRE11 nuclease by pentamidine underlies the radiosensitizing effects of this compound making it an excellent sensitizer for radiotherapy or chemotherapy.</p

Factor Analysis for Land Value Index in Urban Areas Using Agent Analysis Indicator

The factors affecting a land value index such as land assessments are important for the development and growth of urban areas. Ota and Kaneda (2018) conducted a comparative analysis of a land value index in the central Nagoya area of Japan and reported that the factor structure could be explained by three factors: distance from the nearest station as an accessibility factor; the concentration of neighborhood commercial and business uses as a facility volume factor; and the integration value of the entire area as an indicator of the street network centrality of the vis graph analysis of space syntax theory, or “VGA”, as a space configuration factor. In an urban area, a busy street’s land value index is considered to be higher. The integration value of the VGA indicator, which represents the street network centrality as a space configuration, has been used as a busy street factor. However, high street network centrality is not always needed for a busy street. Therefore, it is a possible that simulating actual pedestrians from the space configuration is a stronger factor for a busy street than a high street network centrality. Simulating actual pedestrians from the space configuration can be conducted using agent analysis, or “AA.” In this paper, we examine a multiple regression model for the factors and a land value index of the Kanayama area of Nagoya City using a VGA indicator and then replacing the VGA indicator with the AA indicator as a new factor. By comparing the two models, we explore the potential for using the AA indicator as a land value index factor. In conclusion, the global integration value of the VGA indicator was selected as a factor for a busy street with a multiple correlation coefficient of 0.750, a coefficient of determination of 0.562, and an Akaike information criterion (AIC) of 352.093 with a standard partial regression coefficient of 0.362 in the conventional factor structure. On the other hand, when the number of AA footprints (station occurrences) of the AA indicator was selected as a factor for a busy street, it had a multiple correlation coefficient of 0.830, a coefficient of determination of 0.689, and an AIC of 294.477 with a standard partial regression coefficient of 0.618 in the new factor structure. Thus, we discovered that replacing the VGA indicator with the AA indicator could significantly improve the land value factor structure model

Polarization-artifact reduction and accuracy improvement of Jones-matrix polarization-sensitive optical coherence tomography by multi-focus averaging

Polarization-sensitive optical coherence tomography (PS-OCT) is a promising biomedical imaging tool for differentiation of various tissue properties. However, the presence of multiple-scattering (MS) signals can degrade the quantitative polarization measurement accuracy. We demonstrate a method to reduce MS signals and increase the measurement accuracy of Jones matrix PS-OCT. This method suppresses MS signals by averaging of multiple Jones matrix volumes measured using different focal positions. The MS signals are decorrelated among the volumes by focus position modulation and are thus reduced by averaging. However, the single scattering signals are kept consistent among the focus-modulated volumes by computational refocusing. We validated the proposed method using a scattering phantom and a postmortem medaka fish. The results showed reduced artifacts in birefringence and degree-of-polarization uniformity measurements, particularly in deeper regions in the samples. This method offers a practical solution to mitigate MS-induced artifacts in PS-OCT imaging and improves quantitative polarization measurement accuracy

Expression of Long-form N-Acetylglucosamine-6-O-Sulfotransferase 1 in Human High Endothelial Venules

Two members of the N-acetylglucosamine-6-O-sulfotransferase (GlcNAc6ST) family, GlcNAc6ST-1 and GlcNAc6ST-2, function in the biosynthesis of 6-sulfo sialyl Lewis X-capped glycoproteins expressed on high endothelial venules (HEVs) in secondary lymphoid organs. Thus, both enzymes play a critical role in L-selectin-expressing lymphocyte homing. Human GlcNAc6ST-1 is encoded by a 1593-bp open reading frame exhibiting two 5' in-frame methionine codons spaced 141 bp apart. Both resemble the consensus sequence for translation initiation. Thus, it has been hypothesized that both long and short forms of GlcNAc6ST-1 may be present, although endogenous expression of either form has not been confirmed in humans. Here, the authors developed an antibody recognizing amino acid residues between the first two human GlcNAc6ST-1 methionines. This antibody specifically recognizes the long form of the enzyme, a finding validated by Western blot analysis and immunofluorescence cytochemistry of HeLa cells misexpressing long and/or short forms of human GlcNAc6ST-1. Using this antibody, the authors carried out immunofluorescence histochemistry of human lymph node tissue sections and found endogenous expression of the long form of the enzyme in human tissue, predominantly in the trans-Golgi network of endothelial cells that form HEVs. (J Histochem Cytochem 60:397-407, 2012)ArticleJOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY. 60(5):397-407 (2012)journal articl

