Total Synthesis of Gobiusxanthin Stereoisomers and Their Application to Determination of Absolute Configurations of Natural Products: Revision of Reported Absolute Configuration of Epigobiusxanthin

(3R)-Gobiusxanthin stereoisomers (1a–d) were synthesized by stereoselective Wittig reaction of the (3R)-C₁₅-acetylenic tri-n-butylphosphonium salt 7 with C₂₅-apocarotenal stereoisomers 5a,b and 14a,b bearing four kinds of 3,6-dihydroxy-ε-end groups. The validity of the reported stereochemistry of gobiusxanthin was demonstrated by the fact that the reported spectral data of natural gobiusxanthin were in agreement with those of synthetic (3R,3''S,6''R)-gobiusxanthin (1a). On the other hand, the reported CD spectral data of natural epigobiusxanthin, which has been assigned as (3R,3''R,6''R)-isomer (3''-epigobiusxanthin), were identical with those of synthetic (3R,3''S,6''S)-isomer 1d (6''-epigobiusxanthin) rather than those of the corresponding synthetic 3''-epi-isomer 1b. It was found that the stereochemistry at C3-position has little effect on the shape of their CD spectra. Thus, in order to reinforce the validity of the absolute configurations at C3-position of natural specimens, (3S,3''S,6''R)- and (3S,3''S,6''S)-stereoisomers 1e and 1f were also synthesized and a HPLC analytical method for four stereoisomers was established by using a column carrying a chiral stationary phase. The HPLC analysis has proven that the stereochemistry of the natural epigobiusxanthin is 3R,3''S,6''S.     

Comparison of the glucosinolate-myrosinase systems among daikon (Raphanus sativus, Japanese white radish) varieties

Myrosinase is a cytosolic plant enzyme present in daikon ( Raphanus sativus, Japanese white radish) roots that hydrolyzes 4-methylthio-3-butenyl glucosinolate (MTBGLS) into the natural pungent agent 4-methylthio-3-butenyl isothiocyanate (MTBITC), which possesses antimicrobial, antimutagenic, and anticarcinogenic properties. The concentration of MTBGLS, myrosinase activity, and production of MTBITC in seven daikon varieties (one conventional and six heirlooms) were determined to rank the activity of the glucosinolate-myrosinase system and identify critical factors influencing the production of MTBITC. The six heirloom varieties produced 2.0-11.5 times higher levels of MTBITC as compared to the conventional variety, Aokubi, which is consumed by the present Japanese population. The myrosinase was located exclusively in the outer epidermal layer in Aokubi, and MTBGLS was widely distributed throughout the root tissue. Although the skin is a potentially rich source of myrosinase in Aokubi, the skin is usually peeled off in the current practice of preparing daikon for cooking. New practices are therefore proposed for the preparation of daikon tubers that eliminate the peeling of the skin to avoid removing the enzyme needed to convert MTBGLS to the health-beneficial MTBITC. It is also concluded that the consumption of heirloom daikon varieties in addition to changes in food preparation will optimize the health benefits of daikon.     

Dried green leaf powders of Jew's mellow (Corchorus), persimmon (Diosphyros kaki) and sweet potato (Ipomoea batatas poir) lower hepatic cholesterol concentration and increase fecal bile acid excretion in rats fed a cholesterol-free diet

A cholesterol-free diet containing dried powder of Jew's mellow leaves, persimmon leaves or sweet potato leaves respectively at 5% level as dietary fiber was fed to male Sprague-Dawley rats for about one month. The experiment was conducted twice except for sweet potato leaves. In the groups fed the diet mixed with powders of any of the three different dried green leaves, the hepatic cholesterol concentration significantly decreased. Such lowering was not observed in serum cholesterol concentration compared with the control (cellulose) group. A significant increase in fecal weight was observed in all the groups fed the green leaf samples. All the dried green leaves increased fecal excretion of bile acids per gram or per day compared with the control group in both experiments, but only the dried Jew's mellow leaves showed an increased excretion of neutral sterols. These results suggest that lowering of hepatic cholesterol by powdered green leaves is not necessarily due to the same factor, but to the increased fecal excretion of bile acids due to inhibited enterohepatic circulation in animals given these samples.     

Naturally occurring Tyzzer's disease in a calf.

