Cyclic digene system as a control element of a bacterial biosensor

The Escherichia coil JC1 58(pCIA12/pG FK5) strain carrying a cyclic digene system with a negative feedback on the pCIA12 plasmid reacting to the DNA damage by changing the synthesis level of reporter genes-GFP and beta-galactosidase-was tested. The acquired phenotype was inherited by the next generations after the removal of the genotoxic action when the concentration of the DNA-damaging compounds was above the threshold level. A potential has been shown for the application of bacterial biosensors to monitor the presence of genotoxicants in the environment and to test the consequences of short-term exposures to toxic compounds.     

Noscapinoids with anti-cancer activity against human acute lymphoblastic leukemia cells (CEM): a three dimensional chemical space pharmacophore modeling and electronic feature analysis

We have identified a new class of microtubule-binding compounds—noscapinoids—that alter microtubule dynamics at stoichiometric concentrations without affecting tubulin polymer mass. Noscapinoids show great promise as chemotherapeutic agents for the treatment of human cancers. To investigate the structural determinants of noscapinoids responsible for anti-cancer activity, we tested 36 structurally diverse noscapinoids in human acute lymphoblastic leukemia cells (CEM). The IC₅₀ values of these noscapinoids vary from 1.2 to 56.0 μM. Pharmacophore models of anti-cancer activity were generated that identify two hydrogen bond acceptors, two aromatic rings, two hydrophobic groups, and one positively charged group as essential structural features. Additionally, an atom-based quantitative structure–activity relationship (QSAR) model was developed that gave a statistically satisfying result (R ² = 0.912, Q ² = 0.908, Pearson R = 0.951) and effectively predicts the anti-cancer activity of training and test set compounds. The pharmacophore model presented here is well supported by electronic property analysis using density functional theory at B3LYP/3-21*G level. Molecular electrostatic potential, particularly localization of negative potential near oxygen atoms of the dimethoxy isobenzofuranone ring of active compounds, matched the hydrogen bond acceptor feature of the generated pharmacophore. Our results further reveal that all active compounds have smaller lowest unoccupied molecular orbital (LUMO) energies concentrated over the dimethoxy isobenzofuranone ring, azido group, and nitro group, which is indicative of the electron acceptor capacity of the compounds. Results obtained from this study will be useful in the efficient design and development of more active noscapinoids.     

Metabolic changes of salicylic acid-elicited Catharanthus roseus cell suspension cultures monitored by NMR-based metabolomics

The effect of salicylic acid (SA) on the metabolic profile of Catharanthus roseus suspension cells throughout a time course (0, 6, 12, 24, 48 and 72 h after treatment) was investigated using NMR spectroscopy and multivariate data analysis. When compared to control cell lines, SA-treated cells showed a high level of sugars (glucose and sucrose) up to 48 h after treatment, followed by a dynamic change in amino acids, phenylpropanoids, and tryptamine. Additionally, one compound—2,5-dihydroxybenzoic-5-O-glucoside—was detected solely in SA-treated cells.     

Production of Manoalide and Its Analogues by the Sponge <Emphasis Type="Italic">Luffariella variabilis</Emphasis> Is Hardwired

The Great Barrier Reef sponge Luffariella variabilis (Poléjaeff 1884) produces a range of potent anti-inflammatory compounds as its major metabolites. These major metabolites—manoalide monoacetate, manoalide, luffariellin A and seco-manoalide—were monitored temporally and spatially to quantify the potential yield from wild harvest or aquaculture. Production of the major metabolites was hardwired at the population level with little variation in space and time over meters to tens of kilometers in the Palm Islands, Queensland, Australia. Manoalide monoacetate (35 to 70 mg g⁻¹ dry weight of sponge) was consistently the most abundant compound followed by manoalide (15 to 20 mg g⁻¹ dry weight). Luffariellin A and seco-manoalide were 10 to 70 times less abundant and varied between 0 and 3 mg g⁻¹ dry weight. On a larger spatial scale, L. variabilis from Davies Reef and Magnetic Island contained the same rank order and yields of compounds as the Palm Islands, indicating a generality of pattern over at least 100 km. The “hardwiring” of metabolite production at the population level by L. variabilis was also reflected in the lack of any inductive effect on metabolite production. In addition, individually monitored sponges produced fixed ratios of the major metabolites over time (years). However, these ratios varied between individuals, with some individuals consistently producing high levels of manoalide and manoalide monoacetate, providing the potential for selection of high-yielding stocks.     

