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 ovary lipid from Skipjack tuna (Katsuwonus pelamis) on brain monoamines in rats

In our previous experiments with rats, ovary lipid from Skipjack Tuna (Katsuwonus pelamis) (OLS) was shown to have a mitigating effect on anxiety and/or fear in elevated T-maze tests. This suggests that OLS has some effect on the central nervous system (CNS) of rats. Thus, we performed experiments to examine the status of CNS in rats given OLS. The effect of OLS on chronic stress was also examined at the same time. The feed for control rats used oil and fat that have the same energy percentages for n-6 and n-3 fatty acids and the same n-6/n-3 ratio as OLS. As a result, rats given OLS for 28 days had lower serotonin levels in various brain areas regardless of stress application, showing that OLS affected the serotonin nervous system. From this, it was inferred that the ability of OLS to mitigate anxiety and/or fear resulted from its action on CNS, especially the serotonin nervous system. Substances other than the essential fatty acids may have been responsible for the action of OLS on monoamines and the metabolites. The effect of OLS on CNS, especially the serotonin nervous system, suggests that OLS may suppress anxiety.     

Oleosome-Associated Protein of the Oleaginous Diatom Fistulifera solaris Contains an Endoplasmic Reticulum-Targeting Signal Sequence

Microalgae tend to accumulate lipids as an energy storage material in the specific organelle, oleosomes. Current studies have demonstrated that lipids derived from microalgal oleosomes are a promising source of biofuels, while the oleosome formation mechanism has not been fully elucidated. Oleosome-associated proteins have been identified from several microalgae to elucidate the fundamental mechanisms of oleosome formation, although understanding their functions is still in infancy. Recently, we discovered a diatom-oleosome-associated-protein 1 (DOAP1) from the oleaginous diatom, Fistulifera solaris JPCC DA0580. The DOAP1 sequence implied that this protein might be transported into the endoplasmic reticulum (ER) due to the signal sequence. To ensure this, we fused the signal sequence to green fluorescence protein. The fusion protein distributed around the chloroplast as like a meshwork membrane structure, indicating the ER localization. This result suggests that DOAP1 could firstly localize at the ER, then move to the oleosomes. This study also demonstrated that the DOAP1 signal sequence allowed recombinant proteins to be specifically expressed in the ER of the oleaginous diatom. It would be a useful technique for engineering the lipid synthesis pathways existing in the ER, and finally controlling the biofuel quality.     

Absorption of color additives and settling volume in water of blue-green alga,ishikurage (Nostoc commune)

The properties of ishikurage (Nostoc commune) and other algae were compared. Ishikurage and suizenji nori (Aphanothece sacrum) absorbed much more amaranth than others. They have a high settling volume (SV) in water. The high SV of ishikurage is attributable to its neutral detergent fiber (NDF). The acid detergent fiber (ADF) from the alga had a high amaranth absorption. However the absorption by the ADF did not explain the absorption by ishikurage.The results suggest that ishikurage may have a nutritional significance as a source of dietary fiber.     

Induction of detoxication enzymes in mice by naturally occurring allyl nitrile

Little is known about whether glucosinolate-derived nitriles have the ability to increase phase 2 detoxication enzymes and glutathione (GSH) in vivo. In this study, the ability of allyl nitrile, a hydrolysis product of the glucosinolate sinigrin, to increase tissue levels of the phase 2 detoxication enzymes glutathione S-transferase and quinone reductase and GSH in a variety of mouse tissues was examined. At the lowest dose level (11.8 mg/kg/day), allyl nitrile showed inductive ability in the stomach and lungs. At 23.6 mg/kg/day, the inductive effect was observed in the stomach, rectum, urinary bladder, and lungs, whereas at 47.2 mg/kg/day, it was recorded in the stomach, rectum, urinary bladder, kidneys, and lungs. These results show that allyl nitrile displays its maximum potency in the stomach and lungs, which is of interest in light of epidemiological studies demonstrating an inverse association between crucifer intake and the incidence of stomach and lung cancers.     

Clay minerals of soils from olivine basalt in Kyushu

Soil scientists have been encouraged to investigate soils for clay minerals by very fact that soil characteristics fundamentally depend upon the nature of their clay. It is generally recognized that clays of different species develop under different conditions such as drainage, temperature weathering age, and parent rocks. On Kyushu Island, heavy, reddish soils derived from olivine basalt cover a considerably large area of its northeastern part, and a numbar of small islands belonging to Kyushu. Koga ^* Hiroshi Koga, Clay minerals of Oura basaltic soil, 1954 (unpublished). of this laboratory found a basaltic soil of clay consisting of hydrated halloysite, halloysite, hematite, and gibbsite at the eastern foot of Mt. Tara, by examining its clay separates by X-ray, thermal, chemical, and electron microscopical techniques. Kanno et al⁵ reported that in a basaltic soil at Oura, Saga prefecture, hydrated halloysite developed from plagioclase and ferro-magnesian minerals with gibbsite and hematite which was more abundant than in other Red and Yellow Soils. They also found some illite present in the soil derived from unknown minerals, and halloysite dehydrated in upper layers.     

Evaluation of a sequential extraction procedure for the fractionation of thallium in soils and determination of the content by flame atomic absorption spectrometry

Abstract

The release of carbonyl sulfide (COS) from 7 unamended paddy soils was estimated under aerobic and waterlogged conditions. The addition of organic materials to the soils increased the amount of COS evolved under waterlogged conditions. To assess the sources of the COS released from the soils, the soils were incubated under aerobic and waterlogged conditions after treatment with 17 inorganic or organic S-containing compounds. It was concluded that the following compounds represented sources of the COS released from the paddy soils: cystine, cysteine, thiocyanate, lanthionine and djenkolic acid.     

Exchange acidity of montmorillonite and halloysite

In the previous article (1), montmorillonitic soils were disclosed to have the ratio of total exchange acidity to y¹ less than 3.0 and no ralation of the degree of base saturation to the ratio, although the relation was suggested by AMONINE and WATANABE (2).     

Abundance of sulphate-reducing bacteria in fish farm sediments along the coast of Japan and South Korea

The abundance of sulphate-reducing bacteria (SRB) was investigated in the coastal marine sediments along the coast of Japan and South Korea. Sediment samples were collected from fish and shellfish farms between 2006 and 2008. As non-fish farming reference sites, sediments were also collected from highly eutrophic bays, a highly sulphidogenic saline lake, and the deep sea. A quantitative real-time PCR analysis that targeted the gene coding for a portion of the α-subunit of dissimilatory sulphite reductase (dsrA) was performed to assess the abundance of the SRB in the sediments. Between 2.8 × 10⁷ and 2.5 × 10⁹ copies of the dsrA gene per gram dry sediment were detected. There was no relationship between dsrA gene copy number and total bacterial count in the sediments, whereas organic matter contents (particulate organic carbon and nitrogen, ignition loss and chemical oxygen demand) and acid-volatile sulphide contents were significantly correlated with the dsrA copy number. The data presented demonstrate that organic enrichment of sediment may influence the abundance of SRB communities in coastal marine sediments and that the cell density of SRB may be used as a biological indicator for assessing pollution levels in sediments of marine fish farms.