Contribution of Neuropilin-1 in Radiation-Survived Subclones of NSCLC Cell Line H1299

Non-small cell lung cancer (NSCLC) is an aggressive lung cancer accounting for approximately 85% of all lung cancer patients. For the patients with Stages IIIA, IIIB, and IIIC, the 5-year survival is low though with the combination with radiotherapy and chemotherapy. In addition, the occurrence of tumor cells (repopulated tumors) that survive irradiation remains a challenge. In our previous report, we subcloned the radiation-surviving tumor cells (IR cells) using the human NSCLC cell line, H1299, and found that the expression of neuropilin-1 (NRP-1) was upregulated in IR cells by the microarray analysis. Here, we investigated the contribution of neuropilin-1 to changes in the characteristics of IR cells. Although there were no differences in angiogenic activity in the tube formation assay between parental and IR cells, the cell motility was increased in IR cells compared to parental cells in the cell migration assay. This enhanced cell motility was suppressed by pretreatment with anti-NRP-1 antibody. Although further studies are necessary to identify other molecules associated with NRP-1, the increase in cellular motility in IR cells might be due to the contribution of NRP-1. Inhibition of NRP-1 would help control tumor malignancy in radiation-surviving NSCLC.     

Hemiptelea mikii sp. nov. (Ulmaceae), fossil fruits and woods from the pleistocene of central Japan

A new fossil species,Hemiptelea mikii, is described on the basis of fruits from the Pleistocene of central Japan. It is distinguished fromH. davidii, the only extant species of the genus restricted to Korea and China, in having shorter fruits. Fossil woods discovered from the same horizon differ from the extant one in some anatomical characteristics, and are assigned to the new fossil species. Taken together with earlier records of fossil fruit occurrence in the early and the middle Pleistocene of central Japan, the latest finding of fossil fruits and woods from the last glacial sediments (ca. 50,000 years ago) at Kita-egota site of Tokyo suggests thatH. mikii was widespread in central Japan throughout the Pleistocene and survived until as late as the Last Glacial age.     

Role of tumor necrosis factor-related apoptosis-inducing ligand in immune response to influenza virus infection in mice

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis of various tumor cells but not normal cells. However, various cytokines and virus infection differentially regulate TRAIL and TRAIL receptor expression. It has been demonstrated that virus infection changes the pattern of human TRAIL-receptor expression on normal cells, which were resistant to TRAIL-mediated apoptosis, and makes them susceptible to TRAIL-mediated apoptosis. Since previous studies on the function of TRAIL have been performed mainly in vitro, its physiological role in the immune response to virus infection remains unknown. In the present study, we investigated the expression of TRAIL in the lungs of influenza virus-infected mice and the function of TRAIL in the immune response to infection. Influenza virus infection increased TRAIL mRNA expression in the lung. TRAIL protein expression was induced on NK cells in the lung 4 days after infection. At 7 days after infection, TRAIL protein expression was also detected on CD4(+) and CD8(+) T cells. However, NK cells and T cells in the lungs of uninfected mice did not express a detectable level of TRAIL on their cell surfaces. DR5, which is a mouse TRAIL receptor, was also induced to express after virus infection. Expression of both TRAIL and DR5 mRNAs was reduced to normal level at 6 weeks after virus infection. Administration of anti-TRAIL monoclonal antibody, which blocks TRAIL without killing TRAIL-expressing cells, to mice during influenza virus infection significantly delayed virus clearance in the lung. These results suggest that TRAIL plays an important role in the immune response to virus infection.     

Purkinje cells originate from cerebellar ventricular zone progenitors positive for Neph3 and E-cadherin

GABAergic Purkinje cells (PCs) provide the primary output from the cerebellar cortex, which controls movement and posture. Although the mechanisms of PC differentiation have been well studied, the precise origin and initial specification mechanism of PCs remain to be clarified. Here, we identified a cerebellar and spinal cord GABAergic progenitor-selective cell surface marker, Neph3, which is a direct downstream target gene of Ptf1a, an essential regulator of GABAergic neuron development. Using FACS, Neph3⁺ GABAergic progenitors were sorted from the embryonic cerebellum, and the cell fate of this population was mapped by culturing in vitro. We found that most of the Neph3⁺ populations sorted from the mouse E12.5 cerebellum were fated to differentiate into PCs while the remaining small fraction of Neph3⁺ cells were progenitors for Pax2⁺ interneurons, which are likely to be deep cerebellar nuclei GABAergic neurons. These results were confirmed by short-term in vivo lineage-tracing experiments using transgenic mice expressing Neph3 promoter-driven GFP. In addition, we identified E-cadherin as a marker selectively expressed by a dorsally localized subset of cerebellar Neph3⁺ cells. Sorting experiments revealed that the Neph3⁺ E-cadherin^high population in the embryonic cerebellum defined PC progenitors while progenitors for Pax2⁺ interneurons were enriched in the Neph3⁺ E-cadherin^low population. Taken together, our results identify two spatially demarcated subregions that generate distinct cerebellar GABAergic subtypes and reveal the origin of PCs in the ventricular zone of the cerebellar primordium.     

