Electric field-induced breakdown of lipid bilayers and cell membranes: A thin viscoelastic film model

Summary A simple viscoelastic film model is presented, which predicts a breakdown electric potential having a dependence on the electric pulse length which approximates the available experimental data for the electric breakdown of lipid bilayers and cell membranes (summarized in the reviews of U. Zimmermann and J. Vienken, 1982,J. Membrane Biol. 67:165 and U. Zimmermann, 1982,Biochim. Biophys. Acta 694:227). The basic result is a formula for the time of membrane breakdown (up to the formation of pores): =(/C)/( _m ² ₀ ² U ⁴/24Gh ³+T ²/Gh–1), where is a proportionality coefficient approximately equal to ln(h/2₀),h being the membrane thickness and ₀ the amplitude of the initial membrane surface shape fluctuation ( is usually of the order of unity), represents the membrane shear viscosity,G the membranes shear elasticity modules, _m the membrane relative permittivity, ₀=8.85×10^–12 Fm,U the electric potential across the membrane, the membrane surface tension andT the membrane tension. This formula predicts a critical potentialU _c ;U _c =(24Gh ³/ _m ² ₀ ² )^1/4 (for = andT=0). It is proposed that the time course of the electric field-induced membrane breakdown can be divided into three stages: (i) growth of the membrane surface fluctuations, (ii) molecular rearrangements leading to membrane discontinuities, and (iii) expansion of the pores, resulting in the mechanical breakdown of the membrane.     

Physiological relationship between the temperate phages lambda and phi80

Summary This work deals with the ability of phage 80 to provide defective mutants of with their missing functions. Functions Involved in Recombination. As shown by others, the Int mechanism of 80 cannot excise prophage . However, 80 efficiently excises recombinants from tandem dilysogens, using its Ter mechanism. Likewise, the nonspecific mechanism Red is interchangeable between 80 and . Maturation of DNA by 80. The Ter recombinants excised by 80 from tandem dilysogens are packaged into a 80 protein coat. This contrasts with the fact, already mentionned by Dove, that 80 is extremely inefficient for packaging phage superinfecting a -lysogen. The latter result is also found when the helper phage is a hybrid with the left arm of (80hy4 or 80hy41 — see Fig. 1). However, the maturation of the superinfecting is much more efficient if the 80hy used as a helper has the att-N region of (like 80hy1). Conversely a with the att-N region of 80 (hy6 — see Fig. 1) is packaged more efficiently by 80 or 80hy4 than by 80hy1. It is suggested that the maturation of chromosome superinfecting an immune cell requires a recombination with the helper phage. Vegetative Functions. Among the replicative functoons O and P, the latter only can be supplied by 80. That N mutants are efficiently helped by 80 does not tell that 80 provides the defective with an active N product; the chromosomes are simply packaged into a 80 coat. This shows that 80 is unable to switch on the late genes of . That neither 80 nor any of the 80hy tested can provide an active N product is shown in a more direct way by their complete failure to help N ^-r₁₄; this phage carries a polar mutation which makes the expression of genes O and P entirely N-dependant. The maturation of a N ^- by 80 contrasts with the fact that mutants affected in late genes (A, F or H) are not efficiently helped by 80. This suggests that the products coded by these genes are not interchangeable between 80 and , and that packaging of DNA into 80 coats is possible but inhibited when late proteins are present in the cell. Activation of the Late Genes. Among the im ₈₀ h ⁺ hybrids tested, only 80hy41 is able to switch on the late genes of a N defective mutant. This hybrid differs from the other hybrids studied here, by the fact that it has the Q-S-R region of (see Fig. 1). The results are consistant with the view that the product of Q gene is sufficient for activating the late genes of a DNA. N would thus control the expression of late genes only indirectly by controlling the expression of gene Q (Couturier & Dambly have independantly reached the same conclusion, 1970). Furthermore the failure of 80 and of the 80hy1 and 80hy4 to activate the late genes of would imply that these phages are unable to provide an Q product active on the chromosome Reciprocally, switches on the late genes of prophage 80hy41, but not of prophages 80hy1 and 80hy4. This suggests that the initiation of late genes expression takes place at a main specific site located in the Q-S-R region of the chromosome. The expression of the late genes would thus be sequential, and proceed through the left arm only when steaky ends cohere. Similar conclusions were reached independantly by Toussaint (1969) and by Herskowitz and Signer (1970).

