Volume 60, Issue 9-10

Trichosporon cutaneum R57 and its ʟ-ethionine resistant mutant NZ94 strain were investigated. The amino acid analyses of cell content of both strains were carried out. The pool of free methionine in the mutant strain is enhanced 16.5 times. The total amount of sulphurcontaining amino acids in the mutant cells was significantly increased from 36.8 in the wild strain to 113.4 mg/g protein in the mutant strain. In the process of mutant strain cultivation there was found a high excretion of free methionine (259 μg/ml) in the medium. It was shown that the amino acid content of both wild and mutant strains would be helpful for formulating of new improved animal nutritional diets.

Several species of Nicotiana and Solanum and further members of the Solanaceae have been examined for their exudate flavonoids. Most of the aglycones are widespread flavonols, but rare and unusual flavonols were also found, e.g. in exudates of Physalis and Solanum species. Flavones occur throughout the family, but flavanones are rare. Our data are presented in comparison to previous results. The chemodiversity of the observed structures is discussed in relation to literature reports. Morphological and systematic aspects are briefly addressed.

Three members of the tribe Gnaphalieae and six members of the tribe Inuleae (Asteraceae) were analyzed for their exudate flavonoids. Whereas some species exhibit rather trivial flavonoids, others produce rare compounds. Spectral data of rare flavonoids are reported and their structural identification is discussed. 6-Oxygenation of flavonols is a common feature of two Inula species and Pulicaria sicula. By contrast, flavonoids with 8-oxygenation, but lacking 6-oxygenation, are common in two out of three Gnaphalieae species examined. In addition, B-ring deoxyflavonoids are abundantly present in the leaf exudates of Helichrysum italicum (Gnaphalieae). These distinctive features of the two Asteraceae tribes are in agreement with previous flavonoid surveys of these and related taxa.

From the leaves of Chamaecyparis obtusa several biflavones were isolated and identified, namely: sciadopitysin, ginkgetin, isoginkgetin, podocarpusflavone B, 7,7″-O-dimethylamentoflavone, bilobetin, podocarpusflavone A, and 7-O-methylamentoflavone. The presence of amentoflavone and hinokiflavone was also confirmed. The composition of biflavones in other Chamaecyparis species D Ch. lawsoniana, Ch. thyoides D and cultivar varieties D Ch. pisifera “Squarrosa”, Ch. pisifera “Boulevard” D was compared using the HPLC method. It was stated, that podocarpusflavone A and 7-O-methylamentoflavone in addition to amentoflavone and hinokiflavone may be classified as chemotaxonomic markers of the genus Chamaecyparis

The in vitro antibacterial and antifungal activities of demethoxyviridin and some synthetic analogues were evaluated by the agar diffusion method. The minimum inhibitory concentrations (MIC) of the active compounds were also determined by the agar dilution method. Demethoxyviridin (1) showed moderate antibacterial activity against most of the strains tested. 1α-Hydroxydemethoxyviridin (3) showed antibacterial activity and the most potent in vitro antifungal activity with MIC of 20 μg/ml (0.062 mm) against Aspergillus niger, A. fumigatus, A. flavus, A. parasiticus, Fusarium solani, F. graminarum, Geotrichum candidum whereas 5′-methylfuro-(4′,3′,2′-4,5,6)androst-5-ene-3,17-dione (7) exhibited very weak antifungal activity against Candida albicans only.

The antibacterial activity of coumarin per se and other 45 coumarin derivatives was tested against strains of Bacillus cereus MIP 96016, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Staphylococcus aureus ATCC 25923. The inhibitory effects of coumarins were affected by their substitution patterns. Osthenol (44) showed the most effective antibacterial activity against Gram-positive bacteria with MIC values ranging between 125 and 62.5 μg/ml. These results suggested that the prenyl chain of 44 at position 8 and the presence of OH at position 7 of the benzenic ring are required for the antibacterial activity against these strains.

Simaba ferruginea (Simaroubaceae) is a Brazilian medicinal plant used in traditional medicine to treat several ailments, including gastric ulcers, fever, diarrhea, and dolorous and inflammatory processes. This study examines the chemical composition and antiulcerogenic effects of rhizomes from this plant. Bioassay-guided fractionation led to the isolation of two bioactive indole alkaloids called canthin-6-one (1) and 4-methoxycanthin-6-one (2). The alkaloid fraction and both alkaloids demonstrated potent antiulcerogenic effects when evaluated in gastric lesion-induced animals, as well as significant antinociceptive activity in mice. These results confirm and justify the popular use of S. ferruginea against gastric ulcers and dolorous processes.

