Multigenic system controlling viral systemic infection determined by the interactions between Cucumber mosaic virus genes and quantitative trait loci of soybean cultivars

Soybean 'Harosoy' is resistant to Cucumber mosaic virus soybean strain C (CMV-SC) and susceptible to CMV-S strain D (CMV-SD). Using enzyme-linked immunosorbent assay and Northern hybridization, we characterized the Harosoy resistance and found that CMV-SC did not spread systemically but was restricted to the inoculated leaves in Harosoy. Harosoy resistance was not controlled by either a dominant or recessive single gene. To dissect this system controlling long-distance movement of CMV in soybean, we constructed infectious cDNA clones of CMV-SC and CMV-SD. Using these constructs and the chimeric RNAs, we demonstrated that two viral components were required for systemic infection by the virus. The region including the entire 2b gene and the 5' region of RNA3 (mainly the 5' untranslated region) together were required. By quantitative trait locus (QTL) analysis using an F(2) population and the F(3) families derived from Harosoy and susceptible 'Nemashirazu', we also showed that at least three QTLs affected systemic infection of CMV in soybean. Our study on Harosoy resistance to CMV-SC revealed an interesting mechanism, in which multiple host and viral genes coordinately controlled viral systemic infection.     

Iodine desorption from rice paddy soil

Laboratory experiments on the desorption phenomena of iodine from rice paddy soil under waterlogged conditions, with a special reference to soil redox potential (Eh) and pH, have been conducted. Radioiodine tracer (1251), added to the soil, was readily sorbed on it. At the beginning of the waterlogging, the iodine desorption was low. However, iodine was desorbed into soil solution with time. The iodine desorption was enhanced markedly by the addition of organic substances such as straw pieces and glucose to the soil. Cultivation of rice plants in soil also affected the iodine desorption, suggesting root exudates and/or root autolysis might be participating in the desorption process. Eh dropped considerably after soil was waterlogged due to microbial metabolisms. Particularly low Eh values were observed in soils with plants and also with added organic substances. A negative correlation was seen between the desorption and soil Eh. High desorption was frequently observed when the Eh dropped to about -100 mV or below. Due to the reducing conditions (low Eh) by waterlogging, iodine in soil was leached into the soil solution; consequently total iodine concentration in paddy soil was considerably lower than forest and upland field soils. These iodine desorption phenomena under anaerobic conditions should be considered in assessing transfer of the long-lived radioiodine (129I) in the environment, especially in rice fields and marshland.     

Geographical and Temporal Variations of Chemical Constituents in Winter Precipitation Collected in the Areas along the Coast of the Sea of Japan

The geographical and temporal variations of chemical constituents in winter precipitation collected in the areas along the coast of the Sea of Japan (AASJ) were discussed by analyzing the data obtained in the 1st and 2nd National Acid Deposition Survey by Japan Environmental Laboratories Association. In western Tohoku (WT) and Hokuriku (HR) areas in AASJ, in spite of large amounts of precipitation in winter, concentrations of non sea salt (nss-)SO₄ ^2? are not as low as the other areas, and nss-Ca²⁺ in these areas is lower than the other areas. As a result, H⁺ concentrations of precipitation in these areas are somewhat higher than other areas. From the temporal analysis of daily sampled data and back trajectory analysis of air mass, it was found that the concentrations of nss-SO₄ ^2?, NO₃ ^?, NH₄ ⁺ and nss-Ca²⁺ are correlatively varied when air mass come from the Asian Continent, showing higher concentrations at the western sites in AASJ and depending on the meteorological conditions such as the direction of in flow air mass.     

Precipitation chemistry in Japan 1989–1993

Precipitation chemistry in Japan was discussed on a wet-only sample database obtained in a nationwide survey from April 1989 to March 1993. Wet-only samples were collected at 29 stations over Japan on a biweekly basis. Commonly determined chemical parameters were measured in laboratories. The volume-weighted annual mean pH at each site ranged from 4.50 to 5.83 with a mean of 4.76. Concentration ranges and means (parenthesized) on an equivalent basis for major ions were as follows: nss-SO₄ ^2–; 5.2–58.9 (38.6), NO₃ ^–; 1.8–25.0 (14.1), NH₄ ⁺; 0.55–29.8 (18.3), nss-Ca²⁺; 2.0–34.5(14.2), Na⁺; 6.4–275.3 (49.1), Cl^–; 13.7–322.4 (63.5) eq L^–1. Acid-base relationships for Phase-II records were quantitatively discussed in terms of three measures: pH, fractional acidity, and our proposed pA_i.     

Development of transferred xenogeneic vole embryos in mouse uteri

An experimental model to study interspecific pregnancy using voles, Microtus arvalis, and green fluorescent protein gene‐induced transgenic mice is presented. Xenogeneic blastocysts from the vole were transferred into the uteri of pseudopregnant mice along with allogeneic blastocysts from green fluorescent protein gene‐induced transgenic mice. The uteri containing xeno‐allo combined transfers were examined from day 6 to 13 of gestation. Although the vole embryos implanted, the uteri containing vole embryos were smaller compared with those having allogeneic mouse embryos. On day 8, the uteri containing vole embryos hemorrhaged internally and no vole embryo was found in the pregnant uterus after day 11. Allogeneic mouse embryos developed normally despite the presence and abortion of the vole embryos. In uteri implanted with vole embryos, decidua were formed and numerous blood vessels were distributed around the embryo. Maternal blood cells infiltrated into the celomic cavity of the vole embryo through the discontinuous region of trophoblast. Periodic acid‐Schiff‐positive granulated metrial gland cells were remarkably increased in the decidual sites. These findings suggest that a disorder of embryo–maternal interaction might induce the appearance of numerous granulated metrial gland cells and rejection of the embryos.     

