Fusogenic capacities of divalent cations and effect of liposome size

The initial kinetics of divalent cation (Ca2+, Ba2+, Sr2+) induced fusion of phosphatidylserine (PS) liposomes, LUV, is examined to obtain the fusion rate constant, f11, for two apposed liposomes as a function of bound divalent cation. The aggregation of dimers is rendered very rapid by having Mg2+ in the electrolyte, so that their subsequent fusion is rate limiting to the overall reaction. In this way the fusion kinetics are observed directly. The bound Mg2+, which by itself is unable to induce the PS LUV to fuse, is shown to affect only the aggregation kinetics when the other divalent cations are present. There is a threshold amount of bound divalent cation below which the fusion rate constant f11 is small and above which it rapidly increases with bound divalent cation. These threshold amounts increase in the sequence Ca2+ less than Ba2+ less than Sr2+, which is the same as found previously for sonicated PS liposomes, SUV. While Mg2+ cannot induce fusion of the LUV and much more bound Sr2+ is required to reach the fusion threshold, for Ca2+ and Ba2+ the threshold is the same for PS SUV and LUV. The fusion rate constant for PS liposomes clearly depends upon the amount and identity of bound divalent cation and the size of the liposomes. However, for Ca2+ and Ba2+, this size dependence manifests itself only in the rate of increase of f11 with bound divalent cation, rather than in any greater intrinsic instability of the PS SUV. The destabilization of PS LUV by Mn2+ and Ni2+ is shown to be qualitatively distinct from that induced by the alkaline earth metals.     

Specific recognition by the tripeptide lysyl-tryptophyl-alpha-lysine of structural damage induced in DNA by platinum derivatives

The antitumoral derivative cisPt binds to DNA, as do its inactive analogs, trans- and dienPt. Structural damage introduced into DNA after reaction with the Pt derivatives were probed by using the peptide LysTrpLys. This peptide was used for its preferential binding to single-stranded structures (Brun, F., Toulmé, J.J. and Hélène, C. (1975) Biochemistry 14, 558-563). Phosphorescence lifetime measurements show that the Pt-induced heavy atom effects are quite similar in the three peptide-DNA-Pt complexes whatever the nature of the Pt derivative used. In contrast, fluorescence quenching strongly depends on the nature of the Pt derivatives. This quenching was therefore attributed to the stacking interactions engaged by the tryptophan residue with nucleic acid bases. A comparison of fluorescence quenching data for native and modified DNAs demonstrates that modification by dienPt has no effect on stacking interactions and that high levels of modifications by trans Pt are required to observe a change in stacking efficiency. In contrast modification by cis Pt induces the formation of strong stacking sites. The results strongly suggest the existence of locally opened regions in DNA modified by cis Pt.     

Oligonucleotides covalently linked to intercalating agents. Influence of positively charged substituents on binding to complementary sequences

A pentamethylene chain was used to covalently link the 3'-phosphate of oligothymidylates to the 9-amino group of an acridine derivative. Positively charged substituents were further attached to the 3'-phosphate group to form 3'-phosphotriesters. These molecules form specific complexes with poly(rA) which involve the formation of a number of A X T base pairs equal to that of thymines in the oligonucleotide. Absorption changes induced in the acridine absorption bands are similar to those expected upon intercalation of the acridine dye between A X T base pairs. The acridine covalently linked to the 3'-phosphate strongly stabilizes the complexes formed with poly(rA) as compared with the corresponding unsubstituted oligodeoxynucleotide. The presence of a positively charged substituent on the 3'-phosphate together with the acridine dye further enhances the interaction. The effect of salt concentration on complex stability depends on the number of negatively charged phosphate groups of the oligodeoxynucleotide and on the nature of the substituents borne by the 3'-phosphate group. When the oligothymidylate is substituted by an acridine dye, the stability of the poly(rA) complexes increases when salt concentration increases. If an additional positively charged substituent is present on the 3'-phosphate group, stability decreases when salt concentration increases for the shortest oligonucleotide (trimer) and increases with longer oligonucleotides. Thermodynamic parameters have been calculated from the concentration dependence of melting temperatures.     

The value of provoked expectoration in obtaining sputum samples for cytologic investigation. A prospective, consecutive and controlled investigation of 134 patients

A prospective controlled investigation in 134 consecutive outpatients compared the cytologic adequacy of sputum samples obtained by spontaneous and provoked expectoration. Inhalation of nebulized 10% sodium chloride was used for provoked expectoration. A significantly higher number of adequate samples was produced after provocation, as judged by the presence of alveolar macrophages (X2 = 5.63; p less than 0.02). The improvement in sample adequacy was limited to the nonsmokers and ex-smokers in the study. This result, together with the relatively high cost of cytologic sputum examinations, indicates that provoked expectoration should at least be applied to the collection of sputum samples from nonsmokers and ex-smokers.     

