Expression in Escherichia coli and purification of the functional feline granulocyte colony-stimulating factor

Feline granulocyte colony-stimulating factor (G-CSF) with an N-terminal histidine hexamer tag was expressed as inclusion bodies in E. coli. The G-CSF solubilized in 6 M guanidine solution was absorbed onto a Ni-NTA column and, after washing with decreasing concentrations of guanidine, eluted with imidazole in a soluble and apparently pure form. The activity of the recombinant feline G-CSF was 3×10⁶ U/mg protein, as assayed by its stimulatory effect on NFS-60 cell proliferation. When a low level of purified feline G-CSF was administered once a day for two successive days to cats, the number of neutrophil increased 4-fold while the levels of other blood cell types remained virtually unchanged. Daily administration of G-CSF for a total of 11 days led to a more than 10-fold increase in neutrophils, an 8-fold increase in the number of monocytes and 2-fold increase in lymphocytes. No severe side effects or antibody production was observed in cats after administration of G-CSF.     

Effect of Dietary Inclusion of Blue Mussel Extract on Growth and Body Composition of Japanese Flounder Paralichthys olivaceus

Dietary inclusion of a water-soluble fraction of blue mussel Mytilus galloprovincialis was examined as a feeding stimulant for juvenile Japanese flounder Paralichthys olivaceus . The control diet mainly consisted of fish meal, potato starch, and pollack liver oil. Five, 10, and 20% (weight/weight) of the control diet was exchanged with aqueous extracts of blue mussel meat in experimental groups. Fish of about 10 g in initial body weight were fed each diet to satiation, twice daily, 6 d per wk for 6 wk at 20 C. The final body weight, weight gain, and feed efficiency of fish fed the diets containing blue mussel extracts were significantly higher than those of fish fed the control diet. However, these parameters were not different among experimental groups containing blue mussel extract independent of the inclusion level of extract. A similar trend was shown in protein efficiency ratio as fish fed the control diet had a significantly lower protein efficiency ratio than the other dietary groups. Compared to the control diet, higher plasma protein and lower triglyceride were found in fish fed the diets with the extract, while other blood constituents were relatively similar for the dietary groups tested. On the other hand, whole-body crude lipid content and lipid retention of fish fed the diets with the extract were generally significantly higher than those of fish fed the control diet. Whole body crude protein was identical regardless of the dietary composition; however, protein retention of fish showed a similar trend to lipid retention.     

Active expression of matrix metalloproteinase-13 mRNA in the granulation tissue of equine superficial digital flexor tendinitis

The DNA microarray analysis for matrix metalloproteinase (MMP)-related mRNA expression in equine superficial digital flexor tendinitis indicated that mRNA level of MMP-13 was apparently up-regulated in the tendinitis as compared to normal tendon. In situ hybridization also revealed that fibroblastic cells proliferated in the granulation tissue generated in the tendinitis actively expressed MMP-13 mRNA. On the other hand, in normal tendon, a few fibroblastic cells and vascular components lied in the endotenon barely expressed its mRNA, but other cellular components in the tendon bundle were not positively hybridized. As mentioned above, MMP-13 but not other collagenases or gelatinases, may play an important role in tendon injuries in the racehorses.     

Anesthesia cutoff phenomenon: interfacial hydrogen bonding

Anesthesia "cutoff" refers to the phenomenon of loss of anesthetic potency in a homologous series of alkanes and their derivatives when their sizes become too large. In this study, hydrogen bonding of 1-alkanol series (ethanol to eicosanol) to dipalmitoyl-L-alpha-phosphatidylcholine (DPPC) was studied by Fourier transform infrared spectroscopy (FTIR) in DPPC-D2O-in-CCl4 reversed micelles. The alkanols formed hydrogen bonds with the phosphate moiety of DPPC and released the DPPC-bound deuterated water, evidenced by increases in the bound O-H stretching signal of the alkanol-DPPC complex and also in the free O-D stretching band of unbound D2O. These effects increased according to the elongation of the carbon chain of 1-alkanols from ethanol (C2) to 1-decanol (C10), but suddenly almost disappeared at 1-tetradecanol (C14). Anesthetic potencies of these alkanols, estimated by the activity of brine shrimps, were linearly related to hydrogen bond-breaking activities below C10 and agreed with the FTIR data in the cutoff at C10.     

Potential conflicts for the reuse of rice husk in Thailand

This study aims to investigate the impact of rice-husk power generation on agricultural uses of rice husk and its ash. We conducted case studies at a rice-husk power plant and organic farming groups in northeast Thailand. The case study on a rice-husk power plant showed that rice husk was useful in generating electricity without consuming fossil fuel, while husk ash, the by-product, could be utilized as silicate supplement for farmland soils. The case study on organic farming groups indicated that rice husk was one of the indispensable materials in conducting organic farming with locally available resources. On the basis of these findings, an analytical study on possible husk uses in Thailand was conducted, in which three potential major uses of rice husk were considered: i.e. (1) fuel use inside rice mills followed by husk-ash reuse, (2) fuel use at power plants followed by husk-ash reuse, and (3) direct reuse of husk for compost production. The result indicated that there would be conflicting demands for rice husk among those uses in the future. Therefore, there needs to be a strategy for balancing husk uses among them.     

