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Anaphalis busua (Buch-Ham ex D. Don) is traditionally used to treat cuts and wounds. In the agar dilution method, the hexane extract of the aerial part of A. busua exhibited potent antibacterial activity with a minimal inhibitory concentration of 25 μg/ml against Bacillus subtilis and 500 μg/ml against Staphylococcus aureus. For the isolation of antibacterial compounds, the hexane extract was fractionated by silica gel and Sephadex LH 20 column chromatography. The direct bioautography method was used to determine the antibacterial activity of the fractions. The active fractions were finally purified by semi-preparative HPLC on C18 Phenomenex column under gradient condition. Four flavones derivatives of 3,5-dihydroxy-6,7,8-trimethoxyflavone (1), 3,5,7-trihydroxy-6-methoxyflavone (alnusin) (2), 3,5,7-trihydroxy-8-methoxyflavone (3) and pinocembrin (4) were isolated, and their structures were established by 1H, 13C, DEPT-135, and ESI-MS spectroscopy. These compounds were isolated for the first time from A. busua. The fatty acid profile of the hexane extract was analyzed by gas chromatography-mass spectrometry (GC-MS) by silylation with N-methyl-N-trimethylsilyl-trifluoro-acetamide. Pinocembrin showed antibacterial activity with the minimum inhibitory concentration of 60 μg/mL against Bacillus subtilis and 420 μg/mL against Staphylococcus aureus.

Phenolic metabolite profiling using two dimensional paper chromatographic analysis (2 DPC) was used for assaying the complex mixture of phenolics of an aqueous ethanol aerial part extract of Cuphea ignea (Lytheraceae). A coumarin with a rare structure, namely, 7-hydroxy 3-methoxy coumarin 5-O-β-glucopyranoside was isolated from the investigated extract. The structure was elucidated by conventional methods and spectral analysis, including one and two dimensional NMR (1D and 2D NMR), as well as by interpretation of the spectra obtained by high resolution electrospray ionization mass technique (HRESIMS). The rare coumarin significantly inhibited reactive oxygen species production with an ED50 value of 6.31±1.64 μg/ml and 5.78±0.66 μg/ml as determined by the the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the oxygen radical absorption capacity (ORAC) assay respectively. The isolated coumarin presented a cytotoxic activity assessed by using the neutral red assay (NRU) against lung cancer cell line (H23) with IC50 of 40.38±2.75 μg/ml.

Phenolic metabolite profiling and identification using high performance liquid chromatography (HPLC) coupled to high resolution accurate mass spectrometry (HR-ESI-MS) with detection of negative ions was used for assaying the complex mixture of phenolics of an aqueous ethanol leaf extract of Eugeniajambos L. (Myrtaceae). Eight known polyphenolics were tentatively identified, and, in addition, three hitherto unknown flavonol-O-glycosides were detected in the extract. These unknowns were taken as the targets and isolated by means of consecutive polyamide S6, MCI gel and repeated Sephadex LH-20 column fractionation. The isolation and purification were monitored by HPLC/ESI-MS. The isolates were subsequently identified as quercetin 3-O-xylosyl-(1"' --2")-O-xyloside, myricetin 7-methylether 3-O-xylosyl-(1"' --2")-rhamnoside and myricetin 3',5'-dimethyl ether 3-O-xylosyl-(1"'--* 2")-O-rhamnoside. All known metabolites were also separated by applying the same chromatographic techniques. ESI-MS, ¹H and ¹³C NMR spectra were then recorded, completely interpreted and confirmed by HR-ESI-MS and 2D NMR spectroscopy. In order to get information about biological activities of E. jambos the extract was tested for radical scavenging activity by DPPH and ORAC assay. In addition, its cytotoxicity was assessed by the neutral red assay against non-tumorigenic HaCaT keratinocytes and the human bladder carcinoma cell line 5637.

