Diverse functions of perlecan in central nervous system cells in vitro

Therapeutic treatment targeting one cell type is considered ineffective in remedying any injury to the central nervous system (CNS). Perlecan, a multi‐functional, heparan sulfate proteoglycan, shows diverse effects on distinct cell types, suggesting that it is one of the candidates that can augment the regenerative mechanisms in the injured CNS. Therefore, we examined the functions of perlecan in CNS cells in vitro by using perlecan purified from bovine kidney. Perlecan‐coated cell culture plates, unlike their type I/III collagen‐coated counterparts, did not inhibit the adhesion of neural stem/progenitor cells (NS/PCs) and neurons. The coated perlecan and the perlecan added to the culture medium suppressed astrocyte proliferation; however, perlecan added to the medium promoted NS/PC proliferation. Neurons were promoted to extend their neurites on the perlecan‐coated substrate, and perlecan added to the medium also showed a similar effect. NS/PC proliferation and neurite extension is a major regenerative reaction in CNS injury, whereas excess proliferation of astrocytes cause hypertrophy of glial scars, which repels neurons. Our in vitro study suggests that perlecan is an attractive candidate to promote regenerative mechanisms and to suppress reactions that hamper regenerative processes in cases of CNS injury.     

Disruption of endogenous perlecan function improves differentiation of rat articular chondrocytes in vitro

Heparan sulfate (HS) and heparan sulfate proteoglycans (HSPG) are necessary for normal cartilage development and chondrocyte differentiation. However, recent studies demonstrated that HSPG accelerate dedifferentiation and catabolism in chondrocytes from degenerative cartilage. In this study, we investigated the inhibitory effect of HSPG on chondrocyte differentiation in vitro. Rat articular chondrocytes were cultured at low (0.3 × 10⁴ cells/cm²) and high (1.5 × 10⁵ cells/cm²) density in the presence or absence of heparitinase I, an HS degrading enzyme. Cells cultured at low density dedifferentiated and exhibited an elongated morphology, and treatment with heparitinase I precluded cell elongation. Conversely, populations of chondrocytes cultured at high density exhibited either a dedifferentiated or differentiated phenotype. Glycosaminoglycan accumulation increased in heparitinase I‐treated cells. To determine the function of perlecan, an important HSPG for cartilage development, in chondrocyte differentiation, rat chondrocyte cultures were exposed to an anti‐perlecan antiserum to inhibit perlecan function. Western blotting analysis indicated that preventing perlecan activity increased type II collagen synthesis. Our results suggest that HSPG are negative regulators of chondrocyte differentiation in vitro and that perlecan contributes to chondrocyte dedifferentiation in vitro.     

Assessment of longitudinal tissue Doppler imaging of the left ventricular septum and free wall as an indicator of left ventricular systolic function in dogs

OBJECTIVE: To evaluate tissue Doppler imaging (TDI) of the left ventricular (LV) free wall (FW) and ventricular septum (VS) as an indicator of LV systolic function in dogs. ANIMALS: 7 healthy Beagles. PROCEDURES: Doses of dobutamine (5 and 10 microg/kg/min) and esmolol (50 and 100 microg/kg/min) were infused into the LV of each dog. With each dose, heart rate; myocardial performance index (MPI); transmitral inflow and ejection time (determined via pulsed-wave Doppler [PWD] echocardiography); and FW and VS velocities of the mitral valve annulus (determined via TDI during systole [S'], early diastole [E'], and late diastole [A']) were assessed. RESULTS: With each dose, dobutamine significantly increased heart rate and the first derivatives of LV pressure (+dP/dt and -dP/dt), whereas esmolol significantly decreased the +dP/dt and -dP/dt values, compared with baseline. Esmolol (100 microg/kg/min) significantly decreased the VS-TDI-derived S' velocity and FW-TDI-derived E' velocity; dobutamine significantly increased transmitral inflow and TDI velocities. Regression coefficient between VS-TDI-derived S' velocity and +dP/dt was higher than that between FW-TDI-derived S' velocity and +dP/dt. Compared with baseline, the PWD- and VS-TDI-derived MPI were significantly decreased by dobutamine and significantly increased by esmolol at each dose. Values of FW-TDI-derived MPI were higher than values derived via the other techniques. Correlation between +dP/dt and VS-TDI-derived MPI was greater than that between +dP/dt and FW-TDI- or PWD-derived MPI. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy dogs, the VS-TDI-derived S' velocity and MPI appear to be reliable assessments for evaluating LV systolic function.     