DNA amounts in the nuclei ofParamecium aurelia andTetrahymena pyriformis

DNA amounts have been determined in the micronuclei and macronuclei of 8 strains ofParamecium aurelia and 6 strains ofTetrahymena pyriformis. In the case ofTetrahymena a distribution of values for the amount of DNA in the macronuclei of all the strains was observed but the lowest values were approximately the same, viz. 1.17×10⁻¹¹ g. There are two groups of strains in relation to micronuclear DNA values ofTetrahymena, one giving an average of 0.36×10⁻¹² g and the other 0.815×10⁻¹² g. The ratio of MIC/MAC DNA varies in the two groups.Paramecium again has a range of macronuclear values within each stock—lowest value 2.51×10⁻¹⁰ g—and the micronuclear values are similar in all stocks—approximately 0.613×10⁻¹² g. The ratio of MIC/MAC DNA is similar in each stock.—The haploid genome values calculated from these data show excellent agreement with the values obtained by DNA renaturation studies. Supported by a Research Grant B/SR/8276 from the Science Research Council. The Vickers densitometer was purchased with a grant from the Medical Research Council.     

Winemaking from gaglioppo grapes with hybrid strains of<Emphasis Type="Italic">Saccharomyces</Emphasis>

The fermentative behavior of two hybrid wine yeast strains (first-generation hybrid-strain 12 233×6167—obtained by hybridization of the cryotolerant strainS. bayanus 12 233 with the mesophilic strainsS. cerevisiae 6167, and TT254×6392 arising by hybridization of the thermotolerant strainS. cerevisiae TT254 with the mesophilic strainS. cerevisiae 6392) was compared with that of a commercial wine yeast strainS. cerevisiae K1 in must from black grapes of the Calabrian variety Gaglioppo. The goal was to obtain wines with a high content of ‘polyphenols’ from a grape must with a limited phenolic content such as the Gaglioppo must. The progress of the winemaking was estimated according to residual sugars; at the end of fermentation, the wines were decanted, bottled and principal physico-chemical characteristics determined. Our results point to the possibility to select wine yeasts (significantly differing in the above parameters) by their ability to interact with phenolic compounds.     

Inhibition of lipoxygenase affects induction of both direct and indirect plant defences against herbivorous insects

Herbivore-induced plant defences influence the behaviour of insects associated with the plant. For biting–chewing herbivores the octadecanoid signal-transduction pathway has been suggested to play a key role in induced plant defence. To test this hypothesis in our plant—herbivore—parasitoid tritrophic system, we used phenidone, an inhibitor of the enzyme lipoxygenase (LOX), that catalyses the initial step in the octadecanoid pathway. Phenidone treatment of Brussels sprouts plants reduced the accumulation of internal signalling compounds in the octadecanoid pathway downstream of the step catalysed by LOX, i.e. 12-oxo-phytodienoic acid (OPDA) and jasmonic acid. The attraction of Cotesia glomerata parasitoids to host-infested plants was significantly reduced by phenidone treatment. The three herbivores investigated, i.e. the specialists Plutella xylostella, Pieris brassicae and Pieris rapae, showed different oviposition preferences for intact and infested plants, and for two species their preference for either intact or infested plants was shown to be LOX dependent. Our results show that phenidone inhibits the LOX-dependent defence response of the plant and that this inhibition can influence the behaviour of members of the associated insect community.     