Pilze werden als Nahrungsmittel und zu medizinischen Zwecken verwendet. Auch in der westlichen Hemisphare nimmt der Gebrauch zu. Zur Etablierung von Pilzen als Arzneimittel werden u.a. Standards fur die pharmazeutische Qualitat von Pilzen und deren Zubereitungen benotigt [1]. Agaritin (1), ein aromatisches Hydrazin, ist ein Sekundarmetabolit in Pilzen, z.B. Agaricus bisporus Imbach, dem Champignon, und Agaricus brasiliensis Wasser, dem sog. Mandelpilz, der auch als Medizinalpilz bekannt ist [2]. Agaritin wird verdachtigt, genotoxische Aktivitat zu besitzen [3], Untersuchungen kamen jedoch zu unterschiedlichen Ergebnissen [4, 5]. Der Beitrag beschaftigt sich mit der Validierung einer Methode fur die Quantifizierung von Agaritin in Pilzbiomassen, namentlich Extraktion, Aufreinigung durch Festphasenextraktion (SPE) und Messung mittels LC-MS/MS (Flussigchromatografie mit Tandem-Massenspektrometrie) im positiven Ionisationsmodus. Die Ergebnisse erweisen die Methode als selektiv, richtig und prazise. Sie konnte zur Qualitatskontrolle von pharmazeutischen Zubereitungen, welche Pilze enthalten, verwendet werden und entsprache einer Reinheitsprufung im Sinne einer Pharmakopoe. Was die Agaritin-Gehalte anbelangt, zeigten Weise Champignons mit 2027,4 mg/kg einen leicht hoheren Gehalt als Braune Champignons mit 1416,8 mg/kg. Der Mandelpilz wies den hochsten Gehalt auf (3981,2 mg/kg). In Dosenchampignons war deutlich weniger Agaritin enthalten (29,7 mg/kg). Die Dosenflussigkeit enthielt kein Agaritin. Weitere Untersuchungen zur Toxizitat von Agaritin sollten zu offiziellen Grenzwerten fuhren, falls sich die Bedenklichkeit von Agaritin bestatigt. Ein Dank geht an Prof. Dr. Andreas Link und Dr. Anja Bodtke, beide Greifswald, fur die Akquise und HR-MS-Untersuchung von Agaritin. [1] Lindequist U. Int J Med Mushr 2013; 15: 517 – 523 [2] Wisitrassameewong K et al. Saudi J Biol Sci 2012; 19: 131 – 146 [3] Shephard SE et al. Food Chem Toxicol 1998; 36: 971 – 974 [4] Walton K et al. Carcinogenesis 1997; 18: 1603 – 1608 [5] Roupas P et al. J Funct Foods 2010; 2: 91 – 98

Nine phenolics were isolated from the aqueous ethanol extract of the leaves of Caesalpinia ferrea. The isolates were characterized for the first time from that plant. The structures of all isolates (1-9) were elucidated by conventional methods, spectroscopic analysis, including 1 D and 2D NMR, and by HR-ESIMS as well. The antioxidant capacities using the ORAC method and the cytotoxic activity using the neutral red assay (NRU) for that extract and three major isolates have been evaluated. In addition, the hypolipidemic activity (in vitro and in vivo) of the extract has been assessed.

Chemical and biological investigations of the extract of Ammania auriculata (Lytheraceae) resulted in the identification of eight polyphenols (1 - 8) for the first time from this plant, including the gallotannin, 2,3,6-tri-O-galloyl-(α,β)-4C1-glucopyranose (8), for which 1D and 2D-NMR spectra were recorded and assigned for the first time. The structures of all isolates (1 - 8) were elucidated by conventional methods, spectroscopic analysis, including 1D and 2D NMR, and by HR-ESIMS as well. All of the isolated compounds were evaluated for their antioxidant activities, determined by the DPPH and ORAC methods and for their cytotoxicity against the keratinocyte cell line HaCaT using the neutral red assay (NRU) and cell cycle analysis. Compounds 1, 3, 4, 5, and 6 significantly inhibited reactive oxygen species production with ED50 values between 3.22 and 9.79 μg/ml. Compounds 1, 3, 4, and 5 showed cytotoxic activity against HaCaT cells with IC50 values between 30.7 and 84.1 μg/ml. The new galloyl glucose (8) was found not cytotoxic. Ellagitannins, 2,3-hexahydroxy-((α/β)-glucopyranose (1) and 1 -0-galloyl 2,3-hexahydroxy-(α)-glucopyranose (5) possess remarkable antioxidative and comparably weak cytotoxic activity.

