Comparative anatomical study on the relationships between the vestigial pelvic bones and the surrounding structures of finless porpoises (Neophocaena phocaenoides)

Morphology of the modern cetaceans represents the results of adaptation of the ancestral terrestrial mammals to aquatic life through their evolutional processes. Some of the primitive fossil cetaceans are known to have both fore and hind limbs, whereas the pelvic bones of modern cetaceans are, in general, a pair of slender rod-like structures within the abdominal wall muscles just anterior to the anus with no articulations to the axial skeleton in both sexes. It is interesting and important to consider the causes and processes of how the hind limbs were lost and how the pelvis was reduced during the process of adaptation. In the present study, we tried to evaluate the topography and function of rudimentary pelvic bones of the finless porpoise (Neophocaena phocaenoides), one of the members of the odontocete cetaceans, with special references to the structures around the pelvic bones. Some soft tissues such as M. ischiocavernosus relating to the pelvic bone are transformed following the drastic reduction of the pelvis. This transformation tells us that the cetaceans adapted to the aquatic life during evolutional processes chose the tail flukes driven by the powerful trunk muscles for locomotion, instead of modifying the hind limbs into hind flippers as seen in pinnipeds. On the other hand, it is evident that a function of the pelvic bones of the male finless porpoise was supporting the penis as those of terrestrial mammals. It is noteworthy that the morphological features of the ancestral terrestrial mammals can be traced when they are carefully compared with those of the finless porpoise.     

Effects of body condition score in cows peripartum on the onset of postpartum ovarian cyclicity and conception rates after ovulation synchronization/fixed-time artificial insemination

The aim of this study was to examine whether the nutritional state of cows peripartum was associated with the recovery of ovarian function and conception rates after synchronization of ovulation and fixed-time artificial insemination (OVSYNCH/TAI). The effect of the interval in days from calving to the first ovulation on conception rates after OVSYNCH/TAI was also investigated. Conception rates of cows after OVSYNCH/TAI (n=39) were 43.6%. The conception rates of cows with a body condition score (BCS) of 2.75-3.25 at 30 d postpartum and on the day of OVSYNCH treatment were significantly higher than in cows with a BCS < or =2.5 (P<0.05). The percentage of cows establishing ovarian cyclicity before 55 d postpartum in cows with a BCS of 2.75-3.25 at 30 d postpartum and on the day of OVSYNCH treatment were significantly higher than in cows with a BCS < or =2.5 (P<0.05). The conception rates after OVSYNCH/TAI in cows which recovered ovarian cyclicity within 34 d postpartum were significantly higher than in cows with first ovulation > or =56 d (P<0.05). These results indicated that the nutritional state in cows peripartum influenced the conception rates after OVSYNCH/TAI and the postpartum ovarian cyclicity and also suggested that the conception rates after OVSYNCH/TAI decreased in cows with delayed recovery of ovarian cyclicity.     

Inhibitory effects of oolong tea polyphenols on pancreatic lipase in vitro

Fifty-four polyphenols isolated from tea leaves were evaluated for their inhibitory activities against pancreatic lipase, the key enzyme of lipid absorption in the gut. (-)-Epigallocatechin 3-O-gallate (EGCG), which is one of major polyphenols in green tea, showed lipase inhibition with an IC50 of 0.349 microM. Moreover, flavan-3-ol digallate esters, such as (-)-epigallocatechin-3,5-digallate, showed higher activities of inhibition on lipase with an IC50 of 0.098 microM. On the other hand, nonesterified flavan-3-ols, such as (+)-catechin, (-)-epicatechin, (+)-gallocatechin, and (-)-epigallocatechin, showed zero and/or the lowest activities against pancreatic lipase (IC50 > 20 microM). These data suggested that the presence of galloyl moieties within the structure was required for enhancement of pancreatic lipase inhibition. It is well-known that flavan-3-ols are polymerized by polyphenol oxidase and/or heating in a manufacturing process of oolong tea. Oolonghomobisflavans A and B and oolongtheanin 3'-O-gallate, which are typical in oolong tea leaves, showed strong inhibitory activities with IC50 values of 0.048, 0.108, and 0.068 microM, respectively, even higher than that of EGCG. The oolong tea polymerized polyphenols (OTPP) were prepared for the assay from oolong tea extract, from which the preparation effectively subtracted the zero and/or less-active monomeric flavan-3-ols by preparative high-performance liquid chromatography. The weight-average molecular weight (Mw) and number-average molecular-weight (Mn) values of OTPP were 2017 and 903, respectively, by using gel permeation choromatography. OTPP showed a 5-fold stronger inhibition against pancreatic lipase (IC50 = 0.28 microg/mL) by comparison with that of the tannase-treated OTPP (IC50 = 1.38 microg/mL). These data suggested that the presence of galloyl moieties within their chemical structures and/or the polymerization of flavan-3-ols were required for enhancement of pancreatic lipase inhibition.     

