Development of an on-site plum pox virus detection kit based on immunochromatography

Sharka disease, caused by plum pox virus (PPV), is the most serious viral disease of stone fruit trees. Among the eight known strains of the virus, PPV-D is the most important due to its recent global spread. Although enzyme-linked immunosorbent assay (ELISA) is the most common approach for diagnosing sharka, it involves time-consuming steps and requires expensive equipment and trained technicians. In this study, an on-site PPV detection kit based on immunochromatography was developed using polyclonal antibodies against the coat protein (CP) of a PPV-D isolate. The immunochromatographic (IC) assay kit was as sensitive as a commercial ELISA system for detecting Japanese PPV-D isolates. Moreover, it was easy to use (a one-step procedure), and results could be obtained on-site within 15 min without special laboratory equipment. The IC assay kit detected the virus from every aerial part of symptomatic Japanese apricot trees. In a detailed study of viral localization in leaves, the most suitable plant parts for use in the IC assay were symptomatic mesophyll tissues and the region from the petiole to the main vein. A positive reaction was also observed using the CP of other major (PPV-M and PPV-Rec) and minor (PPV-EA, PPV-W, and PPV-T) strains.     

Growth and maturation in sterlet <Emphasis Type="Italic">Acipenser ruthenus</Emphasis> under high concentrations of dissolved oxygen

This study investigates the effect of high dissolved oxygen concentrations (DO) on sterlet Acipenser ruthenus maturation. Two groups, each comprising 246 individuals of 8-month-old juvenile sterlets, were reared for 37 months under high DO (group H, mean DO of 11.2 mg/L) or normal DO (group C, mean DO of 7.3 mg/L). Significant differences in body weight were observed between group H (638.7?±?191.9 g) and group C (572.2?±?151.9 g) in 12 months (P?<?0.05). Growth until 12 months occurred at a specific growth rate of 0.6%/day for group H and at 0.5%/day for group C. The food conversion rate of groups H and C was 5.2 and 5.8, respectively. The gonadosomatic index of group H was 3.1?±?1.1 for females and 3.1?±?9.7 for males, and showed no significant difference with group C (3.0?±?0.7 for females and 2.8?±?8.9 for males, P?>?0.05) in 24 months. Significant differences in follicle diameter in mature females were also observed. Group H included 68 mature females, with an average follicle diameter of 2318.0?±?175.3 µm, whereas group C included 56 mature females, with a smaller average follicle diameter of 2152.3?±?287.0 µm (P?<?0.001). However, the polarization index in group H (0.194?±?0.049) was higher than that in group C (0.178?±?0.042) (P?<?0.05). These results suggest that, for sterlet, rearing conditions with high DO can affect growth of both the body and follicle diameter of fish compared to normal rearing conditions.     

Decontamination of Cs from Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) via kraft cooking

To investigate the possibility of decontaminating ¹³⁷Cs-contaminated Cryptomeria japonica wood, kraft pulping was conducted and the Cs behavior in the reaction process was examined. ¹³³Cs-treated or ¹³⁷Cs-contaminated bark, sapwood, and heartwood chips of Cryptomeria japonica were digested using an aqueous solution of NaOH and Na₂S. The pulp was washed with ultrapure water and filtered, after which the filtrate (black liquor) was collected. The black liquor was acidified to separate the supernatant and precipitation. The Cs (¹³³Cs and ¹³⁷Cs) concentrations in the chip and reaction products were measured. As for wood samples, the majority of Cs was present in black liquor, while only a minor amount of Cs was retained in the pulp (<1%). In the case of bark, although the majority of Cs was present in the black liquor, the proportion of Cs in the pulp was much higher than that in the wood pulp. In addition, the Cs in the precipitation of the bark was higher than that in the wood, possibly because the Cs in the bark was combined with some components, which is insoluble in alkaline solution. Our results suggest that ¹³⁷Cs-contaminated Cryptomeria japonica wood can be used in the pulp and paper industries.     

