Dystrophin-deficient muscular dystrophy in a Toy Poodle with a single base pair insertion in exon 45 of the Duchenne muscular dystrophy gene

A 10-month-old, intact male Toy Poodle was referred for a postural abnormality. Blood biochemical tests revealed a marked increase in plasma creatine phosphokinase (CPK) concentration. The isoenzyme test showed that 99% of serum CPK consisted of CPK-MM. Histopathological evaluation of muscle biopsy samples confirmed scattered degeneration and necrosis of myofibers. Immunohistochemistry for dystrophin showed an absence of staining in muscle cells. Based on these findings, the dog was diagnosed with dystrophin-deficient muscular dystrophy. Whole genome sequencing using genomic DNA extracted from blood revealed a single base pair insertion in exon 45 of the Duchenne muscular dystrophy (DMD) gene. This is the first report on muscular dystrophy in Toy Poodles and identified a novel mutation in the DMD gene.     

Inhibition of nitrogen uptake by rice after wheat straw application determined by tracer NH4 +-15N

This experiment was carried out to determine whether the delay in rice growth associated with wheat straw application, especially at the early stage, was due to the acceleration of N assimilation or N uptake inhibition.

Tracer ¹⁵N was used for rice plants cultivated in pots. After 24 h of tracer application the plants and soils were sampled for analysis. Seventeen days after transplanting, N uptake of rice decreased and the amount of unavailable tracer remaining in soil increased by wheat straw application. At the booting stage, 6 d before heading, N uptake was larger and the amount of remaining tracer was lower in the plots in which wheat straw was applied than in the control.

It was obvious that the decrease of N uptake by wheat straw application was caused by N uptake inhibition and not by N starvation for a period of time at the early stage. The inhibition was removed at the booting stage.     

Using different versions of the Rothamsted Carbon model to simulate soil carbon in long-term experimental plots subjected to paddy–upland rotation in Japan

We validated the Rothamsted Carbon (RothC) model against 14 experimental plots subjected to different paddy–upland rotation patterns, including continuous paddy rice cropping and different periods (short, medium, and long term) of upland conversion, in Akita prefecture, northern Japan. We ran the model using the original version of the RothC model, the paddy-soil version of the RothC model (which uses a reduced carbon decomposition rate), and alternating the use of the original and the paddy-soil versions for years with summer upland cropping and paddy rice cropping, respectively. The best simulation pattern was provided by the alternate use of the two versions. In comparison with the original RothC model, alternate use of the two versions showed a smaller root mean square error in all experimental plots and a smaller absolute value of the mean difference in 10 plots with continuous paddy rice cropping and short- and medium-term upland conversion. Alternate use of the two versions had a lower root mean square error and lower absolute value of the mean difference in long-term upland conversion plots compared to the values for the paddy-soil version and showed better performance than the paddy-soil version when straw was applied. The model performance from alternating use of the two versions compared well with the performance reported in previous studies. Alternating the use of the two model versions provides a good technique for simulating changes in soil organic carbon with time, even with a paddy–upland rotation pattern. In addition, this technique will enable a more accurate inventory of Japanese greenhouse gases and help land managers better manage soil fertility in a paddy–upland rotation system.     

Status of paddy soils as affected by paddy rice and upland soybean rotation in northeast Japan,with special reference to nitrogen fertility

To investigate the current available nitrogen (N) and chemical properties of paddy soils affected by crop rotation between irrigated paddy rice (Oryza sativa L.) and upland soybean [Glycine max (L.) Merr.] (paddy-upland rotation), topsoils were collected from 22 fields of four different farmers in the northeastern region of Japan. Regardless of organic material application, a significant negative correlation was found between available soil N and an increase in the proportions of upland seasons to total crop seasons after the initiation of paddy-upland rotation. Soil total N and total carbon (C) also tended to decrease with an increase in upland frequency. In fields with repeated applications of cattle manure compost, the soil available N was higher than in fields where only crop residue was applied. A significant negative correlation was also found between the soil available N:total N ratio and upland frequency. This indicates that the part of soil N related to available N was notably lost by the use of paddy fields as upland fields. In order to sustain available soil N over the minimum suitable level of 80?mg?kg^?1, upland frequency should not exceed 65% when only crop residues and no other organic materials are applied. The upland frequency can be raised by the repeated application of organic materials which maintain a higher level of available soil N. The results imply that care should be taken to maintain the N fertility of paddy soil at a suitable level in paddy-upland rotation, and that upland frequency and organic materials applied are important factors to do this.     

