Volume 52, Issue 1-2

For plant seedlings dependent on the breakdown of lipid reserves for gluconeogenesis the onset of lipid mobilization is a critically important process. Until recently, knowledge about the chemical mechanism of the initial breakdown steps has been limited, because the lipolytic activities, characterized by the intracellular localization and the type of substrate, have not always been found. As seedlings that depend on the reserves in lipid bodies may differ in morphology and biochemistry, it is likely that more than one pathway exists. Recent studies with cucumber (Cucumis sativus) cotyledons have shown that a lipid body-associated lipoxygenase, and an O 2 -dependent reaction, initiate both the destabilization of the phospholipid monolayer and the breakdown of triglycerides. Both types of reactions rely on the particular properties of a lipoxygenase isoform exclusively present in lipid bodies.

Seed oil components of wild and cultivated species of Lupinus were analyzed by gas liquid chromatography (GLC). The sterol and fatty acid patterns of Lupinus albescens and L. gibertianus that are considered important germoplasm resources of South America, are reported for the first time and compared with varieties of Lupinus albus, L. angustifolius and L. mutabilis. The taxonomic implication of seed oil composition was evaluated using a multivariate analysis system.

The incorporation of deuterium labeled 1-deoxy-ᴅ-xylulose into isoprene, emitted by three higher plants upon illumination, was analysed by headspace GC-MS. A high extent of incorporation into isoprene was found, indicating that isoprene is synthesized in higher plants via the novel isopentenyl pyrophosphate pathway, of which 1-deoxy-ᴅ-xylulose-5-phosphate is an intermediate. A new chemical synthesis of methyl [ 1- 2 H 1 ]-1-deoxy-α/β-ᴅ-xyluloside from ᴅ-xylulose is presented.

In photosystem II of plants and cyanobacteria, but not in reaction centers of anoxygenic photosynthetic bacteria, formate is known to inhibit electron flow which is reversed fully upon bicarbonate addition. At issue has been an old controversy whether this effect is on the acceptor or the donor side of photosystem II (PS II). We present here data on chloroplast thylakoids for donor side effects, that is accompanied by acceptor side effects, from measurements on chlorophyll a fluorescence yield changes after light flashes 1-6. Further, sensitive differential infrared gas analyser measurements show that bicarbonate is indeed bound in both maize and pea thylakoid suspensions depleted of CO 2 without any inhibitor; here, high rates of electron flow are associated with the presence of a maximum of 0.8 to 1.25 (corrected for residual activity) CO 2 per photosystem II reaction center. It is suggested that bicarbonate bound to the acceptor side is required for photosystem II activity , both on the acceptor and the donor sides in the same experiment and in the same sample.

Biochemical and ultrastructural examinations on the pellicle of autotrophically grown Eu­glena gracilis were carried out after three days under potassium and magnesium deficiency. Cell-surface changes were detected by lectin assay. Compared to cells grown in complete medium, deficient cells become larger in shape, accompanied by rising carbohydrate, chloro­phyll and protein content, bind more and other lectin molecules: an increase of mainly galactose and N-acetylgalactosamine receptors was observed. Investigations with the mucilage stains alcian blue and ruthenium red indicated that mucilaginous material is released under deficient conditions, whereas the control cells show a strong precipitate of these stains well inside the cells beneath the pellicle.

The effects of hexanoic extracts of Viscum cruciatum Sieber parasitic on Crataegus mono­gyna Jacq. (I), Crataegus monogyna Jacq. parasitized with Viscum cruciatum Sieber (II), and Crataegus monogyna Jacq. non-parasitized (III), and of a triterpenes enriched fractions isolated from I, II and III (CFI, CFII, CFIII respectively), on the growth of HEp-2 cells have been evaluated. All the samples demonstrated significant cytotoxic activity against cultured HEp-2 cells, and all of them showed a stronger in vitro activity than 6-mercaptopurine solution used as a positive control. With the hexanoic extracts I, II and III almost similar activity was obtained, but the hexanoic extract I showed comparably better results. Almost complete inhibition was observed with triterpenes-enriched fractions CFI, CFII and CFIII, at the dose 6 μg/ml, after 72 h of treatment. The most intense response was obtained with the triterpenesenriched fraction CFIII (from Crataegus monogyna non-parasitized), where the inhibition was 93%, but the fraction CFI and CFII showed similar inhibition (92% and 83%).