Extractives of muirapiranga (<Emphasis Type="Italic">Brosimun</Emphasis> sp.) and its effects on the vibrational properties of wood

The potential of muirapiranga (Brosimun sp.) as a substitute material for violin bows was estimated in terms of vibrational properties, and the influence of extractives on the vibrational properties was examined. The loss tangent of muirapiranga was somewhat higher and the specific dynamic Young’s modulus was rather lower than the respective values for pernambuco, which is regarded as the best material for violin bows. Therefore, muirapiranga is a poorer bow material in terms of vibrational properties. Impregnation of crude extractives from muirapiranga decreased the loss tangent of other wood specimens. The main compounds of the extractives were identified as xanthyletin and luvangetin. Impregnations of isolated xanthyletin and commercially available methoxsalen, which was tested as an analogue of luvangetin, markedly decreased the loss tangent of other wood specimens. Methoxsalen and xanthyletin differ from conventional loss tangent-decreasing substances, namely protosappanin B and hematoxylin, in terms of water insolubility and the absence of hydroxyl groups. From the similarity in molecular characteristics of loss tangent-decreasing substances found so far, restriction of molecular motion due to an impregnated substance in the wood matrix is suggested as one loss tangent-decreasing mechanism.     

Nebulin and N-WASP cooperate to cause IGF-1-induced sarcomeric actin filament formation

Insulin-like growth factor 1 (IGF-1) induces skeletal muscle maturation and enlargement (hypertrophy). These responses require protein synthesis and myofibril formation (myofibrillogenesis). However, the signaling mechanisms of myofibrillogenesis remain obscure. We found that IGF-1-induced phosphatidylinositol 3-kinase-Akt signaling formed a complex of nebulin and N-WASP at the Z bands of myofibrils by interfering with glycogen synthase kinase-3β in mice. Although N-WASP is known to be an activator of the Arp2/3 complex to form branched actin filaments, the nebulin-N-WASP complex caused actin nucleation for unbranched actin filament formation from the Z bands without the Arp2/3 complex. Furthermore, N-WASP was required for IGF-1-induced muscle hypertrophy. These findings present the mechanisms of IGF-1-induced actin filament formation in myofibrillogenesis required for muscle maturation and hypertrophy and a mechanism of actin nucleation.     

Distribution of CD163-positive cell and MHC class II-positive cell in the normal equine uveal tract

Antigen-presenting cells (APCs) in the uveal tract participate in ocular immunity including immune homeostasis and the pathogenesis of uveitis. In horses, although uveitis is the most common ocular disorder, little is known about ocular immunity, such as the distribution of APCs. In this study, we investigated the distribution of CD163-positive and MHC II-positive cells in the normal equine uveal tract using an immunofluorescence technique. Eleven eyes from 10 Thoroughbred horses aged 1 to 24 years old were used. Indirect immunofluorescence was performed using the primary antibodies CD163, MHC class II (MHC II) and CD20. To demonstrate the site of their greatest distribution, positive cells were manually counted in 3 different parts of the uveal tract (ciliary body, iris and choroid), and their average number was assessed by statistical analysis. The distribution of pleomorphic CD163- and MHC II-expressed cells was detected throughout the equine uveal tract, but no CD20-expressed cells were detected. The statistical analysis demonstrated the distribution of CD163- and MHC II-positive cells focusing on the ciliary body. These results demonstrated that the ciliary body is the largest site of their distribution in the normal equine uveal tract, and the ciliary body is considered to play important roles in uveal and/or ocular immune homeostasis. The data provided in this study will help further understanding of equine ocular immunity in the normal state and might be beneficial for understanding of mechanisms of ocular disorders, such as equine uveitis.     

CHORDOMA OF THE TAIL IN A DEGU (OCTODON DEGUS)

A 6-year-old female degu (Octodon degus) in good body condition was presented with a mass at the base of the tail. Following presentation and the owners’ approval, the tail of the degu was surgically amputated. The mass was submitted for pathological evaluation following the surgical procedure. Histologically, the mass was comprised of lobules and sheets of polygonal cells displaying moderate amounts of basophilic foamy cytoplasm (physaliferous cells). The pathological results of the tail mass were consistent with chordoma. Approximately 3 months following the tail amputation the patient died.