Quantitation of mitochondrial injury in cultured rat heart endothelioid cells with nitroblue tetrazolium

Synopsis A sensitive method is presented for measurement of changes in the permeability of mitchondria in cultured cells. Rat heart endothelioid cells were used to determine the penetration rate of nitroblue tetrazolium (NitroBT) or other reactants into mitochondriain situ. Nitroblue formazan, produced as a consequence of succinate dehydrogenase activity in the mitochondria, was eluted and measured with a spectrophotometer. Prior injury of cells with hypo-osmolar solutions increased the rate of formazan production. Several methods are described or suggested for the statistical analysis of the data.     

“Self” and “Non-Self” in the Control of Phytoalexin Biosynthesis: Plant Phospholipases A2 with Alkaloid-Specific Molecular Fingerprints

The overproduction of specialized metabolites requires plants to manage the inherent burdens, including the risk of self-intoxication. We present a control mechanism that stops the expression of phytoalexin biosynthetic enzymes by blocking the antecedent signal transduction cascade. Cultured cells of Eschscholzia californica (Papaveraceae) and Catharanthus roseus (Apocynaceae) overproduce benzophenanthridine alkaloids and monoterpenoid indole alkaloids, respectively, in response to microbial elicitors. In both plants, an elicitor-responsive phospholipase A2 (PLA2) at the plasma membrane generates signal molecules that initiate the induction of biosynthetic enzymes. The final alkaloids produced in the respective plant inhibit the respective PLA, a negative feedback that prevents continuous overexpression. The selective inhibition by alkaloids from the class produced in the “self” plant could be transferred to leaves of Nicotiana benthamiana via recombinant expression of PLA2. The 3D homology model of each PLA2 displays a binding pocket that specifically accommodates alkaloids of the class produced by the same plant, but not of the other class; for example, C. roseus PLA2 only accommodates C. roseus alkaloids. The interaction energies of docked alkaloids correlate with their selective inhibition of PLA2 activity. The existence in two evolutionary distant plants of phospholipases A2 that discriminate “self-made” from “foreign” alkaloids reveals molecular fingerprints left in signal enzymes during the evolution of species-specific, cytotoxic phytoalexins.     

Ornithologische Notizen aus dem Ebrodelta

Zusammenfassung Im Ebrodelta ist, abgesehen von einigen Seen und Sümpfen, nur noch ein schmaler Küstenstreifen landwirtschaftlich ungenutzt. Hier hat sich ein Teil ursprünglichen Vogelreichtums erhalten, und hier wurde seit einigen Jahren eine Reihe faunistischer Erhebungen durchgeführt.Diese erbrachten mehrere erste Brutnachweise für Spanien oder die spanische Mittelmeerküste:Larus ridibundus, Sterna dougallii, St. sandvicensis, Haematopus ostralegus, Limosa limosa, Asio flammeus, Acrocephalus palustris. Als Rast- und Überwinterungsgebiet ist das Delta von großer Bedeutung, z. B. fürEgretta garzetta undPhoenicopterus ruber. Enge Beziehungen zur Camargue sind vielfach erkennbar, vielleicht sogar hinsichtlich der Ausbreitung vonLarus ridibundus undHaematopus ostralegus. Regelmäßige Wintervorkommen vonMarmaronetta angustirostris sind erwähnenswert.Es wird mit einigen bisher unbekannten Brutplätzen von Laro-Limicolen bekanntgemacht, von denen eine Kolonie vonSterna albifrons mit 300–400 Paaren besonders vermerkt sei. Für weitere Arten können Bestandsangaben gemacht werden.Dieses einzige, bedeutende Brut- und Rastgebiet zwischen Camargue und Coto Doñana ist in höchster Gefahr, durch intensivere landwirtschaftliche Nutzung und touristische Erschließung vernichtet zu werden.