During the last years the use of physical plasma for medical applications has grown rapidly. A multitude of findings about plasma-cell and plasma-tissue interactions and its possible use in therapy have been provided. One of the key findings of plasma medical basic research is that several biological effects do not result from direct plasma-cell or plasma-tissue interaction but are mediated by liquids. Above all, it was demonstrated that simple liquids like water or physiological saline, are antimicrobially active after treatment by atmospheric pressure plasma and that these effects are attributable to the generation of different low-molecular reactive species. Besides, it could be shown that plasma treatment leads to the stimulation of specific aspects of cell metabolism and to a transient and reversible increase of diffusion properties of biological barriers. All these results gave rise to think about another new and innovative field of medical plasma application. In contrast to plasma medicine, which means the direct use of plasmas on or in the living organism for direct therapeutic purposes, this field - as a specific field of medical plasma application - is called plasma pharmacy. Based on the present state of knowledge, most promising application fields of plasma pharmacy might be: plasma-based generation of biologically active liquids; plasma-based preparation, optimization, or stabilization of - mainly liquid - pharmaceutical preparations; support of drug transport across biological barriers; plasma-based stimulation of biotechnological processes.

Extraction methods of fresh plants into aqueous-fermented extracts according to German Homoeopathic Pharmacopoeia (HAB), regulation nos. 33 and 34 were evaluated. In the course of production, the extraction is accompanied by fermentation and the resulting preparation is stored for at least 6 months until further processing. In part 1 of the work, the biochemical reactions proceeding in aqueous-fermented extracts from the fresh flowering herb of Atropa belladonna var. belladonna (L.) were studied and the responsible microorganisms were identified. This second part aimed at shedding light on the phytochemical changes upon manufacture and storage. Additionally, questions were addressed at the robustness of the method by varying production conditions, its reproducibility and the comparison with ethanolic extraction. Studying 110 extracts produced from 2006 to 2009 as well as model experiments on isolated lactic acid bacteria in atropine solutions proved that the active substance atropine was stable under regular pH conditions. Interestingly, no difference between aqueous-fermented and ethanolic extracts could be found with respect to atropine concentration. In contrast, the amounts of scopoletin and kaempferol glycosides from Atropa belladonna differed depending on the extraction procedure.

Extraction methods of fresh plants into aqueous-fermented extracts according to German Homoeopathic Pharmacopoeia (HAB), regulation nos. 33 and 34 were evaluated. In the course of production, the extraction is accompanied by fermentation and the resulting preparation is stored for at least 6 months until further processing. The present work aimed at revealing the underlying biochemical reactions during manufacture and storage. In addition, the responsible microorganisms were isolated and identified. To study the robustness of the preparation method, formulation components as well as production conditions were varied. Additionally, questions were addressed at the reproducibility of the method and a comparison with an ethanolic extract was also performed. From 2006 to 2009, 110 extracts from the fresh flowering herb of Atropa belladonna var. belladonna (L.) were produced and analyzed. The results show that lactic acid bacteria (LAB) are primarily involved in the fermentation process, mainly producing lactic acid besides acetic acid and ethanol. The homofermentative Lactobacillus plantarum and the heterofermentative Lactobacillus brevis were identified as predominant lactic acid bacteria. Finally, factors for a successful fermentation are proposed.

Evaluation of the activity of an aqueous alcoholic extract obtained from the leaves of Heimia myrtifolia (Lythraceae) by determining its stimulating effect on two human osteoblastic cell lines HOS58 and SaOS-2 indicated its potential for use in the prevention and treatment of osteoporosis. In addition, the extract was found to significantly increase the mineralization of cultivated human bone cell SaOS-2, in which a strong dose-dependent increase was observed. A phytochemical investigation of the extract also confirmed that H. myrtifolia is capable of synthesizing and accumulating appreciable amounts of several phenolics, thus leading to the isolation and characterization of sixteen of these constituents. Identified among these isolates were a new natural product, 1,6-di-O-dehydrotrigalloyl-β-D-(4)C(1)-glucopyranose, and a rare natural product (this marks its second report), 5,7,4'-trihydroxy-3-methoxyflavanone (dihydrokaempferol-3-O-methyl ether). Structures of these isolates were fully elucidated on the basis of conventional methods of analysis and confirmed by ESI/MS and (1)H and (13)C-NMR analysis.

