Simultaneous steroids measurement in dogs with hyperadrenocorticism using a column-switching liquid chromatography-tandem mass spectrometry method

We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2–16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.     

Effects of zinc on tissue uptake and toxicity of lead in Sprague Dawley rat

Lead (Pb) exposure occurs together with other metals including zinc (Zn). This study investigated the impact of Zn on Pb tissue accumulation and Pb-induced toxicities. Animals (n=6 rats per group) were exposed to lead acetate (PbAc) or a combination of PbAc and zinc acetate (ZnAc) under the following groups: control (deionized water), low PbAc [12 mg/kg PbAc (3 mg PbAc/rat/day)], low PbAc–ZnAc [12 mg/kg PbAc (3 mg PbAc/rat/day) + 0.2 mg ZnAc/rat/48 hr], high PbAc [120 mg/kg (30 mg PbAc/rat/day)], and high PbAc–ZnAc [120 mg/kg (30 mg PbAc/rat/day) + 1 mg ZnAc/rat/48 hr] for 8 weeks. A significant reduction in body weight gain was observed in the high PbAc group relative to the control group. Muscles and testes both had reduced and increased Pb uptake in low PbAc–ZnAc and high PbAc–ZnAc groups compared to PbAc only groups, respectively. Bone Pb levels in the high PbAc–ZnAc group were lower than the high PbAc group. Zinc co-administration attenuated Pb-induced inhibition of delta aminolaevulinic acid dehydratase enzyme and enhanced catalase enzyme activity at a high level of exposure. Moreover, ZnAc seems to have minimized the effects of Pb-induced mRNA dysregulation in antioxidant and antiapoptotic enzymes encoding genes. Heme oxygenase-1 was downregulated in the kidney and brain in the low PbAc group. Liver glutathione peroxidase and thioredoxin reductase-1 were downregulated in the high PbAc group. These findings suggest that zinc co-administration with lead may partially mitigate against Pb-induced toxicities.     

Diversity in the carotenoid profiles and the expression of genes related to carotenoid accumulation among citrus genotypes

Carotenoids are not only important to the plants themselves but also are beneficial to human health. Since citrus fruit is a good source of carotenoids for the human diet, it is important to study carotenoid profiles and the accumulation mechanism in citrus fruit. Thus, in the present paper, we describe the diversity in the carotenoid profiles of fruit among citrus genotypes. In regard to carotenoids, such as β-cryptoxanthin, violaxanthin, lycopene, and β-citraurin, the relationship between the carotenoid profile and the expression of carotenoid-biosynthetic genes is discussed. Finally, recent results of quantitative trait locus (QTL) analyses of carotenoid contents and expression levels of carotenoid-biosynthetic genes in citrus fruit are shown.     

Secondary chronic respiratory acidosis in a dog following the cervical cord compression by an intradural glioma

An intradural tumor in the upper cervical region was found in a dog with quadriparesis and chronic respiratory acidosis. Surgical removal of the tumor in the atlas and intraoperative radiotherapy were attempted. The tumor was histologically diagnosed as a neural glioma. A preoperative acid-base disturbance was dramatically improved after surgery. The clinical changes appeared in this case suggest that compression of the spinal cord at this region may cause paralysis of the respiratory muscles and secondarily result in chronic respiratory acidosis following the respiratory insufficiency.     

The W‐ and Z‐linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island

The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR‐based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W‐ and Z‐linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR‐based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR‐based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.     

Effect of nitrogen application during ripening period on photosynthesis and dry matter production and its impact on yield and yield components of semidwarf iniica rice varieties under water culture conditions