This study reports the anti-yeast effect of the 4-methoxybenzoic acid hydrazide (pmbah), 4-chlorobenzoic acid hydrazide (pcbah) and their Pt(II) complexes: cis- [PtL 2 X 2 ] and cis- [PtL(NH 3 )Cl 2 ] where L is either pcbah or pmbah and X is Cl, Br or I. MICs of the 4- substituted analogues (20 000-625 μм) are much lower than those of the previously reported benzoic acid hydrazide and 3-methoxybenzoic acid hydrazide. Complex formation results in significant increase of potency which may be due to a change in the mechanism of action, but the MIC (>400-50 μм) and the IC 50 (>400-1 μм) values show that higher activity of the ligands in the free state does not result in enhanced complex activity. Differences in the potency of iodo-, chloro- and bromo complexes suggest MIC and IC 50 values may be in correlation with the stability of the complex, rather than with the activity of the free ligands. Osmotically unstable mutants were more susceptible to the compounds than their parent strains, but differences among the parent strains were greater.

New platinum(II) complexes of cyclohexanecarboxylic acid hydrazide (chcah) were synthesized and characterized by elemental analysis, IR. and 1 H NMR spectra. Their inhibitory effects on cell growth and macromolecular synthesis of Friend leukemia cells in culture as well as the in vivo antitumor activity towards L1210 leukemia in mice were compared with those of complexes containing differently substituted aromatic acid hydrazides. Some of the complexes exhibited antineoplastic activity. No correlation between the in vitro cytotoxicity and the in vivo antitumor activity was found. However, there was a relationship between the in vitro macromolecular synthesis inhibition profile and the in vivo antineoplastic effect, similar to that of cisplatin. On the other hand, only agents containing one ammine ligand were active in vivo. The substitution of the aromatic ring by a cycloalkane residue increased significantly the antitumor effect, with [Pt(NH 3 )(chcah)Cl 2 ] being the most active compound in this study.

We have evaluated the effects of an oral treatment of mice with fraxetin (25 mg/kg for 30 days) on the glutathione system (GSH, GSSG, and GSSG/GSH ratio as stress index), glutathione reductase (GR) and glutathione peroxidase (GPx) in liver supernatants from male C57BL/6J mice (18-month old). A significant antioxidant effect in vivo was found under this treatment by a decrease in the GSSG/GSH ratio and an increased activity of GR compared with the control mice. GSSG rate and GSSG/GSH ratio were correlated with the decline of GPx activity. Our results of increased GR activity could be considered as a supercompensation in glutathione redox status that involves a decrease in the accumulation of GSSG, as well as, in GSSG /GSH ratio. Finally, we suggest that this possible mechanism of supercompensation could lead to an enhancement in the average life span.

Several phenolic constituents of propolis and their synthetic analogs were derivatized with ʟ-lysine. The ability of these complexes to alter complement activity was estimated in vitro in human serum. The influence of selected complexes on C3 hemolytic activity via classical pathway (CP) and alternative pathway (AP) and on zymosan-induced AP activation was determined. The results suppose that the anticomplement effect of the complexes might be related to the interaction with C3 complement component.

The effect of trialkyllead and trialkyltin on pig erythrocyte hemolysis has been studied and compared. The results of experiments showed that the hemolytic activity of organoleads increases with their hydrophobicity and follows the sequence: triethyllead chloride < tri-npropyllead chloride < tributyllead chloride. And similarly in the case of organotins: triethyltin chloride < tri-n-propyltin chloride < tributyltin chloride. Comparison of the hemolytic activity of organoleads and organotins indicates that the lead compounds exhibit higher hemolytic activity. The methods of quantum chemistry allowed to determine the maximum electric potential of the ions R 3 Pb + and R 3 Sn + , and suggest a relationship between the potential and toxicity.

Urine samples of premature babies contain high amounts of thiodiacetic acid (TDA). Since these pre-term infants are exposed to an increased oxygen atmosphere in the incubator, we supposed that these high levels of thiodiacetic acid might be produced from ethylene, generated in the course of lipid peroxidation processes. Considering that conversion of ethylene to ethylene oxide (EO) is well known in biology we investigated whether ethylene oxide is metabolised to thiodiacetic acid or not. Therefore Sprague-Dawley rats and NMRI mice were exposed to ethylene oxide for six hours. Urine specimens were collected after exposure and the amount of thiodiacetic acid was determined by gas chromatography/mass spectrometry. The quantity of excreted TDA increased enormously compared to control samples. So thiodiacetic acid seems to be a metabolite of ethylene oxide in vivo.

