Cloning and sequence analysis of llama cytokines related to cell-mediated immunity

In order to characterize the T helper 1 (Th1) cytokines of llama, we have cloned several llama cytokine genes and compared them to those of other mammalian species. The cDNAs encoding for interleukin (IL)-2, interferon (IFN)gamma, IL-12p35 and IL-12p40 were amplified using specific primers designed from reported sequences of bovine cytokine genes. The cDNAs for llama IL-2, IFN-gamma, IL-12 p35 and IL-12p40 were found to be 465, 501, 669 or 993 bp in length, with open reading frames encoding 154, 166, 222 or 330 amino acids, respectively. Homology analyses of nucleotide and deduced amino sequences of llama IL-2, IFN-gamma, IL-12p35 and IL-12p40 and phylogenetic analysis based on their nucleotide sequences indicated the close relationship in these cytokine genes between llama and eutherian mammalian order Artiodactyla, which includes pig and cattle.     

Detection of urinary luteinizing hormone in Japanese black cows after administration of gonadotropin-releasing hormone

The blood luteinizing hormone (LH) surge in cows is well studied. However, little is known about urinary LH in cows. This study examined urinary LH concentrations after administration of gonadotropin-releasing hormone (GnRH) in six Japanese black cows to induce LH secretion from the pituitary gland into the bloodstream. Abrupt rises in plasma and urinary LH were observed after GnRH administration. Plasma and urinary LH peaked at 2 and 5 hr, respectively. A positive correlation was observed between plasma LH concentrations and urinary LH amounts. Ovulation was confirmed in the cows after 48 hr of GnRH administration. These data strongly suggest that urinary LH is derived from plasma LH, which triggers ovulation in cows.     

Antioxidant activities of black and yellow soybeans against low density lipoprotein oxidation

Several studies have demonstrated that the daily intakes of soy foods were associated with a reduced cardiovascular risk. The aim of our study was to investigate the inhibitory effect of black soybeans on low density lipoprotein (LDL) oxidation in comparison to yellow soybeans. The extract from black soybean had a longer LDL oxidation lag time than that from yellow soybean (205 +/- 16 and 65 +/- 3 min, respectively). When both soybeans were divided into the seed coat and the mixture of the germ and cotyledon, the diluted extract solution from the black soybean seed coat prolonged the lag time significantly more than the original extract of the yellow soybean seed coat. On the other hand, antioxidant effects of the extract from the mixture of germs and cotyledons were similar in both soybeans. Regarding total polyphenol contents, the seed coat of black soybean had a higher polyphenol content than that of yellow soybean (29.0 +/- 0.56 and 0.45 +/- 0.02 mg/g, respectively). Interestingly, the mixture of the germ and cotyledon hydrolyzed by beta-glucosidase in both soybeans showed a stronger inhibitory effect on LDL oxidation than that before being hydrolyzed by beta-glucosidase. These results suggest that black soybeans may be more effective in inhibiting LDL oxidation than yellow soybeans because of total polyphenols contents in its seed coat. In addition, aglycones, which are rich in soybeans fermented or hydrolyzed by beta-glucosidase, may play a crucial role in the prevention of oxidation-related diseases.     

Microbial and chemical properties of <Emphasis Type="Italic">aji-no-susu</Emphasis>, a traditional fermented fish with rice product in the Noto Peninsula,Japan

Aji-no-susu is a Japanese fermented fish product prepared from salted horse mackerel Trachurus japonicus, and cooked rice. We studied the organic acid and free amino acid contents and microflora in 12 aji-no-susu products to clarify their features as a lactic-acid-fermented food. Salinity of the samples was approximately 7.0% (rice portion) and 6.0% (fish portion) (w/w). Water activity was approximately 0.9, and pH was approximately 4.4 and lower. In the rice portions, lactic acid content was very high (57 mg/g sample). The predominant amino acids were alanine (2.3 mg/g rice portion) and lysine (2.1 mg/g). In the case of long-fermented (4 and 12 months) aji-no-susu, a high content of gamma-aminobutyric acid (GABA, 1.5 and 1.4 mg/g) was detected. Total viable counts in rice and fish portions were 7.7 and 7.4 log colony-forming units (cfu)/g, respectively. The number of lactobacilli in the rice and fish portions was 7.3 and 7.1 log cfu/g, respectively. Yeasts were detected in eight samples. Furthermore, acid-tolerant lactic acid bacteria (LAB) (Lactobacillus plantarum), GABA-producing LAB (Lactobacillus sp.), and halophilic or halo-tolerant yeast (Debaryomyces hansenii) were isolated and identified. Results in this study indicate that aji-no-susu is a typical traditional lactic-acid-fermented fish product.     

