Isolation and characterization of nonheterocystous tropical cyanobacteria growing on nitrogen-free medium

Summary Fourteen non-heterocystous cyanobacteria were selected from a continuous waterlogged biotope on a mineral medium free of combined nitrogen. Following morphological characterization, four, nine and one of the strains respectively, were assigned to sections I, II and III of a new cyanobacterial classification. Their phenotypic properties, including growth rates, photoheterotrophy, phycobiliprotein content, antibiotic resistance and toxin production were determined. Three strains were found to harbour one to three plasmids of low molecular weight.

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Resumen Mediante un medio mineral sin nitrogéno combinado se seleccionaron catorce cepas de cianobacterias no heterocísticas a partir de un biotopo permanentemente inundado. Apartir de sus carácterísticas morfológicas cuatro, nueve y una de las cepas fueron asignadas respectivamente a las secciones I, II y III de un nuevo sistema para la clasificación de cianobacterias. Se determinaron sus principales características fenotípicas, incluyendo tasas de crecimiento, fotoheterotrofia, contenido en ficobiliproteína, resistencia a antibióticos y producción de toxinas. Tres de las cepas contenían de uno a tres plásmidos de bajo peso molecular.

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Résumé Quatorze souches de cyanobactéries non-hétérocystées ont été isolées d'un biotope inondé sur un milieu minéral dépourvu d'azote combiné. Après détermination de leurs caractères morphologiques, respectivement quatre, neuf et une des souches ont été assignées aux sections I, II et III de la classification des cyanobactéries. Leurs propriétés phénotypiques telles que taux de croissance, photohétérotrophie, contenu en phycobiliprotéines, résistance aux antibiotiques et la production de toxine ont été déterminées. Trois des souches contiennent de un à trois plasmides de faible masse moléculaire.

     

Cladistic analysis of the Cirripedia Thoracica

We present a cladistic analysis of the Cirripedia Thoracica using morphological characters and the Acrothoracica and Ascothoracida as outgroups. The list of characters comprised 32 shell and soft body features. The operational taxonomic units (OTUs) comprised 26 well-studied fossil and extant taxa, principally genera, since uncertainty about monophyly exists for most higher ranking taxonomic units. Parsimony analyses using PAUP 3.1.1 and Hennig86 produced 189 trees of assured minimal length. We also examined character evolution in the consensus trees using MacClade and Clados. The monophyly of the Balanomorpha and the Verrucomorpha sensu stricto is confirmed, and all trees featured a sister group relationship between the ‘living fossil Neoverruca and me Brachylepadomorpha. In the consensus trees the sequential progression of ‘pedunculate‘sister groups up to a node containing Neolepas also conforms to current views, but certain well-established taxa based solely on plesiomorphies stand out as paraphyletic, such as Pedunculata (= Lepadomorpha); Eolepadinae, Scalpellomorpha and Chthamaloidea. The 189 trees differed principally in the position of shell-less pedunculates, Neoverruca, the scalpelloid Capitulum, and the interrelationships within the Balanomorpha, although the 50% majority rule consensus tree almost fully resolved the latter. A monophyletic Sessilia comprising both Verrucomorpha and Balanomorpha appeared among the shortest trees, but not in the consensus. A tree with a monophyletic Verrucomorpha including Neoverruca had a tree length two steps longer than the consensus trees. Deletion of all extinct OTUs produced a radically different tree, which highlights the importance of fossils in estimating cirripede phylogeny. Mapping of our character set onto a manually constructed cladogram reflecting die most recent scenario of cirripede evolution resulted in a tree length five steps longer than any of our shortest trees. Our analysis reveals that several key questions in cirripede phylogeny remain unsolved, notably the position of shell-less forms and the transition from ‘pedunculate‘to ‘sessile‘barnacles. The inclusion of more fossil species at this point in our understanding of cirripede phylogeny will only result in even greater levels of uncertainty. When constructing the character list we also identified numerous uncertainties in the homology of traits commonly used in discussing cirripede evolution. Our study highlights larval ultrastructure, detailed studies of early ontogeny, and molecular data as the most promising areas for future research.     

