Effect of the ethanol extract of Pleurotus eryngii on bone metabolism in ovariectomized rats.

OBJECTIVE: Postmenopausal bone loss and the possible progression to osteoporosis are a major health concern. Mushrooms have been recognized as functional foods. Pleurotus eryngii extract has been reported to have estrogenic activity, suggesting that its consumption may mitigate postmenopausal bone loss. The aim of the present study was to determine the effect of supplementation with an ethanol extract of P. eryngii on bone metabolism in a postmenopausal osteoporosis rat model. METHODS: Female 12-week-old Wistar rats were subjected to either sham operation or bilateral ovariectomy. The ovariectomized rats were then subdivided into two groups: one fed the extract and the other not. Twelve weeks after surgery, indices of bone mass, bone histomorphometry, and bone mechanics were measured. RESULTS: The right femur bone mineral content and density of the ovariectomized group were significantly lower than in the Sham group, and extract supplementation did not have any significant effect on these differences. Furthermore, ovariectomy significantly increased measures of mineralizing surface and bone formation rates; again, extract supplementation again had no significant effect. CONCLUSION: Our findings suggest that the ethanol extract of P. eryngii does not alter bone metabolism in ovariectomized rats, suggesting that consumption of P. eryngii may not be beneficial in slowing bone loss after menopause.

[Chorioretinal blood flow changes following acupuncture between thumb and forefinger].

PURPOSE: To evaluate the effects of acupuncture stimuli on general circulation and chorioretinal blood flow changes, and to determine the duration of effect and the learning effect. OBJECTS AND METHODS: Twelve healthy young volunteers were divided into two groups. One had no experience of acupuncture (Non-Experience group); the other had experience of acupuncture (Experience group). Hegu (LI 4) between thumb and forefinger was acupunctured. Chorioretinal blood flow was measured via Heidelberg retina flowmeter before, during, and after acupuncture stimuli. RESULTS: In both groups, chorioretinal blood flow increased significantly during stimuli, with continuous bradycardia. The Experience group showed greater changes than the Non-Experience group. CONCLUSIONS: Chorioretinal blood flow was increased through relative parasympathetic reaction by stimulating an acupuncture point. Acupuncture is a promising adjunctive therapy for ischemic ocular diseases.

Characterization of S818L mutation in HERG C-terminus in LQT2. Modification of activation-deactivation gating properties.

We examined the mechanism(s) for HERG channel dysfunction in an S818L mutation in the HERG C-terminus using the heterologous expression system in Xenopus oocytes. Injection of S818L cRNA alone did not produce expressed currents. Coinjection of an equal amount of S818L cRNA with wild-type (WT) cRNA into oocytes did not exhibit apparent dominant-negative suppression. However, coinjection of excess amounts of S818L cRNAs with WT cRNA into oocytes decreased HERG current amplitudes and shifted the voltage dependence of activation to negative potentials, accelerated its activation and deactivation. The data suggest that S818L alone cannot form functional channels, whereas S818L subunits can, at least in part, coassemble with WT subunits to form heterotetrameric functional channels, and imply that the HERG C-terminus may contain a domain involving the activation-deactivation process of the channel. These findings may provide new insights into the structure-function relationships of the HERG C-terminus.

Roselipins, inhibitors of diacylglycerol acyltransferase, produced by Gliocladium roseum KF-1040.

Gliocladium roseum KF-1040, a marine isolate, was found to produce a series of new inhibitors of diacylglycerol acyltransferase (DGAT). Four active compounds, designated roselipins 1A, 1B, 2A and 2B, were isolated from the fermentation broth of the producing strain by solvent extraction, ODS column chromatography and preparative HPLC. The highest production of roselipins was observed when cultured in the medium containing natural sea water. Roselipins inhibit DGAT activity with IC50 values of 15 approximately 22 microM in an enzyme assay system using rat liver microsomes.

Quantitative analysis of Aconitum alkaloids in the urine and serum of a male attempting suicide by oral intake of aconite extract.

A method for the quantitation of diesterditerpene-type Aconitum alkaloids and their hydrolysis products by gas chromatography-selected ion monitoring was applied to a clinical case study. A 45-year-old male attempted suicide by oral intake of Aconitum alkaloids, which are highly intoxicant extracts of Aconitum tubers. It was estimated that he had ingested approximately 11 mg of diesterditerpene-type alkaloids but was saved by intensive gastric irrigation. Mesaconitine, aconitine, hypaconitine, and their hydrolysis products were detected in the serum on the first day only. On the other hand, some alkaloids were still detectable in the urine even six days after intoxication. Aconitum alkaloids are biotransformed, and their hydrolysis products are excreted time-dependently to the urine. The urine was a useful material to identify the toxicants in the case of aconite intoxication.

Genomic structures and chromosomal location of p91, a novel murine regulatory receptor family.

