Season- and Age-related Reproductive Changes Based on Fecal Androgen Concentrations in Male Koalas,Phascolarctos cinereus

The purposes of the present study were to clarify age- and season- related androgen patterns, and to compare the reproductive physiology between Japanese captive koala populations and Australian populations. To measure fecal androgens, feces were collected from male koalas (4.2 to 13.8 years of age) kept in Japanese zoos. Fecal androgens were extracted with methanol from the lyophilized samples and determined by enzyme immunoassay using 4-androstene-3,17-dione antibody. Fecal androgen concentration in male koalas increased after sexual maturation and remained relatively high until old age. In the survey with the Japanese zoo studbook of koalas, copulation (conception) month showed a pyramid shape with a peak in March to June (60.7%) in koalas born and reared in Japanese zoos and from July to April with the highest concentration in September to January (69.7%) in Australian institutes. Japanese zoo koala populations have a characteristic physiological cycle adapted to Japan''s seasonal changes. The suitable month of year for copulation or conception in Japan is diametrically opposed to that in Australia. Mean fecal androgen concentrations by month in the males born and reared in Japan indicated annual changes with the highest concentration in May and the lowest value in November. Fecal androgen analysis may be a noninvasive alternative tool to monitor circulating testosterone and may be helpful in understanding reproductive activity and physiology in male koalas.     

3-(4-Hydroxyphenyl)propionic acid is involved in the biosynthesis of myricanol in Myrica rubra

There is little evidence concerning the biosynthetic pathways for cyclic diarylheptanoids. We previously demonstrated that the cyclic diarylheptanoids myricanol and myricanone were biologically synthesized from two molecules of 4-coumaric acid by the feeding of 4-[8,9-¹³C₂]coumaric acid to young shoots of Myrica rubra. In the present study, using a ¹³C-labeled compound, we revealed that two molecules of 3-(4-hydroxyphenyl)propionic acid could also be a biosynthetic precursor of myricanol in M. rubra. These results indicated that both 4-coumaric acid and its dihydro-derivative were incorporated into myricanol. Competitive feeding experiments with 4-[8,9-¹³C₂]coumaric acid and 3-(4-hydroxyphenyl)-[1-¹³C]propionic acid were performed in M. rubra to determine the preferential incorporation of these two precursors. ¹³C-NMR studies indicated that 3-(4-hydroxyphenyl)-[1-¹³C]propionic acid was preferentially incorporated into myricanol. The data provided evidence for a biosynthetic sequence originating from 4-coumaric acid and leading to myricanol, through 3-(4-hydroxyphenyl)propionic acid, in M. rubra.     

A radio transmission pH measurement system for continuous evaluation of fluid pH in the rumen of cows

We developed a novel wireless radio transmission pH measurement system to continuously monitor ruminal bottom pH in cows, and compared these measurements to pH values determined by a spot-sample method. The wireless system consists of a pH sensor, data measurement receiver, relay unit, and personal computer with special software. The bullet-shaped sensor can be easily administered orally via a catheter into the rumen, without surgery. The glass electrode, using a temperature compensation system, can detect the rumen fluid pH with high accuracy. The ruminal bottom pH in healthy rumen-fistulated cows was measured as 6.52 ± 0.18 by the wireless system and as 6.62 ± 0.20 by the spot-sample method; with a correlation between pH measurements using these different methods (n = 8, 24 samples, r = 0.952, P < 0.01). When measured serially in a cow fed a diet evoking rumen acidosis, the ruminal bottom pH decreased markedly following the morning feeding and then increased gradually by the next morning feeding. This wireless system is a ready-to-use tool for estimating circadian changes in ruminal bottom pH.     

Bromocriptine inhibits salsolinol‐induced prolactin release in male goats

The secretion of prolactin (PRL) is under the dominant and tonic inhibitory control of dopamine (DA); however, we have recently found that salsolinol (SAL), an endogenous DA‐derived compound, strongly stimulated the release of PRL in ruminants. The aim of the present study was to clarify the inhibitory effect of DA on the SAL‐induced release of PRL in ruminants. The experiments were performed from late June to early July. Male goats were given a single intravenous (i.v.) injection of SAL (5 mg/kg body weight (BW)), a DA receptor antagonist (sulpiride, 0.1 mg/kg BW), or thyrotropin‐releasing hormone (TRH, 1 µg/kg BW) before and after treatment with a DA receptor agonist (bromocriptine), and the effect of DA on SAL‐induced PRL release was compared to that on sulpiride‐ or TRH‐induced release. Bromocriptine completely inhibited the SAL‐induced release of PRL (P < 0.05), and the area under the response curve (AUC) for a 120‐min period after the treatment with bromocriptine was 1/28 of that for before the treatment (P < 0.05). Bromocriptine also completely inhibited the sulpiride‐induced release (P < 0.05). The AUC post‐treatment was 1/17 that of pre‐treatment with bromocriptine (P < 0.05). Bromocriptine also inhibited the TRH‐induced release (P < 0.05), though not completely. The AUC post‐treatment was 1/3.8 that of pre‐treatment (P < 0.05). These results indicate that DA inhibits the SAL‐induced release of PRL in male goats, and suggest that SAL and DA are involved in regulating the secretion of PRL. They also suggest that in terms of the regulatory process for the secretion of PRL, SAL resembles sulpiride but differs from TRH.     

