Rapid identification of Ochroconis gallopava by a loop-mediated isothermal amplification (LAMP) method

Ochroconis gallopava is a species of dematiaceous fungi recognized as a causative agent of zoonotic and emerging fungal infections. It affects the central nervous system and respiratory tracts of humans, birds and cats. We designed O. gallopava species-specific primer sets to aid in its identification by a loop-mediated isothermal amplification (LAMP) method based on the D1/D2 domain of the LSU rDNA sequence. The LAMP method successfully detected the gene from both fungal DNA and experimentally infected brains and spleens of mice and will be helpful in the diagnosis of O. gallopava infection.     

Development of a neurosurgical operating table for adult cattle and changes in intracranial pressure and blood pressure in adult cattle undergoing long-time isoflurane anesthesia

We developed a neurosurgical operating table for restraining adult cattle in the sternal recumbent position during long-time inhalation anesthesia, and examined intracranial pressure (ICP), blood pressure and blood gases during isoflurane anesthesia. We confirmed that the maintenance of inhalation anesthesia, the restraint of cattle in the sternal recumbent position and bringing the cattle out of anesthesia could all be carried out safely using the operating table we produced. For the purposes of the present experiment, the cattle were divided into 2 groups: the SR group, which underwent sternal recumbency for 8 hr under isoflurane anesthesia using the neurosurgical operating table, and the RR group, which underwent right lateral recumbency for 3 hr under isoflurane anesthesia on a standard operating table. The mean ICP was found to be significantly lower in the SR group than in the RR group during anesthesia, and PaO2 was significantly higher in the SR group. In the SR group, no complications such as regurgitation or ruminal tympany occurred for 8 hr after the induction of anesthesia, and recovery from anesthesia was uneventful. In contrast, all RR cattle showed ruminal tympany and regurgitated ruminal fluid at 3 hr after the induction of anesthesia. Thus, the neurosurgical operating table developed in the present study may be useful for long-time anesthesia and neurosurgery of adult cattle.     

Analysis of carotenogenic gene expression in petals and leaves of carnation (Dianthus caryophyllus L.)

Carnation (Dianthus caryophyllus L.) belongs to the family Caryophyllaceae in the order Caryophyllales. Plants in this order do not accumulate carotenoids in petals. To understand how carotenoid accumulation is controlled in carnation petals, we analysed the expression of genes related to carotenoid accumulation. Petals at an early stage of development accumulated small amounts of carotenoids. As petals matured, their carotenoid content decreased to extremely low levels. In contrast, carnation leaves contained substantial amounts of carotenoids that are essential for photosynthesis. Most of the carotenogenic genes were expressed in petals at levels similar to those in leaves, and the expression levels of these genes increased during petal development. Genes encoding phytoene synthase and lycopene ε‐cyclase were exceptions. Their expression levels in petals were very low compared with those in leaves. Expression of the gene encoding carotenoid cleavage dioxygenase 4 was detected in neither leaves nor petals. These data suggest that the low levels of carotenoids in carnation petals are caused not by enzymatic degradation but rather by low rates of carotenoid biosynthesis.     

Distribution and sequence of pyknotic cells in rat fetuses exposed to busulfan

Busulfan, an antineoplastic bifunctional-alkylating agent, is known to induce developmental anomalies. In the present study, we examined the distribution and sequence of pyknotic cells in rat fetal tissues exposed to busulfan. Pregnant rats on gestation day 13 were administered intraperitoneally 30 mg/kg of busulfan, and fetal tissues were examined at 6, 12, 24, 36, 48, 72 and 96 hours after treatment (HAT). Pyknosis of component cells was observed markedly in the brain, moderately in the eyes and spinal cord and mildly in the craniofacial tissue, mandible, limb buds, tail bud, ganglions, alimentary tract, lungs, kidneys, pancreas and liver. In the brain, mitotic inhibition was also detected. Most of the pyknotic cells were considered to be apoptotic cells judging from the results of TUNEL staining and electron microscopic examination. Commonly in the above-mentioned tissues, pyknotic cells began to increase at 24 HAT, peaked at 36 or 48 HAT and disappeared at 96 HAT, which is when the histological picture returned to normal in most tissues except for the brain, spinal cord and eyes. The present study clarified the outline of busulfan-induced apoptosis in rat fetuses.     

Canine pemphigus foliaceus antigen is localized within desmosomes of keratinocyte

The target antigen of autoantibodies in human pemphigus foliaceus (PF) is a desmosomal cadherin, desmoglein 1 (Dsg1). It was demonstrated by immunoelectron microscopy (IEM) that the location of the binding sites of PF autoantibodies in the human epidermis was the extracellular regions of the desmosomes. Only a limited number of canine PF sera were shown to react with canine Dsg1, and the target proteins have not yet been identified. The purpose of this study was to demonstrate the ultrastructural binding site of canine PF autoantibodies to the canine skin by two kinds of IEM methods. Three canine PF sera, which were shown to react with the keratinocyte cell surface by immunofluorescence, were tested in this study. Using a technique of immunoprecipitation-immunoblotting, one out of the three canine PF sera were shown to react with recombinant canine Dsg1. By post-embedding IEM using cryofixation technique, one serum, which did not react with canine Dsg1 by immunoprecipitation-immunoblotting, bound broadly to the extra- and intracellular regions of the desmosomes of normal canine skin. By pre-embedding IEM using canine cultured keratinocytes (MCA-B1 cells), the autoantibodies of all three canine PF sera were identified to be bound to the cell-cell contact area of the adjacent cytoplasmic projections. When double stained with human PF serum and canine PF sera, the binding sites of both human and canine autoantibodies were co-localized on the MCA-B1 cells where the cytoplasmic projections contacted each other. Therefore, it may be concluded that the serum antibodies of canine PF targeted desmosomal proteins, regardless of whether or not they react with canine Dsg1 by immunoprecipitation-immunoblotting method.     

