Semiaquatic Heteroptera locomotion: coral treaders (Hermatobates weddi, Hermatobatidae), sea skaters (Halovelia septentrionalis, Veliidae), and water striders (Metrocoris histrio, Gerridae). Usual and unusual gaits

Using high-speed video recordings, we carried out an analysis of the locomotion gaits of the following aquatic Heteroptera: coral treaders Hermatobates weddi (Hermatobatidae), sea striders Halovelia septentrionalis (Veliidae), and water striders Metrocoris histrio (Gerridae), in the Island of Amami Oshima, Kagoshima Prefecture, Japan. Most insects use an alternating double tripod gait for walking, whereas species of Gerridae and some Veliidae use a synchronous rowing gait. We found that H. weddi used a peculiar locomotion gait, a modification of the double tripod gait. In this special gait, two alternating dipods (mid and hind legs) are used, while the forelegs remained inactive. Contralateral mid and hind stroked simultaneously. The mid leg recovered immediately after the stroke; however, the hind leg was delayed and remained extended after the stroke. Next, the following bipod stroked, and when that mid leg finished the stroke, both ipsilateral mid and hind (the one which did not recover after the stroke) legs recovered together. Turning is also unique in H. weddi because the body axis rotation and the course turning (deflection) were clearly separated in two phases. We compared the kinematics of H. weddi pattern with the synchronous rowing pattern found in H. septentrionalis and M. histrio and discussed some biomechanical consequences. We also analyzed phylogenetic implications of this gait, and we posit that the modified double dipod gait is a uniquely derived character of the family Hermatobatidae. The synchronous rowing gait would be an autapomorphy for the clade Gerridae + Veliidae. The modified thorax, with the meso and metacoxae horizontally directed, would be a synapomorphy for the superfamily Gerroidea (Hermatobatidae, Gerridae, and Veliidae). Handling editor: Koen Martens     

Elucidations of the Catalytic Cycle of NADH-Cytochrome b5 Reductase by X-ray Crystallography: New Insights into Regulation of Efficient Electron Transfer

NADH-Cytochrome b₅ reductase (b5R), a flavoprotein consisting of NADH and flavin adenine dinucleotide (FAD) binding domains, catalyzes electron transfer from the two-electron carrier NADH to the one-electron carrier cytochrome b₅ (Cb5). The crystal structures of both the fully reduced form and the oxidized form of porcine liver b5R were determined. In the reduced b5R structure determined at 1.68 Å resolution, the relative configuration of the two domains was slightly shifted in comparison with that of the oxidized form. This shift resulted in an increase in the solvent-accessible surface area of FAD and created a new hydrogen-bonding interaction between the N5 atom of the isoalloxazine ring of FAD and the hydroxyl oxygen atom of Thr66, which is considered to be a key residue in the release of a proton from the N5 atom. The isoalloxazine ring of FAD in the reduced form is flat as in the oxidized form and stacked together with the nicotinamide ring of NAD⁺. Determination of the oxidized b5R structure, including the hydrogen atoms, determined at 0.78 Å resolution revealed the details of a hydrogen-bonding network from the N5 atom of FAD to His49 via Thr66. Both of the reduced and oxidized b5R structures explain how backflow in this catalytic cycle is prevented and the transfer of electrons to one-electron acceptors such as Cb5 is accelerated. Furthermore, crystallographic analysis by the cryo-trapping method suggests that re-oxidation follows a two-step mechanism. These results provide structural insights into the catalytic cycle of b5R.     

Mechanism of NO-mediated oxidative and nitrative DNA damage in hamsters infected with Opisthorchis viverrini: a model of inflammation-mediated carcinogenesis.