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.     

Sympatric distribution of the two morphological types of the common tree shrew in Hat-Yai districts (South Thailand)

The two color types (grayish northern and reddish southern types) of the common tree shrew (Tupaia glis and Tupaia belangeri) were co-distributed in Hat-Yai region (South Thailand). Although the Isthmus of Kra in South Thailand has been considered as distribution barrier of the two types, the sympatric distribution of both types was confirmed in southern side of the Isthmus. In the principal component analysis, the skull measurement character from Hat-Yai region could also be separated into the northern and southern groups according to the skin color identification of corresponding individuals. We could generally distinguish the common tree shrew into two types by skull morphology as well as external skin color.     

A specific and potent inhibitor of brassinosteroid biosynthesis possessing a dioxolane ring

Screening for brassinosteroid biosynthesis inhibitors was performed to find azole derivatives that induced dwarfism, to resemble brassinosteroid-deficient mutants in Arabidopsis, and which could be rescued by brassinosteroid. Through this screening experiment, propiconazole fungicide was selected as a likely inhibitor of brassinosteroid biosynthesis and, thus, propiconazole derivatives with optimized activity and selectivity were synthesized. The biological activity of these compounds was evaluated by examining cress stem elongation. Among the compounds tested, 2RS,4RS-1-[2-(4-trifluoromethylphenyl)-4-n-propyl-1,3-dioxolan-2-ylmethyl]-1H-1,2,4-triazole (12) showed the most potent capability to retard cress stem elongation in the light. The compound-induced hypocotyl dwarfism was restored by the coapplication of 10 nM brassinolide but not by 1 microM gibberellin. These results suggest that 12 should affect brassinosteroid biosynthesis. The potency and specificity of 12 were greater than those of brassinazole, a previously reported brassinosteroid biosynthesis inhibitor.     

Absolute configuration of arylglycerol-β-aryl ethers obtained by asymmetric reduction of the correspondingα-ketonic compound with intactFusarium solani cells

When (±)--oxo-guaiacylglycerol--(vanillic acid) ether (1) is degraded byFusarium solani M-13-1, the-ketone is initially reduced to giveerythro andthreo guaiacylglycerol--(vanillic acid) ethers (2), arylglycerol--aryl ethers, both of which are enantiomerically pure. The absolute configuration in each2 was determined by Mosher's method; the products were converted to,-di-(R)--methoxy--trifluoromethylphenylacetates (MTPA esters) (3) oferythro (-)- andthreo (+)-veratrylglycerol--(methyl vanillate) ethers (3), whose¹H nuclear magnetic resonance (NMR) spectra were examined and compared with those of four di-(R)-MTPA ester (3) diastereomers from chemically synthesizederythro (±)-3 andthreo (±)-3. To assign the- and-MTPA-OCH₃ peaks, the¹H NMR scans of several compounds that have substructures of 3 and their 3,4,5-trimethoxyphenyl analogues were examined. When a racemic alcohol reacts with (R)-MTPA to give a pair of (R)-MTPA ester diastereomers, the value was defined as the absolute value of the difference in the¹H chemical shifts of the peak between the diastereomers. It was found that the values of-MTPA-OCH₃ were larger than those of-MTPA-OCH₃ owing to a shielding effect of the veratryl ring located on the-MTPA-OCH₃, and that the-MTPA-OCH₃ peaks in the 3,4,5-trimethoxyphenyl compounds shifted downfield relative to those in the veratryl compounds. On the basis of the¹h NMR data of (R)-MTPA esters, the absolute configuration of the four chemically prepared diastereomers (3) were determined. The catabolicerythro 3 [fromerythro (-)-3] andthreo 3 [fromthreo (+)-3] were identical to (R, S, R)-erythro 3 and (R, S, S)- threo 3, respectively. An hydrogen species in the fungal reduction would attack the-ketone fromre-face of both (R)-1 and (S)-1, givingerythro (S, R)-2 andthreo (S, S)-2, respectively.Part of this paper was presented at the 33rd Lignin Symposium, Tsukuba, November 1988     