Abstract

A water culture pot experiment was conducted to analyze the effects of N application during the ripening period (RP) on photosynthesis, dry matter production, and its impact on grain ripening and yield in two semidwarf indica type varieties viz. Gui Zhao 2 (GZ) and BR3 (BR) compared with a japonica type variety Koganemasari (KO) under four N rates viz. 0 (N₀), 10 (N₁₀), 20 (N₂₀), and 40 (N₄₀) mg L^?1. Results showed that N application enabled to maintain a higher leaf area and delay leaf senescence in both indica and japonica type varieties but the decrease in the rate, of leaf area was higher in the former than in the latter and the rate was reduced with increasing N rates. Flag leaf photosynthesis and SPAD values of N treated plants were higher throughout the RP, showing the presence of a significant correlation either for each variety or all the varieties together. Higher photosynthetic rate was supported by higher leaf chlorophyll (SPAD value basis) content, stomatai conductance, and N concentration in leaf blades. Top dry matter content increased with increasing N rates mainly due to mean leaf area rather than NAR except for BR during RP but it was higher in KO than in GZ and BR. Reduction of shoot weight due to translocation of dry matter to panicles during RP was suppressed by N rates both in GZ and BR while shoot weight increased in the N-treated plants in KO. The dependency of KO on current photosynthates for panicle weight was found to be almost hundred percent while the contribution of stored carbohydrates in shoot before heading to panicle weight in GZ and BR was in the range of 4-27 and 33-54%, respectively and the rest was contributed by current photosynthates. The percentage increased with increasing N rates. Percentage of ripened grains (PRG) increased with increasing N rates in GZ and BR due to the increase in dry matter production and in the photosynthetic rates of apex leaves, despite the larger spikelet number and larger hull size. However, KO showed almost no variation although it had the highest PRG among the varieties. Brown rice yield followed the same pattern as that of PRG in GZ and BR and the highest yield was produced by BR followed by GZ and KO. These results suggest that N application during RP was more effective in increasing yield in the semidwarf indica type varieties than in the japonica type variety.     

Relationships among the chalkiness,kernel size and endosperm cell morphology of rice kernels at different spikelet positions within a panicle

The occurrence of chalky kernels in rice is causally related to kernel size and endosperm morphology. This study aimed to investigate the occurrence of chalky kernels by analyzing kernel size, and the number and area of endosperm cells. Spikelets were sampled from upper and middle primary branches and lower secondary branches in a panicle, and divided into four categories: upper, middle, and lower perfect (PF) kernels and lower milky-white (MW) kernels. On the lower secondary branches, there was a higher percentage of chalky kernels, with smaller kernel lengths, widths and thicknesses, than the kernels on the upper and middle primary branches. MW kernels were smaller in size than PF kernels even on the same lower secondary branches. Regardless of grain appearance quality traits, the total areas of endosperm cross sections in lower kernels were significantly smaller than in upper kernels owing to the decreased cell area, and there was a significant negative correlation between the number of cells and average cell area. When the numbers and the areas of cells were analyzed using angular 30° intervals from the line connecting the center point and the dorsal vascular bundle, the MW kernels had significantly less cells than PF kernels near the ventral side at 120–180°. Thus, the decrease in the number of cells near the ventral side was a main causal factor in the decrease in MW kernel widths compared with PF kernel widths, and this suggested that cell division in MW kernels was inhibited at the early grain-filling stage.     

First report of ophidiomycosis in Asia caused by Ophidiomyces ophiodiicola in captive snakes in Japan

Ophidiomycosis is an emerging infectious disease caused by the fungus Ophidiomyces ophiodiicola, which has been affecting wild and captive snakes in North America, Europe, and Australia. We report 12 cases of suspected ophidiomycosis in captive colubrid snakes in Japan. Pathological and microbiological examinations were performed, and the results confirmed the diagnosis of ophidiomycosis in two snakes, which indicated that the remaining sympatrically raised snakes also had ophidiomycosis since they exhibited similar lesions. This is the first report of ophidiomycosis in Asia caused by O. ophiodiicola. To prevent the expansion of ophidiomycosis in the natural environment in Japan, there is a need to evaluate the ophidiomycosis carrier status of imported snakes, the pathogenicity of the infection in native snakes, and the prevalence and distribution of O. ophiodiicola in wild and captive snakes. Measures also must be taken to prevent endemicity globally.     

Effect of flexible and rigid roughness elements on aeolian sand transport

In developing countermeasures to reduce the negative effects of strong dust events, the fundamental relationship between surface conditions and sand transport remains problematic. We conducted field observations and wind tunnel experiments to examine the effect on sand transport efficiency of actual frontal area when using flexible roughness elements (artificial grass) and rigid roughness elements (tufts of stiff wire). In the field observations, the sand trap ratio approached a limit as the frontal area of artificial grass, measured as the Actual Frontal Area Index (AFAI), approached 25%, equivalent to a vegetation cover of 20%. The wind tunnel experiments showed that the height of deposited sediment decreased downwind with both roughness elements, due to their reduction of both shear velocity and sand transport rates. The sediment flux decreased as the AFAI increased, and the rigid roughness element trapped more sand particles than the flexible one except at higher wind speeds and lower AFAI values. The sand trapping efficiency was greater for rigid roughness elements than for flexible ones, probably due to their high aerodynamic resistance, and thus wire tufts were more effective for trapping sand than artificial grass at higher AFAI values.