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Ce travail a été réalisé dans le cadre du contrat d'association Euratom-U. L. B. 007-61-10 ABIB et avec l'aide du Fonds de la Recherche Fondamentale Collective.     

Determination of the size of the Azotobacter vinelandii chromosome

The chromosome of Azotobacter vinelandii UW was digested separately with the rape cutter restriction endonucleases Swal (5-ATTTAAAT), PmeI (5GTTTAAAC) and Pacl (5-TTAATTAA) and the products were separated by pulsed-field gel electrophoresis. The size of the chromosome was determined to be approximately 4.5 megabase pairs (Mb) based on the sum of the sizes of the restriction fragments. This is almost the same as the size of the chromosome of Escherichia coli. The inability of the undigested DNA to enter the gel has led us to infer that the chromosome is circular.     

Analysis of the secretions of the subcommissural organs of several vertebrate species by use of fluorescent lectins

Summary The glycoprotein secretions of the subcommissural organ were analyzed with the use of nine fluorescent lectins, specific to different sugar moieties. After exposure to Concanavalin A a bright fluorescence was observed in the ependymal cells of the subcommissural organs of all vertebrates studied (Lampetra planeri, Ameiurus nebulosus, Bufo bufo, Lacerta vivipara, Gallus gallus, Rattus norvegicus, Ovis aries). The fluorescence is abolished by the competitive sugar, -D-mannopyranosyl. The intensity of the lectin fluorescence decreases from the phylogenetically lower to the higher forms, paralleled by a change in polarity of the secretion from a vascular (lower vertebrates) to a ventricular (higher vertebrates) direction. The strong affinity for Concanavalin A suggests the presence of a glycoprotein rich in mannosyl residues in the ependymal cells and a similarity of composition of this glycoprotein among the vertebrates. Lens culinaris agglutinin and wheat germ agglutinin revealed fluorescent rosettes in the hypendymal cells of the sheep. Binding of both these lectins suggests the presence of a glycoprotein rich in N-acetyl-D-glucosamine.In the underlying ventricular cavity, no fluorescence could be observed, suggesting that the Reissner's fiber does not possess the same carbohydrate constitution as the ependymal secretion of the subcommissural organ.     

Development and characterisation of a line of bread wheat,Triticum aestivum,which lacks the short-arm satellite of chromosome 1B and the Gli-B1 locus

Summary About 360 offspring of a tri-parental cross were screened by gel electrophoresis and unexpectedly one of them did not contain chromosome 1B -gliadins derived from either of the primary parents. A line disomic for the -gliadin null was developed from the surviving embryo half of the unique grain. Two dimensional electrophoresis revealed that all the storage protein genes at Gli-B1, coding for -gliadins, -gliadins and low-molecular-weight subunits of glutenin as well as the -gliadin, were not expressed. The nuclei of dividing root-tip cells were shown by light microscopy to lack the normal short-arm satellites of chromosome 1B, indicating that the genes for the missing storage proteins had been lost through a terminal deletion. Using a radioactive ribosomal RNA probe, the deficient 1B chromosomes were shown to contain ribosomal RNA genes demonstrating that at least two-thirds of the short arm was still present. Examination of serial sections of chromosome 1B at metaphase by low-power electron microscopy showed that the point of scission of this chromosome was within the secondary constriction where the ribosomal RNA genes are located. The Gli-B1 locus must therefore be carried on the short-arm satellite. Transmission of the deficient chromosome from female gametes to progeny was normal (i.e., about 50%) but from pollen it was poor (8.8%). Recombination mapping indicated that the distance from the ribosomal RNA genes (Nor1) to Glu-B1 was 22 cM, equivalent to 13 cM from Nor1 to the centromere.     