Persimmon breeding in Japan for pollination-constant non-astringent (PCNA) type with marker-assisted selection

Oriental persimmon (Diospyros kaki) originated in Eastern Asia, and many indigenous cultivars have been developed in China, Japan, and Korea. These cultivars are classified into four groups based on their natural astringency loss on the tree and seed formation: pollination-constant non-astringent (PCNA), pollination-variant non-astringent (PVNA), pollination-constant astringent (PCA), and pollination-variant astringent (PVA). PCNA is the most desirable type because the fruit can be eaten without any postharvest treatment; therefore, one of the goals of our persimmon breeding programs is to release superior PCNA cultivars. The PCNA genotype is recessive to the other three non-PCNA genotypes, and PCNA-type F₁ offspring are obtained exclusively from crosses among PCNA genotypes. Moreover, the number of superior PCNA cross-parents have been limited. In the late 1980s, inbreeding depression became obvious, especially in terms of fruit size, tree vigor, and productivity. To mitigate the inbreeding, a backcross program using PCNA [(non-PCNA × PCNA) × PCNA] was started in 1990. This process, however, was inefficient because only 15% of the offspring were PCNA, and all offspring had to be grown to the fruiting stage. Therefore, molecular markers linked to the PCNA locus were developed for discriminating PCNA offspring. A molecular marker linked to Chinese PCNA has also been developed.     

Assessing sexual maturity of feminized Japanese eel Anguilla japonica by measuring eye size

For efficient production of Japanese eel Anguilla japonica eggs, knowledge of the status of the sexual maturity of potential broodstock females is important because this status directly influences the time required to produce mature eggs by successive hormone doses. Here we apply an eye index (relative eye size) to evaluate the gonadal status of feminized A. japonica, which were induced by administration of estradiol-17β. Examination of gonad somatic and eye indexes of 267 feminized eels, cultivated for 12–56 months, revealed a significant correlation between these two indexes. Artificial maturation of 35 feminized eels showed that the number of hormone injections administered before ovulation was significantly lower as the eye index score increased, indicating availability of this noninvasive method of predicting sexual maturity of feminized eel. There was no correlation between eye index and fertilization rate, hatching rate, or larval survival rate, suggesting that sexual maturity before hormonal treatment does not affect egg quality.     

Mung bean trypsin inhibitor is effective in suppressing the degradation of myofibrillar proteins in the skeletal muscle of blue scad (Decapterus maruadsi)

Mung bean trypsin inhibitor (MBTI) of the Bowman-Birk family was purified to homogeneity with a molecular mass of approximately 9 kDa on tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 8887.25 Da as determined by matrix-assisted laser desorption/ionization-quadrupole ion trap-time-of-flight mass spectrometry (MALDI-QIT-TOF MS). Using blue scad myofibrillar proteins as targets, it was found that, in the absence of MBTI, proteolysis of myofibrillar proteins, especially myosin heavy chain (MHC), could be identified after incubation at 55 °C for 2 h, while in the presence of MBTI, with a final concentration of 25 ng/mL, proteolysis of these proteins was greatly suppressed even after incubation for 3 h. Although cysteine proteinase inhibitor E-64 was also effective in preventing protein degradation, inhibitors for metallo- and asparatic proteinases did not reveal obvious inhibitory effects. Our present results strongly suggested that the naturally occurring legume bean seed protein MBTI can be used as an effective additive in preventing marine fish blue scad surimi gel softening, which is quite possibly caused by myofibril-bound serine proteinase (MBSP).