Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

Effects of mangrove deforestation on fish assemblage at Pak Phanang Bay,southern Thailand

Daytime sampling using a seine net was conducted at Pak Phanang Bay (Nakhon Si Thammarat Province, Thailand) in February and July 2006, to determine differences in fish assemblage structures between a mangrove site and an adjacent site completely cleared of mangroves. The overall numbers of fish species and individuals were significantly higher at the mangrove site than the cleared site in both months. Although benthic crustacean feeders showed more species and individual numbers at the mangrove site in both months, the opposite was found for zooplankton feeders. A cluster analysis, based on the abundance of each species, demonstrated that the fish assemblage structures were distinctly different between the two sites. In addition, significant differences in length frequency distributions for each of the four most abundant species were found between the sites in February and/or July. Small individuals of Scatophagus argus, Ambassis nalua, and Tetraodon nigroviridis were more common at the mangrove site, and of Chelon subviridis at the cleared site. These results suggest that mangrove deforestation exerts marked effects on fish assemblages.     

Seasonal Changes in Rigor Development and Flesh Texture of Wild Japanese Sea Bass (Lateolabrax japonicus)

The season for finding the firmest sashimi made from wild Japanese sea bass was investigated. Flesh rigor tension and contraction were measured by a “new” device with isometric (muscle length kept constant) and isotonic (balance beam moves freely) transducers. Both flesh rigor tension and contraction peaks appeared at the same postmortem times, with attainment of ultimate pH and adenosine triphosphate/inosine monophosphate ratio in four seasons. Seasonal seawater temperature correlated negatively to seasonal maximum rigor contraction and positively to seasonal flesh breaking strength at 72 h postmortem. After rigor peak tension attainment, the tension relaxation was much lower than that of other wild fish species, especially in summer. The sea bass muscle resisted structure disruption in rigor isometric tension generation. Wild sea bass sashimi was firmest in breaking strength in summer.     

Thermal behavior of beta-1 subunits in lignin: pyrolysis of 1,2-diarylpropane-1,3-diol-type lignin model compounds

1,2-Diarylpropane-1,3-diol-type lignin model compounds, 1,2-bis(4-hydroxy-3-methoxyphenyl)propane-1,3-diol (1) and 1-(3,4-diethoxyphenyl)-2-(4-methoxyphenyl)propane-1,3-diol (2), were pyrolyzed at 500 degrees C for 4 s to clarify the thermal behavior of beta-1 subunits in lignin. Products were monitored by gas chromatography/mass spectrometry. The cleavage of the Calpha-Cbeta bond to produce benzaldehydes such as 4-hydroxy-3-methoxybenzaldehyde (9) and phenylethanals as the counterparts such as 4-hydroxy-3-methoxyphenylethanal (10) occurred in pyrolyses of both 1 and 2. In pyrolysis of 1, an oxetane pathway leading to the formation of Z/E-stilbenes without the gamma-CH2OH group such as Z/E-4,4'-dihydroxy-3,3'-dimethoxystilbene (3) was predominant. In pyrolysis of 2, the oxetane pathway was minor, while pathways producing a dimer with a =CgammaH2 group by loss of water and a dimer with an alpha-carbonyl group were predominant. Pyrolysis of Japanese cedar wood provided 3 and 10 in approximately 0.8% and 0.6% yields, respectively, based on the Klason lignin content, while pyrolysis of a guaiacyl bulk dehydrogenation polymer gave them in a very small amount.     

Intratubal insemination with fresh semen in dogs

The number of spermatozoa required to obtain conception by intratubal insemination in dogs was examined. Three groups consisting of 5, 8 and 8 dogs received 0.5 x 10 (6), 2.0 x 10(6) and 4.0 x 10(6) spermatozoa, respectively, into each uterine tube. No conception occurred in the 5 animals inseminated with 0.5 x 10(6) spermatozoa, but conception occurred in 6/8 (75.0%) and 3/8 (37.5%) dogs inseminated with 2.0 x 10(6) and 4.0 x 10 (6) spermatozoa, respectively. Among the pregnant animals, three aborted (33.3%) and the mean number of newborns was small, 2.5 +/- 0.5 (SE). One acardiacus anceps was observed with normal fetus in one animal with a Caesarean delivery.     

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.     

Factors affecting transuterine migration of canine embryos

Dogs, cats, and pigs have a bicornuate uterus, and transuterine migration of embryos occurs in 40% or more of pregnant animals. However, the mechanism of the transuterine migration has not been elucidated in dogs. Thus, we investigated the occurrence of transuterine migration of embryos when embryos were retained in an unilateral uterine tube with more ovulated ova (Experiment 1), when one ovary was excised (Experiment 2), and when ova ovulated from the right and left ovaries were fertilized with sperm from male dogs with different blood types (Experiment 3). Transuterine migration of embryos was observed in 7/8 (87.5%), 10/10 (100%), and 11/17 (67.4%) fertilized animals in Experiments 1, 2, and 3, respectively. In Experiment 3, intrauterine embryo mixing reported in pigs did not occur. These findings suggest that transuterine migration of embryos occurs due to the number of embryos that enter the uterus but that differences in the number of ovulated ova between the right and left ovaries or the number of embryos retained in the uterine tube do not affect the migration.     

Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

We previously reported that the release of O₂ ⁻ from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. The nucleotide sequence data of PsCu/Zn-SOD1 reported are available in the DDBJ/EMBL/GenBank databases under accession number AB189165.     

Performance of using opposing homozygotes for paternity testing in Japanese Black cattle

Genome-wide single nucleotide polymorphism (SNP) markers in Japanese Black cattle enable genomic prediction and verifying parent–offspring relationships. We assessed the performance of opposing homozygotes (OH) for paternity testing in Japanese Black cattle, using SNP genotype information of 50 sires and 3,420 fattened animals, 1,945 of which were fathered by the 50 genotyped sires. The number of OH was counted for each sire–progeny pair in 28,764 SNPs with minor allele frequencies of ≥0.05 in this population. Across all pairs of animals, the number of OH tended to increase as the pedigree-based coefficient of relationship decreased. With a threshold of 288 (1% of SNPs) for paternity testing, most sire–progeny pairs were detected as true relationships. The frequency of Mendelian inconsistencies was 2.4%, reflecting the high accuracy of pedigree information in Japanese Black cattle population. The results indicate the utility of OH for paternity testing in Japanese Black cattle.     

Effects of acorn attack by curculio weevils on the germination and early growth of <Emphasis Type="Italic">Pasania edulis</Emphasis> (Makino) seedlings

We conducted field studies to evaluate the impact that curculio weevil attacks on Pasania edulis (Fagacea) acorns have on the regeneration of this tree. The germination ratio of weevil-attacked acorns was significantly lower than that of sound acorns. The number and position of the attacks on acorns affected the germination ratio. The seedlings from weevil-attacked acorns were shorter and had a smaller leaf area than those from sound acorns. The negative effects on seedling height lasted for at least three years after germination. We also studied the handling of the two types of acorns by granivorous Apodemus mice in a broad-leaved forest dominated by P. edulis. The mice picked up, transported, hoarded, and recovered sound and weevil-attacked acorns similarly. In the study site, the seedlings from weevil-attacked acorns were estimated to account for 1.5–20.4 % of the total seedlings for cohorts of mast years and 0–3.7 % for those of poor or medium acorn production. From these results, we conclude that acorns with the lower part of their cotyledons slightly damaged by weevil larvae might still be able to contribute to the regeneration of P. edulis in the field but that their contribution would be negligible or small even in mast years in this study site.