Glycosphingolipid antigens in neural tumor cell lines and anti-glycosphingolipid antibodies in sera of patients with neural tumors

To characterize biomarkers in neural tumors, we analyzed the acidic lipid fractions of 13 neural tumor cell lines using enzyme-linked immunoabsorbent assay (ELISA) and high-performance thin-layer chromatography (HPTLC) immunostaining. Sulfated glucuronosyl glycosphingolipids (SGGLs) are cell surface molecules that are endowed with the Human Natural Killer-1 (HNK-1) carbohydrate epitope. These glycosphingolipids (GSLs) were expressed in all cell lines with concentrations ranging from 210 to 330 ng per 2 x 10(6) cells. Sulfoglucuronosyl paragloboside (SGPG) was the prominent species with lesser amounts of sulfoglucuronosyl lactosaminyl paragloboside (SGLPG) in these tumor cell lines as assessed by quantitative HPTLC immunostaining. Among the gangliosides surveyed, GD3 and 9-O-acetylated GD3 (OAc-GD3) were expressed in all tumor cell lines. In contrast, fucosyl-GM1 was not found to restrict to small cell lung carcinoma cells. In addition, we have analyzed serum antibody titers against SGPG, GD3, and OAc-GD3 in patients with neural tumors by ELISA and HPTLC immunostaining. All sera had high titers of antibodies of the IgM isotype against SGPG (titers over 1:3,200), especially in tumors such as meningiomas, germinomas, orbital tumors, glioblastomas, medulloblastomas, and subependymomas. Serum in a patient with subependymomas also had a high anti-SGGL antibody titer of the IgG and IgA types (titers over 12,800). The titer of anti-GD3 antibody was also elevated in patients with subependymomas and medulloblastomas; the latter cases also had a high titer of antibody against OAc-GD3. Our data indicate that certain GSL antigens, especially SGGLs, GD3, and OAc-GD3, are expressed in neural tumor cells and may be considered as tumor-associated antigens that represent important biomarkers for neural tumors. Furthermore, antibody titers in sera of patients with these tumors may be of diagnostic value for monitoring the presence of tumor cells and tumor progression.     

Direct activation of mitochondrial apoptosis machinery by c-Jun N-terminal kinase in adult cardiac myocytes.

Although oxidative stress causes activation of c-Jun N-terminal kinase (JNK) and apoptosis in many cell types, how the JNK pathway is connected to the apoptosis pathway is unclear. The molecular mechanism of JNK-mediated apoptosis was investigated in adult rat cardiac myocytes in culture as a model system that is sensitive to oxidative stress. Oxidative stress caused JNK activation, cytochrome c release, and apoptosis without new protein synthesis. Oxidative stress-induced apoptosis was abrogated by dominant negative stress-activated protein kinase/extracellular signal-regulated kinase kinase-1 (SEK1)-mediated inhibition of the JNK pathway, whereas activation of the JNK pathway by constitutively active SEK1 was sufficient to cause apoptosis. Inhibition of caspase-9, an apical caspase in the mitochondrial apoptosis pathway, suppressed oxidative stress-induced apoptosis, whereas inhibition of caspase-8 had no effect, indicating that both the JNK pathway and the mitochondrial apoptosis machinery are central to oxidative stress-induced apoptosis. Both JNK and SEK1 localized on mitochondria where JNK was activated by oxidative stress. Furthermore, active JNK caused the release of apoptogenic factors such as cytochrome c from isolated mitochondria in a cell-free assay. These findings indicate that the JNK pathway is a direct activator of mitochondrial death machinery without other cellular components and provide a molecular linkage from oxidative stress to the mitochondrial apoptosis machinery.     

Characterization of a Heme-Dependent Catalase from Methanobrevibacter arboriphilus

Recently it was reported that methanogens of the genus Methanobrevibacter exhibit catalase activity. This was surprising, since Methanobrevibacter species belong to the order Methanobacteriales, which are known not to contain cytochromes and to lack the ability to synthesize heme. We report here that Methanobrevibacter arboriphilus strains AZ and DH1 contained catalase activity only when the growth medium was supplemented with hemin. The heme catalase was purified and characterized, and the encoding gene was cloned. The amino acid sequence of the catalase from the methanogens is most similar to that of Methanosarcina barkeri.     

