Lactate dehydrogenase (LDH) gene duplication during chordate evolution: the cDNA sequence of the LDH of the tunicate Styela plicata

L-Lactate dehydrogenase (L-LDH, E.C. 1.1.1.27) is encoded by two or three loci in all vertebrates examined, with the exception of lampreys, which have a single LDH locus. Biochemical characterizations of LDH proteins have suggested that a gene duplication early in vertebrate evolution gave rise to Ldh-A and Ldh-B and that an additional locus, Ldh-C arose in a number of lineages more recently. Although some phylogenetic studies of LDH protein sequences have supported this pattern of gene duplication, others have contradicted it. In particular, a number of studies have suggested that Ldh-C represents the earliest divergence among vertebrate LDHs and that it may have diverged from the other loci well before the origin of vertebrates. Such hypotheses make explicit statements about the relationship of vertebrate and invertebrate LDHs, but to date, no closely related invertebrate LDH sequences have been available for comparison. We have attempted to provide further data on the timing of gene duplications leading to multiple vertebrate LDHs by determining the cDNA sequence of the LDH of the tunicate Styela plicata. Phylogenetic analyses of this and other LDH sequences provide strong support for the duplications giving rise to multiple vertebrate LDHs having occurred after vertebrates diverged from tunicates. The timing of these LDH duplications is consistent with data from a number of other gene families suggesting widespread gene duplication near the origin of vertebrates. With respect to the relationships among vertebrate LDHs, our data are not consistent with previous claims that Ldh-C represented the earliest divergence. However, the precise relationships among some of the main lineages of vertebrate LDHs were not resolved in our analyses.      

Plasma proteome analysis in HTLV-1-associated myelopathy/tropical spastic paraparesis

Background

A new subgroup of HIV-1, designated Group P, was recently detected in two unrelated patients of Cameroonian origin. HIV-1 Group P phylogenetically clusters with SIVgor suggesting that it is the result of a cross-species transmission from gorillas. Until today, HIV-1 Group P has only been detected in two patients, and its degree of adaptation to the human host is largely unknown. Previous data have shown that pandemic HIV-1 Group M, but not non-pandemic Group O or rare Group N viruses, efficiently antagonize the human orthologue of the restriction factor tetherin (BST-2, HM1.24, CD317) suggesting that primate lentiviruses may have to gain anti-tetherin activity for efficient spread in the human population. Thus far, three SIV/HIV gene products (vpu, nef and env) are known to have the potential to counteract primate tetherin proteins, often in a species-specific manner. Here, we examined how long Group P may have been circulating in humans and determined its capability to antagonize human tetherin as an indicator of adaptation to humans.

Results

Our data suggest that HIV-1 Group P entered the human population between 1845 and 1989. Vpu, Env and Nef proteins from both Group P viruses failed to counteract human or gorilla tetherin to promote efficient release of HIV-1 virions, although both Group P Nef proteins moderately downmodulated gorilla tetherin from the cell surface. Notably, Vpu, Env and Nef alleles from the two HIV-1 P strains were all able to reduce CD4 cell surface expression.

Conclusions

Our analyses of the two reported HIV-1 Group P viruses suggest that zoonosis occurred in the last 170 years and further support that pandemic HIV-1 Group M strains are better adapted to humans than non-pandemic or rare Group O, N and P viruses. The inability to antagonize human tetherin may potentially explain the limited spread of HIV-1 Group P in the human population.     

Current status of antisense DNA methods in behavioral studies

The antisense DNA method has been used successfully to block the expression of specific genes in vivo in neuronal systems. An increasing number of studies in the last few years have shown that antisense DNA administered directly into the brain can modify various kinds of behaviors. These findings strongly suggest that the antisense DNA method can be used as a powerful tool to study causal relationships between molecular processes in the brain and behavior. In this article we review the current status of the antisense method in behavioral studies and discuss its potentials and problems by focusing on the following four aspects; (i) optimal application paradigms of antisense DNA methods in behavioral studies; (ii) efficiencies of different administration methods of antisense DNA used in behavioral studies; (iii) determination of specificity of behavioral effects of antisense DNA; and (iv) discrepancies between antisense DNA effects on behaviors and those on protein levels of the targeted gene.      