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Resumen Datos ornitologicos sobre el Delta del Ebro. — Sin contar con charcas y marismas, el Delta del Ebro es una franja costera aun no explotada por la agricultura. Gracias a esto una numerosa variedad de aves ha podido mantenerse aprovechando las todavia persistentes condiciones orginarias del medio ambiente. Sobre estas aves se han venido acumulando en los ultimos años un buen acopio de observaciones.Se pudo constatar como nidificantes por primera vez en España o costas mediterranes a las siguientes especies:Larus ridibundus, Sterna dougalli, St. sandvicensis, Haematopus ostralegus, Limosa limosa, Asio flammeus, Acrocephalus palustris. Como lugar de invernada o refugio durante el paso el Delta del Ebro tiene un especial interes, por ejemplo paraEgretta garzetta yPhoenicopterus ruber. Se establecen algunas relaciones con la Camarga principalmente en cuanto a la expansión deLarus ridibundus yHaematopus ostralegus. Se mencionan invernadas regulares deMarmaronetta angustirostris. Se señalan algunos nuevos lugares de cria para Laro-Limicolas, merece destacarse el hallazgo de una colonia deSterna albifrons compuesta por unos 300–400 pares. Para otras especies se dan datos numerieos estimativos.El Delta del Ebro es un lugar unico y de gran importancia, situado entre la Camarga y el Coto de Doñana, que está hoy en peligro de desaparición debido a la colonización intensiva de la agricultura y a la creciente afluencia turistica.

     

Dose-response function for Painter's SCE-model

Summary The replication model for sister chromatid exchange (SCE), when introduced in 1980 by Painter, was claimed to be consistent with the one hit property of SCE. However, the argument offered in favour of the one hit property was based on a defective dose-response function, as shown in this paper, since dose as the independent parameter of any dose-response function was not included in the considerations. This missing part of the model's dose-response function is added and, by using Bessel functions, a formula for the complete dose-response function is presented. A re-examination of the newly derived function shows that, in the model, linearity holds only under certain restricted circumstances.     

Treatment of the Obstructive Sleep Apnea Syndrome

The obstructive sleep apnea syndrome is a disorder of sleep and breathing that is being recognized with increasing frequency. The pathophysiologic consequences range from mild sleepiness to life-threatening cardiovascular and respiratory decompensation. The primary forms of treatment are directed at modifying the upper airway with either an operation or continuous positive airway pressure. Aside from tracheostomy, which is virtually always successful, other forms of treatment have met with varying results. Ancillary therapy, including oxygen, weight loss and drugs, is often helpful but seldom curative. Follow-up sleep studies are necessary to evaluate the effectiveness of treatment. Selecting therapy for a patient with obstructive sleep apnea requires a comprehensive evaluation including polysomnography, special examinations of the upper airway and assessing the cardiopulmonary status. Therapy is based on the severity of disease and must be tailored to each patient.     

Two Agrobacterium tumefaciens genes for cytokinin biosynthesis: Ti plasmid-coded isopentenyltransferases adapted for function in prokaryotic or eukaryotic cells

Summary Tzs and ipt are two Ti plasmid genes coding for proteins with isopentenyltransferase (IPT) activity in vitro. We cloned both genes for protein expression in Escherichia coli and in Agrobacterium tumefaciens, and we investigated differences between the two genes by analysing the properties of the proteins in vitro and in vivo. In vitro, extracts with tzs or ipt-coded proteins had high IPT activity, and the enzymes were identical in most properties. The most important difference was detected in vivo: the tzs-encoded protein was very active in cytokinin production, while the ipt protein required overexpression in order to obtain measurable activity in bacteria. In both cases, rans-zeatin was the major product of the gene activity. Formation of this cytokinin requires a hydroxylase function in addition to the IPT reaction. No such activity could be ascribed to tzs or ipt-encoded proteins in vitro or in vivo, but cytokinin hydroxylase activity was detected in cells and extracts of E. coli, regardless of the presence or absence of the cytokinin genes. Based on these results it is proposed that both genes code for a single enzyme activity (isopentenyltransferase), that the genes and proteins are adapted for function either in bacteria (tzs) or in transformed plant cells (ipt), and that in both prokaryotic and eukaryotic cells hydroxylation to trans-zeatin is a function contributed by host enzymes.Abbreviations DMAPP dimethylallylpyrophosphate - iP isopentenyladenine - iPA isopentenyladenosine - iPMP isopentenyladenosine 5-monophosphate - IPT isopentenyltransferase - trans-Z trans-zeatin