Purification and Characterization of Glucose-6-Phosphate Dehydrogenase from Rat Small Intestine

Glucose-6-phosphate dehydrogenase (G6PD) was purified from rat small intestine with 19.2% yield and had a specific activity of 53.8 units per miligram protein. The pH optimum was determined to be 8.1. The purified rat small intestinal G6PD gave one activity, one protein band on native PAGE. The observation of one band on SDS/PAGE with an Mr of 48 kDa and a specific activity lower than expected may suggest the proteolytically affected enzyme or different form of G6PD in the rat small intestine. The activation energy, activation enthalpy, Q10, and optimum temperature from Arrhenius plot for the rat small intestinal G6PD were found to be 8.52 kcal/mol, 7.90 kcal/mol, 1.59, and 38 degrees C, respectively. The Km values for G6P and NADP+ were 70.1 +/- 20.8 and 23.2 +/- 7.6 microM, respectively. Double-reciprocal plots of 1/Vm versus 1/G6P (at constant [NADP+]) and of 1/Vm versus 1/NADP+ at constant [G6P]) intersected at the same point on the 1/Vm axis to give Vm = 53.8 U/mg protein.     

Cloning of a cDNA for rape chloroplast 3-isopropylmalate dehydrogenase by genetic complementation in yeast

Both insect and mammalian genes have previously been cloned by genetic complementation in yeast. In the present report, we show that the method can be applied also to plants. Thus, we have cloned a rape cDNA for 3-isopropylmalate dehydrogenase (IMDH) by complementation of a yeast leu2 mutation. The cDNA encodes a 52 kDA protein which has a putative chloroplast transit peptide. The in vitro made protein is imported into chloroplasts, concomitantly with a proteolytic cleavage. We conclude that the rape cDNA encodes a chloroplast IMDH. However, Southern analysis revealed that the corresponding gene is nuclear. In a comparison of IMDH sequences from various species, we found that the rape IMDH is more similar to bacterial than to eukaryotic proteins. This suggests that the rape gene could be of chloroplast origin, but has moved to the nucleus during evolution.     

Notes on Convolvulus, Astripomoea, Ipomoea and Merremia (Convolvulaceae) from the Horn of Africa

The six new species Convolvulus scopulatus, a shrub from gypseous semidesert coastal plains in northern Somalia, Astripomoea procera, a woody climber in Acacia-Commiphora bushland on sand in south-central Somalia, Ipomoea hiranensis, a prostrate or climbing shrublet in Acacia-Commiphora bushland in rocky places on sandstone in south-central Somalia, Ipomoea galhareriana, a woody climber in bushland on sand in central Somalia, Ipomoea pogonantha, a woody climber in bushland usually on red soil over limestone in eastern and southern Ethiopia, southern Somalia and northern Kenya, and Ipomoea corrugata, a herb with trailing stems in bushland on sandy soil in south-central Somalia, are described, and illustrations are provided for most of them. The new combinations Ipomoea ovatolanceolata (= I. adenoides var. ovatolanceolata) and Merremia obtusa (= M. ampelophylla subsp. obtusa) are made, and a new synonymy is provided for M. ampelophylla. Merremia ellenbeckii is neotypified and M. lobata is placed in synonymy.     

A cryptic invasion within an invasion and widespread introgression in the European water frog complex: consequences of uncontrolled commercial trade and weak international legislation