The development of the Jamming Avoidance Response (JAR) in Eigenmannia: An innate behavior indeed

Summary In its Jamming Avoidance Response (JAR), the gymnotiform electric fish Eigenmannia shifts its electric organ discharge (EOD) frequency away from similar interfering frequencies. Continual behavioral measurements were carried out in 164 juvenile fish until a correct JAR emerged. Sixty-four of these fish were raised in complete isolation, the remainder in a community of their siblings. A correct JAR emerged in fish of 1.2–1.6 cm in body length, corresponding to a developmental age of 24–32 days. In 6 of 164 fish, the emergence of a correct JAR followed an interim appearance of an incorrect JAR, which involved frequency shifts in the direction opposite to those of a correct JAR. The fish raised in isolation developed the same forms of behavior and showed the same sequence in their appearance as did socially raised fish. This indicates that the JAR and its developmental schedule are innate. The appearance of an incorrect JAR suggests initial errors or incompleteness in the wiring of central nervous connections. A correct JAR ultimately emerged even if a stimulus regimen was offered that rewarded frequency shifts in the direction opposite to those of a correct JAR. This indicates that the development of the JAR is immune to experimental alterations of sensory experience.Abbreviations Df frequency difference between a jamming stimulus and fish's EOD - ELL electrosensory lateral line lobe - EO electric organ - EOD electric organ discharge - JAR Jamming Avoidance Response - nE nucleus electrosensorius - nE subnucleus of nE, causing drop of EOD frequency - nE subnucleus of nE, causing rise of EOD frequency - Pn pacemaker nucleus - PPn prepacemaker nucleus     

A phylogenetic analysis of the Conchostraca and Cladocera (Crustacea, Branchiopoda, Diplostraca)

A computer-assisted cladistic analysis on morphological characters of the Diplostraca (Conchostraca and Cladocera) has been undertaken for the first time. The morphological information has been obtained from literature and transformed into 56 suitable characters. The analysis included 47 ingroup taxa, comprising five conchostracan taxa (four families of the Spinicaudata and the Laevicaudata) and 42 genera of the Cladocera. A detailed character discussion is presented which will be a useful working base for future phylogenetic studies on the group. A number of systematic groups were, with differing degrees of certainty, supported in all 218 equally short trees. These are the Diplostraca, Cladocera, Gymnomera (Onychopoda and Haplopoda), Onychopoda, Podonidae, Cercopagididae, Anomopoda, Daphniidae, Moininae, Scapholeberinae, Chydoridae, Chydorinae and Sididae. The Spinicaudata were only supported on some of the 218 equally short trees while no support was found for the Conchostraca. Two taxa—the Macrothricidae and Aloninae—were relatively strongly indicated to be paraphyletic. A suggested classificatory hierarchy, without indication of absolute rank, is presented.     

The embryonic development of the Drosophila visual system

We have used electron-microscopic studies, bromodeoxyuridine (BrdU) incorporation and antibody labeling to characterize the development of the Drosophila larval photoreceptor (or Bolwig's) organ and the optic lobe, and have investigated the role of Notch in the development of both. The optic lobe and Bolwig's organ develop by invagination from the posterior procephalic region. After cells in this region undergo four postblastoderm divisions, a total of approximately 85 cells invaginate. The optic lobe invagination loses contact with the outer surface of the embryo and forms an epithelial vesicle attached to the brain. Bolwig's organ arises from the ventralmost portion of the optic lobe invagination, but does not become incorporated in the optic lobe; instead, its 12 cells remain in the head epidermis until late in embryogenesis when they move in conjunction with head involution to reach their final position alongside the pharynx. Early, before head involution, the cells of Bolwig's organ form a superficial group of 7 cells arranged in a rosette pattern and a deep group of 5 cells. Later, all neurons move out of the surface epithelium. Unlike adult photoreceptors, they do not form rhabdomeres; instead, they produce multiple, branched processes, which presumably carry the photopigment. Notch is essential for two aspects of the early development of the visual system. First, it delimits the number of cells incorporated into Bolwig's organ. Second, it is required for the maintenance of the epithelial character of the optic lobe cells during and after its invagination.     

