Structure-Activity Relationship Study of the Neuritogenic Potential of the Glycan of Starfish Ganglioside LLG-3

LLG-3 is a ganglioside isolated from the starfish Linchia laevigata. To clarify the structure-activity relationship of the glycan of LLG-3 toward rat pheochromocytoma PC12 cells in the presence of nerve growth factor, a series of mono- to tetrasaccharide glycan derivatives were chemically synthesized and evaluated in vitro. The methyl group at C8 of the terminal sialic acid residue was crucial for neuritogenic activity, and the terminal trisaccharide moiety was the minimum active motif. Furthermore, the trisaccharide also stimulated neuritogenesis in human neuroblastoma SH-SY5Y cells via mitogen-activated protein kinase (MAPK) signaling. Phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 was rapidly induced by adding 1 or 10 nM of the trisaccharide. The ratio of phosphorylated ERK to ERK reached a maximum 5 min after stimulation, and then decreased gradually. However, the trisaccharide did not induce significant Akt phosphorylation. These effects were abolished by pretreatment with the MAPK inhibitor U0126, which inhibits enzymes MEK1 and MEK2. In addition, U0126 inhibited the phosphorylation of ERK 1/2 in response to the trisaccharide dose-dependently. Therefore, we concluded that the trisaccharide promotes neurite extension in SH-SY5Y cells via MAPK/ERK signaling, not Akt signaling.     

Comparison of changes in sugars and ethylene production of cherimoya fruit grown at different seasons

Summary

Changes in fruit weight, total soluble solids (TSS), sugar content and postharvest ethylene production during fruit development were compared on fruit of cherimoya (Annona cherimola Mill., cv. Big Sister) pollinated at different times (early, at the beginning of April, 1995 and late, at the end of May, 1994). Fruit weight increased by growth, independently of pollination times. Changes in TSS and sugar content were strongly affected by pollination time. In late pollinated fruits, both TSS and sugar content increased during the growth period, 110–160 d after pollination, while neither increased with fruit growth in early pollinated fruit. Moreover, changes in TSS and sugar content after ripening depended not only on pollination time but also on the growth period. With late pollination, fruit harvested after 140 d became edible after ripening. The TSS and total sugar contents, (the sum of fructose, glucose and sucrose contents) increased to about 20° Brix and above 16%, respectively. But early pollinated fruits were not edible after ripening, since total sugar content increased only to about 7%, even though they kept growing for 160 d on the tree. The maximum rate of ethylene production declined and the number of days to the ethylene peak from harvest is also reduced with growth in both cases. These changes were closely correlated with days after pollination (r>0.9) regardless of pollination time. Comparisons of accumulated temperature suggest that temperature at pollination time and during the resting period of growth affected fruit development. These results indicate that neither fruit weight nor number of days after pollination is a suitable index for harvesting time.     

A surge-like increase in luteinizing hormone preceding musth in a captive bull African elephant (Loxodonta africana)

This study was conducted to determine the correlation between reproductive hormones and musth in a male African elephant. Changes in circulating luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone and immunoreactive (ir-) inhibin and the degree of musth were evaluated for 4 years. LH increased 4 weeks before musth began. The highest concentrations of testosterone and ir-inhibin were observed from April to October. There were positive correlations among testosterone, ir-inhibin and musth behavior. These findings suggested that the surge-like LH in the pre-musth period might stimulate secretion of testosterone and ir-inhibin and thus initiate the musth behavior. This study also suggested that the high LH level before musth might be a useful biomarker for the beginning of the musth season.     

Early pathophysiologic feature of arthropathy in juvenile dogs induced by ofloxacin, a quinolone antimicrobial agent

Arthropathy in dogs induced by ofloxacin, a quinolone antimicrobial agent, was pathophysiologically investigated. In the in vivo studies, ofloxacin was administered orally once or twice at 20 mg/kg/day to male juvenile (3-month-old, n=3) or adult (36-month-old, n=2) dogs, and the humeral and femoral heads were examined pathologically. Unlike adult dogs, fluid-filled vesicles were macroscopically observed on the articular surfaces of one juvenile dog 24 hours after a single treatment with ofloxacin. These lesions were seen in all juvenile dogs by twice dosing. Microscopically, fissures or cavity formations in the middle zone of the articular cartilage were noted only in juvenile dogs. Furthermore, the cartilage matrix from the abnormal area to the articular surface showed a decreased safranin-O staining intensity, suggesting proteoglycan depletion. Ultrastructurally, chondrocytes in the middle zone of juvenile dogs displayed dilatation of the cisternae in the rough endoplasmic reticulum as an initial hallmark. In the in vitro studies, chondrocytes isolated from the articular cartilage of naive juvenile dogs were exposed to ofloxacin at 6.3-100 microg/ml for 24 hours. Although no changes were noted in the deoxyribonucleic acid synthesis, protein synthesis, or proteoglycan release at concentrations of up to 100 microg/ml, the proteoglycan synthesis was evidently decreased in a dose-dependent manner from 12.5 microg/ml. The results obtained suggest that the inhibitory action of ofloxacin on proteoglycan syntheses in the chondrocytes may largely contribute to the early morphologic features in the articular cartilage of the juvenile dog.     