Concomitant loss of cell surface fibronectin and laminin from transformed rat kidney cells

Both fibronectin and laminin were found by immunofluorescence as a matrix at the surface of normal rat kidney cells. These matrices were absent from the surface of virally transformed rat kidney cells. Soluble fibronectin and laminin were detected in the culture media of the transformed as well as the normal cells. Culture supernates of the transformed cells contained even more fibronectin than the supernates of the transformed cells contained even more fibronectin than the supernates of the normal cells while laminin was present in similar amounts in both culture media. This shows that the loss of fibronectin and laminin from the surface of the transformed cells is caused by failure of the cells to deposit these proteins into an insoluble matrix and not caused by inadequate production. Fibronectins isolated from culture media of the normal and transformed cells were similar in SDS polyacrylamide gel electrophresis. Laminin isolated from culture media by affinity chromatography on heparin-Sepharose followed by immunoprecipitation was composed of three main polypeptides, one with a molecular weight of 400,000 and two with a molecular weight close to 200,000 in both cell types. Fibronectins from both cell types were equally active in promoting cell attachment. Rat fibronectin from transformed cells, like normal cells, when applied to culture dishes coated with fibronectin, readily attached and spread on the substratum, requiring approximately the same amount of fibronectin as the normal cells. On the basis of these results it seem that the failure of the transformed cells to incorporate fibronectin into an insoluble cell surface matix is not a consequence of a demonstrable change in the functional characteristics of the fibronectin molecule or in the ability of the cells to interact with fibronectin. It may depend on as yet unidentified interactions of the cell surface. Similar interactions may be needed for the deposition of laminin into the matrix, because laminin was also absent from the surface of transformed cells, despite its being synthesized by these cells.     

Dissociation of intracellular lysosomal rupture from the cell death caused by silica

The relationship between intracellular lysosomal rupture and cell death caused by silica was studied in P388d(1) macrophages. After 3 h of exposure to 150 μg silica in medium containing 1.8 mM Ca(2+), 60 percent of the cells were unable to exclude trypan blue. In the absence of extracellular Ca(2+), however, all of the cells remained viable. Phagocytosis of silica particles occurred to the same extent in the presence or absence of Ca(2+). The percentage of P388D(1) cells killed by silica depended on the dose and the concentration of Ca(2+) in the medium. Intracellular lyosomal rupture after exposure to silica was measured by acridine orange fluorescence or histochemical assay of horseradish peroxidase. With either assay, 60 percent of the cells exposed to 150 μg silica for 3 h in the presence of Ca(2+) showed intracellular lysosomal rupture, was not associated with measureable degradation of total DNA, RNA, protein, or phospholipids or accelerated turnover of exogenous horseradish peroxidase. Pretreatment with promethazine (20 μg/ml) protected 80 percent of P388D(1) macrophages against silica toxicity although lysosomal rupture occurred in 60-70 percent of the cells. Intracellular lysosomal rupture was prevented in 80 percent of the cells by pretreatment with indomethacin (5 x 10(-5)M), yet 40-50 percent of the cells died after 3 h of exposure to 150 μg silica in 1.8 mM extracellular Ca(2+). The calcium ionophore A23187 also caused intracellular lysosomal rupture in 90-98 percent of the cells treated for 1 h in either the presence or absence of extracellular Ca(2+). With the addition of 1.8 mM Ca(2+), 80 percent of the cells was killed after 3 h, whereas all of the cells remained viable in the absence of Ca(2+). These experiments suggest that intracellular lysosomal rupture is not causally related to the cell death cause by silica or {"type":"entrez-nucleotide","attrs":{"text":"A23187","term_id":"833253","term_text":"A23187"}}A23187. Cell death is dependent on extracellular Ca(2+) and may be mediated by an influx of these ions across the plasma membrane permeability barrier damaged directly by exposure to these toxins.     

Characterization of stable conformational variants of erythrocyte carbonic anhydrases.

Limiting viscosity numbers of bovine and ovine erythrocytes carbonic anhydrase variants were calculated by the objective method of comparing viscosimetric data obtained from low-activity-human erythrocyte carbonic anhydrase and its natural variant. Shifts of mobilities and isoelectric points are shown for all species variants, but variations of limiting viscosity numbers were only detected for human and bovine variants. Results of the study are consistent with the observation that variants arise by deamidation of erythrocyte carbonic anhydrases, and that deamidation is responsible for changes in structure and hydration (i. e. "conformational" modifications). Thus, all the variants so far investigated are stable conformational variants or erythrocyte carbonic anhydrases.     

