Isolation of three high molecular weight polysaccharide preparations with potent immunostimulatory activity from Spirulina platensis, aphanizomenon flos-aquae and Chlorella pyrenoidosa.

This research describes the identification of three new high molecular weight polysaccharide preparations isolated from food-grade microalgae that are potent activators of human monocytes/macrophages: "Immulina" from Spirulina platensis, "Immunon" from Aphanizomenon flos-aquae, and "Immurella" from Chlorella pyrenoidosa. These polysaccharides are structurally complex and have estimated molecular weights above ten million daltons. All three polysaccharides are highly water soluble and comprise between 0.5 % and 2.0 % of microalgal dry weight. Immunostimulatory activity was measured using a transcription factor-based bioassay for nuclear factor kappa B (NF-kappa B) activation in THP-1 human monocytes/macrophages. Using this system the EC(50) values for these microalgal polysaccharides are between 20 and 110 ng/ml (about 10pM). THP-1 activation was confirmed by measuring immune cytokine mRNA induction using reverse transcriptase-polymerase chain reaction (RT-PCR). Each polysaccharide substantially increased mRNA levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha). These polysaccharides are between one hundred and one thousand times more active for in vitro monocyte activation than polysaccharide preparations that are currently used clinically for cancer immunotherapy.

Phenolic compounds from Miconia myriantha inhibiting Candida aspartic proteases.

Assay-guided fractionation of the ethanol extract of the twigs and leaves of Miconia myriantha yielded two new compounds, mattucinol-7-O-[4' ',6' '-O-(S)-hexahydroxydiphenoyl]-beta-D-glucopyranoside (1) and mattucinol-7-O-[4' ',6' '-di-O-galloyl]-beta-D-glucopyranoside (2), along with mattucinol-7-O-beta-D-glucopyranoside (3), ellagic acid (4), 3,3'-di-O-methyl ellagic acid-4-O-beta-D-xylopyranoside, and gallic acid. Complete (1)H and (13)C NMR assignments of compound 1, which possesses a hexahydroxydiphenoyl unit, were achieved using the HMBC technique optimized for small couplings to enhance the four-bond and two-bond H/C correlations. Compounds 1 and 4 showed inhibitory effects against Candida albicans secreted aspartic proteases, with IC(50) of 8.4 and 10.5 microM, respectively.

A new naphthopyrone derivative from Cassia quinquangulata and structural revision of quinquangulin and its glycosides.

A novel naphthopyrone derivative, named quinquangulone (1), has been isolated from Cassia quinquangulata, along with the known compounds quinquangulin (2) and its two glycosides (3 and 4), rubrofusarin (5) and its two glycosides (6 and 7), nor-rubrofusarin (8) and its 6-O-glucoside (9), and three stilbenes (10-12). The structure of quinquangulone was established by spectral interpretation as 5,9-dihydroxy-8-methoxy-2,9-dimethyl-6-oxo-4H,6H,9H-naphtho-[2,3-b]pyran-4-one. Reinvestigation of the NMR spectra of quinquangulin led to revision of its structure as 5,6-dihydroxy-8-methoxy-2,9-dimethyl-4H-naphtho[2,3-b]pyran-4-one (2a). The structures of two quinguangulin glycosides, 3 and 4, were also revised accordingly. Compound 2a exhibited activity against Staphylococcus aureus and methicillin-resistant S. aureus (MIC, 3.125 and 6.25 microg/mL, respectively).

Indolopyridoquinazoline alkaloid from Leptothyrsa sprucei.

The indolopyridoquinazoline alkaloid, 3-hydoxyrutaecarpine, was isolated from Leptothyrsa sprucei, along with 8-methoxypsoralen, 5-methoxypsoralen, imperatorin, isoimperatorin, kaempferol 3-O-alpha-L-rhamnopyranoside, clematine and cnidioside B. The usefulness of the gradient 1H-15N HMBC NMR spectroscopy in the structure elucidation of 3-hydroxyrutaecarpine is noted.

New indole alkaloids from the bark of Nauclea orientalis.

Four new alkaloids, nauclealines A (1) and B (2) and naucleosides A (3) and B (4), together with six known compounds, strictosamide (5), vincosamide (6), pumiloside (7), kelampayoside A, sitosterol, and sitosteryl beta-D-glucoside, were isolated from the bark of Nauclea orientalis. The structures of 1-4 were elucidated using 1D and 2D NMR spectral methods, including COSY, DEPT, HMQC, (13)C-(1)H HMBC, and (15)N-(1)H HMBC.