The smoke of Peganum harmala seeds is traditionally used in Iran as a disinfectant agent. The aim of this study was to determine the antimicrobial activity of two smoke condensates from Peganum harmala seeds. Furthermore the composition of smoke preparations was studied using gas chromatography and mass spectroscopy analysis. The most prevalent compound detected in a dichloromethane extract was harmine. Standard harmine as well as the dichloromethane extract showed antimicrobial activity against all test strains. Harmine was not detected in an n-hexane extract and we did not observe antimicrobial activity from this smoke preparation at the tested concentrations.

Fractionation with n-hexane/ethyl acetate (1:1 v/v) by open column chromatography of the oleoresin from Pinus oocarpa Schiede yielded two diterpenes, pimaric acid (1) and dehydroabietic acid (5), the sesquiterpene longifolene (3) and a diterpenic mixture containing pimaric acid (1), isopimaric acid (4) and dehydroabietic acid (5). Subsequently, the isolated compounds, the mixture of 1, 4 and 5, the oleoresin and the dehydroabietic acid methyl ester (2), were tested in vitro against epimastigotes of Trypanosoma cruzi, the causative agent of Chagas disease. The most active compounds were 1, 3 and the oleoresin, being as active as nifurtimox, a drug effective in the treatment of acute infection by American trypanosomiasis and used in this work as positive control.

The isolation, structure determination, syntheses and biological activity of N b -acetyltryptamine and three proline containing dioxopiperazines, cyclo(S-Pro-S-Leu) (2), cyclo(S-Pro- S-Ile) (3), and cyclo(S-Pro-S-Val) (4), from cultures of Rhizoctonia solani Kuhn are reported here for the first time. Despite the small amounts isolated, the absolute stereochemistry of these naturally occurring dioxopiperazines was established by 1 H NMR using for the first time the chiral solvating agent (R)-(D)-2,2,2-trifluoro-1-(9-anthryl)ethanol.

A new modified stilbene dimer, diptoindonesin D (1), was isolated from the acetone extract of the tree bark of Hopea dryobalanoides, together with seven known compounds, parviflorol (2), (D)-balanocarpol (3), heimiol A (4), hopeafuran (5), (+)-α-viniferin (6), vaticanol B (7) and (D)-hopeaphenol (8). Cytotoxic properties of compounds 1-8 were evaluated against murine leukemia P-388 cells. Compound 8 was found to be the most active with IC 50 of 5.7 μm

The aqueous-ethanolic extract (AEE) of high altitude Podophyllum hexandrum has earlier been reported to render a radioprotective effect against lethal gamma radiation in in vitro model. AEE has also been reported to possess metal chelating and DNA protecting properties. The present study was undertaken to isolate and characterize the bioactive principle present in AEE and investigate its role in radiation protection. A novel molecule was found to be present in AEE and was assigned as 3-O-β-ᴅ-galactoside of quercetin by acid hydrolysis, LC-MS, LC-APCI-MS/MS and 13 C NMR spectra. Various biological activities were investigated at in vitro level. The antioxidant potential of AEE in lipid and aqueous phase was determined against numerous stresses. AEE was found to be significantly (p < 0.05) protective, i.e., against Fe 2+ and Cu 2+ -induced linoleic acid degradation, respectively. Radiationinduced lipid oxidation studies revealed that AEE maximally works at a [lignan]/0.25 kGy ratio 400 (ratio of concentration of AEE divided by the radiation dose, i.e., 0.25 kGy) and no drug-induced lipid oxidation at all concentrations tested was found. In a time-dependent study, total antioxidant activity was maximally exhibited at 1 mg/ml. The site-specific and non-site-specific deoxyribose degradation assay exhibited a dose-dependant hydroxyl scavenging potential of AEE (0.05-500 μg/ml). The anti-lipid peroxidation ability of AEE against radiation (0.25 kGy)-induced lipid peroxidation was higher in case of neural tissue homogenate as compared to kidney homogenate [activity ratio: 0.039 (brain) < 0.24 (kidney)]. The protein protection study using bovine serum albumin was also done for two time intervals (2 h and 4 h) and significant (p < 0.05) protection was observed at 500 μg/ml (> 97%). This study implies that 3-O-β-ᴅ-galactoside present in AEE renders radioprotection by protecting lipids, proteins in renal and neural model system against supra-lethal (0.25 kGy) gamma radiation.