LC-NMR: a new tool to expedite the dereplication and identification of natural products

The rapid identification of known or undesirable compounds from natural products extracts — “dereplication” — is an important step in an efficiently run natural products discovery program. Dereplication strategies use analytical techniques and database searching to determine the identity of an active compound at the earliest possible stage in the discovery process. In the past few years, advances in technology have allowed the development of tandem analytical techniques such as liquid chromatography mass spectrometry (LC-MS), LC-MS-MS, liquid chromatography nuclear magnetic resonance (LC-NMR), and LC-NMR-MS. LC-NMR, despite its lower sensitivity as compared to LC-MS, provides a powerful tool for rapid identification of known compounds and identification of structure classes of novel compounds. LC-NMR is especially useful in instances where the data from LC-MS are incomplete or do not allow confident identification of the active component of a sample. LC-NMR has been used to identify the marine alkaloid aaptamine as the active component in an extract of the sponge Aaptos sp. This extract had been identified as an enzyme inhibitor by a high throughput screening (HTS) effort. Isolated aaptamine exhibited an IC₅₀=120 μM against this enzyme. Strategies for the identification of aaptamine and for the use of LC-NMR in a natural products HTS program are discussed. Journal of Industrial Microbiology & Biotechnology (2000) 25, 342–345. Received 30 March 2000/ Accepted in revised form 03 July 2000     

Response of the mink reproductive system to hormonal stimulation in october as a prognostic criterion of folluculogenesis and fertility

The responsess of the mink reproductive systems in October to a single intramuscular injection of chorionic gonadotropin (CG, Profasi R, Italy) at doses of 10, 20, 50, and 100 IU was studied. The estrous cycle state, folliculogenesis, and reproductive parameters of the female minks were estimated. To evaluate functional polymorphism in the reproductive system of the females in response to the CG injection, a dose of 20 IU was chosen. On the sixth day after administration of this dose of CG, the females (n = 185) were distributed according to the stages of the estrous cycle as follows: anestrus—71 animals (38.4%), anestrus-proestrus—37 (20.0%), proestrus—30 (16.2%), proestrus-estrus—14 (7.6%), estrus—33 (17.8%). In the females of the “anestrus-proestrus” and “proestrus” groups as compared with the other groups as well as with control, a statistically significantly higher fertility was revealed. The study of folliculogenesis in the females with different parameters of the estrous cycle in November demonstrated the high level of fertility to be due to ability of these females to produce, under influence of the CG injection, the maximal number of mature follicles. A high heritable responsiveness of the reproductive system of the females to the CG injection (coefficient of realized heritabilityh = 0.64; the calculation made on the data for the sisters) indicates that this method may be an additional criterion in the selection of females for fertility.     

Two Approaches to the Study of the Origin of Life

This paper compares two approaches that attempt to explain the origin of life, or biogenesis. The more established approach is one based on chemical principles, whereas a new, yet not widely known approach begins from a physical perspective. According to the first approach, life would have begun with—often organic—compounds. After having developed to a certain level of complexity and mutual dependence within a non-compartmentalised organic soup, they would have assembled into a functioning cell. In contrast, the second, physical type of approach has life developing within tiny compartments from the beginning. It emphasises the importance of redox reactions between inorganic elements and compounds found on two sides of a compartmental boundary. Without this boundary, “life” would not have begun, nor have been maintained; this boundary—and the complex cell membrane that evolved from it—forms the essence of life.     

Ackerunkrautgesellschaften Des Südwestlichen Polen Und Die Auswertung Ihrer Untersuchung Für Die Landwirtschaft