Calming effect of orally administered γ‐aminobutyric acid in Shih Tzu dogs

The calming effects of γ‐aminobutyric acid (GABA) by oral administration were investigated in four adult Shih Tzu dogs. Three dosage levels (1, 2 and 4 mg/kg body weight) and non‐administration were tested by an increase and decrease method. Changes in activity (for 1.5 h) and urinary cortisol levels (pre‐administration, 3 and 7 h later) of dogs were monitored after administration. Without reference to dosage level, the mean times spent standing (P = 0.06), sitting (P < 0.05) and walking (P < 0.05) tended to decrease compared to non‐administration. A significant depression in the urinary cortisol level was observed at 7 h after administration (P < 0.05). These results indicate that orally administrated GABA exerts calming effects on dogs as well as humans.     

3-Methylthiopropionic acid ethyl ester, isolated from Katsura-uri (Japanese pickling melon, Cucumis melo var. conomon), enhanced differentiation in human colon cancer cells

The fully ripened fruit of Katsura-uri Japanese pickling melon ( Cucumis melo var. conomon) has rarely been used for food because the midripened fruit is utilized for making pickles, but the fully ripened fruit is no longer valuable for pickles due to the fruit body being too soft. We have considered the utilization of the fully ripened Katsura-uri fruit that may be used for nonpickling products, particularly if the fully ripened fruit demonstrated health benefits such as anticarcinogenic properties. The phytochemical extract from the fully ripened fruit of Katsura-uri Japanese pickling melon was purified via a bioassay-guided fractionation scheme, which was based on the induction of differentiation in a RCM-1 human colon cancer cell line. On the criteria of two differentiation markers (duct formation and alkaline phosphatase activity), the most potent fraction contained a compound identified as 3-methylthiopropionic acid ethyl ester, based on GC retention time, EI-MS, (1)H NMR, and (13)C NMR spectra. Previously, the role of 3-methylthiopropionic acid ethyl ester was considered as an odor producing compound in many fruits, but this study indicates potential medical benefits of this compound.     

Molecular epidemiology of VapA-positive Rhodococcus equi in thoroughbred horses in Kagoshima,Japan

The prevalence of virulent R. equi having 15- to 17-kDa antigens (VapA) in fecal isolates from 13 thoroughbred foals and their dams on 5 farms in Kagoshima, Japan, and the plasmid profiles of VapA-positive isolates by restriction fragment digestion patterns were investigated to compare the genotypic variation among virulence plasmids of R. equi isolates from Japan. In total, 218 (24.6%) of 886 isolates from the feces of the 13 foals and 13 (12.5%) of 104 isolates from the feces of their dams demonstrated VapA-positive R. equi. Plasmid DNA preparations of 231 virulent isolates from foals and dams were analyzed by restriction enzyme digestion with endonucleases EcoRI, EcoT22I and HindIII and were divided into 3 types: 172 isolates contained a 90-kb type I plasmid, 57 contained a 90-kb type III plasmid and 2 contained a 90-kb type IV plasmid. This study demonstrates a geographic character in the distribution of virulence plasmids found in VapA-positive isolates from thoroughbred foals in Kagoshima.     

Studies on herbicidal carbamoyltetrazolinone derivatives: Selection of fentrazamide

1-Phenyl-5(4 H )-4-dialkylcarbamoyltetrazolinones represent a new class of herbicides that are active against barnyardgrass ( Echinochloa spp.) in paddy fields. To select the most favorable compound as a paddy herbicide, herbicidal activity and selectivity of the carbamoyltetrazolinone compounds to transplanted rice were determined in greenhouse tests. Higher herbicidal activities were associated with the compounds having a 2-Cl- or 2-Cl-6-Me-substituted or an unsubstituted phenyl ring, and a combination of C_2–3 linear alkyl group and a C₄ branched-chain alkyl or a C_5–6 cycloalkyl group on the nitrogen atom in the carbamoyl structure. In addition, the compounds carrying a total of seven to nine carbon atoms in the substituents on the carbamoyl nitrogen exhibited a good selectivity between barnyardgrass and transplanted rice plants. From the results of soil thick-layer chromatography, the good crop compatibility of carbamoyltetrazolinones was correlated to the mobility of the compounds in paddy soil. The investigation resulted in the selection of an excellent compound, 4-(2-chlorophenyl)- N -cyclohexyl- N -ethyl-4,5-dihydro-5-oxo-1 H -tetrazole-1-carboxamide (common name: fentrazamide), as a rice paddy herbicide for controlling barnyardgrass.