Gramicidin in chromatophores of Rhodobacter sphaeroides

Chromatophores of Rhodobacter sphaeroides were excited with light flashes to generate a transmembrane electrical potential difference. The electric relaxation was measured by electrochromic absorption changes as a function of added gramicidin. At low gramicidin/bacteriochlorophyll (BChl) molar ratios the decay of the electrochromic absorption changes showed a biphasic behaviour, with a fast phase relaxing at some s, and a slow phase relaxing at more than 100 ms. This was attributable to a mixture of vesicles containing gramicidin dimers with others containing none. The concentration dependence of this effect was linear. This implied full dimerization of gramicidin. The data were interpreted to yield an average bacteriochlorophyll content per chromatophore of 770(±150) and the conductance of a single gramicidin dimer in the chromatophore membrane of 15(±4) pS (in about 115 mM KCl).Abbreviations BChl Bacteriochlorphyll - tricine N-Tris[hydroxymethyllmethylglycine Offprint requests to: W. Junge     

Voltage-dependent channel formation by rods of helical polypeptides

Summary The voltage-dependence of channel formation by alamethicin and its natural analogues can be described by a dipole flip-flop gating model, based on electric field-induced transbilayer orientational movements of single molecules. These field-induced changes in orientation result from the large permanent dipole moment of alamethicin, which adopts -helical conformation in hydrophobic medium. It was, therefore, supposed that the only structural requirement for voltage-dependent formation of alamethicin-type channels might be a rigid lipophilic helical segment of minimum length.In order to test this hypothesis we synthesized a family of lipophilic polypeptides—Boc-(Ala-Aib-Ala-Aib-Ala) _n -OMe,n=1–4—which adopt -helical conformation forn=2–4 and studied their interaction with planar lipid bilayers. Surprisingly, despite their large difference in chain length, all four polypeptides showed qualitatively similar behavior. At low field strength of the membrane electric field these polypeptides induce a significant, almost voltage-independent increase of the bilayer conductivity. At high field strength, however, a strongly voltage-dependent conductance increase occurs similar to that observed with alamethicin. It results from the opening of a multitude of ion translocating channels within the membrane phase.The steady-state voltage-dependent conductance depends on the 8^th–9^th power of polypeptide concentration and involves the transfer of 4–5 formal elementary charges. From the power dependences on polypeptide concentration and applied voltage of the time constants in voltage-jump current-relaxation experiments, it is concluded that channels could be formed from preexisting dodecamer aggregates by the simultaneous reorientation of six formal elementary charges. Channels exhibit large conductance values of several nS, which become larger towards shorter polypeptide chain length. A mean channel diameter of 19 Å is estimated corresponding roughly to the lumen diameter of a barrel comprised of 10 -helical staves. Similar to experiments with the N-terminal Boc-derivative of alamethicin we did not observe the burst sequence of nonintegral conductance steps typical of natural (N-terminal Ac-Aib)-alamethicin. Saturation in current/voltage curves as well as current inactivation in voltage-jump current-relaxation experiments are found. This may be understood by assuming that channels are generated as dodecamers but, while reaching the steady state, reduce their size to that of an octamer or nonamer. We conclude that the overall behavior of these synthetic polypeptides is very similar to that of alamethicin. They exhibit the same concentration and voltage-dependences but lack the stabilizing principle of resolved channel states characteristic of alamethicin.     

Retinotopische Beziehungen und Struktur rezeptiver Felder im Tectum opticum und Praetectum der Katze

Zusammenfassung 1.Von 600 Neuronen des Colliculus superior und Praetectums der Katze wurde mit Stahlmikroelektroden abgeleitet und der Ableitort markiert. Die Lage der rezeptiven Felder wurde mit bewegten und stationären Lichtreizen bestimmt und dem Ableitort zugeordnet.2.Im Colliculus superior und Praetectum fanden sich richtungsspezifische und richtungsunspezifische Bewegungsneurone. Ein Teil der praetectalen Neurone reagierte richtungsspezifisch auf Bewegungen vom Tier weg und auf das Tier zu (S-Neurone).3.Innerhalb einer senkrecht zur Oberfläche des Colliculus verlaufenden Penetrationssäule nahm die Feldgröße bei gleichbleibender Feldposition mit zunehmender Tiefe zu. Zwischen Ableitort und Feldposition bestand eine systematische retinotopische Beziehung. Die Projektion des vertikalen O-Meridians des Gesichtsfeldes verlief im rostralen Drittel des Colliculus von medial nach lateral, die des horizontalen O-Meridians in der Mitte des Colliculus von rostral nach caudal. Das Projektionsschema eines Colliculus enthält einen nasalen Teil der ipsilateralen Gesichtsfeldhälfte.4.Im Praetectum verlief die Projektion vertikaler Meridiane am caudalen Ende von medial nach lateral und überlappte sich teilweise mit dem Projektionsgebiet des vertikalen O-Meridians im Colliculus. Die horizontalen Meridiane verliefen so von caudal nach rostral, daß das Projektionsschema des Praetectums spiegelbildlich zu dem des Colliculus superior angeordnet war. Dieses Projektionsschema galt nur für den Nucleus tractus optici und die Area praetectalis. Die übrigen praetectalen Kerne mit zum Teil sehr großen rezeptiven Feldern und spezifischen Reaktionsweisen erhielten keine retinotopische Projektion.5.Rezeptive Felder der oberflächennahen Schichten waren uniform on-, off-oder on-off strukturiert, Felder tiefergelegener Einheiten waren ungeordnet aus on-, off- und on-off Bezirken zusammengesetzt. Binocular erregbare Neurone zeigten für beide Augen gleiche Position und Struktur der rezeptiven Felder.6.Die Ergebnisse wurden mit den an anderen Tierarten erhobenen Befunden verglichen. Ihre mögliche funktionelle Bedeutung wurde diskutiert.