Chydorus arcticus n.sp., a new cladoceran crustacean (Chydoridae: Chydorinae) from the North Atlantic arctic and Subarctic areas

Chydorus arcticus n.sp. (Cladocera: Chydoridae: Chydorinae) is described, figured, and differentiated from the closely relatedC. sphaericus (O.F. Müller, 1785). The known distribution of the species is given, and some aspects of speciation of arctic crustaceans are pointed out.     

Turkana pastoralism: A case against the Tragedy of the Commons

This paper examines land-use practice among the Turkana pastoralists of Kenya. Drawing on work of both ecologists and anthropologists, it examines the argument that posits that pastoralism is inherently destructive to the environment, commonly referred to as the Tragedy of the Commons. Results of this research suggest that the livestock management practices of East African pastoralists do not degrade the environment, that livestock numbers do not exceed the carrying capacity of the land, and that social institutions successfully function to cope with environmental problems.     

Molecular and enzymatic properties of 1,2-α-d-mannosidase fromAspergillus saitoi

The molecular properties, such as molecular weight, N-and C-terminal amino acids, amino acid composition, and circular dichroism, of 1,2--mannosidase isolated from the culture filtrate ofAspergillus saitoi were determined.The enzyme had aK _m of 0.67 mM andk _cat of 1.27/s with mannobiose at pH 50.0 and 30°C. The anomeric configuration of the reaction products of the enzyme was examined by studying the -anomer. A single Manl2Man linkage in intact Taka-amylase A fromAspergillus oryzae was hydrolyzed, producing free mannose.     

The paracellular pathway in toad urinary bladder: Permselectivity and kinetics of opening

Summary Determination of serosa-to-mucosa fluxes of Na, K, and Cl yields information about the properties of the shunt pathway in toad urinary bladder. We show that measurement of these fluxes at 30-sec intervals following an abrupt increase in mucosal osmolality yields evidence on the rate of opening of the path and of its permselectivity. The relationship between the fluxes of any pair of these ions indicates that the shunt is paracellular both before and after the increase in conductance effected by hyperosmolality and that the transepithelial PD affects the permselectivity properties (at 0 mV,P _K/P _Na/P _Cl= 10.710.57; at +25 mV,P _K/P _Na/P _Cl=10.710.99). The relationship between any of the fluxes and the total transepithelial conductance is linear and yields an estimate of cellular conductance (the intercept of this regression on the conductance axis) which is in accord with that measured electrically. These studies provide information on tight junction permeability to nonelectrolytes, as well. Finally, they provide new information about the role of the shunt path as a controlling influence on transepithelial sodium transport and raise the possibility that, in both leaky and tight epithelia, differences in transepithelial conductance from tissue to tissue, organ to organ, and species to species may be due, in the absence of edge damage, to changes in conductance of the paracellular pathway.     

Ultrastructure of the frontal organ in Convoluta and Macrostomum spp.: significance for models of the turbellarian archetype

Present models of turbellarian evolution depict the organism with a frontal organ — a complex of glands whose necks emerge at the anterior tip of the body — and therefore imply that this organ is homologous throughout the Turbellaria. However, comparisons of representatives of the Acoela and Macrostomida, two putatively primitive orders of the Turbellaria, show that frontal organs in these two are not similar in ultrastructure or histochemistry. The acoel Convoluta pulchra had a prominent cluster of frontal mucous glands whose necks emerged together in a frontal pore at the exact apical pole of the organism, and an array of smaller glands of at least five other types opened at the anterior end, separately from and ventral to this pore. The frontal organs (Stirndrüsen) of two species of Macrostomum on the other hand, comprised an array of discretely emerging necks of at least two gland types including one with rhabdiform (rhammite) and one with globular mucous secretion granules neither of which emerge at the apical pole. In neither species did the organ appear to be sensory. Our findings indicate a low probability of homology between the frontal glands of the Acoela and Macrostomida.     