Liriomyza trifolii (Burgess), the American serpentine leafminer fly, is a well-known serious pest in the world. This insect species attacks plants of more than 21 families including Solanaceae plants. A sweet pepper, Capsicum annuum (Solanaceae), on mature stage, however, shows resistance to this leafminer fly. This resistance is based on the ovipositional deterrent in the sweet pepper leaf against the fly species. Based on the bioassay guided fractionation, phytol [(2E)-3,7,11,15-tetramethyl-2-hexadecen-1-ol] was isolated and identified as an ovipositional deterrent against this insect species. The yield of this compound was 815 μg/g fresh leaf of C. annuum. This compound completely deterred the females from laying their eggs on host plant leaves treated at 35.2 μg/cm2.

Green leaf volatiles (GLV), a series of saturated and monounsaturated six-carbon aldehydes, alcohols, and esters are emitted by plants upon mechanical damage. Evidence is increasing that intact plants respond to GLV by activating their own defense mechanisms, thus suggesting that they function in plant-plant communication. The present paper demonstrates that exposure of maize plants to naturally occurring GLV, including (Z)-3-, (E)-2- and saturated derivatives, induce the emission of volatile blends typically associated with herbivory. Position or configuration of a double bond, but not the functional group of the GLV influenced the strength of the emissions. (Z)-3-Configured compounds elicited stronger responses than (E)-2- and saturated derivatives. The response to (Z)-3-hexen-1-ol increased linearly with the dose between 200 and 1000 nmol per plant. Not only the naturally occurring (E)-2- hexenal, but also (E)-2-pentenal and (E)-2-heptenal induced maize plants, although to a lesser extend. Externally applied terpenoids [(3E)-4,8-dimethyl-1,3,7-nonatriene, β-caryophyllene, and (E)- β-farnesene] did not significantly increase the total amount of inducible volatiles in maize. Of three tested maize cultivars Delprim and Pactol responded much stronger than Attribut. Recovery experiments in the presence and absence of maize plants demonstrated that large proportions of externally applied GLV were assimilated by the plants, whereas (3E)-4,8-dimethyl-1,3,7-nonatriene was recovered in much higher amounts. The results furthermore suggested that plants converted a part of the assimilated leaf aldehydes and alcohols to the respective acetates. We propose that GLV not only can alert neighboring plants, but may facilitate intra-plant information transfer and can help mediate the systemic defense response in a plant.

Algal preparations from Acetabularia crenulata were analyzed for their fatty acid composition to establish the suitability of this alga as a model to study fatty acid oxidation and oxylipin biosynthesis. The work was based on two goals. The first goal of this study was to determine the contribution of fatty acids from contaminating bacteria and how this influenced the total fatty acid composition of cell homogenates of A. crenulata collected in the wild as compared to specimens cultured in sterile conditions. The major fatty acids detected for both specimens were palmitic (C16:0), palmitoleic (C16:1n-7), oleic (C18:1n-9), linoleic (C18:2n-6), linolenic (C18:3n-3), and octadecatetraenoic acid (C18:4n-3). Significant amounts of odd-chain fatty acids common to bacteria were not detected in either sample. Furthermore, branched-chain fatty acids, typical bacterial biomarkers, were not detected in either sample. Data suggest that bacteria do not greatly contribute to the total fatty acid pool of A. crenulata. The second goal was to compare the fatty acid composition of cell homogenates with that of isolated chloroplasts. Comparatively speaking palmitoleic and octadecatetraenoic acid were found at significantly lower concentrations in the chloroplast whereas oleic and linolenic acid were found at significantly higher amounts in this organelle. Furthermore, the amount of hexadecatrienoic acid (C16:3), a fatty acid commonly esterified to monogalactosyldiacylglycerol (MGDG; lipid present at high concentrations inside the chloroplasts of algae), was present at very low concentrations in these plastids (0.7%). Typically green algal follow the “prokaryotic pathway” for MGDG biosynthesis where C18:3 is esterified at the sn-1 position of the glycerol backbone and C18:3 or C16:3 at the sn-2 position, making C16:3 a major fatty acid inside chloroplasts. Interestingly, our results suggest that chloroplasts of A. crenulata appear to follow the “eukaryotic pathway” for MGDG biosynthesis where C18:3 is both at the sn-1 and sn-2 position of MGDG. Taking into account the exceptions noted, the fatty acid composition for A. crenulata is similar to that reported for most chlorophytes.