In Western Europe, many pond owners introduce amphibians for ornamental purposes. Although indigenous amphibians are legally protected in most European countries, retailers are circumventing national and international legislation by selling exotic nonprotected sibling species. We investigated to what extent non‐native species of the European water frog complex (genus Pelophylax) have become established in Belgium, using morphological, mitochondrial and nuclear genetic markers. A survey of 87 sampling sites showed the presence of non‐native water frogs at 47 locations, mostly Marsh frogs (Pelophylax ridibundus). Surprisingly, at least 19% of all these locations also harboured individuals with mitochondrial haplotypes characteristic of Anatolian water frogs (Pelophylax cf. bedriagae). Nuclear genotyping indicated widespread hybridization and introgression between P. ridibundus and P. cf. bedriagae. In addition, water frogs of Turkish origin obtained through a licensed retailer, also contained P. ridibundus and P. cf. bedriagae, with identical haplotypes to the wild Belgian populations. Although P. ridibundus might have invaded Belgium by natural range expansion from neighbouring countries, our results suggest that its invasion was at least partly enhanced by commercial trade, with origins as far as the Middle East. Also the invasion and rapid spread of Anatolian lineages, masked by their high morphological similarity to P. ridibundus, is likely the result of unregulated commercial trade. We expect that Anatolian frogs will further invade the exotic as well as the native range of P. ridibundus and other Pelophylax species elsewhere in Western and Central Europe, with risks of large‐scale hybridization and introgression.     

De novo spatiotemporal modelling of cell-type signatures in the developmental human heart using graph convolutional neural networks

With the emergence of high throughput single cell techniques, the understanding of the molecular and cellular diversity of mammalian organs have rapidly increased. In order to understand the spatial organization of this diversity, single cell data is often integrated with spatial data to create probabilistic cell maps. However, targeted cell typing approaches relying on existing single cell data achieve incomplete and biased maps that could mask the true diversity present in a tissue slide. Here we applied a de novo technique to spatially resolve and characterize cellular diversity of in situ sequencing data during human heart development. We obtained and made accessible well defined spatial cell-type maps of fetal hearts from 4.5 to 9 post conception weeks, not biased by probabilistic cell typing approaches. With our analysis, we could characterize previously unreported molecular diversity within cardiomyocytes and epicardial cells and identified their characteristic expression signatures, comparing them with specific subpopulations found in single cell RNA sequencing datasets. We further characterized the differentiation trajectories of epicardial cells, identifying a clear spatial component on it. All in all, our study provides a novel technique for conducting de novo spatial-temporal analyses in developmental tissue samples and a useful resource for online exploration of cell-type differentiation during heart development at sub-cellular image resolution.     

Ch-Schwellen der Uraninfärbbarkeit des Plasmas einiger Florideen

Ohne Zusammenfassung     

Osmotischer Druck und Ionengleichgewicht beim Blutegelmuskel

Zusammenfassung Es wurde das Verhalten des Blutegelrückenmuskels in Lösungen von verschiedenem osmotischen Druck und verschiedenem Ionengehalt untersucht. In reinen Zuckerlösungen sowie in reinen Kochsalzlösungen zeigt der Muskel einen von der Größe des osmotischen Druckes abhängigen Längszustand oder Tonus. Die Druck-Längenkurve zeigt in beiden Fällen einen Wendepunkt. Der in n/10-NaCl im Gleichgewicht befindliche Muskel erschlafft bei Herabsetzung des osmotischen Druckes bis zu einer Konzentration von n/20–n/30 und beginnt von da ab sich zu kontrahieren. In Na-freier Zuckerlösung liegt der Wendepunkt bei etwa molarer Konzentration, also bei etwa zehnmal so hohem osmotischen Druck. Erhöhung der Zuckerkonzentration führt zu Erschlaffung, Verminderung zu Kontraktion.Zusatz von Kalium zur NaCl-Lösung in einer die Isotonie nicht störenden Menge bewirkt eine wiederum von der Konzentration abhängige Längen- oder Tonusänderung, die gleichfalls einen Wendepunkt zeigt. Dieser liegt bei einer Konzentration von etwa n/250–n/200. Schwächere Konzentrationen bewirken eine Erschlaffung, stärkere eine Erhöhung des Muskeltonus. Kalzium wirkt in allen Konzentrationen erschlaffend. Es erscheint als Antagonist des Kaliums, wenn dieses kontrahierend wirkt, aber als Synergist, bei Kaliumkonzentrationen, die erschlaffend wirken.In O₂-reichem Medium bleibt die elektrische Reizbarkeit über 30 Stunden erhalten. Der Muskel zeigt ein großes Summationsvermögen, auch für einzeln unwirksame Reize. Die Erschlaffungsgeschwindigkeit steigt mit der Steilheit der Kontraktion. O₂-Mangel wirkt tonusvermindernd und erschlaffungsbeschleunigend. Hypotonie erhöht, Hypertonie vermindert die elektrische Reizbarkeit. Kalium hat wenig Einfluß, Kalzium erzeugt eine starke Herabsetzung der Reizbarkeit. Tonus und Reizkontraktion sind mithin von einander völlig unabhängige Erscheinungen.