Campylobacter jejuni is one of the most common bacterial causes of diarrhoea in the industrialised world [1], being associated with the occurrence of Guillain-Barr頓yndrome (GBS) [2] and induces diseases partially through intestinal adherence [3]. With increasing reports of C. jejuni drug resistance against standard antibiotics [4] investigations into anti-adhesive agents for the prevention of bacterial infection [5] are highly significant, particularly given the possibility of avoiding resistance [6]. Different studies have shown anti-adhesive activity of different plant compounds against a variety of bacteria including C. jejuni [5]. Although two studies report on anti-adhesive effects of fungal compounds/extracts against yeast [7] and virus [8], this is the first study that describes anti-adhesive effects of basidiomycetes against any bacteria. Nineteen extracts from 9 edible and medicinal mushroom species (Calvatia gigantea, Flammulina velutipes, Inonotus obliquus, Inonotus hispidus, Inonotus nodulosus, Inonotus dryadeus, Ganoderma lucidum, Ganoderma pfeifferi, Piptoporus betulinus) were screened for anti-adhesive activity against C. jejuni using modifications of previously published anti-adhesion assays [9, 10]. The 2 ethanolic extracts derived from the fruiting bodies of Inonotus nodulosus and Inonotus obliquus showed high anti-adhesive activity (IC50 0.017mg/mL and IC50 0.021mg/mL, respectively). Moreover, the methanolic extracts of the fruiting bodies of Calvatia gigantea and Piptoporus betulinus showed, although not dose dependent, high anti-adhesive activity (81%ᱳ% and 71%ᱰ%, respectively) at 0.002mg/mL. Further investigations need to be conducted to evaluate whether isolated compounds would be candidates for the development of novel drugs against bacterial adhesion.

Four abietane diterpenoids were isolated from the methanolic extract of the roots of Meriandera benghalensis and tested for their biological activity. Cryptotanshinone (2) and 17-hydroxycryptotanshinone (4) are known metabolites however the occurrence of tanshinone IIA (1) and przewaquinone A (3) from Meriandera benghalensis is reported for the first time. The four diterpenoids were identified by MS and one- and two dimensional NMR experiments. The isolated compounds were tested for their in vitro antiproliferative activity against three human cancer cell lines (a lung cancer (A-427), a urinary bladder cancer (5637) and a breast cancer (MCF-7) cell line) and for their antibacterial effect against three Gram-positive bacterial strains. All four abietanes showed potent cytotoxic effect against all cancer cell lines (IC50 between 1 and 8 microM) as well as antibacterial effect against the bacteria tested (MIC values between 33 and 70 microM).

The first glycosylated isoferulic acid, isoferulic acid 3-O-beta-glucopyranoside, together with the new phenolics, tamarixetin 3,3'-di-sodium sulphate and dehydrodigallic acid dimetyl ester have been characterized from a flower extract of Tamarix aphylla. The structures were established on the basis of spectral data. The extract exhibited a distinct radical scavenging effect and to improve the viability of human keratinocytes (HaCaT cells). Also, the known isoferulic acid and ferulic acid which have been determined to be the major components of the investigated extract by HPLC/ESI mass spectrometric screening have been separated, characterized and evaluated as active antioxidants and as cell activity stimulating agents as well.