Ocular lesions in Watanabe heritable hyperlipidemic rabbits

Hyperlipidemic ocular lesions are described for Watanabe heritable hyperlipidemic (WHHL) rabbits. Male WHHL rabbits 8 months old exhibited serum hyperlipidemia and ophthalmoscopically yellowish-white lesions along the corneoscleral junction and in the iris. Histopathologically, foamy macrophages aggregated in the stroma of the cornea, iris, and ciliary body were observed. These findings have been interpreted as lipid keratopathy. In addition, multiple clusters of a large number of foamy macrophages occurred throughout the choroid and sclera in association with the blood vessels. The lesions in the choroid and sclera could not be detected ophthalmoscopy, yet were much more prominent than those in the cornea, iris, and ciliary body, suggesting greater involvement and earlier onset of lipidosis at these sites associated with hyperlipidemia in WHHL rabbits.     

Relative sensitivity of duckweed Lemna minor and six algae to seven herbicides

We investigated the relative sensitivity of duckweed Lemna minor and six species of algae to seven herbicides, using an efficient high-throughput microplate-based toxicity assay. First, we assessed the sensitivity of L. minor to the seven herbicides, and then we compared its sensitivity to that of previously published data for six algal species based on EC₅₀ values. For five herbicides, the most sensitive species differed: L. minor was most sensitive to cyclosulfamuron: Raphidocelis subcapitata was most sensitive to pretilachlor and esprocarb: Desmodesmus subspicatus was most sensitive to pyraclonil; and Navicula pelliculosa was most sensitive to pyrazoxyfen. Simetryn was evenly toxic to all species, whereas 2,4-D was evenly less toxic, with only small differences in species sensitivity. These results suggested that a single algal species cannot represent the sensitivity of the primary producer assemblage to a given herbicide. Therefore, to assess the ecological effects of herbicides, aquatic plant and multispecies algal toxicity data sets are essential.     

Model Intercomparison Study of Long Range Transport and Sulfur Deposition in East Asia (MICS-ASIA)

To help improve the use of models in science & policy analysis in Asia it is necessary to have a better understanding of model performance and uncertainties. Towards this goal an intercomparison exercise has been initiated as a collaborative study of scientists interested in long-range transport in East Asia. An overview of this study is presented in this paper. The study consists of a set of prescribed test calculations with carefully controlled experiments. Models used the same domain, emission inventory, model parameters, meteorological conditions, etc. Two periods (January and May 1993) were selected to reflect long-range transport conditions under two distinct seasons. During these periods measurements of sulfur concentrations and deposition were made throughout the study region using identical sampling and analysis protocols. The intercomparison activity consists of four tasks (Blind Test, Fixed Parameter Test, Source Receptor test, and Tuning Test). All participants were asked to do Task A, and as many of the other tasks as possible. To date seven different models have participated in this study. Results and key findings are presented.     

Inhibition of acetate ester biosynthesis in banana (Musa sapientum L.) fruit pulp under anaerobic conditions

The effect of anaerobic conditions on acetate ester biosynthesis in ripened banana pulp was investigated. Incubation of the pulp in less than 1% O(2) resulted in a significant reduction in the formation of ethyl acetate. Regardless of the presence of a large amount of endogenous ethanol and the remaining exogenous isobutyl alcohol after complete anaerobic incubation with the pulp, the production of acetate ester decreased. The effect of addition of pyruvate, isobutyl alcohol, acetate, and methyl hexanoate on acetate ester formation in 100% N(2) was also investigated. The addition of pyruvate and isobutyl alcohol to the pulp gave lower acetate esters in N(2) than in air, whereas the pulp incubated with acetate and isobutyl alcohol produced more acetate ester in both conditions. Therefore, the lack of acetyl CoA, or more precisely acetate, in the tissue is the main reason for the inhibition of acetate ester formation under anaerobic conditions. The activity of beta-oxidation measured by incubation with methyl hexanoate was detected only in the samples incubated in air. The formation of acetyl CoA, derived from pyruvate through mitochondria and through beta-oxidation, was inhibited by anaerobic conditions, which suggests that mitochondrial activity and/or beta-oxidation are essential for ester biosynthesis.     

EFFECT OF BIODEGRADABLE CHELATING LIGAND ON IRON BIOAVAILABILITY AND RADISH GROWTH

The effect of chelating ligands on iron (Fe) uptake and growth of radish (Raphanus sativus L.) was investigated. The ethylenediaminetetraacetic acid (EDTA) increased ⁵⁵Fe uptake in roots of radish though its subsequent translocation from roots to shoots and leaves did not increase. About 70%—80% of the total ⁵⁵Fe was distributed in the roots while about 5%—15% and 11%—17% were in shoots and leaves, respectively. The EDTA increased iron uptake into the roots of radish, but not in the above ground parts of the plant. The growth of radish (Raphanus sativus L.) decreased drastically in alkaline condition (pH > 9), even though the concentration of iron was sufficient in the growth medium. The growth of radish was enhanced successfully by the addition of hydroxyiminodisuccinic acid (HIDS) and EDTA. This might be because HIDS and EDTA solubilize iron from its precipitation with hydroxides at higher pH, and increase iron bioavailability. The influence of EDTA and HIDS on radish growth was comparable. Increase of radish growth by ethylenediaminedisuccinic acid (EDDS) and methylglicinediacetic acid (MGDA) was less than those by EDTA and HIDS. Considering the reproducibility of the radish growth (biomass production) at pH 10, HIDS is supposed to be more effective compared to EDTA.