Cetacean Toll-like receptor 4 and myeloid differentiation factor 2, and possible cetacean-specific responses against Gram-negative bacteria

Toll-like receptor 4 (TLR4) and myeloid differentiation factor 2 (MD-2) are essential for recognizing the lipopolysaccharides (LPS) of Gram-negative bacteria. We determined the sequences of cDNAs encoding TLR4 and MD-2 from cetaceans and generated three-dimensional (3D) models for a better understanding of their modes of interaction and LPS recognition. The 3D reconstructions showed that cetacean TLR4 and MD-2 formed a horseshoe-like structure comprised of parallel β-strands and a β-cup structure consisting of two anti-parallel β-sheets, respectively. The (TLR4-MD-2)₂ duplex-heterodimer was shown to form a symmetrical structure. Comparison with the interfaces of the complexes in other mammals revealed that cetacean TLR4s have some amino acid residue substitutions involved in duplex-heterodimer formation and in species variation for LPS recognition. These substitutions in the changed amino acid residues may alter the interaction among TLR4, MD-2, and LPS and modify the TLR4/MD-2 immunological responses.     

Spontaneous Erythroid Leukemia in a 7-Week-Old Crl:CD (SD) Rat

A young male Crl:CD (SD) rat with erythroid leukemia that presented with emaciation, abdominal distension and a pale visible mucosal membrane was euthanized at 7 weeks of age. At necropsy, enlargement of liver, spleen and pancreatic lymph node was noted. Analysis of blood smear samples revealed many mono- or binucleated erythroblasts that had PAS-positive vacuoles in the cytoplasm. Histopathologically, neoplastic proliferation of atypical cells was observed in the hepatic sinusoids, splenic red pulp, bone marrow, pancreatic lymph node, kidney and lung. Neoplastic cells showed a round to spindle shape, and some neoplastic cells had deeply stained small nuclei and small cytoplasms and resembled erythroblasts. Immunohistochemically, many neoplastic cells were positive for hemoglobin. To our knowledge, this is the first report of erythroid leukemia in a rat of this age. The observed features were similar to those of pure erythroid leukemia in humans.     

Growth curve analyses in selected duck lines.

1. Growth patterns of male ducks from 4 lines (lines A, B, C and D) selected for market weight were analysed and compared to growth patterns of ducks in the respective line 7 generations earlier. Growth curves were analysed using procedures derived from the Weibull sigmoidal function and the linear-linear relative growth rate model and simple allometry. 2. The ducks were fed ad libitum under 24-h lighting throughout the experiment. At weekly intervals from the time of hatch through 70 d of age, 16 ducks from each line were killed to determine body, carcase, breast-muscle, leg and thigh-muscle, and abdominal fat weights. 3. Line A was the heaviest line, followed by line B, line C and line D. However, body weight, carcase weight and breast-muscle weight at 49 d of age were not significantly different between lines A and B. After 7 generations of selection, the breast-muscle yield was increased to >19% and the abdominal fat percent was reduced to <1.4% in all lines. 4. The Weibull growth curve analysis of body weight showed an increase in the asymptotes during selection, while the age of the inflection point remained constant in all lines (21.3 to 26.0 d). For breast-muscle growth, ducks reached the inflection point 12.8 to 14.3 d later than for body weight. Between line A and line B, asymptotes for body weight, asymptotes for breast-muscle weight and allometric growth coefficients of breast muscle and leg and thigh muscles from 14 to 49 d were not significantly different. 5. The relative growth rate model discriminated body and breast-muscle growth patterns of line A and line B. The initial decline in the relative body growth rate was less and the time to reach the transition was longer in line A than line B. On the other hand, the initial decline in the relative breast-muscle growth rate was greater in line A than line B.     

Cross-reactive antibodies to BLV and HTLV in bovine and human hosts with retrovirus infection

Sera of 22 cattle naturally infected with bovine leukemia virus (BLV), of 2 calves vaccinated with BLV, and of 22 patients with human T cell leukemia virus type I (HTLV-I) infection were tested to BLV and HTLV-I by enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB). Sera of 22 healthy cattle and from 22 healthy persons, and mouse monoclonal antibody to BLV-gp51, to HTLV-I-p24, or to HTLV-I-p19 were also tested. Sera of virus-infected hosts gave significantly higher ELISA values than sera of healthy donors to both BLV and HTLV-I. The correlation between ELISA values of bovine sera to BLV and those to HTLV-I was r = 0.76, whereas that of human sera was r = 0.35. By WB and competitive WB assays, bovine sera that were ELISA-positive to BLV reacted with one or more of p12, p15, and p24 of BLV, and with only p24 of HTLV-I. Human sera that were ELISA-positive to HTLV-I reacted with p12 and p24 of BLV, and with one or more of p12, p15, p19, and p24 of HTLV-I. These results demonstrate that BLV and HTLV-I are capable of evoking cross-reactive immune response in at least some hosts under natural infection as well as by virus vaccination.     

