Quirks of dye nomenclature. 5. Rhodamines

Rhodamines were first produced in the late 19^th century, when they constituted a new class of synthetic dyes. These compounds since have been used to color many things including cosmetics, inks, textiles, and in some countries, food products. Certain rhodamine dyes also have been used to stain biological specimens and currently are widely used as fluorescent probes for mitochondria in living cells. The early history and current biological applications are sketched briefly and an account of the ambiguities, complications and confusions concerning dye identification and nomenclature are discussed.     

Insertional mutagenesis screening identifies the <Emphasis Type="Italic">zinc finger homeodomain 2</Emphasis> (<Emphasis Type="Italic">zfh2</Emphasis>) gene as a novel factor required for embryonic leg development in <Emphasis Type="Italic">Tribolium castaneum</Emphasis>

The genetic control of leg development is well characterized in the fly Drosophila melanogaster. These control mechanisms, however, must differ to some degree between different insect species to account for the morphological diversity of thoracic legs in the insects. The legs of the flour beetle Tribolium castaneum differ from the Drosophila legs in their developmental mode as well as in their specific morphology especially at the larval stage. In order to identify genes involved in the morphogenesis of the Tribolium larval legs, we have analyzed EGFP enhancer trap lines of Tribolium. We have identified the zfh2 gene as a novel factor required for normal leg development in Tribolium. RNA interference with zfh2 function leads to two alternative classes of leg phenotype. The loss of a leg segment boundary and the generation of ectopic outgrowths in one class of phenotype suggest a role in leg segmentation and segment growth. The malformation of the pretarsal claw in the second class of phenotype suggests a role in distal development and the morphogenesis of the claw-shaped morphology of the pretarsus. This suggests that zfh2 is involved in the regulation of an unidentified target gene in a concentration-dependent manner. Our results demonstrate that enhancer trap screens in T. castaneum have the potential to identify novel gene functions regulating specific developmental processes.     

Die Ultrastruktur des Juxtaglomerulären Apparates des Meerschweinchens

Zusammenfassung Der Beweis für das Vorkommen von Sekretgranula in den epitheloiden Zellen des Meerschweinchens ist bisher von keinem Autor erbracht worden. Ihre Abwesenheit ist um so erstaunlicher, als die Renin-Aktivität in der Niere dieses Tieres etwa 1/10 der der Ratte mit ihren stark granulierten Epitheloidzellen beträgt und die des Menschen sogar übertrifft. In unserem Untersuchungsgut finden wir hin und wieder einige wenige, wahrscheinlich Renin enthaltende Granula (Abb. 1–4). Anscheinend erfolgen Synthese und Ausscheidung des Enzyms im gleichen Rhythmus; zu einer geringgradigen Speicherung kommt es offenbar nur unter bestimmten funktionellen Bedingungen.Die Goormaghtighschen Zellen (Abb. 5) zeigen kein besonderes auffälliges artspezifisches Verhalten.An der Macula densa wird erstmals eine starke Erweiterung sowohl der intracytoplasmatischen Einfaltungen des basalen Plasmalemms — des basalen Labyrinthes — als auch der Interzellularspalten beschrieben (Abb. 6–9). Diese oberhalb der Basalmembran gelegenen Pseudovakuolen sind somit extrazellulär und möglicherweise als morphologisches Äquivalent einer starken Reabsorptionstätigkeit zu deuten. Es ist zur Zeit noch nicht entschieden, ob sie mit der von uns angewandten Präparationstechnik auch bei anderen Tieren und beim Menschen darstellbar sind.

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Summary The presence of secretory granules in the epithelioid (juxtaglomerular) cells in the media of the preglomerular portion of the afferent arterioles in the kidney of the Guinea pig has not been proven by any author until now. The absence of these granules is all the more astonishing in view of the fact that the renin activity of this animal is about 1/10 of that of the rat — with its highly granulated cells — and even surpasses that of humans. In our inbread strain rare granules likely containing renin can be found from time to time (Fig. 1–4). Apparently, the synthesis and extrusion of the enzyme takes place with the same rhythm; a very low degree of accumulation occurs probably only under certain functional conditions.The Goormaghtigh Cells (Fig. 5) show no noticeable differences specific for the Guinea pig. In the Macula densa highly developed enlargements of the basal infoldings of the plasma membrane — basal labyrinthe — as well as of the intercellular spaces are described for the first time (Fig. 6–9). These enlargements (Pseudovacuoles), situated above the basement membrane, are extracellular and can possibly be interpreted as the morphological equivalent of a high degree of absorption activity. At the present time it has not been decided, if, using the authors' technique, these enlargements can be demonstrated in other animals and humans.

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In Zusammenarbeit mit dem Pharmakologischen Institut der Universität Lausanne und mit Unterstützung durch die Fritz Hoffmann-La Roche-Stiftung zur Förderung wissenschaftlicher Arbeitsgemeinschaften in der Schweiz.     