Some aspects of the natural history and food selection ofAvahi laniger

Two groups ofAvahi laniger were studied in the Forêt de Analamozoatra near Perinet in the eastern rainforest of Madagascar from August to October 1984. Overlap between the home ranges of neighbouring groups ofA. laniger was minimal. Group size ranged from one to four individuals with a median group size of two. In four out of ten groups a baby was born between August and September.A. laniger were most active after dusk and before dawn. They had an extended resting period around midnight. Their diet consisted mostly of leaves from at least 17 different plant species. They also ate flowers. Fruit eating was recorded twice. Leaves eaten had high contents of protein and sugar but did not contain alkaloids. The concentration of condensed tannins did not differ between food items and non-food items. There was no indication of competition with other prosimians that might explain their nocturnality.     

Ultrastruktur und Bildungsweise penialer Hartstrukturen bei freilebenden Plathelminthen

Zusammenfassung Ultrastruktur und Differenzierung von Penisstiletten bzw. Stilettnadeln wurden an Vertretern verschiedener Taxa freilebender Plathelminthen untersucht. Die Ausdifferenzierung der Hartstrukturen erfolgt auf unterschiedliche Weise, jedoch stets intrazellulär. Die Stilettnadeln von Philocelis cellata (Acoela) bestehen aus Mikrofibrillen und werden sukzessiv gebildet, mit der Spitze beginnend und basalwärts fortschreitend. Eine ebenfalls sukzessive Bildungsweise zeigen die Stilettapparaturen des Taxons Paromalostomum (Macrostomida); doch bestehen die Hartstrukturen hier aus Mikrotubuli mit angelagertem elektronendichtem Material und nicht aus Mikrofibrillen. Die sukzessive Ausdifferenzierung des Stiletts von Ciliopharyngiella intermedia erfolgt ähnlich wie bei den Hartstrukturen bestimmter Proseriaten mit der Anlage von Mikrofibrillen, die mit elektronendichtem Material verkleidet werden, jedoch weist C. intermedia zusätzlich eine innere und äußere Glättungsschicht auf. Dagegen erfolgt die Stilettbildung bei Adenorhynchus balticus (Typhloplanoida), Marirhynchus longasaeta (Kalyptorhynchia) und Provortex psammophilus und P. tubiferus (Dalyellioida) simultan und ohne Anlage einer Rohform mit einem fibrillären oder tubulären Gerüst. Strukturell sehr ähnlich wie bei A. balticus und M. longasaeta, jedoch sukzessiv erfolgt die Bildung der langen Stilette bei verschiedenen Species des Taxons Promesostoma (Typhloplanoida).Die bisher bekannten feinstrukturellen Organisationsmerkmale und die verschiedenen Bildungsmodi von penialen Hartstrukturen werden unter phylogenetischen Gesichtspunkten diskutiert.

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Ultrastructure and differentiation of penial hard structures in free-living plathelminths

Summary Ultrastructure and differentiation of penis stylets and stylet needles have been investigated in several representatives of different groups of the free-living plathelminths. The formation of the hard structures occurs in different ways, but always intracellularly. The stylet needles of Philocelis cellata (Acoela) consist of microfibrils and are formed successively, beginning with the distal tip. The species of the taxon Paromalostomum (Macrostomida) have stylet apparatuses which also show a successive formation mode; however, the hard elements are built of microtubules and enveloped by electron-dense material. The successive differentiation of the stylet of Ciliopharyngiella intermedia occurs similarly to the hard structure formation of certain proseriates by building a framework of microfibrils which becomes enveloped by electron-dense material; in addition to this rough-form, in C. intermedia an intracellular smooth layer is formed on the inner and outer side of the stylet. In contrast, the stylet formation of Adenorhynchus balticus (Typhloplanoida), Marirhynchus longasaeta (Kalypthorhynchia), Provortex psammophilus and P. tubiferus (Dalyellioida) occurs synchronously and without a roughform, containing a fibrillar or tubular framework. The stylets in several species of the taxon Promesostoma (Typhloplanoida) are built in a way very similar to that in A. balticus and M. longasaeta, but successively.The fine structural properties known at present and the various formation modes of penial hard structures are discussed from the phylogenetic aspect.

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Abkürzungen ae Atriumepithelzelle - ag Atrium genitale - am Atriummuskulatur - ba Bakterium - bl Basallamina oder vergleichbare Interzellularsubstanz - bm Bulbusmuskulatur - bz Stilettbildungszelle - cw Cilienwurzel - de Ductus ejaculatorius - dr Drüsenrohr - dz Ductuszelle - fz Füllzelle - hd Hemidesmosom - hz Hüllzelle - k Kern einer Stilett- bzw. Nadelbildungszelle - m entstehende Muskulatur - ma Macula adhaerens - mc Muskelzylinder - mf Mikrofibrillen - mv Mikrovilli - n Nerv - nz Nadelbildungszelle - pm äußere Penismuskulatur - rm Ringmuskulatur - s Stilett - sd Septatdesmosom - sg Sekretgangzelle - sgz Sekretgangzelle - skg Spermakornsekretgang - skr Spermakornsekretrohr - sm stilettumhüllende Muskulatur - sn Stilettnadel - sna Stilettnaht - sp Spermium - sz Sekretzelle - vg Vesicula granulorum - vgm Muskulatur der Vesicula granulorum - za Zonula adhaerens     