Accumulation of gamma-oryzanol in teleost

Japanese people consume crop bran, which contains relatively high amounts of gamma-oryzanol (ORZ), as foodstuffs and food materials. We have recently confirmed that ORZ inhibits NF-κB activation, activates peroxisome proliferator-activated receptor gamma (PPARγ), and increases plasma adiponectin levels. ORZ is therefore expected to improve lipid and carbohydrate metabolisms and various inflammatory diseases, i.e., metabolic syndrome including type 2 diabetes. ORZ administration also allowed fish, such as rainbow trout, yellowtail, and red sea bream, to accumulate protein through enhancement of lipid and carbohydrate metabolisms. In the present study, we have investigated ORZ accumulation levels in mouse and rainbow trout administered ORZ-containing feed. Although mouse muscle and liver hardly contained ORZ, muscle tissues of every fish species accumulated higher amounts of ORZ. These findings suggest that rainbow trout, red sea bream, and yellowtail accumulate ORZ in muscle.     

Distribution of adipocyte-related cells in skeletal muscle of rainbow trout Oncorhynchus mykiss

Adiponectin is one of the adipokines secreted mainly from adipocytes in mammals. In rainbow trout, however, adiponectin is highly expressed in skeletal muscle. Although it has been suggested that fish skeletal muscle contains adipocytes, their endocrine function and distribution are poorly understood. Recently, an EST analysis of rainbow trout found that heart-type fatty acid binding protein (H-FABP), a member of the intracellular fatty acid binding protein family, encodes an adipose-specific gene. In this study, we produced anti-adiponectin and H-FABP antibodies and investigated the distribution of adipocytes and related cells in skeletal muscle of rainbow trout. The adiponectin signal was detected at around 75 kDa in muscle in Western blotting. Since the molecular mass of rainbow trout adiponectin is around 25 kDa, this 75 kDa band would be a trimer. For H-FABP, the signal band was detected at around 15 kDa. Immunohistochemistry of skeletal muscle sections indicated that adiponectin and H-FABP signals were present outside of muscle cells and throughout the muscle tissues, suggesting the existence of adipocyte-related cells in these regions. These results will contribute to our understanding of energy metabolism in fish skeletal muscle.     

New Azaphilones,Seco-Chaetomugilins A and D,Produced by a Marine-Fish-Derived Chaetomium globosum

Seco-chaetomugilins A and D were isolated from a strain of Chaetomium globosum that was originally isolated from the marine fish Mugil cephalus, and their absolute stereostructures were elucidated on the basis of spectroscopic analyses, including 1D and 2D NMR techniques, along with the chemical transformation from known chaetomugilins A and D. Seco-chaetomugilin D exhibited growth inhibitory activity against cultured P388, HL-60, L1210, and KB cells.     

A canine case of otitis media examined and cured using a video otoscope

Otitis media of the left ear was diagnosed by video otoscopic examination in a 7-year-old, intact male Shih-tzu dog (weight, 5.1 kg), that also had three complex ceruminous adenomas and a Pseudomonas aeruginosa infection in the left ear canal. In such cases, total ear canal ablation is usually required. However, a complete cure was achieved in the present case without total ear canal ablation. The complex ceruminous adenomas were excised using a diode laser, and repeated cleansing of the tympanic cavity and ear canal was implemented using a video otoscope. As a result, the ear canal was closed in a U-form, and the otitis media was cured.     

Growth hormone secretion during longterm incubation of the pituitary of the Japanese eel,Anguilla japonica

Growth hormone (GH) secretion from organ-cultured pituitaries of the eel (Anguilla japonica) was studied during incubation in a defined medium for 2 weeks, using a homologous radioimmunoassay which does not distinguish between the two molecular forms of eel GH. The total amount of GH secreted increased gradually during the incubation period; so that the amount of GH released on day 14 was about 30 times greater than that on day 1. On day 14, the proportion of GH released relative to the total amount of GH present (the sum of GH released into the medium and residual content in the pituitary) was 96% and the amount produced on day 14 was 4 times greater than the content in the unincubated pituitary. Somatostatin (SRIF, 1.8 × 10^-7 M) inhibited the increase in GH release. On day 7, the proportion of GH released by pituitaries treated with SRIF (28%) was less than that released by the control pituitary (91%). There was no significant difference in GH release between the pituitaries incubated in isotonic medium (300 mOsm) and those in hypotonic medium (240 mOsm) for 2 weeks except for the first 3 days, when the pituitaries in hypotonic medium secreted significantly greater amounts of GH than those incubated under isotonic condition. Hypertonic medium (350 mOsm) had no effect on GH release except for significant inhibition on days 6 and 14. When secretion of the two forms of GH (GH I and II) was examined after separation by polyacrylamide gel electrophoresis followed by densitometry, slightly more GH I tended to be secreted than GH II during the culture period, although the effects of SRIF and osmolality of the media on GH I release were similar to those on GH II. It is concluded that GH secretion and production in the eel is mainly under the inhibitory control of hypothalamus, and that osmolality has a minimum influence on the GH release.