Nitrogen and oxygen isotope enrichment factors of nitrate at different denitrification rates in an agricultural soil

ABSTRACT

Quantitative evaluation of denitrification by the dual isotope approach, which measures the stable isotope ratios of nitrogen (δ¹⁵N) and oxygen (δ¹⁸O) in nitrate, has been hampered by the wide range of values reported for the ratio of enrichment factors for ¹⁵N and ¹⁸O (¹⁵ε and ¹⁸ε, respectively) during denitrification. The objectives of this study were to determine ¹⁵ε and ¹⁸ε values at different denitrification rates under controlled conditions, and to infer possible mechanisms by which the ¹⁸ε/¹⁵ε ratio is influenced under different conditions. Column experiments were conducted at 25, 15, and 10°C, which enabled determination of ¹⁵ε and ¹⁸ε at different denitrification rates, in the absence of nitrate replenishment from ammonium oxidation and other sources. The values of ¹⁵ε and ¹⁸ε ranged from ?11.8 to ?14.9‰ and from ?8.4 to ?15.9‰, respectively, with ¹⁵ε less sensitive to changes in the denitrification rates. The resultant ¹⁸ε/¹⁵ε ratio, ranging from 0.70 to 1.17, was close to the values reported for sediment incubations, and larger than those for groundwater systems. These results are consistent with the explanations that ¹⁸ε/¹⁵ε value itself is close to unity during denitrification, and that at smaller denitrification rates, concurrent reactions including re-oxidation of nitrite to nitrate lead to smaller apparent fractionation of ¹⁸O and smaller ¹⁸ε/¹⁵ε ratios. This suggests that while linear relationships between δ¹⁸O and δ¹⁵N give a strong evidence of denitrification, apparent ¹⁸ε/¹⁵ε values are site specific and depend on the ambient conditions. In evaluating denitrification in such systems, we suggest the use of ¹⁵ε in preference to ¹⁸ε because ¹⁵ε is less sensitive to denitrification rates.     

Contribution of enhanced efflux to reduced susceptibility of Salmonella enterica serovar Choleraesuis to fluoroquinolone and other antimicrobials

We examined antimicrobial susceptibility and efflux systems in laboratory-derived mutants of Salmonella enterica serovar Choleraesuis selected by culture on fluoroquinolone-containing plates. The mutants exhibited decreased susceptibilities to quinolones and several other antimicrobials. Mutations in the gyrA gene were not always found in the mutants. Accumulation assays revealed that intracellular enrofloxacin concentrations were significantly lower in the mutants compared with parent isolates. Increased expression of acrB mRNA can explain the decreased susceptibilities to several antimicrobials but not in the case of carbonyl cyanide m-chlorophenylhydrazone (CCCP). Decreased susceptibility to CCCP may result from the increased expression of emrA mRNA. These results suggest that the enhancement of multiple efflux pumps is responsible for decreased susceptibilities to several antimicrobials in the laboratory-derived mutants.     

Fetal and neonatal goiter in cynomolgus monkeys following administration of the antithyroid drug thiamazole at high doses to dams during pregnancy

To evaluate morphologic alterations in the thyroid gland in the second generation in cynomolgus monkeys, pregnant dams were exposed to high doses of thiamazole. In Experiment A, dams received thiamazole intragastrically via a nasogastric catheter from gestation day (GD) 50 to GD 150 or on the day before delivery. Initially, the dose level was 20 mg/kg/day (10 mg/kg twice daily); however, the dose level was subsequently decreased to 5 mg/kg/day (2.5 mg/kg twice daily), since deteriorated general conditions were observed in two dams. Six out of seven neonates died on the day of birth. The cause of neonatal death was tracheal compression and suffocation from goiter. The transplacental exposure to thiamazole affected the fetal thyroid glands and induced goiter in all neonates. The surviving neonate was necropsied 767 days after discontinuation of thiamazole exposure and showed reversibility of the induced changes. In Experiment B, dams were intragastrically administered thiamazole at 5 mg/kg/day (2.5 mg/kg twice daily) for treatment periods from GDs 51 to 70, 71 to 90, 91 to 110, 111 to 130 and 131 to 150. All fetuses showed enlarged thyroid glands but were viable. Histopathologically, hypertrophy and/or hyperplastic appearance of the follicular epithelium of the thyroid gland was observed at the end of each treatment period. The most active appearance of the follicular epithelium, consisting of crowded pedunculated structure, was demonstrated at end of the treatment period from GD 131 to 150. This is the first report on the morphology of fetal and neonatal goiter in the cynomolgus monkey.     

Reciprocal interference between specific CJD and scrapie agents in neural cell cultures

Infection of mice with an attenuated Creutzfeldt-Jakob disease agent (SY-CJD) interferes with superinfection by a more virulent human-derived CJD agent (FU-CJD) and does not require pathological prion protein (PrPres). Using a rapid coculture system, we found that a neural cell line free of immune system cells similarly supported substantial CJD agent interference without PrPres. In addition, SY-CJD prevented superinfection by sheep-derived Chandler (Ch) and 22L scrapie agents. However, only 22L and not Ch prevented FU-CJD infection, even though both scrapie strains provoked abundant PrPres. This relationship between particular strains of sheep- and human-derived agents is likely to affect their prevalence and epidemic spread.