Recently, we found a novel murine cell-surface glycoprotein, designated as p91, expressed mainly in myeloid cells such as macrophages and mast cells. The molecule has six immunoglobulin-like extracellular domains, a transmembrane segment, and a cytoplasmic tail containing four immunoreceptor tyrosine-based inhibition motif (ITIM) or ITIM-like sequences, resembling the structural features of human killer-cell inhibitory receptors (KIR). Here we show that p91 comprises a polymorphic gene family, harboring one potent inhibitory-type p91 and at least two other p91 genes. Tyrosine-phosphorylated, but not nonphosphorylated, synthetic peptides matching the third ITIM and the fourth ITIM-like sequences, respectively, found in the cytoplasmic portion of p91A, the sole inhibitory-type p91, were associated with the tyrosine phosphatases, SHP-1 and SHP-2. In addition, the phosphotyrosyl peptide matching the third ITIM sequence also bound the inositol 5-phosphatase, SHIP. These results support the notion that p91A may function as an inhibitory cell-surface molecule against cell activation. The p91 genes were shown to be clustered in the proximal region of mouse chromosome 7, a syntenic position of human chromosome 19 where the genes for the KIR family are found. A human cDNA clone cross-hybridizing to a murine p91 probe was isolated from a human spleen cDNA library, and was found to code for a molecule quite similar to members of the immunoglobulin-like transcript (or ILT) family. The gene was found to be located on human chromosome 19q13.3-13.4. These results establish the existence of a novel set of potent regulatory receptors in mouse and man, similar but different from the KIR family.

1alpha,25-dihydroxyvitamin D3-24-hydroxylase (CYP24) hydroxylates the carbon at the end of the side chain (C-26) of the C-24-fluorinated analog of 1alpha,25-dihydroxyvitamin D3.

The sequential oxidation and cleavage of the side chain of 1alpha, 25-dihydroxyvitamin D3 (1alpha,25(OH)2D3) initiated by the hydroxylation at C-24 is considered to be the major pathway of this hormone in the target cell metabolism. In this study, we examined renal metabolism of a synthetic analog of 1alpha,25(OH)2D3, 24, 24-difluoro-1alpha,25-dihydroxyvitamin D3 (F2-1alpha,25(OH)2D3), C-24 of which was designed to resist metabolic hydroxylation. When kidney homogenates prepared from 1alpha,25(OH)2D3-supplemented rats were incubated with F2-1alpha,25(OH)2D3, it was mainly converted to a more polar metabolite. We isolated and unequivocally identified the metabolite as 24,24-difluoro-1alpha,25,26-trihydroxyvitamin D3 (F2-1alpha,25,26(OH)3D3) by ultraviolet absorption spectrometry, frit-fast atom bombardment liquid chromatography/mass spectroscopy analysis, and direct comparison with chemically synthesized F2-1alpha,25,26(OH)3D3. Metabolism of F2-1alpha,25(OH)2D3 into F2-1alpha,25,26(OH)3D3 by kidney homogenates was induced by the prior administration of 1alpha,25(OH)2D3 into rats. The C-24 oxidation of 1alpha,25(OH)2D3 in renal homogenates was inhibited by F2-1alpha,25(OH)2D3 in a concentration-dependent manner. Moreover, F2-1alpha,25,26(OH)3D3 was formed in ROS17/2.8 cells transfected with a plasmid expressing 1alpha,25(OH)2D3-24-hydroxylase (CYP24) but not in the cells transfected with that expressing vitamin D3-25-hydroxylase (CYP27) or containing inverted CYP27 cDNA. These results show that CYP24 catalyzes not only hydroxylation at C-24 and C-23 of 1alpha,25(OH)2D3 but also at C-26 of F2-1alpha,25(OH)2D3, indicating that this enzyme has a broader substrate specificity of the hydroxylation sites than previously considered.

Method for the simultaneous determination of Aconitum alkaloids and their hydrolysis products by gas chromatography-mass spectrometry in human serum.

This paper describes a newly developed method using gas chromatography-selected ion monitoring (GC-SIM) for the simultaneous determination of diester-diterpene type Aconitum alkaloids and their hydrolysis products in human serum. When the alkaloids were converted into their TMS derivatives, they produced a well defined peak on selected ion recording (SIR). Good linear response over the range of 100 pg to 7.5 ng was demonstrated for each alkaloid. When the alkaloids were spiked into human serum, good quantitative recovery was observed.

Determination of Aconitum alkaloids in the tubers of Aconitum japonicum using gas chromatography/selected ion monitoring.

A rapid, specific and precise method using GC/SIM was applied to separate and quantify Aconitum alkaloids in Aconitum tubers. The TMS derivatives of each alkaloid produced a well defined peak on selected ion recording (SIR). Good linear response over the range of 100 pg - 7.5 ng was demonstrated for each alkaloid. Furthermore, we investigated the changes in the contents of Aconitum alkaloids, i.e., hypaconitine, mesaconitine, aconitine, and jesaconitine, in Aconitum tubers paying special attention to the harvesting season. Mesaconitine had the highest value in all seasons. Mesaconitine and aconitine showed the highest value in the samples harvested in May 1991 while hypaconitine had the highest value among those harvested in December 1990. As to the total amount of the alkaloids, the highest value, 4190 microg/g, and the lowest value, 1881 microg/g, were observed in the samples harvested in May and September, respectively. GC/SIM was very useful for the determination of Aconitum alkaloids in the tubers.