The nucleosome contains of 200 base pairs of DNA complexed with four core histone complex: H2A , H2B, H3, and H4. The fifth histone species, the H1 histone, interacts with linker DNA connecting neighbouring nucleosomes. We have studied the influence of the phosphorylation on the interactions of a repeating unit 15 residues long, containing the SPKK motif, the motif thought to induce turn along peptides sequences, enclosed within the trout testis H1 C-terminal domain with octanucleotide by means of the thermal denaturation and CD technique. The results indicate that the peptide preferentially binds to a single stranded oligonucleotide. It has been shown further that there is no β structure present but a distorted helical structure has been detected.

In all DNA-DNA in situ hybridization (ISH) procedures described so far in the literature, the production of single-stranded target DNA sequences plays a decisive role. This can be achieved either by enzymatic treatment at physiological temperatures or by the separation of double-stranded DNA sequences. Denaturation by heat and chemical agents (e.g. formamide) is regarded as a prerequisite for the non-enzymatic ISH process. However, additional mechanisms of a non-enzymatic ISH procedure are conceivable which do not require high temperature treatment combined with formamide. Here, we report on a non-enzymatic, non-formamide, low temperature, fluorescence in situ hybridization (FISH) procedure which allowed a microscopic visualization and quantitative fluorescence analysis of the binding sites of a repetitive DNA probe. Following only probe denaturation at 94 °C, hybridization was performed at 52 °C for 30 min, i.e., at nearly physiological temperatures. Moreover, increasing the hybridization time to 3 hours indicated that hybridization sites became also visible at 37 °C. Since the protocols are based on recently described Fast FISH developments, the technique will be called Low Temperature Fast-FISH (LTFF).

By computer simulations -molecular mechanics and molecular dynamics with the amber force field (Weiner et al., (1986), J. Comp. Chem. 7, 2 30-252) -we have determined the stabilities of oligoribotide strands built with ᴅ -and ʟ-riboses, and of peptide chains with ᴅ -and ʟ-amino acid residues. In particular, complementary double-chains of oligoribotides were studied, since they are an important feature of the growing mechanism of modern nucleic acids. Peptide chains on the other hand, grow without need of a template. We found that mixed oligoribotides are less stable than homochiral ones, and that this chiral effect is less noticeable in peptide chains. The results support the interpretation that ʟ-riboses act as terminators to the template-assisted growth of oligo-r-Gᴅ (enantiomeric cross-inhibition; Joyce et al., (1987), Proc. Natl. Acad. Sci. USA 84, 4398-4402). Based on this effect, a chemical pathway is proposed which could, under assumed prebiotic conditions, bypass the hindrance of homochiral growth.

Specific and unspecific binding and uptake (internalization) by macrophages of 125 iodine -labelled, copper-oxidized human low density lipoprotein is differently influenced by the anti­ oxidants α-tocopherol (α-Toc), probucol (Prob), pyridoxal-5'-phosphate (PP) and the magnesium-pyridoxal-5'-phosphate glutamate complex (MPPG). Binding as well as internalization, mediated by the so-called "scavenger receptor" is lower in the presence of MPPG whereas both specific binding and internalization are enhanced. The comparison of the effects in vitro allows a rating of the potentially anti-atherogenic and thus protective effects of the tested substances as follows: MPPG > PP > α-Toc > Prob.

Two hundred isolates of Lactobacillus were obtained from seven indigenous fermented foods namely: 'fufu' (fermented cassava), 'iru' (fermented African locust bean), 'kenkey' and 'ogi' (fermented maize), 'kunu-zarki' (fermented millet), 'ugba' (fermented African oil bean) and 'wara' (fermented skimmed cow milk). Lactobacillus species identified were Lacto­ bacillus casei (17.0%), L. lactis (4.5%), L. plantarum (41.5%), L. brevis (11.5%), L. jensenii (5.5%), L. acidophilus (3.5%) L. cellobiosus (5.0%), L. delbrueckii (2.5%), L. salivarius (3.5%), L. leichmannii (3.5%) and L. fermentum (2.0%). The commonest species isolated was L. plantarum. This paper highlight the importance of Lactobacillus in the processing of African fermented foods.