Interspecies Transmission of Feline Immunodeficiency Virus from the Domestic Cat to the Tsushima Cat (Felis bengalensis euptilura) in the Wild

Feline immunodeficiency virus (FIV) was isolated from a wild-caught Tsushima cat (Felis bengalensis euptilura), an endangered Japanese nondomestic subspecies of leopard cat (F. bengalensis). Phylogenetic analysis of the env gene sequences indicated that the FIV from the Tsushima cat belonged to a cluster of subtype D FIVs from domestic cats. FIVs from both the Tsushima cat and the domestic cat showed similar levels of replication and cytopathicity in lymphoid cell lines derived from these two species. The results indicated the occurrence of interspecies transmission of FIV from the domestic cat to the Tsushima cat in the wild.     

Distribution and temperature tolerance of the Antarctic free-living mite Antarcticola meyeri (Acari,Cryptostigmata)

The ecology and physiology of a free-living mite species Antarcticola meyeri, Cryptostigmata, rarely discovered in the Continental Antarctic Zone, were studied near the Japanese Antarctic Base, Syowa Station. The distribution of this mite species correlated noticeably with the distribution of its food, the imperfect lichen species, which grow in moss carpets. Within a limited habitat, this mite species preferred dry carpets to wet carpets. Low humidity appeared disadvantageous to the species at high temperatures (ca. 30°C), but at low temperatures (ca. -25°C), low humidity appeared advantageous. This may explain why this species preferred dry carpets. The mean supercooling point of starved individuals was -30.6°C (nymphs) to-33.9°C (eggs). When fed with lichens, however, it was significantly raised, probably because the gut contents functioned as ice nucleators or contained ice-nucleating agents. Acclimation to low temperatures significantly lowered the supercooling point of larvae but not of other growth stages.     

Taxonomic character of plant species in absorbing and accumulating alkali and alkaline earth metals grown in temperate forest of Japan

Summary Absorption and accumulation of alkali (Li, Na, K, Rb, Cs) and alkaline earth (Mg, Ca, Sr, Ba) metals were investigated as taxonomic characteristics (in 62 plant species). Leaf and soil samples were collected from 9 sites in temperature forest in Japan and the above mentioned elements were analyzed. Considerable differences were found among species in their ability to accumulate alkali and alkaline earth metals. Very high concentrations of Li (45 ppm, D.W.), K (37×10³ ppm), Rb (159 ppm) and Cs (8.2 ppm) were detected inLastrea japonica which were about 412, 12, 27 and 6 times higher than those of the species with the lowest concentrations. Na content was high inAcer micranthum (358 ppm) which was 16 times higher than species with the lowest concentration. Other species containing high levels of alkali metals wereHydrangea macrophylla, Struthiopteris niponica, Clethra barbinervis. Mean discrimination ratio (D.R.) for all investigated plant species for Li, Na, Rb, and Cs to K were 1.7, 0.44, 0.9 and 1.8 respectively. High concentrations of alkaline earth metals Ca (36×10³ ppm), Sr (345 ppm), and Ba (241 ppm) were found in the leaves ofHydrangea paniculata which were about 31, 84, and 72 times higher than those for the species with the lowest concentration. Mg was very high inStruthiopteris niponica (83×10² ppm). Other species with high concentrations of alkaline earth metals belonged to the genus Viburnum. Mean D.Rs. for Mg, Sr, and Bavs Ca were 1.0, 0.7 and 0.08. Principal component analysis of interrelationships between the mineral content in leaf tissues indicated that these elements could be classified into 2 groups with respect to their accumulation behavior in plants. The alkali metals K, Li, Rb, and Cs behaved similarly in their accumulation in leaves but Na behaved independently. Alkaline earth metals Ca, Mg, Sr, and Ba were also found to behave similarly in their accumulation. Factors scores of 1st and 2nd components revealed three groups of plant species: alkaliphilic, alkaline earthphilic, and neutral (non-accumulators).     

Entomological and ecological surveys on Mt. Usu in 1984 VI: Faunal make-up and ecological distribution of springtails (Collembola) on Volcano Usu

A survey of springtails was conducted in 1984 on Volcano Usu which suffered damage to it's ecosystem from eruptions in 1977 and 1978. The summit area, almost completely deforested by the eruptions, was still inhabited by many forest species such asDesoria notabilis, Willowsia sp. 2,Pseudachorutes sp. 2,Isotoma sp. 4,Tullbergia yosii, Onychiurus sp. 1,Entomobrya sp. 2,Isotomiella minor, Proisotoma sp. 2,Xenylla sp.,Neanura frigida, etc. These species preferred standing or fallen dead trees, mosses, bark chips, former topsoils and soil around the root of plants. The fauna in the summit area was most similar to that of coniferous or broad-leaved forests surveyed on the foot of the mountain. This suggests that the 1977–78 eruptions did not exterminate these springtail species which probably survived on the standing trees or in the former topsoil.