Capacités reproductrices et descendance de poulets ayant subi un transfert de cellules germinales primordiales durant la vie embryonnaire

Résumé Des cellules germinales primordiales de Dindon, transférées par injection intravasculaire à des embryons de Poulet préalablement stérilisés, peuvent subir une maturation complète dans les gonades de l'hôte et fournir des gamètes plus ou moins aptes à la fécondation.Les spermatozoïdes résultants ont fécondé des oeufs de poule avec une fréquence plus grande que les spermatozoïdes normaux de Dindon, mais sans permettre un développement embryonnaire plus important ou plus normal. Cependant, on n'est pas parvenu à leur faire féconder des oeufs de dinde.Les ufs résultants ont montré parfois un vitellus anormal et n'ont été pondus que durant 7 mois. Mis en présence de spermatozoïdes de coq, ils ont été quelquefois fécondés (ou peutêtre simplement activés), mais ils n'ont jamais fourni d'embryons. Fécondés par des spermatozoïdes de Dindon, ils ont fourni des embryons, parfois anormaux, qui ont atteint dans le meilleur cas le 15è jour d'incubation (stade 38 HH). Certainespraepennae de l'embryon qui a atteint ce stade montraient une pigmentation rouge brun qui ne peut pas être déterminée par le génotype du zygote (celui d'un Dindon de race blanche) et qui rappelle le phénotype de la mère nourricière (une poule rouge brun).A la suite de transferts intraspécifiques de cellules germinales primordiales, on a pu obtenir la maturation d'ovocytes de Rhode Island Red à l'intérieur d'un ovaire de Wyandotte Blanche (chez deux poules) et vice versa (chez une poule). La ponte a également été possible, mais à une fréquence souvent inférieure à la normale.Lorsqu'une poule Wyandotte Blanche, portant des ovocytes de Rhode Island Red a été accouplée à un coq Rhode Island Red normal, le duvet de leurs descendants s'est montré plus clair que celui des poussins Rhode Island Red dans un cas, mais il a été remplacé ensuite par un plumage rouge brun conforme au génotype.Lorsqu'une poule Rhode Island Red, portant des ovocytes de Wyandotte Blanche a été accouplée à un coq Wyandotte Blanche normal, le duvet de leurs descendants n'a jamais été conforme au génotype. Il présentait toujours du pigment noir plus ou moins étendu et, dans un cas, du pigment rouge brun, qui sont tous deux présents chez la mère nourricière. On ne connaît ni l'origine ni le mécanisme d'un tel transfert de pigments, qui ne représente peut-être qu'un effet transitoire affectant le duvet de la première génération. La crête, quant à elle, s'est toujours montrée conforme au génotype d'origine.

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Reproductive capacity and offspring of chickens submitted to a transfer of primordial germ cells during embryonic life

Summary Turkey primordial germ cells transfered by intravascular injection to previously sterilized chick embryos can undergo complete maturation inside the host's gonads and can give rise to gametes which are more or less suitable for fertilization.The resulting spermatozoa fertilized hen eggs at a higher frequency than normal turkey spermatozoa, but without allowing a longer or a more normal development. However, it was impossible to fertilize turkey eggs with them.The resulting eggs sometimes had an abnormal-looking yolk and were laid during the first 7 months only. Brought in contact with chicken spermatozoa, they were fertilized (or perhaps merely activated), but they never gave rise to embryos. Fertilized by turkey spermatozoa, they developed into embryos, sometimes abnormal, which in the best case reached the 15th day of incubation (stage 38 HH). Somepraepennae of the latter embryo showed a red-brown pigment which cannot be determined by the genotype of the zygote (a white turkey's) and which resembled the phenotype of the foster mother (a red-brown hen).After intraspecific transfer of primordial germ cells, maturation of Rhode Island Red oöcytes inside a Wyandotte White ovary (in two hens) and vice versa (in one hen) was achieved. Laying was also possible but often at a lower frequency than normal.When a Wyandotte White hen bearing Rhode Island Red oöcytes was mated with a normal Rhode Island cock, the down of their offspring looked brighter than Rhode Island Red chicken's in one case, but it was subsequently replaced by red-brown feathers according to the genotype.When a Rhode Island Red hen bearing Wyandotte White oöcytes was mated with a normal Wyandotte White cock, the down of their offspring was never in agreement with the genotype. It always showed a black pigment over more or less large areas and, in one case, a red-brown pigment, both of which were present in the foster mother. The origin and the mechanism of such a transfer of pigments are not understood. It might represent merely a temporary effect acting upon the down of the first generation. As far as the comb is concerned, it was always in agreement with the original genotype.