Zusammenfassung Es wird berichtet über eine vegetationskundliche Untersuchung der Ackerunkrautgesellschaften des Südwestlichen Polen, Fünf Regionen werden unterschieden: Flachland-, Hochland-, Flu?tal-, Gebirgs- und Gebirgskessel-Region. Auf 100 nach der ‘Methode der Goldene Punkte’ bestimmten Punkten im Gebiet wurden ca 1500 pflanzensoziologische Aufnahmen gemacht. Drei Assoziationen:Euphorbio-Melandrietum, Aphano-Matricarietum undTeesdaleo-Arnoseridetum mit verschiedenen Untereinheiten werden beschrieben. Es werden zwei Beispiele der Kartierung dieser Gesellschaften gezeigt. Es wird einiges über die Auswertung dieser Untersuchungen in der Landwirtschaft mitgeteilt. Die Verteilung der Arten der verschiedenen für das Gebiet aufgestellte soziologischen Gruppen über 12 Ackerbenützungskomplexe wird dargestellt: 1 — sehr guter Weizenkomplex, 2 — guter Weizenkomplex, 3 — schwacher Weizenkomplex, 4 — sehr guter Roggenkomplex, 5 — guter Roggenkomplex, 6 — schwacher Roggenkomplex, 7 — Roggen-Lupinekomplex, 8 — Starker Korn-Futterkomplex, 9 — Schwacher Korn-Futterkomplex, 10 — Gebirgs-Weizenkomplex, 11 — Gebirgs-Kornkomplex, 12 — Gebirgs-Hafer-Kartoffelkomplex. Für die Nomenklatur: s. Tabelle 1.     

Ontogenesis of the cricket Gryllus argentinus Sauss. (Orthoptera, Gryllidae)

The development of Gryllus argentinus Sauss. was studied under stable laboratory conditions: the temperature of 26°C, the air humidity of 60%, and the photoperiod of 12h light: 12 h dark. The life cycle of Gryllus argentinus includes four stages: egg, pronymph, nymph, and adult. The duration of embryonic development is 18 days. The depth of egg bedding in the peat is 9.63 ± 0.12 mm (n =145), the clutch containing 2–4 eggs. A female can lay over 1100 viable eggs during the entire oviposition period. Nymphal development includes 9 instars and lasts 97 days. The duration of nymphal instars (days) is: I—5; II—6; III—6; IV—6; V—8; VI—10; VII—13; VIII—14; IX—29. The duration of the adult life is 51 days in males and 69 days, in females. In the imaginal ontogenesis of males and females, three periods can be distinguished: pre-reproductive, reproductive, and postreproductive. Males start to emit the aggressive signal on the 6th (5–8th) day (the pre-reproductive period). They enter the reproductive period (start to emit the calling song) on the 9th (8–13th) day. Females enter the reproductive period (become capable of responding to the calling song and of copulation) on the 9th (8–10th) day. Oviposition starts on the day after the first copulation. The reproductive period lasts about 40 (15–59) days in males and 58 (21–70) days in females. The post-reproductive period starts in females at the moment of finishing the egg laying period and in males, with disappearance of reproductive behavior. The period ends in the animal’s death.     

Post-dispersed pine seed removal and its effect on seedling establishment in a Mexican Temperate Forest

In order to assess the impact of seed removal and seedling establishment for four species of pines, we designed experiments with and without the exclusion of seed removers in three vegetation types (pine forest, oak-alder forest, and subalpine grassland). Seed removal was significantly different between species of pines (P < 0.01), as was the interaction between the vegetation types and category of exclusion (remover) (P < 0.0001). The significant difference in terms of seed removal was between P. patula and P. montezumae (33.0 and 17.9%, respectively). Seed removal was significant between pine forest—without exclusion (20.0%) versus oak-alder forest—without exclusion (7.0%). There was significant interaction between seedling establishment of the pine species and vegetation type (P < 0.0001). Highlighting differences between the pine forest—P. teocote (84.1%) versus oak-alder forest—P. patula (19.0%), oak-alder forest—P. pseudostrobus (45.0%), pine forest—P. patula (20.2%), pine forest—P. pseudostrobus (45.6%), subalpine grassland—P. montezumae (24.3%), subalpine grassland—P. patula (27.9%), and subalpine grassland—P. pseudostrobus (17.5%). The impact of the food preferences of rodents and other granivores for P. patula and P. pseudostrobus seeds, as well as the poor survival of seedlings of these species in the pine and oak-alder forest are both factors which may explain the dominance of P. teocote in the study region.     