Dihydrochalcones from Piper longicaudatum.

Bioactivity-guided fractionation of an ethanolic extract of the leaves and twigs of Piper longicaudatum Trelease & Yunker (Piperaceae) resulted in the isolation of one new (1) and three known (2-4) dihydrochalcones. The known compounds are: 2',6'-dihydroxy-4'-methoxydihydrochalcone (2), 2',6',4-trihydroxy-4'-methoxydihydrochalcone (asebogenin) (3), and 2'-hydroxy-4'-methoxy-2'-[1-hydroxy-1-methylethyl]-2",3"-dihy- drofurano[4",5":5',6"]-3"-[2-hydroxy-5-methoxycarbonylphe- nyl]dihydrochalcone (piperaduncin B) (4). The new compound is 2'-hydroxy-4'-methoxy-2"-[2-hydroxy-5-methoxycarbonyl- phenyl]-furano[4",5":5',6']-dihydrochalcone (longicaudatin) (1). Compounds 1-4 were tested for antibacterial activity against S. aureus and methicillin-resistant S. aureus (MRSA); only compound 3 showed inhibitory activity (IC50 of 10 and 4.5 micrograms/ml, respectively).

Resolution and absolute configuration of naturally occurring auronols.

Resolution of racemic 2-benzyl-2-hydroxy-1-benzofuran-3(2H)-ones (auronols) and CD data of the ensuing enantiomers permit assessment of the absolute configuration at the single stereocenter.

Antifungal chalcones from Maclura tinctoria.

Five prenylated flavonoids, including one new natural product, were isolated from an ethanol extract of the leaves of Maclura tinctoria (L.) Gaud. The new compound has been characterized as 2',4',4,2''-tetrahydroxy-3'-[3''-methylbut-3''-enyl]chalcone (1). The known compounds were identified as 2',4',4-trihydroxy-3'-[3''-methylbut-3''-enyl]chalcone (isobavachalcone) (2), 4,2'-dihydroxy-2''-[1-hydroxy-1-methylethyl]-2'',3''-dihydrofurano[4'',5'':3',4']chalcone (bakuchalcone) (3), 4,4',5''-trihydroxy-6'',6''-dimethyldihydropyrano[2'',3'':5',6'']chalcone (bavachromanol) (4), and 5,7,3',4'-tetrahydroxy-6,8-diprenylisoflavone (6,8-diprenylorobol) (5). All the isolated compounds were evaluated against the AIDS-related opportunistic fungal pathogens, Candida albicans and Cryptococcus neoformans. Compound 2 was active against both yeasts.

7-Caffeoylsedoheptulose from Nyssa sylvatica.

7-Caffeoylsedoheptulose has been isolated from the wood of Nyssa sylvatica, along with two ellagic acid derivatives and scopoletin. Its structure was established by chemical and spectral evidence.

New antimycobacterial saponin from Colubrina retusa.

A new jujubogenin saponin was isolated from the stems of Colubrina retusa and identified as jujubogenin 3-O-alpha-L-arabinofuranosyl (1-->2)-[3-O-(trans)-p-coumaroyl-beta-D-glucopyranosyl (1-->3)]-alpha-L-arabinopyranoside (4) on the basis of chemical and spectroscopic data. The antimycobacterial activity expressed as minimum inhibitory concentration (MIC) for compound 4 was 10 microg/mL.

Antifungal jujubogenin saponins from Colubrina retusa.

Antifungal assay-guided isolation of the 95% ethanol extract of the stems of Colubrina retusa yielded jujubogenin 3-O-alpha-L-arabinofuranosyl(1-->2)-[beta-D-glucopyranosyl (1-->3)]-alpha-L-arabinopyranoside (1), which showed modest growth-inhibitory effects against Candida albicans, Cryptococcus neoformans, and Aspergillus fumigatus (MICs, 50 microg/mL). In addition, two new minor saponins, jujubogenin 3-O-alpha-L-arabinofuranosyl(1-->2)-[2-O-(trans, cis)p-coumaroyl-beta-D-glucopyranosyl(1-->3)]-alpha-L-arabinopy ranosi de (2), and jujubogenin 3-O-(5-O-malonyl)-alpha-L-arabinofuranosyl (1-->2)-[beta-D-glucopyranosyl(1-->3)]-alpha-L-arabinopyranoside (3), were obtained. Saponin 2 was marginally active against only C. neoformans, with a MIC of 50 microg/mL, while 3 was inactive. NMR spectroscopy was used extensively for the structure determination of these compounds. The previously reported ambiguity of the NMR assignments of jujubogenin saponins for carbons -26 to -29 was clarified by a comprehensive analysis of the NMR spectra of 1.