[A case of recurrent hepatocellular carcinoma after hepatic resection surviving over five years by hepatic arterial infusion of lipiodol-anticancer drug suspension].

A 63-year-old male with four intrahepatic recurrences of surgically resected hepatocellular carcinoma was admitted to our hospital in June 1985. He underwent lateral segmentectomy of the liver in November 1983. Pathologic finding of Edmondson II with liver cirrhosis had been confirmed by the operative specimen. Sizes of four recurrent tumors were assessed by CT as 3.5 x 2.2 cm, 2.6 x 2.2 cm, 2.2 x 2.2 cm and 2.2 x 2.2 cm, respectively. During five years until July 1990, the patient was treated with hepatic arterial infusion of Lipiodol-anticancer drug suspension eight times (total 5-FU 900 mg, ADM 77 mg, MMC 73 mg, and Lipiodol 36 ml) and hepatic arterial chemoembolization of MMC microcapsules one time. In addition, two hepatic arterial infusions of CDDP (total 70 mg) were given and 5-FU (total 10 g) was administered intravenously. Partial response (PR) was obtained for 19 months. Hepatic arterial infusion of Lipiodol-anticancer drug suspension was given only once every 6 months, and he maintained a good quality of life for over four and half years. The man died in July 1990. In general, multiple intrahepatic recurrence of surgical resected hepatocellular carcinoma has a poor prognosis. Therefore it was considered that hepatic arterial infusion of this drug brought about the relatively long survival of more than five years.

[The development of immunoreactive corticotropin-releasing factor (IR-CRF) in the hypothalamus, pancreas and gastrointestinal tract of immature rats].

Corticotropin-releasing factor (CRF), a hypothalamic releasing hormone, is now known to be widely distributed in extrahypothalamic tissues such as the pancreas and gastrointestinal tract. There are, however, no data concerning the development of CRF in the gastrointestinal tract in immature animals. Using a specific RIA technique for rat CRF, we have determined the IR-CRF concentrations in acid extracts of hypothalamic, pancreatic and gastrointestinal tissues in immature rats. Gel filtration of the extracts was also undertaken in order to investigate the molecular characteristics of the IR-CRF in the digestive system. Pregnant rats were decapitated at 20 days of gestation and the male fetuses (1 day before birth) were removed by hysterotomy. The male pups were also killed by decapitation immediately after birth and at 1, 3, 7, 14, 21, 28 and 42 days postnatally. The rats older than 3 days were divided into fed and 15-h-fasted groups. Pups were weaned at 21 days. The hypothalamus, pancreas, gastric antrum, duodenum and proxymal jejunum were removed immediately after decapitation. The individual tissue fragments were homogenized in 2N acetic acid and boiled for 5 min. The homogenates were centrifuged at 15,000g at 4 degrees C for 30 min and supernatants were removed, lyophilized and stored frozen until assay. Gel filtration was performed at 4 degrees C on a Sephadex G-50 (fine) column (1.6 X 82 cm). A 3 ml of sample was applied to the column and eluted with 0.2N acetic acid containing 0.25% BSA. Fractions of 3 ml were collected and stored at -20 degrees C until assay. The reagents for RIA were purchased from Peninsula Lab., INC. (Belmont, CA.). 125I-Tyr degrees-rat-CRF was labelled by chloramine-T method. Rabbit anti-CRF serum was used at the final dilution of 1:33,000. The antigen bound antibody was separated from the free antigen by the double antibody method using goat anti-rabbit IgG serum. The sensitivity of this RIA was 2-4 pg/tube. The average coefficients of variation for interassay and intraassay were 11.2% and 9.0%, respectively. Extracts of the pancreas, duodenum and jejunum of immature rats gave dilution curves that were parallel to the rat CRF standard curve as well as that of the hypothalamus.(ABSTRACT TRUNCATED AT 400 WORDS)

[Immunological study of the actions of auxiliary antitumor agents--effects on lymphocyte transformation reaction and natural cytotoxicity activity in nude mice].

P-aminobenzoic acid-N-xyloside (K-247) and dimethyl-2- (tetrahydro-2-furanyl) ethylsulfonium-p-toluene sulfonate (GT-101) were tested their in vivo effects on both mitogen-induced lymphoproliferative reactions and natural cell-mediated cytotoxicities in BALB/c nude mice (homozygous and heterozygous) spleen lymphocytes. The animals were injected i.p. either 400 mg/kg of K-247 or 5 mg/kg of GT-101 (for 7 days consecutively). GT-101 caused a positive increase in lymphoproliferations by PHA and SPA, while the administration of K-247 had no effect on PHA-and SPA-induced lymphoproliferations. Furthermore, in a 12-hour 51Cr release assay, both drugs had no effect on the natural cell-mediated cytotoxicity against YAC-1 cells.