Phytoestrogens as natural prodrugs in cancer prevention: a novel concept

It has been generally accepted that regular consumption of fresh fruits and vegetables is linked with a relatively low incidence of cancers (e.g. breast, cervix, and colon). A number of plant-derived compounds have been identified that are considered to play a role in cancer prevention. However, at present there is no satisfactory explanation for the cancer preventative properties of the above-mentioned compound groups. The current review is an effort to develop a consistent and unambiguous model that explains how some plant-derived compounds can prevent tumour development. The model is based on recent discoveries in the fields of genomics and drug-metabolism; notably, the discovery that CYP1 genes are highly expressed in developing tumour cells but not in the surrounding tissue, and that a variety of plant-derived compounds are substrates for the CYP1 enzymes. Our hypothesis is that some dietary compounds act as prodrugs, i.e. compounds with little or no biological effect as such, but become pharmaceutically effective when activated. More specifically, we state that the abovementioned prodrugs are only activated in CYP1-expressing cells—i.e. cells in the early stages of tumour development—to be converted into compounds which inhibit cell growth. Thus, the prodrugs selectively kill precancerous cells early in tumour development. The review focuses on the identification of naturally-occurring prodrugs that are activated by the tumour-specific CYP1 enzymes and aims to assess their role in cancer prevention.     

Role of protein kinases of human red cell membrane in deformability and aggregation changes

The proteomic analysis has shown that the red cell membrane contains several kinases and phosphatases. Therefore the aim of this study was to investigate the role of protein kinases of human red cell membrane in deformability and aggregation alterations. The exposure of red blood cells (RBCs) to some chemical compounds has led to a change in the RBC microrheological properties. When forskolin (10 μM), an adenylyl cyclase (AC) and a protein kinase A (PKA) stimulator were added to RBC suspension, the RBC deformability (RBCD) was increased by 20% (p<0.05). Somewhat more significant deformability rise appeared after RBC incubation with dB-AMP (by 26%; p<0.01). The red cell aggregation (RBCA) was significantly decreased under these conditions (p<0.01). Markedly less changes of deformability were found after RBC incubation with protein kinase stimulator C (PKC)—phorbol 12-myristate 13-acetate (PMA). This drug reduced the red cell aggregation only slightly. The red cell tyrosine phosphotase activity was changed by N-vanadat and a significant RBCD rise and RBCA lowering were obtained. The similar effect was found when the cells were incubated with cisplatin as a tyrosine protein kinase (TPK) activator. It is important to note that a selective TPK inhibitor—lavendustin eliminated the above mentioned effects.     

Genome structures in a lysogenic Rhizobium meliloti system

Summary The genome structure of the temperateRhizobium meliloti phage and the attachment site of this phage on the host chromosome were examined by genetic means. The heat-sensitive mutants used in 2 and 3 point crosses gave a linear chromosome map. There was no evidence for map circularity. The immunity region has a distal position on the phage chromosome. The functional grouping of the used 23 phage mutants was made byin vivo andin vitro complementation tests and 20 cistrons were obtained. The cistrons, near to the immunity region, were identified as early genes, the remaining ones as morphogenetic cistrons. The latter inin vitro complementation tests gave two complementing groups, presumably as head and tail donors. The attachment site of the prophage on the host chromosome was localized by pulse mutagen treatments in synchronously replicating cultures. The sequence of markers are O-str — hs — att ¹⁶⁻³ — T.     

Epicatechin Inhibits Human Plasma Lipid Peroxidation Caused by Haloperidol In Vitro

