Establishment and characterization of a canine sebaceous epithelial cell line derived from an eyelid mass

Little is known about the pathological roles of sebaceous glands in canine skin diseases, as most examinations have been conducted with cultured human sebaceous epithelial cell lines. To our knowledge, there is no available canine sebaceous epithelial cell line. The purpose of this study was to establish a canine sebaceous epithelial cell line and characterize it. An eyelid mass in a dog was surgically resected for treatment, and it was histologically diagnosed as sebaceous epithelioma. Collected tissue was conducted for culture, and the growing epithelial-like cells were passaged. The cells showed continuous proliferation for over 6 months. After 40 passages, the cells were named CMG-1. Lipid droplets in the cytoplasm of CMG-1 cells were confirmed by Oil Red O staining. As reported in studies with human sebaceous epithelial cell lines, lipogenesis in CMG-1 cells was promoted by linoleic acid, whereas transforming growth factor-β (TGF-β) suppressed it. Additionally, real-time PCR revealed that the expression levels of chemokines and cytokines, including CC chemokine ligand (CCL)-2, CCL-20, CXCL-10, Tumor necrosis factor-α (TNF-α), Interleukin (IL)-1α, IL-1β, and IL-8, were significantly increased in CMG-1 cells following treatment with lipopolysaccharide. In conclusion, we successfully established a new canine sebaceous epithelial cell line. Our data indicated that lipogenesis and inflammatory responses were quantitatively evaluable in this cell line. CMG-1 cells could be useful for the pathological analysis of sebaceous gland diseases in dogs.     

Differences in food composition between territorial and aggregative juvenile crimson sea bream Evynnis japonica

ABSTRACT:   Food composition, fork length and condition factor of juvenile crimson sea bream Evynnis japonica were examined for two behaviorally distinct types of fish inhabiting a nursery ground. Studies were carried out from March to September 1996 at Morode Cove, Ehime Prefecture, Japan. One type of fish is solitary and territorial and the other type is aggregative. The food compositions of the two types of fish were different. Solitary fish foraged mainly on Gammaridea and Caprellidea (benthic organisms), and Copepoda (planktonic organisms); while aggregative fish foraged mainly on Copepoda, Appendiculata and Cladocera (planktonic organisms). These findings suggest that when we study food composition of E. japonica , it is imperative to consider whether the samples used for the analyses are from solitary (territorial) fish. Solitary fish showed significantly greater fork length and condition factor than aggregative fish, suggesting that the former will have a greater fitness value than the latter.     

Effects of medium‐chain fatty acid‐cyclodextrin complexes on ruminal methane production in vitro

The purpose of the present study was to evaluate effects of medium‐chain fatty acid‐cyclodextrin (CD) complexes on ruminal methane and volatile fatty acid production, and protozoal activity in vitro. Medium‐chain fatty acid‐CDs used in this study were caprylic acid (C8)‐αCD or ‐βCD, capric acid (C10)‐αCD or ‐βCD, and lauric acid (C12)‐αCD or ‐βCD. A 60‐mL of diluted rumen fluid was incubated anaerobically at 38°C for 6 h with the addition of the complex (10–40 mg as fatty acid). Each of the fatty acid‐CDs reduced the number of protozoa, with the order C10 > C12 > C8, and βCD complexes were more effective than αCD complexes. Molar proportions of acetic acid remained unchanged with the addition of fatty acid‐CD, while that of propionic acid increased, being significant for C8‐αCD and βCD, and C10‐αCD and βCD (P < 0.05). Hydrogen production decreased by about 70% of control with the addition of 40 mg of C8 and C10‐CD, on the other hand, it tended to increase with the addition of C12‐CD in both αCD and βCD. Methane production decreased by about 20% with the addition of 40 mg of complexes, except for C10‐βCD, which significantly reduced methane production by about 60%. In conclusion, the addition of C8 or C10‐CD to ruminant diets may be effective in reducing methane production.     

A comparative study of the effects of glycerol and hydroxyethyl starch in canine red blood cell cryopreservation