     

In Situ Fe and S isotope analyses in pyrite from the 3.2 Ga Mendon Formation (Barberton Greenstone Belt,South Africa): Evidence for early microbial iron reduction

On the basis of phylogenetic studies and laboratory cultures, it has been proposed that the ability of microbes to metabolize iron has emerged prior to the Archaea/Bacteria split. However, no unambiguous geochemical data supporting this claim have been put forward in rocks older than 2.7–2.5 giga years (Gyr). In the present work, we report in situ Fe and S isotope composition of pyrite from 3.28‐ to 3.26‐Gyr‐old cherts from the upper Mendon Formation, South Africa. We identified three populations of microscopic pyrites showing a wide range of Fe isotope compositions, which cluster around two δ⁵⁶Fe values of ?1.8‰ and +1‰. These three pyrite groups can also be distinguished based on the pyrite crystallinity and the S isotope mass‐independent signatures. One pyrite group displays poorly crystallized pyrite minerals with positive Δ³³S values > +3‰, while the other groups display more variable and closer to 0‰ Δ³³S values with recrystallized pyrite rims. It is worth to note that all the pyrite groups display positive Δ³³S values in the pyrite core and similar trace element compositions. We therefore suggest that two of the pyrite groups have experienced late fluid circulations that have led to partial recrystallization and dilution of S isotope mass‐independent signature but not modification of the Fe isotope record. Considering the mineralogy and geochemistry of the pyrites and associated organic material, we conclude that this iron isotope systematic derives from microbial respiration of iron oxides during early diagenesis. Our data extend the geological record of dissimilatory iron reduction (DIR) back more than 560 million years (Myr) and confirm that micro‐organisms closely related to the last common ancestor had the ability to reduce Fe(III).     

Does Optic Nerve Head Surface Topography Change Prior to Loss of Retinal Nerve Fiber Layer Thickness: A Test of the Site of Injury Hypothesis in Experimental Glaucoma

Purpose

To test the hypothesis that optic nerve head (ONH) deformation manifesting as changes in its mean surface height precedes thinning of the peripapillary retinal nerve fiber layer (RNFL) in experimental glaucoma (EG).

Methods

68 rhesus macaque monkeys each had three or more baseline imaging sessions under manometric intraocular pressure (IOP) control to obtain average RNFL thickness (RNFLT) and the ONH surface topography parameter mean position of the disc (MPD). Laser photocoagulation was then applied to the trabecular meshwork of one eye to induce chronic, mild-to-moderate IOP elevation and bi-weekly imaging continued. Event analysis was applied to determine for each parameter when an ‘endpoint’ occurred (signficant change from baseline) for eight different endpoint criteria. Specificity was assessed in the group of 68 fellow control eyes. Classical signal detection theory and survival analysis were used to compare MPD with RNFLT.

Results

Regardless of the endpoint criterion, endpoints were always more frequent for MPD than for RNFLT. The discriminability index (d’) was 2.7 ± 0.2 for MPD and 1.9 ± 0.2 for RNFLT (p<0.0001). Endpoints were reached by MPD an average of 1-2 months earlier than by RNFLT (p<0.01). At the onset of the first specific, detectable MPD change in EG eyes, there was still no significant change in RNFLT on average (p=0.29) and only 25% of individual eyes exhibited signficant reduction. In contrast, at onset of signficant RNFLT change, MPD had already changed an average of 101 µm from baseline (p<0.0001) and 71% of the individual eyes had exhibited significant change. The magnitude of MPD change was more than could be explained on the basis of axon loss alone.

Conclusions

This study demonstrates that the average surface height of the ONH changes prior to any detectable loss of average peripapillary RNFL thickness in non-human primate eyes with experimental glaucoma.