Solid-phase synthesis of mimosine tetrapeptides and their inhibitory activities on neuraminidase and tyrosinase

Neuraminidase is a rational target for influenza inhibition, and the search for neuraminidase inhibitors has been intensified. Mimosine, a nonprotein amino acid, was for the first time identified as a neuraminidase inhibitor with an IC(50) of 9.8 ± 0.2 μM. It was found that mimosine had slow, time-dependent competitive inhibition against the neuraminidase. Furthermore, a small library of mimosine tetrapeptides (M-A(1)-A(2)-A(3)) was synthesized by solid-phase synthesis and was assayed to evaluate their neuraminidase and tyrosinase inhibitory properties. Most of the tetrapeptides showed better activities than mimosine. Mimosine-FFY was the best compound, and it exhibited 50% neuraminidase inhibition at a low micromolar range of 1.8 ± 0.2 μM, whereas for tyrosinase inhibition, it had an IC(50) of 18.3 ± 0.5 μM. The kinetic studies showed that all of the synthesized peptides inhibited neuraminidase noncompetitively with K(i) values ranging from 1.9 -to 7.2 μM. These results suggest that mimosine could be used as a source of bioactive compounds and may have possibilities in the design of drugs as neuraminidase and tyrosinase inhibitors.     

Kinetics of 15N -labelled nitrogen from co-compost made from cattle manure and chemical fertilizer in a paddy field

In order to produce an effective organic fertilizer, cattle manure was cocomposted with chemical fertilizer. And the kinetics of nitrogen uptake by rice plants from the co-compost was investigated using the ¹⁵N labelled co-composts on either cattle manure or chemical fertilizer. As a control, nitrogen kinetics from the mixture of cattle manure and chemical fertilizer without co-composting was investigated. At the early stage, rice growth may have been promoted by co-composting, while, it may have been promoted by the larger N-content of cattle manure at the harvesting stage. The ratios of nitrogen uptake by rice plants and residual nitrogen in soil from the cattle manure and chemical fertilizer were determined by measuring ¹⁵N -atom%. The N -uptake ratios by rice plants from the cattle manure in the co-composted plot were about 2–4 times higher than those from the cattle manure without co-composting. However, the N -uptake ratios from the chemical fertilizer in the co-composted plot were lower than those from the chemical fertilizer without co-composting. The N -content of the rice plants derived from chemical fertilizer without co-composting decreased consistently after 28 d. The nitrogen from chemical fertilizer in the co-compost was absorbed again in the latter period of rice growth. The total nitrogen uptake by rice plants from cattle manure and chemical fertilizer was similar regardless of co-composting. However, co-composting would be advantageous at least· in terms of the following aspects: increase of the N -uptake by rice plants from cattle manure, slow-release ability of nitrogen from chemical fertilizer, decrease of nitrogen loss by denitrification.     

Physico-chemical characteristics of the soils from the coastal plain in Shanghai,China

Characterization of the physico-chemical properties of the soils collected from the coastal plain covering a total area of about 1,908 km² in the Changjiang Delta in Shanghai, China, has been conducted in order to investigate the current status of agricultural soils, with special reference to the effect of alluvial deposits on the development and utilization of the soils. The experimental results indicated that the soils of the coastal plain displayed a sandy (loam) texture. The soils from Qingcun and Guangming Towns in the Fengxian District, Shanghai, contained the highest amount of clay, showing the highest capacity to remain fertile. The soils from Xinghuo Town in the Fengxian District contained the highest amount of salts due to the strong mineralization degree from the groundwater and the effect of the marine deposits from the Hangzhou day, which resulted in the highest exchangeable sodium percentage in the soils. The soils from greenhouses (Sunqiao, Huinan, and Qingcun Towns) showed a higher salt content than others, in contrast to the paddy and orchard soils. The analysis of the soil fertility indicated that the soils from Guangming and Qingcun Towns in Fengxian were rich in organic matter, total nitrogen, and total potassium, and were fertile soils. In contrast, though the soil from Xinghuo Town was rich in potassium, it lacked other nutrients. The phosphorus content in the soils was mainly determined by the application of fertilizers, and the total and available phosphorus contents in the greenhouse soils were higher than those in the soils with other land use types.     

Detection of a quantitative trait locus for intramuscular fat accumulation using the OLETF rat

The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type 2 diabetes. We showed that the OLETF rat exhibits higher levels of intramuscular fat content in Musculus longissimus as compared to the Fischer-344 (F344) rat. Our investigation was designed to identify quantitative trait loci (QTLs) contributing to the increased levels of intramuscular fat content by performing a whole-genome search using 108 F2 intercross obtained by mating the OLETF and the F344 rats. We identified one QTL responsible for intramuscular fat accumulation on rat chromosome 1 with a maximum lod score of 3.4, which accounts for 5% of the total variance. As expected, the OLETF allele corresponds to the increased levels of intramuscular fat content.     