Effects of neonicotinoid insecticides on the bionomics of twospotted spider mite (Acari: Tetranychidae)

Previous reports indicate that applications of imidacloprid, a neonicotinoid insecticide, can lead to population buildups of twospotted spider mite, Tetranychus urticae Koch, in the field. Moreover, laboratory studies showed enhanced fecundity of T. urticae after an imidacloprid treatment. In this study, experiments were conducted in the greenhouse to investigate the potential effects of imidacloprid and several other neonicotinoid insecticides on fecundity, egg viability, preimaginal survivorship, and sex ratio of T. urticae (German strain WI) on French beans, Phaseolus vulgaris L. Four insecticides, i.e., imidacloprid (Confidor 200SL), thiacloprid (Calypso 480 SC), acetamiprid (Mospilan 70 WP), and thiamethoxam (Actara 25 WG), were tested at field-relevant (100, 120, 125, and 95 ppm) and sublethal doses (10, 12, 12.5, and 9.5 ppm), respectively. Both spray and drench applications were tested. At field-relevant doses, fecundity of T. urticae decreased and was lower in the treatments compared with the untreated control, whereas preimaginal survivorship and proportion of female offspring (i.e., sex ratio) were lower compared with the control. At sublethal doses, no significant differences were found among the treatments. Data on egg viability, preimaginal survivorship, and sex ratio at sublethal doses followed the same trends as at field-relevant doses. In an additional experiment, the metabolism of imidacloprid into monohydroxy-imidacloprid, olefine, guanidine, and 6-chloronicotinic acid was compared with the oviposition pattern of T. urticae. These findings are discussed with regard to previous laboratory and field observations of imidacloprid-induced fertility increases in T. urticae.     

Identification of the ryanodine receptor mutation I4743M and its contribution to diamide insecticide resistance in Spodoptera exigua (Lepidoptera: Noctuidae)

Insect ryanodine receptors (RyRs) are the targets of diamide insecticides. Two point mutations G4946E and I4790M (numbering according to Plutella xylostella, PxRyR) in the transmembrane domain of the insect RyRs associated with diamide resistance have so far been identified in three lepidopteran pests, P. xylostella, Tuta absoluta and Chilo suppressalis. In this study, we identified one of the known RyR target site resistance mutations (I4790M) in a field‐collected population of Spodoptera exigua. The field‐collected WF population of S. exigua exhibited 154 fold resistance to chlorantraniliprole when compared with the susceptible WH‐S strain. Sequencing the transmembrane domains of S. exigua RyR (SeRyR) revealed that the resistant WF strain was homozygous for the I4743M mutation (corresponding to I4790M in PxRyR), whereas the G4900E allele (corresponding to G4946E of PxRyR) was not detected. The 4743M allele was introgressed into the susceptible WH‐S strain by crossing WF with WH‐S, followed by three rounds of backcrossing with WH‐S. The introgressed strain 4743M was homozygous for the mutant 4743M allele and shared about 94% of its genetic background with that of the recipient WH‐S strain. Compared with WH‐S, the near‐isogenic 4743M strain showed moderate levels of resistance to chlorantraniliprole (21 fold), cyantraniliprole (25 fold) and flubendiamide (22 fold), suggesting that the I4743M mutation confers medium levels of resistance to all three diamides. Genetic analysis showed diamide resistance in the 4743M strain was inherited as an autosomal and recessive trait. Results from this study have direct implications for the design of appropriate resistance monitoring and management practices to sustainably control S. exigua.     

Skeletal growth in the echinoderm Asterias rubens L. (Asteroidea,Echinodermata) estimated by 45Ca-labelling

The skeletal dry weight of the 4.4 ± 0.2 cm size class of Asterias rubens L. from Kiel Bay in the western Baltic is 0.34 ± 0.08 g. The sum of calcium and magnesium carbonates in the skeleton amounts to 94.0 ± 1.3% while the individual concentrations are 86.9 ± 1.3% CaCO₃ and 7.1 ± 0.7% MgCO₃. The MgCO₃ is 4% lower than expected for a magnesium calcite precipitated under Baltic temperature conditions (8°C). Strontium was not determined but strontium carbonate is known to be in the region of 0.4%. The remainder is organic matter and this gives rise to 2.9 ± 1.3% organic carbon.Animals studied were at the ‘waiting stage’ and their actual growth was minimal. Uptake of ⁴⁵Ca in the skeleton consists of a fast step followed by a slow step. The fast step is attributed to saturation of exchangeable skeletal pools while the slow step is due to net deposition of CaCO₃. Skeletal growth at the waiting stage calculated from the second rate constant was found to be 0.76 μg CaCO₃j-mg skeleton^?1 · day^?1 or 0.09% · day^?1 compared with 9.3 μg CaCO₃ · mg skeleton^t1̄ · day^?1 or 1.1% · day^?1 at log phase. The isotope method is considered superior to size-frequency analysis in that it is capable of detecting differences in growth rate in individuals of the same size class and thus provides an insight into asteroid population structure.