Streptococcus pneumoniae heat shock protein 70 does not induce human antibody responses during infection

Mouse monoclonal antibodies (mAbs) were developed against Streptococcus pneumoniae in search for potential common pneumococcal proteins as vaccine antigens. mAb 230,B-9 (IgG1) reacted by immunoblotting with a 70-kDa protein which was isolated by immunoaffinity chromatography and subsequent preparative electrophoresis. N-terminal amino acid sequencing showed homology to that of heat shock protein 70 (hsp70). The hsp70 epitope reactive with mAb 230,B-9 was found in all the pneumococci examined as well as in other streptococci and enterococci. The epitope was not expressed in several other examined Gram-positive or -negative bacteria. Pneumococcal hsp70 has by other investigators been proposed to be a vaccine candidate. Binding experiments using flow cytometry showed that the epitope was not surface-exposed on live exponential phase grown S. pneumoniae. Human patient sera did not react with affinity-purified pneumococcal hsp70. Therefore the pneumococcal hsp70 does not seem to be of special interest in a vaccine formulation. The human sera contained antibodies to high molecular proteins co-purified with hsp70. Some of these proteins could be the pneumococcal surface protein A.     

Specific gangliosides increase rapidly in rat liver following partial hepatectomy

Rat liver gangliosides (sialic acid containing glycosphingolipids) were analyzed by HPTLC and HPLC following either partial hepatectomy or sham operation. Analysis of whole liver gangliosides by HPTLC demonstrated that within 6 h after partial (68%) hepatectomy, there was a significant increase in GM1 compared to both sham and control animals. By 48 h, GM1 was further increased and the polysialylgangliosides GD1a, GD1b and GT1b had also risen significantly, whereas changes in GM3 were negligible. Gangliosides associated with the plasma membrane were increased up to 3.5-fold in regenerating liver compared to sham-hepatectomized controls as assessed by HPLC. Although elevations in membrane gangliosides were associated with hepatocyte proliferation, they did not closely follow the growth curve. The time course of changes in ganglioside biosynthesis suggests differential upregulation of GM3 synthase and GD3 synthase in regenerating livers.     

Application of an optimized marker amplification protocol in immunohistochemistry — a valuable tool for visualizing antigenic sites of low signal strength or sparse distribution

Amplification of immunohistochemical markers received considerable attention during the 1980s and 1990s. The amplification approach was largely abandoned following the development of antigen retrieval and reporter amplification techniques, because the latter were incorporated more easily into high throughput automated procedures in industrial and diagnostic laboratories. There remain, however, a number of instances where marker amplification still has much to offer. Consequently, we examined experimentally the utility of an optimized marker amplification technique in diagnostically relevant tissue where either the original signal strength was low or positive sites were visible, but sparsely distributed. Marker amplification in the former case not only improved the visibility of existing positive sites, but also revealed additional sites that previously were undetectable. In the latter case, positive sites were rendered more intense and therefore more easily seen during low magnification examination of large areas of tissue.     

Beitrag zum funktionellen Bau des hyalinen Knorpels (auf Grund von Untersuchungen im polarisierten Lichte)

Zusammenfassung Es wurde der Verlauf der interterritorialen kollagenen Fibrillenzüge in verschiedenen Knorpeln des erwachsenen Menschen, einiger Säugetiere (Pferd, Rind, Hund, Kaninchen) und eines Haifisches mittels des Polarisationsmikroskopes untersucht.Alle untersuchten Knorpel zeigen das gleiche Konstruktionsprinzip: zwei Systeme von S-förmigen Fibrillenzügen (vgl. Schema Abb. 1b, S. 282 und Abb. 4a, S. 288), welche, sich unter einem Winkel von durchschnittlich 30–45° kreuzend, den Knorpel auf Quer- und Längsschnitten schräg durchziehen und beiderseits in das Perichondrium einbiegen.Dieses Verhalten entspricht demjenigen, welches zuerst von H. Bormuth für verschiedene Selachierknorpel beschrieben worden ist. Im übrigen werden die Ansichten verschiedener Autoren über den funktionellen Bau des Knorpels diskutiert, und besonders wird auf die Arbeiten von A. Benninghoff eingetreten.Das architektonische Prinzip mit den schräg verlaufenden, sich überkreuzenden Fibrillensystemen ist vielseitiger als das ältere Schema mit den senkrecht zur Oberfläche angeordneten Fibrillenzügen; zudem vermittelt es mit einem geringeren Materialaufwand eine größere Festigkeit.     

Age and growth of mangrove red snapper Lutjanus argentimaculatus at its cool‐water‐range limits

This study investigates the age and growth of Lutjanus argentimaculatus at its southern (cooler) range limits in eastern Australia. Specimens were collected from New South Wales and southern Queensland between November 2011 and December 2013. Fork lengths (L_F) ranged from 190 to 1019 mm, and ages ranged from 2+ to 57+ years. Growth was described by the von Bertalanffy growth function with coefficients L_∞ = 874·92 mm, K = 0·087 year^?1 and t₀ = ?2·76 years. Estimates of the instantaneous natural mortality rate (M) ranged from 0·072 to 0·25. The L_F (mm) and mass (W; g) relationship was represented by the equation: . The maximum age of 57+ years is the oldest reported for any lutjanid and comparisons with tropical studies suggest that the age‐based demography of L. argentimaculatus follows a latitudinal gradient. High maximum ages and low natural mortality rates indicate considerable vulnerability to overexploitation at the species' cool‐water‐range limits. These results demonstrate the need to identify underlying processes driving latitudinal gradients in fish demography.