Primary production data from the south-eastern Weddell Sea

Summary Phytoplankton production for three size classes (<20 m, 20–100 m, >100 m), total primary production and qualitative composition of phytoplankton populations were recorded from 18 stations in the south-eastern Weddell Sea in February/March 1983. Total primary production ranged between 80 and 1670 mg C m^-2 d^-1 with an average of 670 mg C m^-2 d^-1, nearly 70% of which was contributed by the <20 m size fraction (usually pennate and/or centric diatoms). Production of phytoplankton was in the higher range of values reported by other authors for the same region. Variations in primary production could not be attributed to composition of populations, ambient light levels or concentrations of macronutrients (N, P, Si). Phytoplankton populations had a higher diversity in the deeper parts of the Weddell Sea and coincided with different oceanographic situations. Three zones (along the shelf-ice edge from Atka Bay to Halley Bay, west of Halley Bay and off the Filchner/Rønne Ice Shelf) with different communities could be clearly distinguished.     

Stoichiometric binding of diacylglycerol to the phorbol ester receptor

The major phorbol ester receptor is the Ca++-activated, phospholipid-dependent protein kinase C. Diacylglycerol stimulates protein kinase C in a fashion similar to the phorbol esters. Likewise, it inhibits phorbol ester binding competitively. Both results suggest that diacylglycerol is the/an endogenous phorbol ester analogue. Alternatively, the diacylglycerol might simply be acting to modify the phospholipid environment of the protein. If diacylglycerol were indeed functioning as an analogue, it should interact with the receptor stoichiometrically. This interaction can be quantitated by measuring the perturbation in apparent diacylglycerol binding affinity as a function of the ratio of diacylglycerol to receptor. We report here that 1,2-dioleoylglycerol interacts with the receptor with the predicted stoichiometry.     

Neither an enhancement of autolytic wall degradation nor an inhibition of the incorporation of cell wall material are pre-requisites for penicillin-induced bacteriolysis in staphylococci

In contrast to what has been postulated, penicillin G at its optimal lytic concentration of 0.1 g per ml did not lead to a detectable activation of autolytic wall processes in staphylococci in terms of the release of uniformly labelled wall fragments from cells pretreated with the drug for 1 h. Rather a considerable inhibition of this release was observed. A similarly profound inhibition of the release of peptidoglycan fragments occurred when staphylococci pretreated for 1 h with 0.1 g penicillin per ml acted as a source of crude autolysins on peptidoglycan isolated from labelled normal cells of the same strain. This clearly demonstrated that the overall inhibition of autolytic wall processes caused by penicillin was mainly due to a decreased total autolysin action rather than to an altered wall structure. Furthermore, no substantial penicillin-induced inhibition of the incorporation of ¹⁴C-N-acetylglucosamine into the staphylococcal wall could be observed before bacteriolysis started, i. e., approximately during the first 80 min of penicillin action. These results are not consistent with any of the models hitherto proposed for the action of penicillin.Dedicated to Prof. Dr. Gerhart Drews on the occasion of his 60th birthday     

Spike synchronization of tympanic receptor fibres in a grasshopper (Chorthippus biguttulus L., Acrididae)

In recordings from single tympanic receptor fibres in C. biguttulus, the response to synthesized sounds (rectangularly modulated white noise) interrupted by very brief (a few milliseconds) gaps was examined. In behavioral tests, females of the species respond very differently to such 'model syllables' at moderate intensities, depending on the gap width. If the gaps (in a moderate-intensity syllable) are larger than 2 ms, the stimulus fails to elicit a response, whereas stimuli with gaps smaller than 1 ms are as effective as uninterrupted syllables (D. von Helversen 1972; O. von Helversen 1979). Neither the mean spike count nor the interspike-interval distribution of the single receptor response contains the information sufficient to distinguish uninterrupted syllables from syllables with gaps. On the other hand, examination of the temporal distribution of the spikes reveals that gaps (or the pulse onsets following the gaps) cause spike synchronization. An index of synchronization (IS) was defined as a measure of this gap-induced effect. Analysis of the receptor responses based on IS revealed differences that correspond quantitatively to the abrupt abolition of the behavioral response at a gap-width between 1 and 2 ms. From the hypothesis that such brief gaps are detected by the nervous system by way of spike synchronization in the tympanic nerve, one can predict certain features of the behavioral response to high-intensity stimuli. The gap-induced spike synchronization was more pronounced at higher temperatures. This effect was demonstrated in both summated recordings from the tympanic nerve and single fibre recordings. Experiments with primary auditory fibres of Locusta migratoria showed that the receptors in this species respond very similarly to the same stimuli. That is, the receptors of C. biguttulus are not specially adapted for detecting very brief gaps. Synchronization of the spikes in parallel receptor fibres of the tympanal nerve is probably a general feature of acridids; we infer that in C. biguttulus this gap-induced synchronized activity is detected by special processing in higher auditory centres.