Quality evaluation of health foods containing licorice in the Japanese Market.

Focusing on licorice, a highly used raw material in health foods, quantitative analysis of functional/medicinal components and a safety and functional evaluation was carried out for herbal medicines, health food ingredients, and so-called health foods. A functional component, glabridin, was detected in herbal medicines from Glycyrrhiza glabra and G. inflata, health food ingredients, and in commercially available health foods that contain licorice. Likewise, glycyrrhizin, a medicinal component, was detected in these sources, except in licorice oil extract. Estrogen activity in vitro was detected in some of the herbal medicines, health food ingredients, and in health foods containing licorice. In the in vivo study, liver weight in ovariectomized (OVX) mice treated with licorice oil extract was significantly higher than that in OVX and sham mice in a dose dependent manner. These results suggest that excessive intake of licorice oil extract from health foods should be avoided, even though these ingredients might be beneficial for medical use in order to maintain bone health in postmenopausal women. Measurement of hepatic cytochrome P-450 (CYP) activity, reproductive organ weight, and fat and bone mass in OVX mice was considered useful for evaluating the safety and efficacy of estrogenic health food ingredients derived from herbal medicines.

Evaluation of the safety and efficacy of Glycyrrhiza uralensis root extracts produced using artificial hydroponic and artificial hydroponic-field hybrid cultivation systems.

Glycyrrhiza uralensis roots used in this study were produced using novel cultivation systems, including artificial hydroponics and artificial hydroponic-field hybrid cultivation. The equivalency between G. uralensis root extracts produced by hydroponics and/or hybrid cultivation and a commercial Glycyrrhiza crude drug were evaluated for both safety and efficacy, and there were no significant differences in terms of mutagenicity on the Ames tests. The levels of cadmium and mercury in both hydroponic roots and crude drugs were less than the limit of quantitation. Arsenic levels were lower in all hydroponic roots than in the crude drug, whereas mean lead levels in the crude drug were not significantly different from those in the hydroponically cultivated G. uralensis roots. Both hydroponic and hybrid-cultivated root extracts showed antiallergic activities against contact hypersensitivity that were similar to those of the crude drug extracts. These study results suggest that hydroponic and hybrid-cultivated roots are equivalent in safety and efficacy to those of commercial crude drugs. Further studies are necessary before the roots are applicable as replacements for the currently available commercial crude drugs produced from wild plant resources.

Purification and characterization of two metalloproteinases from squid mantle muscle, myosinase I and myosinase II.

Metalloproteinases, myosinase I and myosinase II, that hydrolyze the heavy chain of myosin, were purified from squid mantle muscle. Myosinase I does not hydrolyze other muscle proteins, casein, haemoglobin, or MCA-substrates, while II hydrolyzes tropomyosin. Both myosinase I and myosinase II gave a single protein band on SDS-PAGE with a molecular mass of 16 and 20 kDa, respectively. Their activities were inhibited by EDTA and 1,10-phenanthroline, and II was also inhibited by EGTA. They could be reactivated with some divalent cations, I was especially reactivated with Co2+ and II especially with Zn2+. The optimum pH of both activities was 7.0; the optimum temperature for both was 40 degrees C. Myosinase I hydrolyzes myosin heavy chains to produce 130 and 90 kDa fragments. The N-terminal amino-acid sequence of the 90 kDa fragment indicates that myosinase I splits the myosin heavy chain between Ala-1161 and Thr-1162 in subfragment 2. Myosinase II hydrolyzes myosin heavy chain to produce 158 and 65 kDa fragments, and it splits between Glu-1381 and Thr-1382 in LMM. Myosinases I and II are most likely related to the metabolism of myosin in vivo.

Carnosol Is a Potent Lung Protective Agent: Experimental Study on Mice.

INTRODUCTION: Oxidative stress has been implicated in various disease states and ischemia/reperfusion injury is a direct consequence of oxidative stress in lung transplantation. Because the success rate of organ transplantation in which ischemia/reperfusion is inevitable is highly influenced by oxidative stress, development of strategies to control oxidative stress would be beneficial. Here we identified natural compounds to reduce oxidative stresses in isolated mouse lungs. METHODS: We screened compounds associated with antioxidative stress in 200 plant extracts by monitoring the activities of nuclear factor erythroid 2-related factor 2 (NRF2). Compounds found to ameliorate antioxidative stress were enriched and mice were administered the extract orally every day for 1 week. Then, the lungs were isolated and cultured in the culture medium at 37 °C. Lung damage was monitored by lactate dehydrogenase (LDH) released in the culture medium. Arterial (left ventricle) blood gas levels were also monitored after hilar clamping. RESULTS: We found that Callicarpa longissima extract was rich in NRF2 activators. The responsible compounds were carnosic acid and its oxidative product, carnosol. Carnosol induced heme-oxygenase 1 (HO-1) expression, which is downstream of NRF2, more efficiently than carnosic acid. CONCLUSIONS: Lungs from mice treated with C longissima extract were less damaged than those from control mice and accompanied by HO-1 induction. These results suggest that carnosol is a candidate compound to increase the success rate of lung transplantation.

