Isolation of Olive latent virus 1 from Tulip in Toyama Prefecture

A virus whose coat protein gene had a high sequence homology with the coat protein gene of Olive latent virus 1 was isolated from diseased tulip in Toyama Prefecture. Received 1 June 2001/ Accepted in revised form 1 August 2001     

Efficacy of doramectin against naturally acquired nematode infection in Iberian swine

Studies were carried out to determine the therapeutic efficacy of doramectin, administered intramuscularly at a dose of 300 microg/kg live weight, against naturally acquired helminths of extensively farmed Iberian pigs. The first study (slaughter study) evaluated, through necropsy of the study animals, the product's efficacy against gastrointestinal and pulmonary nematodes (Ascaris suum, Oesophagostomum dentatum and Metastrongylus sp.) whilst the second, faecal egg count reduction study, (FECR study) evaluated the drug's efficacy only against gastrointestinal helminths (A. suum, Trichuris suis and Oesophagostomum sp.).The first study used 20 animals divided into two equal groups of 10 on the basis of body weight and faecal egg count. One group constituted saline treated controls and the other was doramectin treated. On Day 14 post treatment half of the animals in each group were necropsied and the number of parasites present counted. On Day 15 the remaining half of each group underwent the same procedure. The second study was carried out with 40 animals divided equally into two groups of 20. This study determined the effect of doramectin treatment on faecal egg counts as an indicator of parasite burden.The first study demonstrated an efficacy of 100% against adult Metastrongylus sp. and A. suum, whilst the efficacy against O.dentatum was 96.3%. The second study indicated that at Day 21 post treatment there was a 100% reduction in egg counts in faeces in comparison to untreated controls.     

Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.     

Determination of primary structure of amberjack myosin heavy chain and its relationship with structural stability of various fish myosin rods

The structural stability of fish myosin depends upon species and temperatures of water in which fish live. Primary, secondary, and quaternary structures of myosin heavy chain (MyHC) from three species of fish living at different temperature ranges have been compared with those of rabbit MyHC in order to investigate the differences in stability. Primary structure of MyHC, although being accessible for warm-water and cold-water fish (carp and walleye pollack), was not available in previous for tropical-water fish literature; so in this study primary structure of MyHC of the tropical-water fish amberjack has been determined by cloning and sequencing its cDNA. The MyHC has 1938 amino acid residues (AA), which are almost as much as as those of carp and walleye pollack. The amberjack MyHC is 91–95% homologous with other fish and rabbit MyHCs. There is a discernible difference between animal species with stable myosin rod (amberjack, carp, and rabbit) and walleye pollack with unstable rod. Stable rod species have a high probability of forming coiled-coil around the COOH-terminal end of the rod, while the pollack has a low coiled-coil formation probability. In addition, the average scores of the coiled-coil for myosin rod were rabbit (1.738) > amberjack (1.691) > carp (1.680) > walleye pollack (1.674) which correlated exactly with the observed stability. The results suggest that coiled-coil forming ability, particularly around the COOH-terminal end, directs structural stability of fish myosin rod.     

Low reproductive success of leatherback turtles, Dermochelys coriacea, is due to high embryonic mortality

We examined the mechanism responsible for low reproductive success in leatherback turtles (Dermochelys coriacea) at Playa Grande, Costa Rica: low egg fertilization versus high rates of embryonic death. Leatherbacks at this beach had a high rate of fertility (=93.3%±2.5%, n=819). We incubated 10 eggs from every clutch encountered of 19 females during 3 months of the 1998-1999 nesting season. Fertility rate of some females decreased during the nesting season, but overall was high. Detection of fertility was difficult using standard methods because fertility rates cannot be determined accurately from nests excavated after hatching because of egg decomposition. Removal and incubation of eggs from nests provided a better estimate. Embryonic death, particularly in the beginning of incubation before embryos are visible to the unaided eye, was the cause of low hatching success in this population. Hatching success increased with increasing fertility and differed between females, with some mothers having 71-81% success and others 23-32%. Embryonic death and not low egg fertility drives poor recruitment at Playa Grande. Improved conservation of this species at Playa Grande will require a better understanding of the mechanism behind embryonic death.     

Bioactive cinchona alkaloids from Remijia peruviana

Three known Cinchona alkaloids of the quinine type, quinine (1), cupreine (2), cinchonine (3), and the possible artifact cinchonine-HCl (3-HCl), along with two new ones, acetylcupreine (4) and N-ethylquinine (5), have been isolated from the bark of Remijia peruviana (Rubiaceae). Their stereochemical structures were established by high resolution NMR spectroscopy. Alkaloids 2-4 had antifeedant effects on Leptinotarsa decemlineata with varying potencies. Compound 4 was cytotoxic to both insect Sf9 and mammalian CHO cells after 48 h of incubation, while 3-HCl had stronger and selective cytotoxicity to Sf9. Quinine 1 had a moderate to low effect on Trypanosoma cruzi. Tumoral cells were also affected by these alkaloids, with 4 and 3-HCl being the most cytotoxic to all the cell lines tested. Overall, the 8R, 9S configurations, as in 3 and 3-HCl, as well as the C-6'acetylated alkaloid 4, with an 8S, 9R configuration, showed stronger biological effects.     

Screening of 170 Peruvian plant species for allelopathic activity by using the Sandwich Method

Peru is one of the 20 botanically extremely diverse countries in the world, with >17 000 flowering plants, of which 30% are endemic. So far, no systematic research has been conducted on the screening of the allelopathic plants. In this study, the allelopathic activity of 170 species from 61 families of Peruvian plants that were collected from the three main regions of Peru – the Costa (Pacific coastline), the Sierra (Andean mountains), and the Selva (Amazonian rainforest) – was evaluated. The allelopathic activity was determined by the Sandwich Method, which can evaluate the activity of leaf leachates. The species that were found to be highly inhibitory in this screening, under the criterion of >90% inhibition of the radicle of lettuce (Lactuca sativa) seedlings, were Aristeguietia ballii and Diplostephium foliosissimum (Asteraceae) and Spondias mombin (Anacardiaceae). All of these species are native plants from Peru. This study gives a strong clue regarding the potential of isolating potent allelochemicals from these plants in the future.     

Eucalyptus camaldulensis density and fiber length estimated by near-infrared spectroscopy

Density and fiber length belong to the parameters that are used by the pulping industry as indicators of wood quality for different industrial processes and final paper products. The feasibility of Fourier transform near-infrared (FT-NIR) spectroscopy for the non-destructive evaluation of fiber length and air-dry density of fast-growing E. camaldulensis from Thailand was investigated using 50 samples. NIR spectra taken from solid wood and air-dry density as well as fiber length were used for partial least squares (PLS) regression analyses. It is the first time that the fiber length of E. camaldulensis solid wood could be predicted with high accuracy and precision and that the ratios of performance to deviation (RPD) obtained are the first that fully fulfill the requirements of AACC Method 39-00 (AACC 1999) for screening in breeding programs (RPD?≥?2.5). The RPDs for cross-validation (test set validation) of the NIR-PLS-R models of 3.3 (3.8) for air-dry density and 3.5 (3.9) for fiber length allow drawing the conclusion that the models are at least applicable for screening in breeding programs as they lie in-between screening (RPD?≥?2.5) and quality control (RPD?≥?5). Even when 40% of the samples were removed in cross-validation of the air-dry density model, the RPD is 3.2, which confirms that the model is robust, stable, and well qualified for prediction. The good model statistics obtained in this study might be due to the fact that measurement sites for the measurement of NIR spectra, air-dry density, and fiber lengths were strictly coincided.