Using Ca 2+ -dependent photoprotein aequorin-transformed tobacco BY-2 cell suspensions, the sugar-induced increase in cytosolic free Ca 2+ concentration ([Ca2+] cyt ) was investigated by measuring the luminescence intensity. When 0.5 ᴍ sucrose or some other sugars were fed to the cells, strong and transient luminescence was observed. Salts or sugar analogues didn’t show this effect. In addition, the intensity of sucrose-induced aequorin luminescence was gradually enhanced when cells were exposed to sugar-starvation. This was observed with the concurrent expression of the sucrose/H + co-transporter, NtSUT1A. The [Ca 2+ ] cyt increase may initiate Ca 2+ -signaling leading to the expression of genes related to biosynthesis of storage carbohydrates in a sink organ. The sugar-signaling may play an important role in the conversion on nutritional stage of plant tissue, source organ to sink organ.

Protein kinases are involved in signal transduction for environmental stress responses. In response to drought and salinity, a 48-kDa protein kinase (AAPK; abscisic acid-activated protein kinase (AAPK) in guard cells is activated by abscisic acid (ABA) and phosphorylates several targets such as the carboxy-terminus of inward-rectifying K + channel and heterogeneous mRNA binding protein to adopt to the changing environment. The AAPK expressed specifically in guard cells, and recombinant AAPK was phosphorylated only with the extract from ABA-treated guard cells but not from untreated cells. This indicates the presence of an AAPK kinase (AAPKK), which is activated by ABA and phosphorylates AAPK preceding the activation of AAPK. Both AAPK and AAPKK are involved in the protein kinase cascade for the rapid ABA-signaling.

Economical breeding is important to obtain maximum gain from the breeding in the animal sector. The economic loss has to be eliminated or should be minimized. The liver fluke, Fasciola hepatica, present mostly in sheep and dairy cattle affect the yield of animals and even cause their death. To eliminate or minimize the impact of these parasites on the animals, it is important to understand the genetic diversity of the liver fluke populations and the relationship between parasite and host at regional bases. This research was carried out to determine diversity by sequence analysis of the mitochondrial ND1 gene and ribosomal ITS1 region.

The shoulder gland secretions of captive males of the Indian flying fox (Pteropus giganteus), the little golden-mantled flying fox (P. pumilus), the island flying fox (P. hypomelanus), and the large flying fox (P. vampyrus) were examined by gas chromatography-mass spectrometry. Sixty-five compounds, including hydrocarbons, carboxylic acids, alcohols, aldehydes, ketones, esters, and amides, were identified among the four species. Many of these compounds, such as squalene, cholesterol, and C 5 -C 16 straight- and branched-chain carboxylic acids, are typical of tetrapod epidermal products. Aldehydes, which were detected in all four Pteropus species, and some straight- and branched-chain ketones, which were detected in P. hypomelanus and P. pumilus, are known from other mammalian skin glands. Acetophenone, 4-acetoxyacetophenone, and 4-hydroxyacetophenone were observed in P. pumilus; the last compound comprised 37.1% of the total ion current. 2,3-Butanediol, a prominent component (5.2-19.3%) in the secretions of P. giganteus, P. hypomelanus, and P. pumilus, and C 10 and C 12 isopropyl esters and C 10 -C 14 1-methylbutyl esters, observed in P. hypomelanus and P. vampyrus, have not previously been reported from vertebrates. α-Methyl-4-methoxybenzyl alcohol and dihydro-5-phenyl-2(3H)-furanone, from P. giganteus and P. pumilus, are new natural products. 1-Chloro-3-methyl-2-butene, another new natural product, and five C 5 compounds exhibiting a similar isoprenoid structure were observed in P. giganteus. Striking contrasts were observed in the chemical profiles of the species we examined, with even general chemical classes differentially represented among them.