Excision and transposition of two Ds transposons from the bronze mutable 4 derivative 6856 allele of Zea mays L

Summary The regulatory mutation bronze mutable 4 Derivative 6856 (bz-m4 D6856) contains a complex 6.7 kb Dissociation (Ds) element tagged with a duplication of low copy bz 3 flanking sequences (Klein et al. 1988). This creates a unique opportunity to study the transposition of a single member of the repetitive family of Ds elements. Eighteen full purple revertants (Bz alleles) of bz-m4 were characterized enzymatically and by genomic mapping. For 17 of the Bz alleles, reversion to a wild-type phenotype was caused by excision of the 6.7 kb Ds transposon. Nine of these Bz alleles retained the transposon somewhere in their genome. In this study we show that like Ac (Schwartz 1989; Dooner and Belachew 1989), the 6.7 kb Ds element can transpose within a short physical distance, both proximal and distal to its original position. Additional bz sequences have been mapped immediately distal to the mutant locus in bz-m4 D6856; genetic evidence suggests these are flanked by two additional Ds elements. The remaining Bz revertant, Bz :107, arose from excision of a more complex 13 kb Ds element.     

An evaluation of bias in k-factor analysis

Summary Randomization and simulation are used to detect bias in k-factor analysis. In nine previously published data sets there is strong evidence of bias. This may result from either non-independence of observations or the arithmetic relationship used to estimate k-factors, which can generate spurious correlations. Randomization can be used to test for density dependence without bias. This procedure confirms the existence of densitydependent effects in 8 of the 9 populations and 11 of the 16 k-factors previously thought to have density-dependent effects.     

Erratum: Cloning and sequencing of the phycocyanin gene from Spirulina maxima and its evolutionary analysis

The genes were cloned for the two apoprotein subunits, and ,of phycocyanin from the cyanobacterium Spirulina maxima = Arthrospiramaxima) strain F3. The - and -subunit gene-coding regionscontain 489 bp and 519 bp, respectively. The -subunit gene is upstreamfrom the -subunit gene, with a 111-bp segment separating them.Similarities between the -subunits of S. maxima and nine othercyanobacteria were between 58% and 99%, as were those between the -subunits. The maximum similarity between the - and -subunits from S. maxima was 27%.     

The distribution and abundance of krill faecal material and oval pellets in the Scotia and Weddell Seas (Antarctica) and their role in particle flux

Summary The abundance and depth distribution of zooplankton faeces in spring to early summer were investigated along meridional transects (47°W and 49°W) that extended from the Scotia Sea (57°S) across the Weddell-Scotia Confluence and into the Weddell Gyre (62°S). The sea ice edge retreated from 59°30S to 61°S during the study. Faeces were sampled with nets, Niskin bottles and sediment traps and subsequently analysed by light and electron (SEM) microscopy. Krill faecal strings and oval faecal pellets of unknown origin were by far the most important zooplankton faeces and highest concentrations were always found in the Confluence often close to the ice border. Krill faeces were usually more abundant in the uppermost layer (0–50m) where they contributed an average of 130 g dry weight m^–3. There was an exponential decrease with depth, with a minimum of 0.6 g dry weight m^–3 in the 500–1000 m stratum. Oval pellets were more evenly distributed in the upper 1000 m of the water column, with an average of 9 g dry weight m ^–3, although there was a small peak (20 g dry weight m^–3) in the subsurface layer (50–150 m depth). Consecutive collections (day-night) of krill faeces using drifting sediment traps showed that only the larger strings sank from 50 to 150 m depth. Peritrophic membranes appeared to deteriorate during sinking. Diatoms (in particular Nitzschia and Thalassiosira spp.) contributed by far the bulk of material in krill and oval faeces. In samples collected near or under the pack ice, remains of crustaceans in both krill- and oval faeces were also found.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation     

A new isolate of the rhodospirillum fulvum group and its photosynthetic pigments

Summary The present paper reports the isolation of an obligate phototrophic bacterium which belongs to the Rhodospirillum fulvum-group on the basis of its colour, morphology, nutritional requirements and strictly anaerobic nature. p-Aminobenzoic acid was shown to be the only growth factor required. Rhsp. fulvum synthesizes bacteriochlorophyll a as its only chlorophyllous pigments. The carotenoid composition comprises lycopene (I) and rhodopin (II) as the major components; traces of 1,2,1,2-tetrahydro-1,1-dihydroxy-lycopene (III) were also present.

