Ex vivo tissue discrimination by visible and near-infrared spectra with chemometrics

The risk of infections from zoonotic pathogens of tissues and/or tissue-derived products has been increasing. One preventive approach in reducing infection risk is tissue decontamination, where selection and screening of highly infectious tissues are strictly followed. Therefore, the development of reliable analytical methods for rapid tissue discrimination is essentially important. In the present study, a procedure has been developed for intact tissue discrimination on the basis of multivariate analysis of visible and near-infrared (Vis-NIR) spectra of certain tissues such as brain, liver, kidney and testis of mice without any pretreatment. Transmittance spectra in the 600- to 1000-nm regions were subjected to a principal component analysis (PCA), and leave-out cross-validation was employed to develop multivariate models for tissue discrimination. The plot of PCA scores against Vis-NIR spectra of brains, kidneys, livers and testes from 11 mice portrayed reliable tissue discrimination. This result suggests that Vis-NIR spectroscopy combined with chemometrics analysis may provide a potentially useful approach for rapid non-destructive discrimination of tissues.     

Determination of antioxidant activity and characterization of antioxidant phenolics in the plum vinegar extract of cherry blossom (Prunus lannesiana)

Sakura-cha (salted cherry blossom tea) is a Japanese tea that is traditionally served at celebrations such as wedding ceremonies. The production of Sakura-cha includes the immersion of cherry blossom flowers in Japanese plum vinegar, and through this process, the byproduct (plum vinegar extract of cherry blossom) is obtained. In this study, the antioxidant activity of the plum vinegar extract of cherry blossom was examined. The plum vinegar extract of cherry blossom had a greater superoxide anion scavenging activity compared with red wine, which is a well-known strong antioxidant. Liquid chromatography-mass spectrometry analysis showed that cyanidin-3-glucoside, cyanidin-3-rutinoside, and caffeic acid were the major components in the phenolic extract prepared from plum vinegar extract of cherry blossom, and they possessed superoxide anion scavenging activity. Caffeic acid is mainly responsible for the scavenging activity of phenolic extract; the contributions of cyanidin-3-glucoside and cyanidin-3-rutinoside were minor.     

Exophthalmos Caused by Chronic Sinusitis in a Horse: A Case of Exophthalmos Caused by Sinusitis

A 12-year-old Thoroughbred gelding exhibited a unilateral exophthalmos on the left side. There had been no other clinical symptoms, such as rhinorrhea, before the first consultation. Clinical, hematologic, plasma biochemical, cytologic, and computed tomography (CT) examinations and complete necropsy were performed. These examinations did not reveal neoplastic lesions. CT images suggested that maxillary and frontal sinuses may have been filled with fluid. Cytologic and pathologic examinations revealed that the maxillary and orbital bones were pushed up with a large amount of the inflammatory fluid effused by chronic sinusitis. The CT and cytologic examinations were of great use for differential diagnosis in this case. This was an unusual case of exophthalmos induced by deformation of the orbital bones resulting from chronic sinusitis.     

Changes in expression and localization of GPRC5B and RARalpha in the placenta and yolk sac during middle to late gestation in mice

The mRNA expression of GPRC5B, an orphan G protein-coupled receptor, is induced by retinoic acid (RA). Because RA plays critical roles in embryonic development, reproductive functions, metabolism and homeostasis, GPRC5B is also considered crucial in these physiological events. We investigated the changes in expression of GPRC5B and RA receptor (RAR) alpha mRNAs and immunohistochemical localization of their proteins in the murine placenta and yolk sac at 13.5, 15.5 and 17.5 days post coitus. Stable levels of GPRC5B and RARalpha mRNAs were detected in the placenta and yolk sac. In the placenta, GPRC5B was present in maternal and fetal vascular endothelial cells, stromal cells, fibroblast-like cells and glycogen cells. A strong reaction to RARalpha was detected in maternal and fetal vascular endothelial cells and stromal cells. The levels of GPRC5B and RARalpha proteins in maternal and fetal vascular endothelial cells decreased with gestation. In the yolk sac, GPRC5B and RARalpha proteins were detected in vascular endothelial cells, but their levels did not change during the gestation period. These findings indicate that GPRC5B is involved in RA-dependent morphogenesis/angiogenesis and regulation of extracellular matrix synthesis in the murine placenta and yolk sac.     

Effects of twice harvesting on total dry matter yield of rice

We examined the effect of twice harvesting of forage rice with the first harvest at the full heading stage on total dry matter yield in the cultivars Taporuri, Mohretsu, and Hinohikari and in line Saikaishi 253 in southwestern Japan. Taporuri produced the highest total dry matter yield, with a value 60% higher than that of Hinohikari. The first crop of Taporuri had a longer duration of vegetative growth than those of Mohretsu and Hinohikari, and had the highest dry matter yield because of its greater weight per tiller than Hinohikari and its more tillers per square meter than Saikaishi 253, which had a similar duration of vegetative growth. The second crops of Mohretsu, Taporuri, and Saikaishi 253 had longer vegetative growth and higher dry matter yields than Hinohikari. Dry matter yields of both crops were closely related to the duration of vegetative growth and the weight per tiller, but not to tillers per square meter. These results suggest that cultivars with a long duration of vegetative growth, high weight per tiller, and adequate tillers per square meter can produce high dry matter yield in both crops. There was a significant interaction (P < 0.05) in total dry matter yield between cultivar or line and cultivation type. The total dry matter yield of Taporuri in twice harvesting was about 10% higher than that in once harvesting, whereas that of Hinohikari was about 10% lower. The suitability for twice harvesting ([total dry matter yield in twice harvesting]/[total dry matter yield in once harvesting]) was significantly positively correlated (r = 0.875) with the increase in dry weight before the full heading stage, but significantly negatively correlated (r = −0.903) with the increase in dry weight between full heading and the yellow ripe stage. Thus, cultivars with a high increase in dry weight before full heading appear to be more suitable for twice harvesting than those with a high increase between full heading and the yellow ripe stage. In addition, our study showed that twice harvesting can prevent lodging in Taporuri, thereby increasing the suitability of this cultivar as a forage rice. Our results suggest that twice harvesting can be performed in southwestern Japan and warmer regions.     