Inflammation mediated by infection is an important factor causing carcinogenesis. Opisthorchis viverrini (OV) infection is a risk factor of cholangiocarcinoma (CHCA), probably through chronic inflammation. Formation of 8-nitroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), and expression of inducible nitric oxide synthase (iNOS) and heme oxygenase-1 (HO-1) were assessed in the liver of hamsters infected with OV. We newly produced specific anti-8-nitroguanine antibody without cross-reaction. Double immunofluorescence staining revealed that 8-oxodG and 8-nitroguanine were formed mainly in the same inflammatory cells and epithelium of bile ducts from day 7 and showed the strongest immunoreactivity on days 21 and 30, respectively. It is noteworthy that 8-oxodG and 8-nitroguanine still remained in epithelium of bile ducts on day 180, although amount of alanine aminotransferase activity returned to normal level. A time course of 8-nitroguanine was associated with iNOS expression. Furthermore, this study demonstrated that HO-1 expression and subsequent iron accumulation may be involved in enhancement of oxidative DNA damage in epithelium of small bile ducts. In conclusion, nitrative and oxidative DNA damage via iNOS expression in hamsters infected with OV may participate in CHCA carcinogenesis.     

Accumulation of 8-nitroguanine in human gastric epithelium induced by Helicobacter pylori infection

Helicobacter pylori infection causes chronic inflammation, which can lead to gastric carcinoma. A double immunofluorescence labeling study demonstrated that the level of 8-nitroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) apparent in gastric gland epithelium was significantly higher in gastritis patients with H. pylori infection than in those without infection. A significant accumulation of proliferating cell nuclear antigen, a prognostic factor for gastric cancer, was observed in gastric gland epithelial cells in patients with H. pylori infection as compared to those without infection, and its accumulation was closely correlated with the formation of 8-nitroguanine and 8-oxodG. These results suggest that nitrosative and oxidative DNA damage in gastric epithelial cells and their proliferation by H. pylori infection may lead to gastric carcinoma. 8-Nitroguanine could be not only a promising biomarker for inflammation but also a useful indicator of the risk of gastric cancer development in response to chronic H. pylori infection.     

Mechanism of metal-mediated DNA damage induced by a metabolite of carcinogenic acetamide

Acetamide is carcinogenic in rats and mice. To clarify the mechanism of carcinogenesis by acetamide, we investigated DNA damage by and acetamide metabolite, acetohydroxamic acid (AHA), using 32P-5'-end-labeled DNA fragments. AHA treated with amidase induced DNA damage in the presence of Cu(II) and displayed a similar DNA cleavage pattern of hydroxylamine. DNA damage was inhibited by both catalase and bathocuproine, suggesting that H2O2 and Cu(I) are involved. Carboxy-PTIO, a specific scavenger of nitric oxide (NO), partially inhibited DNA damage. The amount of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) by amidase-treated AHA was similar to that by hydroxylamine. ESR spectrometry revealed that amidase-treated AHA as well as hydroxylamine generated NO in the presence of Cu(II). From these results, it has been suggested that AHA might be converted into hydroxylamine by amidase. These results suggest that metal-mediated DNA damage mediated by amidase-catalyzed hydroxylamine generation plays an important role in the carcinogenicity of acetamide.     

Isolation and Mapping of a Family of Putative Zinc-finger Protein cDNAs from Rice

To understand the functions of rice homologues of the Arabidopsisflowering-time gene CONSTANS (CO) and salt-tolerance gene STO,we performed a similarity search of the single-run sequencedata of cDNA clones accumulated by the Rice Genome ResearchProgram, and isolated seven rice cDNA clones (S3574, C60910,S12569, R2931, R1479, R1577, and E10707) coding for proteinscontaining one or two zinc-finger-like motifs. Comparison ofthe deduced amino acid sequences between these cDNAs and theCO gene revealed significant similarities (46%-;61%) in theregion of zinc-finger motifs. A domain having a high contentof basic amino acids at the C-terminus of the CO protein wasfound in the corresponding region of proteins predicted fromcDNAs S3574, C60910, and S12569. Two amino acid sequences, "CCADEAAL"and "FCV(L)EDRA," which were present inside each zinc-fingerin the Arabidopsis regulatory protein STO, were also found ineach of the two zinc-finger regions of proteins predicted fromcDNAs R2931, R1479, R1577, and E10707. Using restriction fragmentlength polymorphism (RFLP) linkage analysis, we determined thechromosomal location of the seven cDNA clones. The positionof R2931 on the RFLP linkage map was closely linked to Hd-3,one of the putative quantitative trait loci (QTL) controllingheading date in rice.     