Two auronols from Pseudolarix amabilis.

Two new auronols, amaronols A (1) and B (2), were isolated from the bark of Pseudolarix amabilis, along with pseudolaric acid B (3), pseudolaric acid C (4), demethoxydeacetoxy-pseudolaric acid B (5), pseudolaric acid B-beta-D-glucoside (6), pseudolaric acid A-beta-D-glucoside (7), and myricetin (8). The structures of amaronols A and B were established by spectral data interpretation as 2,4,6-trihydroxy-2-[(3',4',5'-trihydroxyphenyl) methyl]-3(2H)-benzofuranone and 2,4,6-trihydroxy-2-[(3', 5'-dihydroxy-4'-methoxyphenyl) methyl]-3(2H)-benzofuranone, respectively. Antimicrobial testing results of the eight compounds indicated that only pseudolaric acid B was active against Candida albicans (MIC, 3.125 microg/mL; MFC, 6.25 microg/mL), while myricetin was marginally active against Trichophyton mentagrophytes (MIC, 50 microg/mL).

Effect of drying Taxus needles on their taxol content: the impact of drying intact clippings.

Evaluation of various methods of drying yew (Taxus) biomass was accomplished. The effect of the separation of the needles from clippings on the taxol content of Taxus plants was noted. Drying clippings intact preserves their taxol content.

Taxol content of stored fresh and dried Taxus clippings.

The taxol content of dried Taxus biomass was monitored monthly for 15 months. Intact and finely ground biomass was stored at room temperature (22 degrees-24 degrees) as well as under refrigeration (2 degrees-4 degrees). In addition, intact fresh clippings stored under refrigeration in sealed plastic bags for up to 10 weeks were evaluated for changes in taxol content. Analysis indicates that properly dried Taxus clippings can be stored either intact or powdered at room temperature or under refrigeration with no apparent loss of taxol content. The taxol content in fresh intact clippings was also stable for at least 10 weeks when stored under refrigeration.

Structure elucidation and thermospray high-performance liquid chromatography/mass spectroscopy (HPLC/MS) of the microbial and mammalian metabolites of the antimalarial arteether.

Microbial metabolism studies of the antimalarial drug arteether (1) have shown that arteether is metabolized to six new metabolites in addition to those previously reported (3). Large-scale fermentations with Cunninghamella elegans (ATCC 9245) and Streptomyces lavendulae (L-105) have resulted in the characterization of these metabolites primarily by two-dimensional nuclear magnetic resonance (2D-NMR) methods as 9 beta-hydroxyarteether (2), a ring rearrangement metabolite (3), 3 alpha-hydroxy-11-epi-deoxydihydroartemisinin (4), 9 alpha-hydroxyarteether (5), 2 alpha-hydroxyarteether (6), and 14-hydroxyarteether (7). Thermospray mass spectroscopy/high-performance liquid chromatographic analyses have shown that four of these metabolites (2, 5, 6, 7) are also present in rat liver microsome preparations.

Poppy seed ingestion and opiates urinalysis: a closer look.

Review of scientific literature shows that ingestion of poppy seed containing products can result in a positive urinalysis test for opiates. In many cases the amount of seeds ingested is unrealistically high or is not specified. This study is designed to correlate the amount of seeds ingested with the urinary concentration of total morphine as a function of time. Two males and two females were involved in all four protocols, which were separated by at least one week. Subjects ingested one, two, or three poppy seed rolls, each containing 2 g of Australian seeds (108 micrograms morphine/g seed) in three protocols. In the fourth protocol subjects ingested two rolls per day for four consecutive days. Urine specimens were collected for 48 h after ingestion, analyzed by RIA, EMIT, and TDx, and selected samples were confirmed by GC/MS. The data show that the highest concentrations of total morphine in urine were found 3-8 h after ingestion or in the first-void samples. Of the 264 samples collected, there were only 16 specimens that exceeded 300 ng/mL by any of the methods used for analysis with only three samples exceeding 400 ng/mL by GC/MS (406, 611, and 954 ng/mL). In all cases, the total opiates level was less than 150 ng/mL 24 h after ingestion. Following these studies, one of the subjects ingested a poppy seed cake containing 15 g seed obtained from a bakery which analyzed for 169 micrograms morphine/g seed. Urine specimens were collected over 48 h, and all specimens were analyzed by GC/MS.(ABSTRACT TRUNCATED AT 250 WORDS)