The present research study deals with the evaluation of sixty four methanolic and aqueous extracts of thirty Yemeni plants used in traditional medicine for their in vitro antiproliferative activity against three human cancer cell lines in a microtiter plate assay based on cellular staining with crystal violet, for their antimicrobial activity against antibiotic susceptible three Gram-positive, three Gram-negative bacterial and one fungal stains and three multiresistant Staphylococcus strains by the agar diffusion method and the determination of MIC against three Gram-positive bacteria with the broth micro-dilution assay, as well as for their antioxidant activity using the DPPH radical scavenging method. Furthermore the chemical composition of the methanolic extracts was determined by using chromatographic methods. As a result of this work, 12 Yemeni herbs namely Centaurothamus maximus, Costus arabicus, Cupressus sempervirens, Dichrocephala integrifolia, Euphorbia schimperi, Gomphocarpus fruticosus, Kanahia laniflora, Meriandera benghalensis, Pulicaria inuloides, Solanum glabratum, Tarconanthus camphoratus and Vernonia leopoldii demonstrated a noteworthy growth inhibitory effect against all cancer cell lines with IC50 values50 microg/ml. Pronounced antimicrobial activity was observed only against Gram-positive bacteria among them multiresistant bacteria with inhibition zones15 mm and MIC values500 microg/ml, by 9 plants especially Centaurothamus maximus, Cupressus sempervirens, Enicostemma verticillare, Meriandera benghalensis, Nepeta deflersiana, Pulicaria inuloides, Tarconanthus camphoratus, Teucrium yemense and Vernonia leopoldii. Moreover, the methanolic extracts of Cupressus sempervirens, Meriandera benghalensis, Pulicaria inuloides and Rhus retinorrhaea showed a remarkable radical scavenging effect at low concentrations.

The capability of an aqueous methanol extract obtained from the leaves of Feijoa sellowiana Berg. on possible prevention and treatment of osteoporosis has been examined by evaluating its stimulating effect on the two human osteoblastic cell lines HOS58 and SaOS-2. The extract was found to increase significantly the mineralization of cultivated human bone cell, whereby a clear increment (15.3 +/- 2.7%) in von Kossa positive area was determined when administering 25 microg/ml leaf extract. A phytochemical investigation of the extract has demonstrated the high phenolic content and led to the isolation and identification of twenty three of them, among which the new 3-methoxyellagic acid 4-O-beta-glucopyranoside was fully identified. All structures were elucidated on the basis of conventional analytical methods and confirmed by FTMS, 1D- and 2D-NMR data. The new compound was found to cause a significant increase of mineralized area at 20 microg/mL, while at lower concentrations the effect was not significant. However, an increase of the number of mineralized spots (nodules) at all tested concentrations of the compound was observed.

Two new bisdesmosidic triterpenoid saponins, 3-O-[alpha-L-2-O-sulphonylarabinopyranosyl]-pomolic acid-28-O-[beta-D-glucopyranosyl] ester (zygoeichwaloside H) and 3-O-[beta-D-2-O-sulphonylglucopyranosyl]-pomolic acid-28-O-[beta-D-glucopyranosyl] ester (zygoeichwaloside K) were isolated from the roots of Zygophyllum eichwaldii. The structures were established primarily on the basis of NMR spectroscopy and chemical transformations.

From the mushroom species Tricholoma populinum LANGE which shows interesting pharmaceutical effects the steroid 5α,8α-epidioxyergosta-6,22-dien-3β-ol has been isolated. The acetate of this compound crystallizes in the monoclinic space group P 21 with two molecules in the unit cell of dimensions a = 19.788, b = 7.241, c = 9.914 A, β = 103,97°. The final R-value is 0.079 for 1082 reflections. Aus Pilzen der Art Tricholoma populinum LANGE, die interessante pharmazeutische Wirkungen zeign, wurde das Steroid 5α,8α-Epidioxyergosta-6,22-dien-3β-ol isoliert. Das Acetat dieser Verbindung kristallisiert in der monoklinen Raumgruppe P 21 mit zwei Molekulen in der Elementarzelle. Die Gitterkonstanten sind a = 19.788, b = 7.241, c = 9.914 A, β = 103,97°. Der abschliesende R-Wert betragt 0,079 fur 1082 Reflexe.

