Chlorogenic acid supplementation during in vitro maturation improves maturation,fertilization and developmental competence of porcine oocytes

Chlorogenic acid (CGA) is a quinic acid conjugate of caffeic acid, and a phytochemical found in many fruits and beverages that acts as an antioxidant. The present study investigated the effects of CGA supplementation during in vitro maturation (IVM), on in vitro development of porcine oocytes, to improve the porcine in vitro production (IVP) system. Oocytes were matured either without (control) or with CGA (10, 50, 100 and 200 μM). Subsequently, the matured oocytes were fertilized and cultured in vitro for 7 day. The rates of maturation, fertilization and blastocyst formation of oocytes matured with 50 μM CGA were significantly (p < .05) higher than those of the control oocytes. Hydrogen peroxide (H₂O₂) is one of the reactive oxygen species and induces DNA damage in porcine oocytes. When oocytes were matured with 1 mM H₂O₂ to assess the protective effect of CGA, 50 μM CGA supplementation improved the maturation rate and the proportion of DNA‐fragmented nuclei in oocytes compared with control oocytes matured without CGA. Moreover, when oocytes were matured with either 50 μM CGA (control) or caffeic acid (10, 50 and 100 μM), the rates of maturation, fertilization and the blastocyst formation of oocytes matured with 50 μM CGA were similar to those of oocytes matured with 10 and 50 μM caffeic acid. Our results suggest that CGA has comparable effects to caffeic acid, and IVM with 50 μM CGA is particularly beneficial to IVP of porcine embryos and protects oocytes from DNA damage induced by oxidative stress. Supplementation of CGA to the maturation medium has a potential to improve porcine IVP system.     

The <Emphasis Type="Italic">Arabidopsis NST3/SND1</Emphasis> promoter is active in secondary woody tissue in poplar

Wood biomass is one of the promising future materials for biofuels with no competing food uses. However, the higher cost to produce bioethanol from wood feedstocks is regarded as a priority issue. Genetic engineering techniques have been proposed to enhance the quality and quantity of wood materials to overcome the cost problem. Although many genetically engineered trees with applicable traits such as low lignin, a high syringyl to guaiacyl ratio and high cellulose content are generated, ectopic expression of an effector gene under a constitutive promoter can sometimes induce untoward side effects on plant growth and development. Our recent study demonstrated that AtNST3/SND1 promoter of Arabidopsis thaliana is a candidate tool for driving a potent activator to enhance wood biomass production in poplar without any growth retardation. However, the tissue- and cell-dependent activity of the promoter remains to be elucidated. In the present study, we generated transgenic poplar expressing AtNST3/SND1promoter::GUS to examine in detail the activity of the AtNST3/SND1 promoter. Histochemical analysis revealed that the promoter was predominantly active in secondary woody tissue. Our result indicates that the AtNST3/SND1 promoter is an option for expressing an effector gene to modify secondary cell wall components and wood biomass.     

Overwintering of <Emphasis Type="Italic">Taphrina wiesneri</Emphasis> within cherry shoots monitored with species-specific PCR

Taphrina wiesneri, the pathogen of witches’ broom of cherry, is highly pathogenic to Cerasus × yedoensis, the most widely planted ornamental cherry species in Japan. For adequate control of this disease, it is necessary to understand the life history of T. wiesneri. However, sites inhabited by T. wiesneri within infected trees are little understood, except during flowering and leafing periods in spring. Therefore, we attempted to detect the location of T. wiesneri in shoots of witches’ broom before flowering and leafing in spring using PCR with a T. wiesneri-specific primer pair that was designed from 69 sequences in rDNA-internal transcribed spacer region of 32 Taphrina species. DNA extracted from symptomatic and asymptomatic C. × yedoensis sampled before leafing was amplified by PCR. T. wiesneri was detected in every bud and 5-mm stem segment of symptomatic shoots, except for one stem segment, and locally inside buds and the inner bark of stem segments. These results indicate that T. wiesneri overwinters inside symptomatic shoots. Fungal hyphae were observed with an epifluorescence microscope in intercellular spaces of young leaves in symptomatic buds but not in asymptomatic ones in thin sections stained with fluorescein-isothiocyanate-conjugated concanavalin A. This observation supports the results of PCR detection.     

Time dependence of Poisson’s effect in wood II: volume change during uniaxial tensile creep

To determine the viscoelasticity of wood three-dimensionally, a longitudinal tensile creep test was conducted on 12 species of wood to examine the change in the rate of volume increase (ΔV/V) with time. Immediately after the beginning of creep, ΔV/V was positive, and during creep, ΔV/V decreased rapidly, then more gradually. The decrease in tangential strain was considered to mainly contribute to the decrease in ΔV/V during creep. Immediately after the removal of the load, ΔV/V decreased to a negative value; thereafter, it decreased slowly and finally reached a certain value. The value of ΔV/V during creep tended to decrease with increasing density of wood. Also, there was a negative correlation between wood density and the rate of increase in ΔV/V.     