Substrate kinetic properties of cytochrome oxidase in rat liver, kidney, brain and heart mitochondria were examined using ascorbate + N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD) as the electron donor system. Analysis of the substrate kinetics data revealed tissuespecific expression of kinetic components exhibiting differences with respect to Km, Vmax and Kcat/Km values. Regression analysis data suggest that the enzyme activity may be regulated in a tissue-specific manner.

We compare the adsorption behavior of high density lipoproteins (HDL) and low density lipoproteins (LDL) on “fibrogenic” and “nonfibrogenic” mineral dusts. The adsorption tests with bovine lipoprotein concentrate and human serum produced the following results: 1) All seven examined fibrogenic dusts (SiO 2 DQ12, SiO2 F600, silica, graphite, TiC, kaolin, talc) adsorbed significantly more high density lipoproteins (HDL), than the five examined nonfibrogenic (inert) dusts (TiO 2 , SnO 2 , Al 2 O 3 , Fe 2 O 3 , Fe 3 O 4 ). This different behavior was particularly conspicuous in the presence of competing adsorbates (serum proteins). 2) In contrast, the adsorption of LDL did not correlate with the fibrogenicity of the mineral dusts. 3) The known silicosis-protective substance polyvinylpyridine-N-oxide inhibits the HDL adsorption of α-quartz. These results indicate that the adsorption of HDL could have a causal relationship with the triggering of a fibrotic reaction. The adsorption on the surface of fibrogenic dust particles provides an exceptional opportunity for the intake of HDL by macrophages. During the phagocytosis of the inhaled dust particles, the HDL adsorbed on the surface of the particles could be taken up by macrophages regardless of the receptor. There the HDL particles and/or compounds associated with them, such as lecithin-cholesterol-acyltransferase, could stimulate the macrophages to release fibrogenic mediators by some yet unknown mechanism.

During a co-precipitation of cholesterol (Chol) and slight amounts of polycyclic aromatic hydrocarbons (PAHs) or aminoazo dyes (AZOs) in aqueous albumin solution, complex particles are formed; on their surfaces apolipoproteins with an amphipathic α-helix (e.g. apoA-I) are more or less firmly adsorbed. An efficacy index can be calculated from the strength of the hydrophobic interactions between apoA-I and the [Chol/PAH]- or [Chol/AZO]-complex, and the solubility of the PAH or AZO in an aqueous medium, which correlates to the carcinogenicity of these compounds. A short-term test for PAHs and AZOs is described, in which the efficacy index can be determined in the simplest manner without any great expenditure on equipment. The previous results suggest that the parent compounds of the PAHs and AZOs can be involved in a specific interaction with cholesterol-domains of the plasma membrane of a cell. The changes in membrane fluidity and architecture caused by these specific interactions could modulate the distribution and/or activity of membrane proteins which are critical to the regulation of cellular proliferation.

The water proton relaxation rate enhancement of Mn(II) bound to bovine serum albumin (BSA) and the association constant for manganese to BSA have already been determined, but such determinations have not been done for human serum albumin (HSA) and other human serum proteins and also for human serum. In this work, NMR T 1 values in aqueous solutions of serum proteins and serum were measured versus increasing concentration of Mn(II). Proton relaxation rate enhancements (ε*) caused by different manganese concentrations were determined for each solution and 1/ε* was fitted against concentrations of Mn(II). Proton relaxation rate enhancements (ε b ) of Mn(II) bound to albumin, γ-globulin, (α+β)- globulins and serum were found to be 13.69, 3.09, 8.62, and 10.87, respectively. Free and bound manganese fractions, resulted from each addition of Mn(II) to the sample, were determined by using corresponding (ε*) and the ε b values. Association constants for Mn(II) to HSA and γ-globulin were calculated as 1.84 x 10 4 ᴍ -1 and 2.35 x 10 4 ᴍ -1 , respectively. Present data suggest that the proton relaxation rate enhancement of Mn(II) in serum is caused by Mn(II) bound to various serum constituents. Data also suggest that association constants for Mn(II) to γ-globulin are nearly the same as that to HSA.