Pollen tube attraction by the synergid cell

In flowering plants, guidance of the pollen tube to the embryo sac (the haploid female gametophyte) is critical for successful fertilization. The target embryo sac may attract the pollen tube as the final step of guidance in the pistil. We show by laser cell ablation that two synergid cells adjacent to the egg cell attract the pollen tube. A single synergid cell was sufficient to generate an attraction signal, and two cells enhanced it. After fertilization, the embryo sac no longer attracts the pollen tube, despite the persistence of one synergid cell. This cessation of attraction might be involved in blocking polyspermy.     

Changes in spatial patterns of sika deer distribution and herbivory of planted seedlings: a comparison before and after deer population control by culling

Sika deer (Cervus nippon) sometimes cause extensive damage to planted tree seedlings. To evaluate the effects of culling on the spatial distribution patterns of sika deer and browse damage to planted seedlings, we compared the data collected before and after experimental culling in a cool, temperate, mixed forest on Kyushu Island, Japan. Experimental culling, conducted in an area of 1 km², removed five, four and two sika deer in April, June and October 2011, respectively. During the year before culling, the spatial pattern of the number of sika deer caught on camera corresponded to the predicted sika deer density. Sika deer immediately browsed planted seedlings after the initial planting. The cumulative number of browsed seedlings increased over time, especially in winter. The spatial pattern of the cumulative number of sika deer caught on camera corresponded to that of browsed seedlings at the year’s end. During the year when culling was conducted, the number of sika deer caught on camera decreased around the center of the study site where the culling was conducted and the number of browsed seedlings decreased. During the year following culling, the cumulative number of browsed seedlings was very similar to that in the year before the culling, while the same low number of sika deer was caught on camera. These results indicate that the effects of deer culling resulted in decreased levels of sika deer appearance and browse damage for more than 1 year and for several months, respectively.     

A non-arthropathic dose and its disposition following repeated oral administration of ofloxacin, a new quinolone antimicrobial agent, to juvenile dogs

A non-arthropathic dose and disposition of ofloxacin, a potent new quinolone antimicrobial agent, were assessed in male juvenile (3-month-old) dogs, when administered orally at 5, 10 and 20 mg/kg/day once daily for 8 consecutive days. Ofloxacin concentrations in sera and articular cartilages were analyzed by high-performance liquid chromatography (HPLC). Macroscopically, arthropathy characterized by fluid-filled vesicles in articular surface of the humerus and femur was observed in animals receiving 10 and 20 mg/kg/day of ofloxacin, but not in those given 5 mg/kg/day. At 20 mg/kg/day, arthropathy of comparable severity also occurred on day 2. Microscopically, the cavity formation in the middle zone of the articular cartilage was first identified and then necrotic chondrocytes were found numerous around the cavity, followed by appearance of chondrocyte clusters. In pharmacokinetics, peak serum concentration (Cmax) and area under the concentrations (AUC0-24) were increased in a dose-dependent manner. However, no remarkable differences in these two parameters were noted between a single and repeated treatments, suggesting no accumulation of the drug. The articular ofloxacin concentration 2 hr after treatment was approximately 1.8 (day 2) to 2.0 times (day 8) higher than the serum concentration. Based on these results, a non-arthropathic dose of ofloxacin in male juvenile dogs following an 8-day treatment is considered to be 5 mg/kg/ day, and its Cmax, AUC0-24 and articular cartilage concentrations 2 hr after treatment were 3.4 microg/ml, 35.1 microg-hr/m/ and 7.0 microg/g, respectively, under these experimental conditions. Thus, arthropathy due to ofloxacin may be predicted by monitoring serum drug concentration.     

Absorption and Toxicity of Copper and Zinc in Bean Plants Cultivated in Soil Treated with Chicken Manure

The interaction between copper, zinc, and chicken manure, whenadded to soil, was studied in a greenhouse setting. Coppersulfate and of zinc chloride at concentrations of 0.5 to 2.0mmol kg^-1 were added separately to pots containing 3.0 kgof dystrophic dark red latosoil (LEd) plus 1,0% (m/m) pH 4.2chicken manure. IAPAR 57 beans were used as test plant andcultivated in the experimental soils for 30 days. Plant growthin terms of dry weight and the concentrations of copper and zincin the aerial part of the plants were determined. Bean plantingswere repeated four times, at four-month intervals. A 20%decrease in dry weight was observed when amounts of copper orzinc equal to or geater than 1.0 or 2.0 mmol kg^-1, respectively, were applied to the soil over an average of fourplantings. For these concentrations, the average amount of metalfound in the plant was 13.4 mg kg^-1 for copper and 224.8 mgkg^-1 for zinc. Chlorosis of the leaves in the bean plantswas also observed. The copper and zinc concentrations in thetreated soil were determined using different extractants. Amongthe solutions evaluated for the extraction of available zinc,1.0 M ammonium acetate, pH 4.8, showed a better correlation withthe metal concentration in the plant. As for copper, the bestcorrelation was found with the Mehlich-1 extractant.