Effects of glucose and amino acids on ghrelin secretion in sheep

Two experiments were conducted to elucidate the effects of post‐ruminal administration of starch and casein (Exp. 1), plasma amino acids concentrations (Exp. 2), and plasma glucose and insulin concentrations (Exp. 2) on plasma ghrelin concentrations in sheep. In Exp. 1, plasma ghrelin concentrations were determined by four infusion treatments (water, cornstarch, casein and cornstarch plus casein) in four wethers. Abomasal infusion of casein increased plasma α‐amino N (AAN) concentrations. Infusion of starch or casein alone did not affect plasma ghrelin concentrations, but starch plus casein infusion increased plasma levels of ghrelin, glucose and AAN. In Exp 2, we investigated the effects of saline or amino acids on ghrelin secretion in four wethers. Two hours after the initiation of saline or amino acid infusion into the jugular vein, glucose was also continuously infused to investigate the effects of blood glucose and insulin by hyper‐glycemic clump on plasma ghrelin concentrations. Infusion of amino acids alone raised plasma levels of ghrelin, but the higher plasma glucose and insulin concentrations had no effect on plasma ghrelin concentrations. These results suggest that high plasma levels of amino acids can stimulate ghrelin secretion, but glucose and insulin do not affect ghrelin secretion in sheep.     

Localization of eNOS in the olfactory epithelium of the rat

Nitric oxide (NO) is a free radical and produced from L-arginine by nitric oxide synthase (NOS). Since NO is recently suggested to be involved in olfactory perception, the expression of eNOS, an isoform of NOS, was examined in the rat olfactory epithelium. The activity of NADPH-diaphorase was also examined as a marker of NOS. In the dorsomedial region of the nasal cavity, intensely positive reactions for NADPH-diaphorase were observed in the entire cytoplasm of sensory cells (olfactory cells). By immunohistochemistry, intensely positive reactions for eNOS were also found in the dorsomedial region of the nasal cavity. These reactions were observed on the free border of the olfactory epithelium. By immunoelectron microscopy, positive reactions for eNOS were found in the cilia of olfactory cells. In addition, in situ hybridization analysis of the olfactory epithelium revealed the expression of eNOS mRNA in the olfactory cells. These results indicate the presence of eNOS in the olfactory cells of the rat, and differential expression of eNOS in the olfactory epithelium depending on the regions of the nasal cavity. In addition, NO produced by eNOS may be involved in olfactory perception in the cilia of olfactory cells.     

Fecal progestagens to detect and monitor pregnancy in captive female cheetahs (Acinonyx jubatus)

The purposes of the present study were to establish a noninvasive monitoring assay of fecal progestagen measurement to detect pregnancy and to identify the components of fecal progestagens in early, middle and late pregnancy in cheetahs. Feces were collected from 7 female cheetahs and analyzed from 30 days before the last copulation to parturition in 9 pregnancies. Blood was collected from one cheetah. Fecal progestagen and serum progesterone concentrations were determined by enzyme immunoassay (EIA). The profiles of the fecal progestagen concentrations were similar to the serum progesterone profile. Fecal progestagen and serum progesterone concentrations remained at the baseline until copulation. In the mean fecal progestagen profile during pregnancy (92.8 ± 0.4 days; from the last copulation to parturition), the concentrations increased 3-4 days after the last copulation and remained high until parturition. To investigate changes in the components of progestagen metabolites in the tripartite periods of gestation, fecal progestagens were analyzed by HPLC-EIA. Marked immunoreactive peaks consistent with 5α-pregnan-3α/β-ol-20-one and 5α-pregnan-3,20-dione and small peaks consistent with 5β-pregnan-3α/β-ol-20-one were detected. There were no distinct difference in the components of progestagens among the first, second and third trimesters of pregnancy. The hormone assay, as an indicator of fecal 5α-reduced pregnanes, is useful for detecting pregnancy and monitoring pregnant luteal activity in cheetahs.     

Epibiotic microorganisms associated with microcrustaceans in overlying water of Philippine paddy fields

Epibiotic microorganisms associated with micro crustaceans in the overlying water of Philippine paddy fields were examined by scanning electron microscopy, and their associations were compared with those observed in Japanese paddy fields (Soil Sci. Plant Nutr., 43, 633-641, 651-664, 1997). Various types of microorganisms, including bacteria, protozoa, and algae, colonized microcrustaceans and their kinds and colonizing patterns varied depending on the kinds of microcrustaceans. Common associations between epibionts and respective micro crustaceans were observed in the Philippines and in Japan: e.g., rodshaped bacteria on Cyclopidae and Cypridopsidae; no microorganisms on Simocephalus spp.     

K/Ca and NH4/Ca selectivity of hydroxyaluminum-interlayered vermiculite and montmorillonite: Contribution from regular and frayed edge exchange sites

Cation distribution in adsorbed and solution phases and their mobility in the soil profile are largely influenced by the cation exchange capacity and selectivity coefficient of the soil. Vermiculites (Vt) and montmorillonites (Mt) in acid soils and sediments are frequently interlayered by hydroxyaluminum (HyA). Such interlayering causes significant changes in various properties of Vt and Mt. We evaluated the contribution of HyA-induced frayed edge sites (FES) in Vt and Mt to the overall selectivity of K/Ca and NHJCa. A more preferential adsorption and exchange of K to Ca was observed in the case of Vt in contrast to Mt. The overall K/Ca selectivity of Vt increased markedly by HyA-interlayering, whereas that of Mt was slightly affected. An increased overall K/Ca selectivity, especially of Vt was also observed due to the larger amount of HyA adsorption at higher initial basicity of the HyA ionic solution. Selective adsorption of K and NH₄ predominantly occurred on the FES. Interlayering substantially enhanced such FES contribution. The overall K/Ca and NH₄/Ca selectivity coefficients did not indicate any striking competitiveness of NH₄ with K, unlike that for the natural micaceous clay minerals. However, the parameter of FES contribution suggested that NH₄ could be slightly more competitive with K for the FES. The relative abundance of FES on different clay specimens determining K/Ca and NH₄/Ca selectivity and a slightly higher preference of NH₄ to K on FES played a major role at low K or NH₄-saturation.