Evaluation of bone toxicity in various bones of aged rats

The aim of the present study was to provide a method for evaluating bone toxicity induced by drugs in various bones in aged rats. Male Crl:CD (SD) rats at 46 weeks of age were administered 15 mg/m(2) body surface area of doxorubicin, which effects the growth plate in weanling rats, weekly for 9 weeks by intravenous injection, and the femur, sternum, humerus and tibia were examined histopathologically. In the doxorubicin-treated group, thinning of the growth plate was remarkably observed in the proximal tibia and humerus; however, these changes were not observed in other regions. In addition, the osteoclast number per bone perimeter in the proximal tibia was significantly higher than others in control aged rat. Thus, recognizing the various histological reactions related to the time of epiphyseal closure is important for evaluating bone toxicity in aged rats.     

Alleviation of manganese toxicity and manganese-induced iron deficiency in barley by additional potassium supply in nutrient solution

Barley plants were grown hydroponically at two levels of K (3.0 and 30 mm) and Fe (1.0 and 10 μm) in the presence of excess Mn (25 μm) for 14 d in a phytotron. Plants grown under adequate K level (3.0 mm) were characterized by brown spots on old leaves, desiccation of old leaves, interveinal chlorosis on young leaves, browning of roots, and release of phytosiderophores (PS) from roots. These symptoms were more pronounced in the plants grown under suboptimal Fe level (1.0 p,M) than in the plants grown under adequate Fe level (10 μm). Plants grown in 10 μm Fe with additional K (30 mm) produced a larger amount of dry matter and released less PS than the plants grown under adequate K level (3.0 mm), and did not show leaf injury symptoms and root browning. On the other hand, the additional K supply in the presence of 1.0 μM Fe decreased the severity of brown spots, prevented leaf desiccation, and increased the leaf chlorophyll content, which was not sufficient for the regreening of chlorotic leaves. These results suggested that the additional K alleviated the symptoms of Mn toxicity depending on the Fe concentration in the nutrient solution. The concentration (per g dry matter) and accumulation (per plant) of Mn in shoots and roots of plants grown in 10 μm Fe and 30 mm K were much lower than those of the plants grown in 10 μm Fe and 3.0 mm K, indicating that additional K repressed the absorption of Mn. The concentration and accumulation of Fe in the shoots and roots of the plants grown in 10 μm Fe and 30 mm K were higher than those of the plants grown in 10 μm Fe and 3.0 mm K, indicating that the additional K increased the absorption of Fe under excess Mn level in the nutrient solution. The release of PS, chlorophyll content, and shoot Fe concentration were closely correlated.     

Diurnal variations in absorption and translocation of a ferrated phytosiderophore in barley as affected by iron deficiency

The release of phytosiderophore (PS) from roots of Fe-deficient graminaceous plants follows a distinct diurnal rhythm with maximum release rates occurring usually 3 to 4 hours after the onset of light. However, it remains to be determined whether absorption of the PS-Fe³⁺ complex shows a diurnal rhythmicity similar to that of PS release, Barley plants grown with or without 10 µM FeEDTA for 7 days were fed with ferreted PS (10 µM labelled with ⁵⁹Fe) at 4-h intervals to study the diurnal variations in the absorption and transloca tion of ⁵⁹Fe, The absorption of ⁵⁹Fe, irrespective of the Fe nutritional status of the plants, was higher during the day and lower during the night but did not show any peak throughout the day-night cycle. On the other hand, the translocation of ⁵⁹Fe into shoots of Fe-deficient plants was lower than that of Fe-sufficient plants, while the Fe nutritional status of the plants did not affect the absorption of ⁵⁹Fe by roots, The formation of root apoplastic ⁵⁹Fe was lower during the day and higher during the night, regardless of the Fe nutritional status of plants. Our results showed that the absorption of the PS-Fe³⁺ complex by roots did not follow the PS release pattern.     

Effect of soil solarization on subsequent nitrification activity at elevated temperatures

Soil solarization is a nonchemical method of soil disinfection achieved by covering the soil surface with sheets of vinyl plastic to generate elevated soil temperature, generally over 45°C. Such elevated temperatures may be detrimental to some nitrifying microorganisms and favorable to others. However, little information exists to indicate how nitrification activity in soil is affected after solarization. We performed several experiments to investigate the effects of soil solarization on nitrification activity. We found that: (1) if a soil was subjected to pretreatment of 45 or 50°C for as little as 1 d, nitrification activity in a subsequent incubation at 30°C was less than that of a soil that did not receive any high-temperature pretreatment. However, if a soil received pretreatments of 45 or 50°C for more than 7 d, nitrification activity in a subsequent incubation at 45 or 50°C was greater than that of soil that did not receive high temperature pretreatment. (2) Nitrification activity in three kinds of soil taken from 0–5 cm depth after solarization treatment was greater at 45°C than 30°C. (3) Nitrification activity at 45°C in soil that had received solarization in the preceding year was greater than that in soil that had not been subjected to solarization. This was consistent with the fact that the population densities of ammonia oxidizers were greater in soils that had been subjected to solarization. These results suggest that soil solarization induces nitrifying microorganisms that are more active at 45–50°C than they are at 30°C, and that the effect of solarization on nitrification persists until the next crop season.     