Nucleotide Sequence of a 28-kbp Portion of Rice Mitochondrial DNA: the Existence of Many Sequences That Correspond to Parts of Mitochondrial Genes in Intergenic Regions

The nucleotide sequence of a 27,588-bp region of rice mitochondrialDNA was determined. This sequence contains putative genes thatencode initiator methionine tRNA (trnfM), subunits III (nad3)and IV (nad4) of the NADH dehydrogenase complex, and ribosomalproteins S3 (rps3), S12 (rps12) and L16 (rp116). An open readingframe that contains sequences homologous to parts of rps2 andatpA is also present. In addition to these regions, there aremany short sequences with homology to fragments of mitochondrialDNAs from rice or other plants. These sequences may be remnantsof multiple rearrangements of the genome and their presenceseems to explain, in part, the large sizes of the mitochondrialgenomes of higher plants. (Received July 15, 1994; Accepted September 26, 1994)     

The importance of allochthonous litter input on the biomass of an alien crayfish in farm ponds

To evaluate the effects of allochthonous litter input on the population density of invasive red swamp crayfish (Procambarus clarkii) in Japanese farm ponds, we analyzed gut contents, stable isotope ratios, and the correlation between crayfish biomass and environmental factors in the ponds. For our correlation analysis, we used Akaike’s information criterion (AIC) corrected for small sample size (AIC_C) to select appropriate models within the generalized linear model. Allochthonous litter input was the most influential variable affecting crayfish biomass, followed by pond area. Gut content analysis demonstrated a positive correlation between the percentage of litter in the crayfish gut and the amount of litter input into the pond from which animals were collected. Crayfish δ¹³C became increasingly similar to litter δ¹³C as litter input into ponds increased. Nitrogen isotope signature analysis suggested that microorganisms attached to litter may contribute to crayfish diet. The above results obtained by three complementary approaches demonstrated an important influence of allochthonous litter input on crayfish biomass in farm ponds. We propose that the appropriate management of surrounding forests may be effective in controlling the abundance of exotic crayfish with minimized impacts on native communities.     

Expression of Na+, K+-ATPase α-Subunit in Animalized and Vegetalized Embryos of the Sea Urchin, Hemicentrotus pulcherrimus.

A marked increase in the Na⁺, K⁺-ATPase activity of sea urchin embryos occurred following an elevation of its mRNA level, revealed by Northern blotting analysis, in developmental period between the swimming blastula and the late gastrula stage. cDNA clone of Na⁺, K⁺-ATPase α-subunit, obtained from γgt10 cDNA library of sea urchin gastrulae, was digested with EcoRl ad Hindlll. The obtained 268 bp cDNA fragment, hybridized to a 4.6 Kb RNA, was used as probe for Northern blotting analysis. The level of Na⁺, K⁺-ATPase mRNA was higher in embryo-wall cell fraction isolated from late gastrulae (ectoderm cells) than the level in the bag fraction, containing mesenchyme cells (mesoderm cells) and archenteron (endoderm cells). The activity of Na⁺, K⁺-ATPase and the level of its mRNA were higher in animalized embryos obtained by pulse treatment with A23187 for 3 hr, starting at the 8–16 cell stage and were considerably lower in vegetalized embryos induced by 3 hr treatment with Li⁺ than that in normal embryos at the post gastrula corresponidng stage. Augmentation of Na⁺, K⁺-ATPase gene expression can be regarded as a marker for ectoderm cell differentiation at the post gastrula stage, which results from determination of cell fate in prehatching period.