Epicatechin belongs to flavonoids protecting cells against oxidative/nitrative stress. Oxidative/nitrative stress observed in schizophrenia may be caused partially by the treatment of patients with various antipsychotics. The aim of our study was to establish the effects of epicatechin and antipsychotics action (the first generation antipsychotic (FGA)—haloperidol and the second generation antipsychotic (SGA)—amisulpride) on peroxidation of plasma lipids in vitro. Lipid peroxidation in human plasma was measured by the level of thiobarbituric acid reactive species (TBARS). The properties of epicatechin were also compared with the action of a well characterized antioxidative commercial polyphenol—resveratrol (3,4′,5-trihydroxystilbene) and quercetin (3,5,7,3′,4′-pentahydroxyflavone). Amisulpride, contrary to haloperidol (after 1 and 24 h) does not significantly influence the increase of plasma TBARS level in comparison with control samples (P > 0.05). After incubation (1 and 24 h) of plasma with haloperidol in the presence of epicatechin we observed a significantly decreases the level of TBARS (P < 0.001, P < 0.001, respectively). In our other experiments, we found that epicatechin also decreased the amount of TBARS in human plasma treated with amisulpride. In conclusion, the presented results indicate that epicatechin—the major polyphenolic component of green tea reduced significantly human plasma lipid peroxidation caused by haloperidol. Moreover, epicatechin was found to be a more effective antioxidant, than the solution of pure resveratrol or quercetin.     

Faunistic similarity and endemism of earthworms in east mediterranean region

The analysis presents earthworm distribution in eight designated tectonic units (C—Cyprus, EP—East Pondites, ET—East Taurus block, Sc—Sakarya continent, Kb—Kırşehir block, L—the Levant, TM—Taurus–Menderes block, WP—West Pondites) in the East Mediterranean region (EM). It represents a full list of earthworm species (N = 81) of the region and reveals significant faunal similarities between Sc + WP, and between Kb + TM. The new Sc + WP unit is characterized by the presence of archaic (Spermophorodrilus) as well as modern earthworm fauna. In contrast, the second newly established Kb + TM unit is characterized by poorly presented earthworm autochthonous fauna and by a lack of endemic species. The analysis done on the species list (N = 73) representing the autochthonous family Lumbricidae showed statistically supported separation of L and EP between themselves and from other designated units. The Levant, part of the Arabian tectonic plate, is characterized by a high level of endemism in ancient (Healyella) and modern (Dendrobaena, Perelia) genera, and by the lack of archaic genera. The EP harbors unique species representing archaic (Eophila), ancient (Healyella), and modern (Allolobophora, Aporrectodea, Dendrobaena and Eisenia) earthworm genera in the EM. The analysis also indicates origin of earthworm fauna in TM and faunistic relationships among the designated units. Clearly, the present-day division of the EM earthworm fauna shows recognizable influences of tectonic history.     

Quantitative relationships between structure and antimicrobial activity of new “Soft” bisquaternary ammonium salts

New surface-active bisquaternary ammonium salts derived from bis-(2-dimethylaminoethyl) ester of glutaric acid are highly effective against representatives of Gram-positive, Gram-negative bacteria and yeasts. Relationships between structure, lipophilicity and antimicrobial effectiveness were demonstrated by quantitative structure-activity methodology. The nonlinear dependence of biological activity on the structure as well as lipophilicity (expressed as critical micelle concentration—CMC) was shown using Kubinyi’s bilinear model. The most effective compounds were those with the alkyl chain of 11–12 carbon atoms and with the CMC values around 0.7−1.0 mmol/L. These derivatives possessed higher antimicrobial activity particularly to Gram-negative bacteria. Part 45 of the seriesQuaternary Ammonium Salts; part 44: Brádlerová A.et al., Pharmazie, in press.     

Synthesis and pharmacological activity of amides and the ozonolysis product of maleopimaric acid

The synthesis of a new group of maleopimaric acid amides containing fragments of methyl ethers of amino acids, aliphatic amines, imidazole, and N-methylpiperazine was carried out. The ozonolysis of methylmaleopimarate occurs via the cleavage of the double bond C18(19) and the opening of an anhydrous ring with the formation of secotriacid. As a result of the screening of the anti-inflammatory and antiulcer activity of maleopimaric acid derivatives, new effective compounds such as maleopimaric acid and its methyl ether, a product of ozonolysis—diterpenic secotriacid—and maleopimaric acid amide with L-leucine were found. An important advantage of the studied compounds is the low toxicity and the presence of bidirectional activity in the absence of adverse effects on the animal.