Proteomic analysis of the G-layer in poplar tension wood

Angiosperm trees bend their stems by forming tension wood at the upper side of leaning stems. Most tension wood has a cellulose-rich G-layer in the innermost surface of the fiber cell wall. Strong tensile stress is considered to occur in the G-layer. This study undertook to identify the proteins involved in G-layer formation and function through a proteomic analysis of G-layer-localized protein. G-layers of poplar were loosened by sonication and isolated as doughnut-shaped pieces of thinly sliced transverse sections. The proteins, once extracted with urea/detergent solution, were separated by two-dimensional polyacrylamide gel electrophoresis, and 110 spots were subjected to liquid chromatography tandem mass spectrometry (LC/MS/MS). A database search for these spots’ mass spectrum patterns identified 72 proteins. In addition, all peptide digestion mixtures of G-layer proteins were separated by strong cation exchange chromatography and 39 proteins were identified using LC/MS/MS analysis. Proteins involved in wall formation, such as lignin biosynthesis-related protein, xyloglucan endotransglucosylase, and fasciclin-like arabinogalactan protein, were notably detected in the G-layer.     

Variation in Amylase Activities in Radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis>) Cultivars

The radish (Raphanus sativus) is a root vegetable of the Brassicaceae family which shows amylolytic activity in the taproot. However, there is little information about differences in these amylolytic activities among radish cultivars. We analyzed the amylase activities and starch contents of 7 kinds of radish cultivars. The Koshin cultivar showed the highest amylase activity, with a level approximately 6 times higher than that of the Sobutori cultivar, which had the lowest. Cultivars with higher amylase activities showed higher starch contents. These results suggest that there are intraspecies variations in amylolytic activities in radishes, and positive correlations between amylase activity and starch content.     

Optimal stocking density of juvenile red king crabs Paralithodes camtschaticus under cannibalism consideration

Juvenile red king crabs (RKCs) Paralithodes camtschaticus have a strong tendency for cannibalism, which is an obstacle for seedling production. We investigated the effects of density-dependent cannibalism on the survival rate and on growth, and estimated the optimal stocking density. Four different stocking densities of juvenile first instar crabs were maintained in tanks (50, 100, 200 and 400 individuals per 100-l tank: bottom area was 0.25 m²) during the 209 or 216 days of rearing. The survival rate was affected by stocking density, elapsed days, and the interaction between these variables. Although carapace length, an index of body size, was also affected by density, elapsed days, and the interaction between these variables, the effect associated with density was quite small, suggesting that cannibalism did not intensely affect growth of juvenile crabs. We then estimated the initial stocking density that maximizes the number of survivors using parameters of analyses. Optimal initial stocking density was predicted as 416, 197 and 149 individuals per tank when the target size was assumed as 5.0, 10.0 and 15.0 mm, respectively. This estimation provides a baseline for seedling production in RKCs and could be further improved by considering other factors that affect survival and growth of RKCs.     

Characteristic computed tomographic myelography findings in 23 Thoroughbred horses

The purpose of this observational study was to determine the characteristic computed tomographic (CT) myelography findings of cervical vertebral stenotic myelopathy (CVSM) lesions in Thoroughbred horses. A total of 23 Thoroughbred horses (age range, 155–717 days on CT examination; mean, 410.9 days) were analyzed. All 23 Thoroughbred horses underwent unenhanced radiography, radiographic myelography, and CT myelography. Unenhanced radiographs were observed the presence of cervical vertebral malalignment and osseous lesions. Radiographic myelograms were observed for signs of cervical spinal cord compression; additionally, CT myelograms were used to detect cervical vertebral osseous lesions. Ventral compressions were frequently observed in the cranial cervical vertebrae (C2–C4), whereas dorsal compressions were frequently observed in the caudal cervical vertebrae (C5–C7). Furthermore, osseous lesions of the caudal articular process developed more frequently than those of the cranial articular process. CT myelography in Thoroughbred horses is a useful method for detecting CVSM changes.     

Embryo and fetal position during pregnancy by ultrasonographic examinations in bottlenose dolphin (Tursiops truncatus)

From 2012 to 2017, serial ultrasonographic evaluation of 5 healthy bottlenose dolphins, Tursiops truncatus, were performed over the course of 6 pregnancies in Enoshima Aquarium. A total of 98 ultrasonographic examinations were included in the study. In three out of six cases, two embryos were observed between umbilicus and genital slit, and side of umbilicus in the dam’s body. All embryos were located in right below the peritoneum and observed from 308 to 325 days pre-partum. These days were corresponding to from 58 to 61 days after copulation respectively. The diameter of the embryo sac was approximately 4 cm. In three cases, the fetal head was located in the dam’s left lateral between umbilicus and genital slit from approximately 90 days pre-partum to the parturition. A snout of fetus is at the top of the uterine horn, and the tail lies close to the cervix. All six calves were fluke-first births (breech presentation). It was conjectured that the contraction of the dam’s uterus during parturition forced the fetus to invert, and the fetal tail fluke was expelled from the dam’s body. In three cases, judging from orientation of fetal tail fluke faced towards left side of the dam, a fetal position might be RSL (Right Sacrum-Lateral) within the birth canal. In the other three cases, the left and right positions of the fetus and the dam’s body are reversed during pregnancy and parturition.