Effectiveness of Phytosiderophore in Absorption and Translocation of 59Iron in Barley in the Presence of Plant-Borne,Synthetic, and Microbial Chelators

The mechanisms of iron (Fe) absorption and translocation in plants have received much study because they are the key processes in the supply of Fe to plants. The objective of this research was to study the effectiveness of phytosiderophore (PS) in the absorption and translocation of ⁵⁹Fe in Fe-deficient barley (Hordeum vulgare L. cv. ‘Minorimugi’) plants in the presence of plant-borne, synthetic, or microbial chelators. Plants grown under Fe-deficient conditions in a phytotron at pH 5.5 for 7–18 d were fed with Fe³⁺ (10 μ M labeled with ⁵⁹Fe) in the presence of 10 μ M of different chelators with or without 10 μ M PS for 4 h starting at 2 p.m. (6 h after the onset of light period). The absorption and translocation of ⁵⁹Fe in plants treated with PS and Fe^{3 +} were increased relative to plants fed solely with Fe^{3 +} (control). There was no effect found on absorption and translocation of ⁵⁹Fe in plants treated with EDTA or p-coumarate relative to the control, but a differential increase was observed in ⁵⁹Fe absorption and translocation in plants treated with EDTA or p-coumarate in the presence of PS. In comparison with the control, a decrease in ⁵⁹Fe absorption and translocation was observed in plants treated with HEDTA or EDDHA or FOB, but this decrease was avoided in plants treated with HEDTA or EDDHA or FOB in the presence of PS. The enhancement of ⁵⁹Fe absorption and translocation in plants treated with citrate, and the highest ⁵⁹Fe absorption and translocation in plants treated with citrate and PS, indicated that citrate had an additive effect on Fe absorption and translocation in plants. Our results showed that PS effectively played a role in Fe absorption and translocation in plants in the presence of other chelators. Plants treated with any chelators had lower extracellular ⁵⁹Fe in the roots compared with the control.     

A growth inhibitor specific to upland rice produced by Pyrenochaeta SP.

An attempt was made to isolate a growth inhibitor of upland rice produced by Pyrenochaeta sp. Partially purified pink substance having absorption maxima at 481, 502, 512,537 and 548 nm was obtained from mycelia of Pyrenochaeta sp. This substance inhibited the growth of upland rice at a lower concentration than p-coumaric acid. It inhibited the growth of upland rice and sorghum, but stimulated the growth of Chinese cabbage and cucumber.     

Relationships between body size and secondary sexual characters,and sperm characters in male Dolly Varden char (Salvelinus malma)

In Salmonidae, subordinate males are exposed to higher risks of sperm competition than dominant males and thus are expected to improve the sperm characteristics (sperm concentrations, sperm velocity and sperm longevity). In this study, we investigated the relationships between body size and secondary sexual characters (breeding colour, hump height and snout length), and sperm characteristics of one‐year‐old (newly matured) Dolly Varden char. Small males displayed higher sperm concentrations than large males. Moreover, males with dull breeding colours, but not with lesser snout length and hump height, displayed an increased sperm velocity compared to males with bright colours, suggesting a trade‐off between sperm quantity and the investment in breeding colour. In addition, sperm longevity decreased as sperm swimming velocity increased. These findings indicate that small males with dull breeding colours improve the quantity and quality of their sperm to a great extent to enhance their chances of reproductive success.     

Hair cortisol levels of lactating dairy cows in cold‐ and warm‐temperate regions in Japan

We compared the hair cortisol levels of lactating dairy cows in a cold‐ and a warm‐temperate region out of four climatic zones in Japan. We simultaneously investigated the effects of calving number, lactation period and month of hair sampling. Hair of nine Holstein lactating cows chosen from each region (i.e. 18 cows per sampling) was sampled in March, June, September and December. Number of calvings (1, 2, ≥3) and lactation duration (early: <100, middle: 101–200, and late: >201 days) were balanced between regions. Cortisol was extracted from hair by methanol, and its level was determined with a cortisol immunoassay kit. A multi‐way analysis of variance revealed that the effects of month of hair sampling (P < 0.001) and its combination with region (P < 0.05) were significant. In a multiple comparison test, significant differences (P < 0.01) in hair cortisol level (pg/mg of hair) were found between June (13.0 ± 1.0) and the other 3 months, and between September (1.6 ± 0.2) and December (4.5 ± 0.3). The rise in cortisol level from March to June was more intense in the cold‐temperate region. These results demonstrate the necessity of considering seasonal variations in each climatic region when we use hair cortisol level as an indicator of stress.     

A novel culture method of canine peripheral blood lymphocytes with concanavalin a and recombinant human interleukin-2 for adoptive immunotherapy

This study was designed to develop a novel culture method for the efficient proliferation of canine peripheral blood lymphocytes (cPBL) for adoptive immunotherapy. When cPBL were cultured in the presence of concanavalin A (Con A), proliferation of cPBL was induced and expression of interleukin-2 receptor (IL-2R) which enables to respond to exogenously added IL-2 was upregulated. And then, when cPBL were cultured with recombinant human interleukin-2 (rhIL-2) in addition to Con A, proliferation was accelerated and increased to about 10-fold after 1 week. The phenotypic analysis showed that the main population of the cultured cPBL was consisted of CD8+ positive lymphocytes. Among them, CD4+CD8+ double positive (DP) lymphocytes had significantly increased, and the ratio of CD4+ single positive (SP) lymphocytes to CD8+ SP lymphocytes (CD4+SP/CD8+SP) was decreased as compared to before culturing. To evaluate the cytotoxic activity of cPBL cultured with Con A and rhIL-2, furthermore, cytotoxic assay was carried out against xenogeneic melanoma cell line (MeWo), which resulted in MHC-unrestricted cytokilling. These results suggest that the culture method of cPBL by the use of Con A and rhIL-2 may be useful for generating lymphokine activated killer cells, and also this may be beneficial for adoptive immunotherapy of tumor-bearing dogs.