QTL mapping of fruit nutritional and flavor components in tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis>) using genome-wide SSR markers and recombinant inbred lines (RILs) from an intra-specific cross

Fruit nutritional and flavor components are important targets for breeding new cultivars of tomato (Solanum lycopersicum L.). We developed 108 recombinant inbred lines (the K39 RILs) in the F₆ generation from a cross between two phenotypically different breeding lines, K03 and K09. A linkage map was constructed using 172 genome-wide simple sequence repeat markers, 3 single-nucleotide polymorphism markers, and 2 phenotypic markers. The K39 RIL map consists of 12 linkage groups (LGs) and covers a genetic distance of 1089 cM. We measured the fruit soluble solids content (SSC), titratable acidity (TA), glutamic acid content (GLU), and lycopene content (LYC) of each line in four generations (F₆, F₈, F₁₀, F₁₁), β-carotene content (CAR) in two generations, and pH in one generation. By composite interval mapping that considered yearly variations in components as non-genetic effects, we detected three quantitative trait loci (QTLs) for SSC, four for TA, two for CAR, and one each for GLU, LYC, and pH. Among them, we found two QTLs for TA in LGs 6 and 11, those for GLU and LYC were candidates for novel QTLs. QTLs detected in this study were clustered in five LGs, but we observed no apparent trade-off relationships among the QTLs in each LG. Being derived from an intra-specific cross of tomato breeding materials, these QTLs can be used in practical breeding for improving fruit quality with low risk of linkage drag.     

Residual strain analysis with digital image correlation method for subsurface damage evaluation of hinoki (<Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis>) finished by slow-speed orthogonal cutting

A digital image correlation (DIC) method was applied to measure strain which arose and remained beneath the finished surface in slow-speed orthogonal cutting of hinoki (Chamaecyparis obtusa), to evaluate the damage in the subsurface cell layers. While the quarter-sawn surface was cut parallel to the grain, the side surface, flat-sawn surface, was captured by a high-speed camera. The images were analyzed to calculate strain in a region of 0.67 × 0.22 mm allocated beneath the finished surface. Almost no strain normal to the cutting direction was detected for the depth of cut and cutting angles, 0.05 mm and smaller than 60°, respectively. For the depths of cut and cutting angles, larger than 0.1 mm and smaller than 60°, respectively, the fore-split induced tensile strain normal to the cutting direction, although it hardly remained after the cutting. The compression strain normal to the cutting direction clearly remained for the cutting angles larger than 70°, regardless of the depths of cut employed in this study. The subsurface damage assumed from the residual strain distribution corresponded to the appearance of the subsurface layer in the X-ray computed tomography (CT) images. It was also revealed, and the DIC program could not always measure excessively large strain correctly.     

An electric field strongly deters whiteflies from entering window-open greenhouses in an electrostatic insect exclusion strategy

Dual functions (insect repelling and capturing) of a single-charged dipolar electric field screen were evaluated to successfully exclude whiteflies from a window-open greenhouse. The screen consisted of three parts: 1) insulated conductor wires (ICWs) arrayed in parallel at 5?mm intervals, 2) two earthed stainless nets placed within 3?mm of both sides of the ICW layer, and 3) a voltage generator for the negatively charged ICWs. The screen formed two electric fields between the ICW-layer and the ICW-side surface of the earthed net and between the ICWs. At negative charging of 1.5–2.5?kV, all whiteflies reaching the outer surface of the screen net avoided entering the electric field and flew away from the screen. This avoidance was disturbed by 3?m s^?1 wind, as the insects were compulsorily blown inside. However, almost all whiteflies (99.4?%) were captured with the ICW. These results indicate that the insect-capturing function is effective to complement a failure to repel. A greenhouse assay was conducted in the screen-attached and non-screened parts in which a greenhouse was divided with a partition. During the 3-month operation, the screen was durable and functional for excluding pests, and better air ventilation changed the climate conditions in the greenhouse. Thus, the present study demonstrated that our electric field screen can provide an airy condition for tomatoes in a window-open greenhouse and successfully exclude whiteflies using dual screen functions.     

A case of congenital hyperostosis in a newborn piglet

One female newborn piglet showed prominent thickening of both forelimbs and died soon after birth. Histopathologically, thin and woven trabeculae of bone was extending out from the edge of cortical bone in the affected forelimbs, and diagnosed as congenital hyperostosis. The extent of radially proliferated trabeculae was most prominent in radioulna. Many round- to spindle-shaped cells were observed in periosteum, which were considered to be osteoblasts. Around the periosteum, the mesenchymal proliferation was extensive with abundant mucus, and cartilaginous metaplastic changes were observed mainly around the radioulna and humerus. Dilatation of vessels with fibrin deposition in vessel walls was often observed, which were considered to reflect the localized circulatory disturbance.