Preservation of feline anti-Toxoplasma gondii antibody activity using blood absorbed on filter paper stored under different conditions

The present study deals with the successful long-term preservation of feline anti-Toxoplasma gondii antibody activity on filter paper strips stored with silica gel. When filter paper strips were kept at 25 degrees C with silica gel, antibody activity in blood samples was preserved for at least six months.     

The first record of sucking louse, Neohaematopinus callosciuri, infesting pallas squirrels in Japan

A species of sucking louse, Neohaematopinus callosciuri, was found for the first time in Japan. The species was found on an invasive species of squirrel, Pallas squirrel, Callosciurus erythraeus, in the Kamakura district, Kanagawa Prefecture, Japan. A total of 52 lice were obtained from 22 of 104 squirrels captured. The lice were about three times more prevalent in male squirrels than in females and were detected most frequently in the winter. As N. callosciuri has never been reported on wild animals in Japan, this species probably was introduced into Japan along with their host, Pallas squirrels.     

Molecular cloning of canine mucosal addressin cell adhesion molecule-1 cDNA and its expression in normal tissues

A cDNA encoding canine mucosal addressin cell adhesion molecule-1 (MAdCAM-1) was cloned. The entire open reading frame of canine MAdCAM-1 cDNA comprises 1137 bp, corresponding to 378 amino acid residues. The deduced amino acid sequence of canine MAdCAM-1 was 55.2%, 53.7%, and 52.4% identical to rat, mouse, and human MAdCAM-1, respectively. Canine MAdCAM-1 appeared to contain two immunoglobulin-like domains at the N-terminus, followed by a mucin-like domain and a third immunoglobulin-like domain. The structures of the dog, rat, and mouse proteins are likely similar because all of the cysteine residues in the immunoglobulin-like domains were conserved. Canine MAdCAM-1 mRNA was confirmed to express extremely in the mesenteric lymph node by RT-PCR.     

Cloning of canine Toll-like receptor 7 gene and its expression in dog tissues

Toll-like receptor 7 (TLR7) is activated by single strand RNA and imidazoquinoline compounds, and induces interferon production. In this study, canine TLR7 cDNA was cloned and sequenced. The full-length cDNA of canine TLR7 gene was 3419bp, encoding 1032 amino acids. The similarities of canine TLR7 with human and mouse TLR7 were 84 and 80% at the nucleotide sequence level, and 86 and 79% at amino acid sequence level, respectively. Further, the expression of TLR7 mRNA was investigated in canine normal tissues by semiquantitative RT-PCR analysis. The common expression level of TLR7 mRNA in tissues from three dogs examined was in large intestine, lung, pancreas, small intestine and skin, though the expression level in each tissue was varied among these healthy dogs. In other tissues (kidney, liver, lymph node, spleen, adrenal gland, and PBMCs), the level of TLR7 mRNA expression was different in individuals.     

Effects of Increasing Temperatures on Spikelet Fertility in Different Rice Cultivars based on Temperature Gradient Chamber Experiments

Spikelet sterility in rice (Oryza sativa L.) induced by high temperatures is a major concern given global warming predictions. We studied differences among eight rice cultivars in spikelet fertility at five different temperature levels in temperature gradient chamber (TGC) experiments. Six japonica and two indica cultivars were exposed to high‐temperature gradients in TGCs during the 2005 flowering season. Spikelet sterility increased with temperature in TGCs and differed among cultivars because of both variations in temperature tolerance and timing of heading. The correlation between spikelet fertility of individual panicles and both air temperature and panicle temperature during flowering was analyzed to compare tolerances among cultivars. The temperature (T₇₅) at which spikelet fertility was 75 % of maximum ranged from 34 to 39 °C air temperature and differed significantly among cultivars. Indica varieties had higher T₇₅ values than japonica varieties. The T₇₅ values based on panicle temperature also differed among cultivars, but the difference between indica and japonica varieties were less significant. We concluded that the higher temperature tolerances of indica cultivars in our experiments could be attributed to lower spikelet temperatures, and cultivars with similar spikelet temperatures still had different heat tolerances due to differences in pollination ability.     

An economic evaluation of Kanazawa and Shichika irrigation water’s multi-functional roles using CVM

In order to confirm that the regional function of Kanazawa’s irrigation water is representative of the urban type, and that Shichika’s irrigation water is representative of the rural, the contingent valuation method (CVM) was applied to determine the economic value of the functions aside from agricultural production, that is, the multi-functional roles of the irrigation water. The economic value of the regional function in Kanazawa was significantly lower than in the Shichika area, reflecting a difference in charge for irrigation water and people’s preference or consciousness in urban and rural areas. In urban areas, people tend to place a higher value on ground view, peace of mind and environmental friendliness. In contrast, in rural areas, irrigation water is presently used for domestic purposes. In other words, in urban areas people see water as public goods, while in rural areas the people see water as private goods. In addition, differences in the economic value of each item that was statistically significant for CVM were estimated using the Turnbull model comparing the items approved or rejected by the respondent.