Late Quaternary vegetation and climatic history of eastern Nepal

The late Quaternary vegetation and climatic history at high altitude in eastern Nepal was studied through stratigraphy, radiocarbon dating and pollen analysis. The Thulo Pokhari lake (27°41′ N, 87°43′ E, ca. 3980 m a.s.l.) is just above the subalpine Abies spectabilis–Betula utilis forest zone and surrounded by Rhododendron scrub. The climate in the studied area is very humid under the influence of the monsoon. Silty sediments suggesting lake conditions prevailed at the lower part of the sediment samples, whereas the upper part consisted mainly of herbaceous peat indicating a mire condition. The pollen record extends back to ca. 11000 yr bp . Although the absence of major changes between the pollen assemblages of the Pleistocene and those in the Holocene was attributed to the situation of the sampling plot above the forest limit, the results suggest the following history. (1) The abundance of Pinus pollen around 11000 yr bp represents expansion of the area unoccupied by late successional trees and/or a drier condition. Significant occurrence of Picea pollen is noteworthy, since this genus is absent in eastern Nepal at present in spite of its wide distribution throughout the neighbouring areas of the Himalayas. (2) The increase in abundance of Quercus pollen and the decline in Pinus pollen in the latest Pleistocene represent climatic amelioration favourable to late successional mesophytes. (3) The climate was most moist and mild in the SP-III, although the absolute age of the pollen zone is not distinct. The percentages of broadleaved trees such as Alnus, Betula, Carpinus and Corylus were increased at the expense of Quercus and coniferous pollen types. Sedimentation was interrupted by lake level decline and/or glacial advance during the period from ca. 11000–1600 yr bp . (4) The increase in abundance of coniferous pollen types such as Pinus, Abies and Tsuga indicates that the climate became drier and cooler, and the present subalpine forest zone composed of A. spectabilis, T. dumosa and Betula utilis was established around 1600 yr bp . The formation of bog or alpine meadow vegetation during the period was shown by the ¹⁴C dates of the peat and the increase in Rosaceae and herbaceous pollen. (5) The increase in abundance of Ericaceae and Alnus pollen in the SP-V indicates that the present alpine Rhododendron scrub zone has been established since ca. 940 yr bp as a result of human influence on the original vegetation. This interpretation is supported by the decline in the proportion of arboreal pollen. Since the age of the initiation of human influence varies with the altitude and region, further study will be necessary to determine the accurate age.     

Cutting edge: Fyn is essential for tyrosine phosphorylation of Csk-binding protein/phosphoprotein associated with glycolipid-enriched microdomains in lipid rafts in resting T cells

In resting T cells, Csk is constitutively localized in lipid rafts by virtue of interaction with a phosphorylated adaptor protein, Csk-binding protein (Cbp)/phosphoprotein associated with glycolipid-enriched microdomains, and sets an activation threshold in TCR signaling. In this study, we examined a kinase responsible for Cbp phosphorylation in T cell membrane rafts. By analyzing T cells from Fyn-/- mice, we clearly demonstrated that Fyn, but not Lck, has its kinase activity in membrane rafts, and plays a critical role in Cbp phosphorylation, Cbp-Csk interaction, and Csk kinase activity. Naive CD44(low)CD62 ligand(high) T cells were substantially reduced in Fyn-/- mice, presumably due to the inhibition of Cbp phosphorylation. Thus, Fyn mediates Cbp-Csk interaction and recruits Csk to rafts by phosphorylating Cbp. Csk recruited to rafts would then be activated and inhibit the kinase activity of Lck to keep resting T cells in a quiescent state. Our results elucidate a negative regulatory role for Fyn in proximal TCR signaling in lipid rafts.