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Retinotopic relationship and structure of receptive fields in the optic tectum and pretectum of the cat

Summary 1.600 neurons of the cat's superior colliculus and pretectum were recorded and marked with stainless-steel microelectrodes. The position and structure of receptive fields were tested with stationary flickering and moving stimuli. The position of the stimuli in the visual field was determined by the direction of the lamp projecting the light-points because animal and lamp were arranged in a fixed relationship to the screen. The positions of the stimuli were described in a coordinate system based on the horizontal-and vertical zeromeridean of the retina.2.About 55% of tectal neurons are directionally selective and signal mainly movements directed to the periphery of the visual field. Neurons of the pretectum have the same response characteristics as neurons of the superior colliculus but in addition some are selectively responsive to movements towards the animal or away from it (S-neurons).3.Neurons in one functional column (diameter 0.5 mm, length 3.6 mm) perpendicular to the surface of the superior colliculus react to the same position and preferred direction of a moving stimulus. The size, complexity and directional selectivity of the receptive fields increase with the depth of the recorded neurons. The projection of the vertical zero-meridean passes across the rostral part of the colliculus but does not form the rostral border of the superior colliculus. The nasal part of the ipsilateral visual field projects to the most rostral part of the superior colliculus. The projection of the horizontal zeromeridean passes rostro-caudally in a nearly sagittal plane down the middle of the colliculus. Along this projection-line the resolving power is 13°/column in the caudal part and 6°/column in the rostral part of the superior colliculus. The size of the receptive fields increase with their excentricity in the visual field. (Average of field diameters: 26±13°).4.The diameter of receptive fields in the pretectum was 21±11°, except for a few very large fields (70° and larger). Along the medio-lateral axis of the pretectum there was a retinotopic organization identical to that in the colliculus. Along the caudo-rostral axis, the retinotopic organization was the mirror image of that in the colliculus. No retinotopic organization was observed in the so-called deep pretectal nucleus or in the nucleus of the posterior commissure. Neurons of these nuclei may represent more complex levels in the visual pathway.5.The more superficial neurons of the colliculus (0.1–1.8 mm deep) react mainly with uniform on-, off- or on-off responses to stationary flickered stimuli, i.e. their receptive fields (7–20° in diameter) are uniformly on-, off- or on-off. The deeper neurons (2 mm and deeper) have receptive fields (20–40° in diameter) with compound but not antagonistic structure. No receptive fields showed on- or off-inhibition. Binocularly driven neurons have the same position and structure of their receptive fields for both eyes.6.A survey of the literature reveals that all vertebrates so far investigated show small differences in the destination and retinotopic organization of their retinofugal fibre projections and in the types of tectal receptive fields. These differences seem to indicate an adaption to the development of binocular representation of the center of the visual field, of a specialized area of the retina and of a retino-cortical system.

     

AdaptivSearch: A Novel Iterative Algorithm for the Generation of n-dimensional Hypersurfaces Using an Adaptive Numerical Strategy

AdaptivSearch is the first adaptive strategy based algorithm for the rational and economical construction of n-dimensional hypersurfaces. AdaptivSearch works iteratively: At each step it parcels out the definition range into several triangles, evaluates the worst according to a built-in error criterion, and refines the approximation to the unknown function by choosing the barycenter of this partial area as the node to be calculated next. Based upon the error criterion, AdaptivSearch selectively approaches those parts of the hypersurface in which the curvature exhibits the strongest changes. Some examples of AdaptivSearch applications for both analytical functions and chemical model surfaces are given in order to demonstrate the behavior of the algorithm. These show its broad applicability and the usefulness, especially for chemical problems.     

The effects of tail autotomy on survivorship and body growth of Uta stansburiana under conditions of high mortality

We examined the effects of tail autotomy on survivorship and body growth of both adult and juvenile Uta stansburiana by directly manipulating tail condition. Tail loss decreased neither survivorship nor rate of body growth for individuals in two natural populations. Lack of an influence of tail loss on survivorship in these two populations may be the result of high mortality. Under high mortality any differential effects of tail loss will be lower than in populations facing lower mortality. Growth experiments in the laboratory demonstrated that, under conditions of minimal environmental variation and social interactions, there is no tradeoff between body growth and tail regeneration as has been suggested for other species of lizards. One possible reason for this difference is that U. stansburiana does not use the tail as a storage organ for lipids. The original and regenerated tails are composed mainly of protein. In general, any differential body growth between tailed and tailless individuals may be due to social interactions and not a diversion of limited energy into tail regeneration.     