Influence of dietary protein intake on whole-body protein turnover in chicks

1. Experiments were conducted to investigate whether or not varying dietary protein intake affects whole-body protein turnover rates in young chicks. 2. Seven-d-old single comb White Leghorn male chicks were fed on diets with protein concentrations of 0, 100, 200 or 400 g/kg diet under conditions of ad libitum or equalised feeding. At the end of the experiments, the rate of protein synthesis and protein degradation in the whole body were measured in vivo. 3. The results showed that both fractional and absolute rates of protein synthesis increased with increasing dietary protein up to 200 g/kg; above this concentration they remained almost constant when feeding was ad libitum. 4. Similar responses were found with equalized feeding except that a significant reduction in protein synthesis was found when dietary protein was increased from 200 to 400 g/kg diet. 5. Less sensitive and almost parallel changes in protein degradation rates were found. 6. It was concluded that adaptation to varied dietary protein intake occurred primarily through changes in protein synthesis, accompanied by parallel alterations in protein degradation in the whole body.     

Reliability of using reagent test strips to estimate blood urea nitrogen concentration in dogs and cats

OBJECTIVE: To evaluate the clinical accuracy of reagent test strips used to estimate BUN concentration in dogs and cats. DESIGN: Prospective study. ANIMALS: 116 dogs and 58 cats. PROCEDURE: Blood samples were collected at the time of admission to the hospital. Estimates of BUN concentration obtained with reagent test strips (category 1 [5 to 15 mg/dL], 2 (15 to 26 mg/dL], 3 [30 to 40 mg/dL], or 4 [50 to 80 mg/dL]) were compared with SUN concentrations measured with an automated analyzer. For dogs, category 1 and 2 test strip results were considered a negative result (nonazotemic) and category 3 and 4 test strip results were considered a positive result (azotemic). For cats, category 1, 2, and 3 test strip results were considered a negative result (nonazotemic) and category 4 test strip results were considered a positive result (azotemic). RESULTS: On the basis of SUN concentration, 40 of the 174 (23%) animals (20 dogs and 20 cats) were classified as azotemic. One dog and 2 cats had false-negative test strip results, and 1 dog had a false-positive result. Sensitivity and specificity were 95% (20/21) and 99% (94/95), respectively, for dogs and 87% (13/15) and 100% (43/43), respectively, for cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that reagent test strips are a reliable method for rapidly estimating BUN concentrations in dogs and cats. Because test strip results are only semiquantitative and there remains a potential for misclassification, especially in cats, urea nitrogen concentration should ultimately be verified by means of standard chemistry techniques.     

Elevated endothelin-1 expression in dogs with heartworm disease

We explored the involvement of endothelin-1 (ET-1) in the pathophysiology of dog dirofilariasis (heartworm disease caused by Dirofilaria immitis) by analyzing mRNA levels of preproendothelin-1 (PPET-1), the precursor form of ET-1, in cardiopulmonary organs as well as ET-1 peptide levels in plasma. To determine the cDNA sequence and primary protein structure of dog PPET-1, we performed molecular cloning of the full-length cDNA. Based on the determined sequence information, comparative expression analysis of PPET-1 mRNA was carried out by real-time polymerase chain reaction on cardiopulmonary organs from healthy (n=5) and filarial (n=5) dogs. Filarial dogs showed a significantly (p<0.05) higher mRNA expression level in the heart (about one hundred times) and lung (about ten times) than healthy dogs. Analysis of plasma ET-1 levels in healthy (n=10) and filarial (n=10) dogs showed that filarial dogs (6.9+/-2.7 pg/ml) have significantly (p<0.01) increased plasma ET-1 levels compared with healthy dogs (1.4+/-0.3 pg/ml). To assess the pathophysiological significance of ET-1 in dirofilariasis relative to other cardiopulmonary disorders, plasma ET-1 levels determined in dogs diagnosed with mitral regurgitation (n=10), tricuspid regurgitation (n=5), ventricular septal defect (n=5), and patent ductus arteriosus (n=5) were compared to plasma ET-1 levels in filarial dogs. Filarial dogs, which commonly develop serious pulmonary hypertension, exhibited by far the highest ET-1 levels of the disease states examined. Based on the fact that ET-1 is a potent bioactive mediator that induces vasoconstriction and promotes vascular remodeling, these findings suggest that ET-1 plays an important role in the pathophysiology of dog dirofilariasis as an aggravating factor by inducing pulmonary hypertension.     

Establishment and characterization of eight feline mammary adenocarcinoma cell lines

Eight new feline mammary adenocarcinoma cell lines derived from either primary or metastatic lesions were established. The morphology of all the cell lines was epithelioid and round to spindle in shape, with cell growth occurring in a monolayer fashion. On immunohistochemistry, these cells reacted with anti-keratin and anti-vimentin antisera. The doubling time of these cells was between 19 and 54 hr. Tumor masses were developed in nude mice by subcutaneous inoculation of the cells that were histologically identical to their original mammary tumor lesions. Telomerase activities measured using the telomeric repeat amplification protocol assay revealed high telemetric activity in all of the cells.