Tricholoma populinum is widely distributed in Europe and North America. In the GDR 110 localities have been recorded, ranging from sea level to 470 m s.m. It is associated with many Populus spp. on acidic and neutral ground; its frequency has locally increased by recultivation of slap-dumps and waste land with poplars. Maximum of fructification is in the first decade of October. Investigations, stimulated by empiric observations on immunosuppressive effects of the mushroom in allergic diseases of man, lead to isolation of ergosterol peroxide as one substance responsible for these effects.

Maharanga bicolor, Boraginaceae, is used in the Nepalese ethnomedicine for the treatment of several diseases. In the course of screening investigations the dichloromethane extract of the roots of Maharanga bicolor was found to inhibit the growth of gram positive bacteria. Bio-assay directed fractionation led to the isolation of five active naphthazarins, deoxyalkannin (1), alkannin (2), acetylalkannin (3), alkannin beta-hydroxyisovalerate (4) and alkannin beta-acetoxyisovalerate (5). Compounds 2-5 showed antibacterial activity against multi resistant human pathogenic Staphylococcus and Enterococcus species and 1, 4 and 5 showed antiviral activity against herpes simplex virus type-1.

The present work evaluated the anticancer activity of methanol extracts from 24 plants used in Yemeni traditional medicine. To evaluate the in vitro cytotoxic potency of the investigated extracts, an established microtiter plate assay based on cellular staining with crystal violet was used with 5 human cancer cell lines: two lung cancer (A-427 and LCLC-103H), two urinary bladder carcinoma (5637 and RT-112) and one breast cancer (MCF-7) line. The methanolic extracts of Dendrosicyos socotrana, Withanina aduensis, Withania riebeckii, Dracena cinnabari and Buxus hildebrandtii exhibited the highest toxicity on all tumor cell lines with IC50 values ranging between 0.29 and 5.54 microg/ml. The extracts of Jatropha unicostata and Punica protopunica showed a moderate potency on the most tumor cell lines.

Fractions of the aqueous alcohol extracts of the rind and kernel of Brahea aramata fruits have been investigated for their activity against 5alpha-reductase type II, which is expressed predominantly in the prostate. This isozyme represents a major target for drugs against benign prostate hyperplasia (BPH) and prostate cancer. Also, a structural analysis of the phytophenolics, present in both aqueous alcohol extracts as the major constituents, has led to the isolation of five phenolics, including the new natural product, 4',6'-dimethoxy beta,4,2'-trihydroxy chalcone from the rind extract and three phenolics, including the new natural product, 1-p-hydroxybenzoyl glycerol from the kernel extract. All structures were confirmed by ESI-MS and NMR analysis.

The methanolic extracts of 20 medicinal plants from the island Soqotra/Yemen were screened with respect to their inhibitory potency against angiotensin converting enzyme (ACE), neutral endopeptidase (NEP) and aminopeptidase N (APN). Eight extracts did not show significant inhibitory activity against the enzymes tested, only Kalanchoe farinacea, Boswellia elongata and Cissus hamaderohensis inhibited all three enzymes. The most active extract was prepared from Kalanchoe farinacea characterized by low IC50 values especially for NEP and APN.

The carpophores of Paecilomyces tenuipes are known in the Orient for their strong antitumor activity. In continuation of our study on acetoxyscirpendiol (ASD, 4beta-acetoxyscirpene-3alpha,15-diol) as a cytotoxic component from this fungus, we report particularly on the mode of action of ASD in inducing apoptosis in human MOLT-4, THP-1 and Jurkat T cell leukaemia in vitro. The antiproliferative effects of ASD seem attributable to its induction of apoptosis in the cells, as it blocked the cell cycle, induced hypodiploidity and bound annexin V and also cleaved poly-(ADP-ribose) polymerase (PARP) in these cell lines. The 50% inhibitory concentrations (IC50) of ASD on MOLT-4, THP-1 and Jurkat T cells were found to be 60, 85 and 60 ng/ml, respectively. ASD arrested the cell cycle at the G1/S transition and showed hypodiploidity due to the accumulation of sub-G0 population. Annexin V binding was increased in the presence of ASD in the MOLT-4 cell line in a time-dependent manner. ASD and three of its derivatives also induced cleavage of PARP in both MOLT-4 and Jurkat T cell lines. From these data, it is suggested that ASD exerts its cytotoxic activity by inducing apoptosis in leukaemia cell lines in vitro.