Naturally occurring Clostridium piliforme infection (Tyzzer's disease) was found in a calf. Light microscopic examination revealed multifocal coagulative necrosis in the liver, catarrhal gastroenteritis, tracheitis and pneumonia, and thymic atrophy. Warthin-Starry staining clearly showed large filamentous bacilli in bundles or criss-cross patterns within the hepatocytes and epithelium and smooth muscle cells of the ileum and cecum. Immunohistochemistry using anti-C. piliforme RT and MSK strain antisera showed positive reaction against the bacilli. Electron microscopic examination revealed bacilli within the hepatocytes that demonstrated a characteristic vegetative form, with peritrichous flagella, and spores. The polymerase chain reaction (PCR) study using the paraffin-embedded liver sections, the 196-bp DNA fragment specific to 16S ribosomal RNA of C. piliforme was amplified. The characteristics of these bacilli are consistent with those of of C. piliforme. The PCR technique using paraffin-embedded sections should be useful for confirming C. piliforme infection in spontaneous cases.     

Clay mineralogy of the B horizons of two hydrandepts,a Torrox and a Humitropept in Hawaii

The mineralogy of the clay fractions separated from the B horizons of two Hydrandepts (Hilo and Akaka soils), a Torrox (Molokai soil) and a Humitropept (Kolekole soil) was determined by a combination of methods.The Hydrandept B horizons were characterized by predominance of hydrous non-crystalline alumina and iron oxides associated with considerable amounts of humus and with very small amounts of silica. Allophane, allophane-like constituents and imogolite were present but in minor amounts. Gibbsite, goethite, chlorite and illite were also present as accessory minerals.The Torrox and Humitropept B horizons were characterized by predominance of kaolinite, hematite and goethite. The iron oxide minerals were present as fine particles (40–80 A diameter) often clustered to form larger aggregates. Neither imogolite nor allophane and allophane-like constituents were detected. Considerable amounts of dithionite-citrate soluble Al and humus were, however, present in the Humitropept B horizon, which may reflect the effect of an admixture of volcanic ash to the parent material.     

Immune potency of lyophilized, killed vaccine for equine viral arteritis and its protection against abortion in pregnant mares

Immune potency test was conducted in horses by inoculating a killed vaccine for equine viral arteritis (EVA) which had been freeze-dried and contained aluminum hydroxide adjuvant. Serum neutralizing (SN) antibody to equine arteritis virus (EAV) was detected at maximal titers of 1:80 to 1:640, 1 to 2 weeks after 2-dose vaccination of 6 female horses. However, 6 pregnant mares inoculated with the vaccine which had been kept in storage for 1 year at 4°C produced much higher titers ranging from 1:320 to 1:1280. A maximal mean titer of 1:199.5 occurred in the 1st and 2nd week after 2-dose inoculation with the nonpreserved vaccine, whereas a maximal mean titer of 1:794.3 occurred in the 2nd week using the preserved vaccine. The horses showed no systemic or local adverse reactions clinically or hematologically after vaccination. Four of the 6 vaccinated pregnant mares were exposed to the Bucyrus strain of EAV but resisted challenge exposure, while 3 nonvaccinated control pregnant mares revealed acute EVA causing abortion and death. Isolation of EAV was positive from the body tissues of the aborted and dead fetuses and their dams, but was negative from the vaccinated mares. No significant rise of SN antibody titers was detected in the vaccinated mares following challenge exposure, suggesting that the vaccine can protect against EAV infection in pregnant mares and prevent abortion or death.     

Clinical and virological observations on swine experimentally infected with Getah virus

The pathogenicity of Getah virus for swine was examined. All 8 pigs (4 adults and 4 piglets) inoculated with Strains MIP-99 and MI-110 developed pyrexia ranging from 39.4 to 40.7 degrees C and anorexia. Mild depression and diarrhea were observed in 2 of the 4 piglets. These clinical signs were transient. Viremia occurred 1-2 days post-inoculation (p.i.) and the maximum titer was 10(3.0) TCID50 0.1 ml-1. The virus was recovered from a piglet autopsied on Day 3 p.i. from spleen and various lymph nodes. The maximum titer of virus (10(3.75) TCID50 0.1 g-1) was detected in the inguinal lymph node. Seroconversion was demonstrated in all the pigs on Day 6 p.i. These results suggest that Getah virus is mildly pathogenic for swine, which may play a role as an amplifying host in nature.