[Chemical and chemotaxonomical studies of ferns. LXXXV. Constituent variation of Microlepia marginata (2)].

A new chemotype of Microlepia marginata, P-type strain, was found in the Central districts of Japan. The two main constituents were characterized to be 2 beta,15(R),16-trihydroxy-ent-pimar-7-en-3-one (fumotoshidin A) and 3 alpha-alpha-L-arabinofuranosyloxy-15(R),16-dihydroxy-ent-pimar+ ++-7-ene (fumotoshidin arabinoside). The young fronds of this strain have reddish stripes, which is a common feature to Y-type strains also containing ent-pimarane-type glycosides.

New sesquiterpene lactones from Elephantopus mollis and their leishmanicidal activities.

The leishmanicidal compounds isolated from whole plants of Elephantopus mollis H.B.K. were identified as follows. Three new sesquiterpenoid lactones, 2,5-epoxy-2beta-hydroxy-8alpha-(2-methylpropenoyloxy)-4(15),10(14),11(13)-germacratrien-12,6alpha-olide, (4betaH)-8alpha-(2-methylpropenoyloxy)-2-oxo-1(5),10(14), 11(13)-guaiatrien-12,6alpha-olide and (4betaH)-5alpha-hydroxy-8alpha-(2-methylpropenoyloxy)-1(10),11(13)-guaiadiene-12,6alpha-olide, were isolated from Peruvian and Brazilian collections together with four known sesquiterpenoids, molephantin, elephantopin, isoelephantopin and 2-deethoxy-2beta-methoxyphantomolin. They exhibited potent in vitro leishmanicidal activities against Leishmania major. The alpha-methylene-gamma-butyrolactone moiety was found to be essential to the potent leishmanicidal effect observed.

Chemical and chemotaxonomical studies on Dicranopteris species.

Clerodane glycosides and flavonoids in Dicranopteris pedata and three varieties of D. linearis were investigated. All the ferns contained a new glycoside, (6S,13S)-6-[6-O-acetyl-beta-D-glucopyranosyl-(1-->4)-alpha-L-rhamnopy - ranosyloxy]-13-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-fucopyra nosyloxy]- cleroda-3,14-diene, as a chemical marker of this group. Flavonoids were limited to flavonol 3-O-glycosides. The ferns and isolated flavonoids are as follows; D. pedata: afzelin, quercitrin. D. linearis var. brevis: afzelin, quercitrin. D. linearis var. tenuis: quercitrin, isoquercitrin. D. linearis var. sebastiana: astragarin, isoquercitrin, rutin, kaempferol 3-O-(4-O-p-coumaroyl-3-O-alpha-L-rhamnopyranosyl)-alpha-L-rhamn opy ranosyl- (1-->6)-beta-D-glucopyranoside.

Bactericidal action of a glycoprotein from the body surface mucus of giant African snail.

1. Bactericidal action of a glycoprotein, Achacin, purified from the giant African snail, Achatina fulica Férussac, has been studied. 2. Achacin kills both gram-positive and gram-negative bacteria, but only in their growing states. 3. Achacin does not have any bacteriolytic activity. 4. The strain which has no cell wall is a little more sensitive than the native strain and the cell membrane-damaged strain. 5. Achacin was observed on the cytoplasmic membrane and on the cell wall of treated Escherichia coli by immunoelectron microscopy. 6. Achacin attacks the cytoplasmic membrane of the cell.

Molecular cloning of the antibacterial protein of the giant African snail, Achatina fulica Férussac.

An expression cDNA library was constructed with poly(A)-rich RNA extracted from the collar of the giant African snail, Achatina fulica Férussac. A 1.9-kbp cDNA clone encoding a precursor of antibacterial glycoprotein of the snail, achacin, was isolated from the cDNA expression library. The cDNA sequence contains an open reading frame with 1593-nucleotide residues. The deduced amino acid sequence of this achacin precursor starts with a 29-residue leader peptide followed by a 502-residue mature peptide (56 kDa) with four possible N-glycosylation sites, Asn-Xaa-Ser or Asn-Xaa-Thr. The Northern-blot analysis proved that the achacin precursor was specifically expressed in the tissue of snail collar and processed to mature achacin. cDNA inserts encoding achacin precursor were subcloned into expression plasmids. Three kinds of expressed polypeptides were cross-reacted with rabbit antiserum raised against achacin. The largest polypeptide (M(r) 63,000) should be the achacin precursor.

Morphological aspects of Achacin-treated bacteria.

1. The morphology of bacteria treated with the bactericidal glycoprotein, Achacin, purified from the giant African snail, Achatina fulica Férussac, has been studied. 2. Achacin lengthens the bodies of Escherichia coli by three to seven times. 3. Achacin damages the surface of Staphylococcus aureus and sinks the cytoplasmic membranes into the cytoplasm. 4. Achacin causes neither the leakage nor the destruction of cells.