Hydroxyethyl starch (HES) is a nonpenetrating extracellular cryoprotectant. In contrast to glycerol, it does not require labor-intensive removal from thawed red blood cells (RBCs) prior to transfusion. In this study, we compared glycerol and HES, and assessed HES as a substitute for glycerol in cryopreserved canine RBCs. The RBCs were preserved for 2 months in liquid nitrogen using a 20% (w/v) glycerol solution, and variable concentrations of HES solution. We evaluated the two cryoprotectants by the percentage of post-thaw hemolysis from the total free hemoglobin, saline stability, osmotic fragility, and by observing the erythrocyte morphology using a scanning electron microscope after thawing. The optimal concentration of HES was 12.5% (w/v) for the cryopreservation of canine RBCs. The thaw hemolysis, saline stability, and osmotic fragility index were 25.6 +/- 4.7%, 87.8 +/- 6.9%, and 0.445 +/- 0.024% NaCl respectively. These parameters resemble the results of RBCs frozen in a 20% (w/v) glycerol solution, which are 24.7 +/- 5.2%, 99.2 +/- 0.1%, and 0.485 +/- 0.023% NaCl respectively. From a morphological point of view, 12.5% (w/v) HES showed the best cryoprotection of RBCs compared to the other concentrations of HES. These results suggest that HES could be a possible substitute for glycerol for the cryopreservation of canine RBCs.     

Effects of warming basal ends of Carolina poplar (Populus × canadensis Moench.) softwood cuttings at controlled low-air-temperature on their root growth and leaf damage after planting

We investigated the effects of warming the basal ends of Carolina poplar (Populus × canadensis Moench.) softwood cuttings at controlled low-air-temperature on their root growth and leaf damage after planting. The warming treatment was applied to the cuttings by soaking 10 mm beyond the cut end in warmed water maintained at 30 °C in a cold chamber maintained at an air temperature of 10 °C and a photosynthetic photon flux density (PPFD) of 10 μmol m^?2 s^?1 (near the light compensation point at 10 °C) until rooting was observed. The warmed cuttings were then grown in a growth chamber at an air temperature of 30 °C, relative humidity 85–90 %, and a PPFD of 100 μmol m^?2 s^?1. Control cuttings were grown in the growth chamber throughout the experiment. Rooting occurred simultaneously for both warmed and control cuttings, irrespective of air temperature. Root development was greater and leaf damage, evaluated on the basis of extent of necrosis, was less for warmed cuttings than for control cuttings. The reduction of leaf damage for warmed cuttings probably resulted from reduced post-planting water stress and leaf senescence, because of improved root development as a result of the pre-planting warming treatment. This technique could improve the propagation of cuttings of woody plants, because it would ensure that the cuttings are ready to develop roots with minimum loss of carbohydrates, irrespective of weather conditions.     

Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Effect of boiled barley-rice-feeding in hypercholesterolemic and normolipemic subjects

Barley contains approximately 10% dietary fiber and is easily cooked with rice, the dominant cereal in Japan, to increase the intake of dietary fiber. This research involved three experiments to examine the influence of barley on blood lipids in human subjects. All subjects received a boiled barley-rice (50/50 w/w mix) supplement two times per day in place of rice for 2 or 4 weeks. In the normolipemic subjects, serum lipids were unaffected by the ingestion of barley for 4 weeks. In twenty hypercholesterolemic men aged 41 ± 5 years, the ingestion of barley was associated with a significant fall in serum total cholesterol, LDL-cholesterol, phospholipids and LDL and VLDL-lipoproteins. In seven mildly hypercholesterolemic women aged 56±7 years, a significant improvement of serum lipid profiles was observed. The present study suggests the possibility that the ingestion of barley-rice could lower serum lipids in hypercholesterolemic subjects.     

寒地水稻生长发育动态研究初探

以黑龙江省主栽的两个水稻品种为试验材料,通过调查试验材料的生育期、干物质积累、叶面积变化和分蘖消长动态的测定,明确了拔节至抽穗期是水稻产量形成的关键时期;根据试验材料干物质积累的变化规律建立了干物质随时间变化的生长曲线。经检验拟合较好。     

Anatomical characterization of decayed wood in standing light red meranti and identification of the fungi isolated from the decayed area

To further our understanding of wood decay in living light red meranti (Shorea smithiana) trees, microscopic characteristics of the cell and cell wall degradations of S. smithiana wood in the presence of the decay fungi, the identity of the causal fungi, and the decay potential and pattern by an isolated fungus were investigated. Cell wall degradations, including cell wall thinning, bore holes formation, rounded pit erosion, and eroded channel opening were clearly observed under light and scanning electron microscopy. In transverse view, many large voids resulting from a coalition of degraded wood tissue appeared in the decayed canker zone. All these observations suggest the well-known simultaneous decay pattern caused by white-rot fungi. By phylogenetic analysis based on the sequences of internal transcribed spacer region of ribosomal DNA, a basidiomycete fungus isolated from the decayed wood was identified as Schizophyllum commune. The degradation caused by this fungus on sound S. smithiana wood in an in situ laboratory decay test was classified as the early stage of simultaneous decay, and showed a similar pattern to that observed in the wood samples naturally decayed.