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Summary The behaviour of the dorsal muscle of leech in solutions of different osmotic pressure and different ions has been investigated. In pure sugar solutions as well as in pure solutions of sodium chloride the permanent state of length (tonus) of the muscle depends on the osmotic pressure of the solution. The curve representing the relation between pressure and length reveals in both cases a turning point. The muscle beeing in equilibrium in n/10-NaCl solution relaxes with diminishing osmotic pressure untill a concentration of n/20–n/30 is reached. With further diminution starts a contraction.In sodium free sugar solutions the turning point lies at approximatly molar concentrations that is at osmotic pressure ten times higher. Increase of sugar concentration leads to relaxation and its diminution to a contraction.Addition of potassium to the NaCl solution without disturbing the isotonic condition produces also a change of the state of length or the tonus. This change too depends on the concentration and shows a turning point at a K-concentration of about n/250–n/200. Lower concentrations produce a relaxation and higher ones a contraction. Calcium has a relaxing effect in all concentrations. It is an antagonist of potassium as far as the latter one produces a contraction, but acts as its Synergist in K-concentrations leading to a relaxation.In a medium rich in oxygen the electrical excitability of the muscle may be preserved for more than 30 hours. The muscle shows a great power of summation even for single shocks not effective by itself. The velocity of relaxation increases with the steepness of the contraction. Lack of oxygen diminishes the tonus and accelerates the relaxation. Hypotonic solutions increase while hypertonic ones diminish the electrical excitability. Potassium has little influence whereas calcium produces a strong diminution of excitability.Therefore tonus and contractions by stimulation are entirely independent phenomena.

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Karl von Frisch zum 60. Geburtstag gewidmet.     

Polyploids and aneuploids of crepis capillaris produced by treatment with nitrous oxide

Summary When plants ofCrepis capillaris are treated with nitrous oxide of ten atmospheres pressure for four to six hours, at the time when the first or second zygotic divisions are passing in their pollinated flowers, a fair yield of polyploid and aneuploid plants is obtained in their progeny.     

A Real-Time PCR Array for Hierarchical Identification of Francisella Isolates

A robust, rapid and flexible real-time PCR assay for hierarchical genetic typing of clinical and environmental isolates of Francisella is presented. Typing markers were found by multiple genome and gene comparisons, from which 23 canonical single nucleotide polymorphisms (canSNPs) and 11 canonical insertion-deletion mutations (canINDELs) were selected to provide phylogenetic guidelines for classification from genus to isolate level. The specificity of the developed assay, which uses 68 wells of a 96-well real-time PCR format with a detection limit of 100 pg DNA, was assessed using 62 Francisella isolates of diverse genetic and geographical origins. It was then successfully used for typing 14 F. tularensis subsp. holarctica isolates obtained from tularemia patients in Sweden in 2008 and five more genetically diverse Francisella isolates of global origins. When applied to human ulcer specimens for direct pathogen detection the results were incomplete due to scarcity of DNA, but sufficient markers were identified to detect fine-resolution differences among F. tularensis subsp. holarctica isolates causing infection in the patients. In contrast to other real-time PCR assays for Francisella, which are typically designed for specific detection of a species, subspecies, or strain, this type of assay can be easily tailored to provide appropriate phylogenetic and/or geographical resolution to meet the objectives of the analysis.