Surface lipids obtained from the silverfish by short-term solvent extraction contain ali­ phatic hydrocarbons, monoester waxes, cholesteryl esters, triglycerides, free cholesterol, and free fatty acids. Together, cholesteryl esters and free cholesterol account for >30% of the total lipids. As found for other arthropods living in aquatic or moist environments unsaturated homologues predominate among the acidic constituents. The cuticular lipid composition o f silverfish resembles that of other more primitive arthropod forms such as stoneflies and dragonflies.

A biologically active principle has been isolated from the uropygial gland secretion of the gannet (Sula bassana) exhibiting growth inhibitory activity for Gram-positive bacteria, yeasts, moulds and, in particular, for dermatophytes.The feather lipids of this species consist of monoester waxes predominantly composed of 3,x-dimethylalkanols esterified with fatty acids belonging to homologous series of 2-and 3-methyl-, 2,x-and 3,x-dimethyl-as well as 2,x,y-and 3,x,y-trimethyl-substituted alkanoic acids. For chemosystematic reasons other pelecani­ form species (Pelecanus crispus, Pelecanus onocrotalus, Phaeton lepturus) and one gaviiform species (Gavia stellata) have also been analysed. The pattern of their preen wax constituents suggest to assume that the Pelecaniformes are polyphyletic with gannets and cormorants being closely related, whereas pelicans and tropic birds are different, both from the former and from each other. Pelicans resemble very much the grebes, while tropic birds seem to be more closely related to the charadriiform, alciform, and possibly to the gaviiform birds.

The spider Cupiennius salei Keys. (Araneae, Ctenidae) is a nocturnal hunter living on monocotyledones. It does not build webs for prey capture. During the day it remains in its retreat and during dusk it begins to hunt for prey or to search for mates. C. salei is well equipped with mechanosensory systems to detect air- or substrate- borne vibrations elicited by prey or predators. If none of them produce either air movements or substrate vibrations, and the light intensity is below the threshold (0.1 lx), the animal is virtually " blind" . There­ fore a hypothetical, additional sensory input should exist, which is used only in complete darkness. The animal was tested on a locomotion compensator were it performs constant walks towards a visual target. Three different light intensities were used (bright 200 lx, dim 0.1 lx, and dark at 950 nm, which is outside the spectral sensitivity range of the animal). At bright and dim illumination the animal walked in the alternating tetrapod gait towards the target. In complete " darkness" the walk was no longer directed and the animal changed its gait and continued walking on only six legs using the first pair as guide-sticks. If the first pair is missing, the second cannot replace this function. This shows a twofold use of the first pair o f legs as ordinary walking legs and as guide-sticks or " antennae" . Therefore one can assume that visual input causes a behavioral change, which can not be explained by a fixed locomotion pattern but by adaptive changes caused by visual inputs.

13 Alkaloids were identified in leaves and twigs of Platycelphium vöense (a monotypic genus in the tribe Sophoreae) by capillary GLC and GLC-MS. Cytisine, N-methylcytisine, anagyrine and 5,6-dehydrolupanine figured as major and N-formylcytisine, N-acetylcytisine, baptifoline, thermopsine, isolupanine, and rhombifoline as minor alkaloids in both leaves and twigs. Lupanine occurred in considerable amount in the leaves and as a minor component in the twigs. The rare alkaloid, 6β-hydroxylupanine, was detected only in the leaves while N-ethylcytisine was shown to be a minor constituent of the twigs only.

The sensitivity of pig cytochrome P450cl7 (CYP17), an endoplasmic reticulum membrane-bound enzyme, towards heat denaturation (48 °C) was measured by the P450-to-P420 spectral transition indicating conforma­tional labilization of the protein. Both sucrose and glucose have comparable and increasingly protective effects at concentrations ranging from 100 to 800 mм, while ectoine, a novel zwitterionic compatible solute which regulates bacterial osmoadaptation and stabilizes cytoplasmic enzymes, has a strong labilizing effect on CYP17 and favours its proteolytic inactivation possibly by electrostatic derangements. Sucrose or glucose, but not ectoine, can therefore eventually be proposed as compatible stabilizers of cytochrome P450 structures.