Multi log-normal density structure in Cygnus-X molecular clouds: A fitting for N-PDF without power-law

We studied the H$_2$ column density probability distribution function (N-PDF) based on molecular emission lines using the Nobeyama 45-m Cygnus X CO survey data. Using the DENDROGRAM and SCIMES algorithms, we identified 124 molecular clouds in the $^{13}$CO data. From these identified molecular clouds, an N-PDF was constructed for 11 molecular clouds with an extent of more than 0.4 deg$^2$. From the fitting of the N-PDF, we found that the N-PDF could be well-fitted with one or two log-normal distributions. These fitting results provided an alternative density structure for molecular clouds from a conventional picture. We investigated the column density, dense molecular cloud cores, and radio continuum source distributions in each cloud and found that the N-PDF shape was less correlated with the star-forming activity over a whole cloud. Furthermore, we found that the log-normal N-PDF parameters obtained from the fitting showed two impressive features. First, the log-normal distribution at the low-density part had the same mean column density ($\sim$ 10$^{21.5}$ cm$^{-2}$) for almost all the molecular clouds. Second, the width of the log-normal distribution tended to decrease with an increasing mean density of the structures. These correlations suggest that the shape of the N-PDF reflects the relationship between the density and turbulent structure of the whole molecular cloud but is less affected by star-forming activities.Comment: 14 pages, 7 Figures, Accepted in MNRA

WRN participates in translesion synthesis pathway through interaction with NBS1.

Werner syndrome (WS), caused by mutation of the WRN gene, is an autosomal recessive disorder associated with premature aging and predisposition to cancer. WRN belongs to the RecQ DNA helicase family, members of which play a role in maintaining genomic stability. Here, we demonstrate that WRN rapidly forms discrete nuclear foci in an NBS1-dependent manner following DNA damage. NBS1 physically interacts with WRN through its FHA domain, which interaction is important for the phosphorylation of WRN. WRN subsequently forms DNA damage-dependent foci during the S phase, but not in the G1 phase. WS cells exhibit an increase in spontaneous focus formation of poleta and Rad18, which are important for translesion synthesis (TLS). WRN also interacts with PCNA in the absence of DNA damage, but DNA damage induces the dissociation of PCNA from WRN, leading to the ubiquitination of PCNA, which is essential for TLS. This dissociation correlates with ATM/NBS1-dependent degradation of WRN. Moreover, WS cells show constitutive ubiquitination of PCNA and interaction between PCNA and Rad18 E3 ligase in the absence of DNA damage. Taken together, these results indicate that WRN participates in the TLS pathway to prevent genomic instability in an ATM/NBS1-dependent manner

Induction of Excess Centrosomes in Neural Progenitor Cells during the Development of Radiation-Induced Microcephaly

The embryonic brain is one of the tissues most vulnerable to ionizing radiation. In this study, we showed that ionizing radiation induces apoptosis in the neural progenitors of the mouse cerebral cortex, and that the surviving progenitor cells subsequently develop a considerable amount of supernumerary centrosomes. When mouse embryos at Day 13.5 were exposed to γ-rays, brains sizes were reduced markedly in a dose-dependent manner, and these size reductions persisted until birth. Immunostaining with caspase-3 antibodies showed that apoptosis occurred in 35% and 40% of neural progenitor cells at 4 h after exposure to 1 and 2 Gy, respectively, and this was accompanied by a disruption of the apical layer in which mitotic spindles were positioned in unirradiated mice. At 24 h after 1 Gy irradiation, the apoptotic cells were completely eliminated and proliferation was restored to a level similar to that of unirradiated cells, but numerous spindles were localized outside the apical layer. Similarly, abnormal cytokinesis, which included multipolar division and centrosome clustering, was observed in 19% and 24% of the surviving neural progenitor cells at 48 h after irradiation with 1 and 2 Gy, respectively. Because these cytokinesis aberrations derived from excess centrosomes result in growth delay and mitotic catastrophe-mediated cell elimination, our findings suggest that, in addition to apoptosis at an early stage of radiation exposure, radiation-induced centrosome overduplication could contribute to the depletion of neural progenitors and thereby lead to microcephaly