Der Einfluß von Objektart und früher Erfahrung auf die Diskriminationsleistungen von Haushühnern (Gallus domesticus)3

Main subject of the reported investigation is the question in which way the acquisition of a conditional discrimination is modified on the one hand by the kind of objects presented, on the other hand by early experience. 40 newly hatched chickens grew up either with all the wooden eggs or cubes used for training and test, or without these objects, or in a ‘natural’ (enriched) environment. The results prove an influence of the kind of objects — cubes are discriminated twice or three times as fast as wooden eggs — but no effects of the different environments on acquisition (number of trials for reaching learning criterion). Results are discussed under aspects of species-specific constraints on learning.     

Serologische Untersuchungen zur Gliederung derBrassicaceae

The serological investigations support the opinion ofJanchen (1942) to combine the generaBunias, Isatis, andSisymbrium in the tribeSisymbrieae; Cheiranthus, Erysimum, andMatthiola in the tribeHesperideae; andBrassica, Crambe, Sinapis, andSuccowia in the tribeBrassiceae. They further underline the central position of theSisymbrieae and the isolated position of theHeliophileae. In accordance withEigner (1973) theBrassiceae are placed closer to theSisymbrieae than inJanchen; the same holds for thePringleeae. No serological justification could be found to uniteArabis andBarbarea in the tribeArabideae, andAlyssum andLunaria in theAlysseae. From the antigen-systems used among the representatives ofJanchen's Lepidieae the generaLepidium andNeslia show remarkable correspondence both toCamelina andThlaspi, but not toCochlearia which appears distant fromCamelina andThlaspi also.

Teil 1/Part 1.     

Biliprotein assembly in the disc-shaped phycobilisomes of Rhodella violacea electron microscopical and biochemical analyses of C-phycocyanin and allophycocyanin aggregates

C-phycocyanin and allophycocyanin from the red alga Rhodella violacea were investigated by electron microscopy and biochemical methods using samples taken from the same fractions.The molecular weights of the native biliprotein aggregates C-phycocyanin and allophycocyanin are about 139,000 (140,000) and 130,000 (145,000) as revealed by calibrated gel chromatography, gradient gel electrophoresis and morphological measurements on the basis of an average protein packing density. These molecular weights are direct evidence for a trimeric aggregation form ()₃ of these biliproteins. Independently, their monomers were determined to be about 34,400 (C-phycocyanin) and 33,900 (allophycocyanin).C-phycocyanin and allophycocyanin are ringshaped, six-membered, biliprotein aggregates with dimensions of about 10.2×3.0 nm and 10.0×3.0 nm, respectively. The aggregates are made up of six subunits, 3 and 3, which are assumed to be associated in alternating positions. They are arranged in regular hexagons in C₆ symmetry. Hexameric aggregates ()₆, so far only isolated for C-phycocyanin, originate by face to face association of two trimeric aggregates.     

Neuraminidase-associated plasminogen recruitment enables systemic spread of natural avian Influenza viruses H3N1

Repeated outbreaks due to H3N1 low pathogenicity avian influenza viruses (LPAIV) in Belgium were associated with unusually high mortality in chicken in 2019. Those events caused considerable economic losses and prompted restriction measures normally implemented for eradicating high pathogenicity avian influenza viruses (HPAIV). Initial pathology investigations and infection studies suggested this virus to be able to replicate systemically, being very atypical for H3 LPAIV. Here, we investigate the pathogenesis of this H3N1 virus and propose a mechanism explaining its unusual systemic replication capability. By intravenous and intracerebral inoculation in chicken, we demonstrate systemic spread of this virus, extending to the central nervous system. Endoproteolytic viral hemagglutinin (HA) protein activation by either tissue-restricted serine peptidases or ubiquitous subtilisin-like proteases is the functional hallmark distinguishing (H5 or H7) LPAIV from HPAIV. However, luciferase reporter assays show that HA cleavage in case of the H3N1 strain in contrast to the HPAIV is not processed by intracellular proteases. Yet the H3N1 virus replicates efficiently in cell culture without trypsin, unlike LPAIVs. Moreover, this trypsin-independent virus replication is inhibited by 6-aminohexanoic acid, a plasmin inhibitor. Correspondingly, in silico analysis indicates that plasminogen is recruitable by the viral neuraminidase for proteolytic activation due to the loss of a strongly conserved N-glycosylation site at position 130. This mutation was shown responsible for plasminogen recruitment and neurovirulence of the mouse brain-passaged laboratory strain A/WSN/33 (H1N1). In conclusion, our findings provide good evidence in natural chicken strains for N1 neuraminidase-operated recruitment of plasminogen, enabling systemic replication leading to an unusual high pathogenicity phenotype. Such a gain of function in naturally occurring AIVs representing an established human influenza HA-subtype raises concerns over potential zoonotic threats.