Detection of equine Babesia spp. gene fragments in Dermacentor nuttalli Olenev 1929 infesting mongolian horses,and their amplification in egg and larval progenies

Babesia equi (EMA-1) and Babesia caballi (BC48) gene fragments were amplified by polymerase chain reaction (PCR), in blood samples, and partially fed-females and egg and larval progenies of Dermacentor nuttalli, collected from horses in Altanbulag, Tuv Province, Mongolia. While Babesia parasite DNA was detected in some horse blood samples during the first PCR, all positive cases in partially fed-female ticks, eggs and larvae were confirmed by nested PCR. Present study reinforces earlier similar findings in unfed D. nuttalli ticks collected from an open space vegetation in Bayanonjuul, Tuv Province in Central Mongolia, pointing to the most likely important role of D. nuttalli in the transmission of equine babesiosis in Mongolia. The detection of parasite DNA in eggs and larval progenies is likewise suggestive of transovarial parasite transmission in this tick species.     

Plant growth inhibitors in turmeric (Curcuma longa) and their effects on Bidens pilosa

Turmeric (Curcuma spp.) has numerous biological activities, including anticancer, antibacterial, antifungal and insecticidal properties. Here, we evaluated the plant growth‐inhibitory activities of two cultivars of Curcuma longa (C. longa ; Ryudai gold and Okinawa ukon) against radish, cress, lettuce and Bidens pilosa (B. pilosa ). The methanol extracts of both turmeric varieties inhibited the seed germination and seedling growth of all the tested plants. Ryudai gold had a significantly higher inhibitory effect on the seed germination and root and shoot growth of the plants than Okinawa ukon. Therefore, Ryudai gold was chosen for the isolation of plant growth‐inhibitory compounds using a silica gel column and high‐performance liquid chromatography. The structural identification of the compounds was carried out using ¹H NMR, ¹³C NMR and liquid chromatography–tandem mass spectrometry. The growth inhibitors were identified as four curcuminoids; dihydrobisdemethoxycurcumin ( 1 ), bisdemethoxycurcumin ( 2 ), demethoxycurcumin ( 3 ) and curcumin ( 4 ). The IC₅₀ of the curcuminoids against the root and shoot growth of B. pilosa ranged from 8.7 ± 1.7 to 12.9 ± 1.8 and from 15.5 ± 1.8 to 38.9 ± 2.8 μmol L^?1, respectively. Compound 1 showed the lowest IC₅₀ against the root and shoot growth of B. pilosa . These results suggested that Ryudai gold has a potential growth‐inhibitory effect against B. pilosa .     

EGFP-Rhm51 foci enable the visualization and enumeration of DNA double-strand breaks in <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

To detect and count DNA double-strand breaks (DSBs) during the life cycle of Magnaporthe oryzae, we constructed an expression vector for EGFP-Rhm51 fusion protein and introduced it into strain Ina86-137. EGFP-Rhm51 foci were detected, and the number of foci in mycelia treated with mitomycin C, which induces DSBs, was higher than in untreated samples, indicating that the foci enabled the visualization of DSBs during the life cycle. EGFP-Rhm51 foci were observed at all stages of the asexual cycle including when invasive hyphae formed in an intact rice leaf sheath, demonstrating that M. oryzae undergoes DSBs during vegetative and parasitic growth.     

Genetic characterization of GRA6 genes from Toxoplasma gondii from pigs in Okinawa, Japan

Toxoplasma gondii from pigs in Okinawa Prefecture was characterized by nested PCR-restriction fragment length polymorphism (RFLP) and DNA sequence analysis of the dense granule antigen GRA6 gene. By nested PCR, parasite DNA was detected in 33 out of 91 lymph node samples with lesions similar to those found in toxoplasmosis samples that had been collected from pigs at an abattoir. RFLP analysis with MseI was successfully conducted in 29 of 33 PCR-positive samples to group the isolates into one of the three genotypes of T. gondii. Genotyping of the 29 studied samples rendered the following results: 13 of type I (44.8%), 14 of type II (48.3%), and 2 of type III (6.9%). The GRA6 genes of 12 Okinawa isolates were cloned and sequenced. Nine new nucleotide sequences were found, and nucleotide substitutions specific for the Okinawa isolates were found at 13 positions. Phylogenetic analysis indicated that all GRA6 sequences were divided into one of the 3 main groups, and Okinawa isolates of GRA6 genotypes II and III seemed to be closely related to the Beverley strain and the NED strain, respectively. The results from this study may provide basic and useful information for the analysis of the molecular epidemiology of T. gondii infection within Japan.