Lectin-binding characteristics and capacitation of canine epididymal spermatozoa

Cross sections of the testes and the caput, corpus and cauda epididymides removed from 12 dogs were stamped on glass slides, and the sperm on the slides were stained with 6 different FITC-lectins (Con A, DBA, PNA, PSA, SBA, and WGA) to examine the characteristics of the surface glycoproteins (GPs) on canine epididymal sperm. The corpus epididymal sperm were washed three times by centrifugation, and their lectin-binding characteristics were investigated. The washed sperm from the corpus and cauda epididymides were incubated for 24 hr, and the fertilizing capacity of the sperm was evaluated by calculating the percentages of actively motile sperm (%MO), hyperactivated sperm (%HA), and acrosome-reacted sperm (%AR), and the number of canine zona-pellucida (ZP)-binding sperm. The testicular sperm did not stain with SBA lectin, but the SBA lectin fluorescence was observed on the surface of the entire heads of the caput epididymal sperm. Although all of the entire heads or acrosomal regions of the corpus epididymal sperm stained with all 6 FITC-lectins, the heads and acrosomal regions of the cauda epididymal sperm did not stain with DBA or SBA lectins. Washing the sperm from the corpus epididymis resulted in loss of the fluorescence of the FITC-DBA and -SBA lectins. The mean %MO, %HA, %AR, and ZP-binding number of the cauda epididymal sperm after 24 hr of incubation were higher than the values for the corpus epididymal sperm. All of the mean values for the washed sperm from the corpus and cauda epididymides were higher than the values for the unwashed sperm from the corpus and cauda, and with the exception of %AR, the values from the washed sperm from the corpus epididymis were significantly higher (P<0.05, 0.01). The results indicate that DBA- and SBA-lectin-binding GPs on the surface of canine epididymal sperm are associated with the fertilizing capacity and may be decapacitation factors.     

Effects of extracellular lactate on production of reactive oxygen species by equine polymorphonuclear leukocytes in vitro

Objective-To evaluate effects of extracellular lactate on viability, shape change, lactate metabolism, and reactive oxygen species (ROS) production in equine polymorphonuclear leukocytes (PMNs). Sample-PMNs isolated from equine venous blood samples. Procedures-PMNs were incubated with 0 to 300mM lactate for 30 minutes before each experiment. Viability was assessed via trypan blue exclusion. Shape change was assessed via flow cytometry and light microscopy. Relative quantification of monocarboxylic acid transporter and lactate dehydrogenase lactate dehydrogenase (LDH) isotype mRNAs was performed with a real-time PCR assay. Effects of lactate at a pH of 7.4 to 6.0 on ROS production in response to phorbol 12-myristate 13-acetate, opsonized zymosan, or N-formyl-methionyl-leucyl-phenylalanine was assessed by luminol-dependent chemiluminescence. Results-Lactate had no effect on viability of PMNs but did alter their size and density. Monocarboxylic acid transporter 1 and lactate dehydrogenase B mRNA values were not altered. Monocarboxylic acid transporter 4 and lactate dehydrogenase A mRNA values were significantly decreased. Lactate incubation of cells significantly decreased PMN-derived luminol-dependent chemiluminescence and induced different sensitivities to stimulants (phorbol 12-myristate 13-acetate, opsonized zymosan, and N-formyl-methionyl-leucyl-phenylalanine). The response ratio to N-formyl-methionyl-leucyl-phenylalanine revealed that PMNs were primed by incubation with up to 50mM lactate, significantly increasing the production of ROS. Incubation with lactate and acidic pH caused a synergistic effect on ROS production. Conclusions and Clinical Relevance-Extracellular lactate potentially has a direct effect on the capacity to produce ROS by equine PMNs, which may be associated with alterations in innate immune functions within a short period after high-intensity exercise.     

An additional life-history tactic of masu salmon: Migration of parr to coastal habitats

Alternative life-history tactics of masu salmon Oncorhynchus masou are well documented. Subsequent to the freshwater parr stage (age ≥ 1 + years), an anadromous form migrates to the sea after smolting, while a resident form matures without seaward migration. In addition to this typical migratory dimorphism, anecdotal reports based on field observations have indicated that some underyearling masu salmon use estuarine waters. However, no empirical evidence indicates saltwater utilisation and subsequent survival in the early parr stage. Here, we used otolith microchemistry to examine whether a portion of masu salmon parr in northern Japan enters coastal habitats. The otolith Sr:Ca ratios of most juveniles collected from six rivers had consistently low values, indicating that masu salmon parr inhabiting these rivers stay only in freshwater. In contrast, in individuals from a steep-gradient river the Sr:Ca ratios increased at about a 200-µm distance from the otolith core. These results suggest that some masu salmon parr might use brackish water or sea water temporarily. In addition, three masu salmon parr were found in another steep river where a culvert located only ten metres from the river mouth completely blocked upstream migration for spawning. The Sr:Ca ratios in these fish increased at about >200 µm from the otolith core, indicating the parr had immigrated to the non-natal river from the sea. Such flexible behaviour at an early life stage may contribute to the spatial expansion of masu salmon, and the movement could moreover help to stabilise its population dynamics.     

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.