The influence of the two anthracyclin antibiotics daunomycin and adriamycin on the proteome of Bacillus subtilis was investigated. They intercalate in the double helix causing strand breaks. Both compounds induce proteins related to DNA damage and oxidative stress as indicated by the induction of some members of the PerR and the DinR-regulon. The mild induction of some members of the HrcA- and the CtsR-regulon may indicate protein oxidation as well. Furthermore, an influence on the sigmaB-dependent general stress response was shown. These data show that the proteomic signature is a valuable experimental tool for a comprehensive evaluation of the action of different drugs.

Methanol water extract from rhizomes of Bergenia ligulata, a plant used in Nepalese ethnomedicine, inhibited in vitro the replication of influenza virus in a dose dependent manner and did not show virucidal activity at effective concentration. Pretreatment of cells with B. ligulata extract was shown to be most effective to prevent cell destruction. The extract inhibited viral RNA synthesis and reduced viral peptide synthesis at 10 microg/ml. The virus inhibitory effect is related to the presence of condensed tannins in the extract.

The investigation of new biological active substances from limited sources for example from marine organisms needs sensitive and evident test systems. Such a system is the glucose biosensor controlled perfusion cell culture. The glucose consumption of cells is a very sensitive parameter which allows the continuous measurement of external effects of test substances on the cells under in vivo-like conditions. Cytotoxic concentrations of active substances as well as a virus infection lowers the glucose consumption of continuously perfused cells. This effect can be monitored using a glucose biosensor. The influence of drugs and the virus infection can be observed simultaneously in the same system continuously over several days. With two substances isolated from the marine fungus Kirschsteiniothelia maritima investigations for cytotoxic and for antiviral effects are demonstrated.

The antimicrobial efficacy as well as the content of preservative agents of six commercially available grapefruit seed extracts were examined. Five of the six extracts showed a high growth inhibiting activity against the test germs Bacillus subtilis SBUG 14, Micrococcus flavus SBUG 16, Staphylococcus aureus SBUG 11, Serratia marcescens SBUG 9, Escherichia coli SBUG 17, Proteus mirabilis SBUG 47, and Candida maltosa SBUG 700. In all of the antimicrobial active grapefruit seed extracts, the preservative benzethonium chloride was detected by thin layer chromatography. Additionally, three extracts contained the preserving substances triclosan and methyl parabene. In only one of the grapefruit seed extracts tested no preservative agent was found. However, with this extract as well as with several self-made extracts from seed and juiceless pulp of grapefruits (Citrus paradisi) no antimicrobial activity could be detected (standard serial broth dilution assay, agar diffusion test). Thus, it is concluded that the potent as well as nearly universal antimicrobial activity being attributed to grapefruit seed extract is merely due to the synthetic preservative agents contained within. Natural products with antimicrobial activity do not appear to be present.

Extracts of the culture medium from the marine fungus Corollospora maritima exhibited concentration dependent antibacterial activity against Staphylococcus aureus and other microorganisms. Bioactivity-guided fractionation and purification afforded the new isobenzofuranone or phthalid type compound corollosporine.

The effects of two phenolic compounds, hispolon and hispidin isolated from fruit bodies of the basidiomycete Inonotus hispidus (Bull. ex Fr.) Karst, were investigated on the chemiluminescence response by LPS- or zymosan-activated human mononuclear cells (MNC) and on the concanavalin A-induced proliferation of spleen lymphocytes of mouse in vitro. Both compounds showed inhibitory activity in the chemiluminescence-test with an IC50 (the concentration of test compound causing 50% effect) ranging from 4.4 to 4.6 micrograms/ml (20.3 to 21.2 microM) for hispolon and0.1 to 1.5 micrograms/ml (from0.4 microM to 6.0 microM) for hispidin. Antiproliferative effects have been achieved in the lymphocyte transformation test (LTT) by hispolon with an IC50 of 3.4 micrograms/ml (15.5 microM).