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Zusammenfassung Es wird die Isolierung eines obligat phototrophen Bakteriums beschrieben, welches auf Grund seiner Farbe, Morphologie, Nährstoff-Bedürfnisse und streng anaeroben Lebensweise in die Rhodospirillum fulvum-Gruppe eingeordnet wird. Als einzigen Wachstumsfaktor benötigt der Stamm p-Aminobenzoesäure. Rhsp. fulvum bildet Bacteriochlorophyll a als einzigen chlorophyllartigen Farbstoff. Die Hauptmenge der Carotinoide besteht aus Lycopin (I) und Rhodopin (II); es sind außerdem Spuren von 1,2,12-Tetrahydro-1,1-dihydroxy-Lycopin (III) nachgewiesen.

     

Similarities of omega gliadins from<Emphasis Type="Italic"> Triticum urartu</Emphasis> to those encoded on chromosome 1A of hexaploid wheat and evidence for their post-translational processing

The -gliadins encoded on chromosome 1 of the A genome were purified from Triticum aestivum L. (2n=6x=42, AABBDD) cv. Butte86, nullisomic 1D-tetrasomic 1A of cv. Chinese Spring (CS N1DT1A), and the diploid T. urartu (2n=2x=14, AA). Reverse-phase high-performance liquid chromatography combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis of gliadin extracts from CS nullisomic-tetrasomic (NT) lines confirmed the assignment to chromosome 1A. The purified -gliadins were characterized by mass spectrometry and N-terminal sequencing. The 1A-encoded -gliadins were smaller than 1B- or 1D-encoded -gliadins. The N-terminal amino acid sequences for 1A -gliadin mature peptides were nearly identical to those for the T. urartu -gliadins and were more similar to 1D -gliadin sequences than to sequences for T. monococum -gliadins, barley C-hordeins, or rye -secalins. They diverged greatly from the N-terminal sequences for the 1B -gliadins. The data suggest that T. urartu is the A-genome donor, and that post-translational cleavage by an asparaginyl endoprotease produces those -gliadins with N-terminal sequences beginning with KEL.Communicated by J. Dvorak     

Inheritance of resistance to potyviruses in Phaseolus vulgaris L. II. Linkage relations and utility of a dominant gene for lethal systemic necrosis to soybean mosaic virus

Summary A single dominant factor, Hss, that conditions a rapid lethal necrotic response to soybean mosaic virus (SMV) has been identified in Phaseolus vulgaris L. cv. Black Turtle Soup, line BT-1. Inoculated plants carrying this factor developed pinpoint necrotic lesions on inoculated tissue followed by systemic vascular necrosis and plant death within about 7 days, regardless of ambient temperature. BT-1 also carries dominant resistance to potyviruses attributed to the tightly linked or identical factors, I, Bcm, Cam, and Hsw, so linkage with Hss was evaluated. No recombinants were identified among 381 F₃ families segregating for potyvirus susceptibility, thus if Hss is a distinct factor, it is tightly linked to I, Bcm, Cam, and Hsw. BT-1 was also crossed reciprocally with the line Great Northern 1140 (GN 1140) in which the dominant gene, Smv, for systemic resistance to SMV was first identified. Smv and Hss segregated independently and are co-dominant. The (GN 1140 x BT-1) F₁ populations showed a seasonal shift of the codominant phenotype. Evaluation of the (GN 1140 x BT-1) F₂ population under conditions where Smv is partially dominant allowed additional phenotypic classes to be distinguished. Pathotype specificity has not been demonstrated for either Smv or Hss. Genotypes that are homozygous for both dominant alleles are systemically resistant to the virus and in addition show undetectable local viral replication or and no seed transmission. This work demonstrates that a gene which conditions a systemic lethal response to a pathogen may be combined with additional gene(s) to create an improved resistant phenotype.