New chromosome 21 DNA markers isolated by pulsed field gel electrophoresis from an ETS2-containing Down syndrome chromosomal region

To generate new chromosome 21 markers in a region that is critical for the pathogenesis of Down syndrome (D21S55-MX1), we used pulsed field gel electrophoresis (PFGE) to isolate a 600-kb NruI DNA fragment from the WA17 hybrid cell line, which has retained chromosome 21 as the only human material. This fragment, which contains the oncogene ETS2, was used to construct a partial genomic library. Among the 14 unique sequences that were isolated, 3 were polymorphic markers and contained sequences that are conserved in mammals. Five of these markers mapped on the ETS2-containing NruI fragment and allowed us to define an 800-kb high-resolution PFGE map.     

Variants of the anti-Müllerian hormone gene in a compound heterozygote with the persistent Müllerian duct syndrome and his family

Summary The human genome contains a large number of interspersed simple repeat sequences that are variable in length and can therefore serve as highly informative, polymorphic markers. Typing procedures include conventional multilocus and single locus probing, and polymerase chain reaction aided analysis. We have identified simple sequences in a cosmid clone stemming from the human Y chromosome and consisting of (gata)_n repeats. We have compared these with two equivalent simple repeat loci from chromosome 12. After amplifying the tandemly repeated motifs, we detected between four and eight different alleles at each of the three loci. Codominant inheritance of the alleles was established in family studies and the informativity of the simple repeat loci was determined by typing unrelated individuals. The polymorphisms are suitable for application in linkage studies, practical forensic case work, deficiency cases in paternity determination, and for studying ethnological questions. The mutational mechanisms that bring about changes in simple repeats located both on the autosomes and on the sex chromosomes, are discussed.Professor Dr. Otto Prokop (Humboldt-Universität Berlin) on the occasion of his 70th birthday     

The potential use of silicon compounds as oxygen carriers for free and immobilized cells containing l-amino acid oxidase

Summary The relatively low solubility of oxygen in water presents a problem in particular when immobilized cells are used or enzymes are applied for oxygen dependent reactions. The other main purpose is the requirement of oxygen for the increase of biomass. In this investigation the usefulness of silicon emulsions as oxygen carriers is demonstrated.In case of l-amino acid oxidase activity of immobilized cells, an increase by a factor of four was found in the presence of silicon emulsions. Likewise, growth medium enriched with silicon emulsions showed a significantly increased growth of cells inside alginate beads compared to normal growth medium.     

Bukettförmige Strukturen im Ependym der Regio hypothalamica des III. Ventrikels beim Kaninchen

Zusammenfassung Die Wand des III. Ventrikels (Regio hypothalamica) vom erwachsenen Kaninchen wurde elektronenmikroskopisch untersucht. — Unterschiedliche keulenförmige portsätze, deren Perikaryen in nahen und entfernten Regionen liegen, dringen in den Ventrikel ein. Sie heben eine Ependymzelle ab und bilden eine bukettförmige Struktur. Einige Fortsätze enthalten eine spärliche feingranulierte Masse, andere Bläschen (600–1500 Å) oder Bläschen mit dichtem Kern (450–1200 Å). Inmitten des Buketts kann eine Nervenzelle liegen, die einen dicken Fortsatz gegen das Hypendym schickt. Unterhalb des Buketts findet man eine andere Nervenzelle, die Fortsätze ins Bukett sendet. Zahlreiche in das Bukett eintretende Fortsätze stammen aus entfernteren Kerngebieten. Die funktionelle Bedeutung der Strukturen (Neurosekretion ? Resorption ? Rezeptor ?) wird diskutiert.

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Bouquet-like ependymal structures in the hypothalamic region of the 3rd Ventricle (rabbit)The question of neurosecretion or receptor function

Summary The wall of the III. ventricle (Regio hypothalamica) of the adult rabbit has been investigated electron microscopically. —Various bulblike processes of nerve cells, having their perikarya in close or distant regions, penetrate into the ventricle. They lift an ependymal cell and form a bouquet-like structure. Some of the processes contain a sparse granulated material and others include vesicles (600–1500 Å) or dense-cored vesicles (450–1200 Å). In the middle of the bouquet a perikaryon may be seen extending a thick process towards the hypendyma. Below the bouquet another perikaryon may be found extending several processes into the bouquet. The functional significance of these structures (neurosecretion ? resorption? receptor?) is discussed.

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Die Untersuchung wurde mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Serology of Rubella—Comparison of Fluorescent Antibody,Complement Fixation and Neutralization Tests for Diagnosis of Current Infections and Determination of Sero-immunity

Neutralization, complement fixation (CF) and indirect fluorescent antibody (FA) assays for rubella virus were compared for sensitivity in the serologic diagnosis of infection, for demonstrating antibody in the sera of infants with suspected rubella syndrome, and in the detection of antibody elicited by past infection (determination of immunity status). The combination of CF and FA tests was shown to be the most useful for serologic diagnosis of infection, largely eliminating the need for the slower and more cumbersome interference neutralization test.Neutralizing antibodies were found to appear rapidly in the course of infection, antibodies demonstrable by immunofluorescent staining appeared slightly later, and CF antibodies were rarely demonstrable in sera collected earlier than 14 days after onset of illness. Antibodies detected by all three techniques showed good correlation in infants with clinical evidence of rubella syndrome and corresponding maternal sera. The indirect FA technique compared favorably with the neutralization test for the detection of antibody elicited by past infection (determination of immunity status) and offered distinct advantages in ease of technical performance and more rapid results. In both current and past infections, FA titers tended to be higher than neutralizing antibody titers.     

Intraventrikuläre markhaltige Nervenfasern nahe der Apertura lateralis ventriculi quarti des Kaninchengehirns

Zusammenfassung In der Gegend der Apertura lateralis ventriculi quarti des Kaninchengehirns wurden intraventrikuläre markhaltige Nervenfasern der Fasergruppe B gefunden. Ihre Markscheide beginnt oder endigt innerhalb des Ventrikellumens. Besonderheiten des Ependyms dieser Gegend werden mitgeteilt.

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Summary In the 4th ventricle of the brain near the apertura lateralis in rabbit myelinated nerve fibers occur. These nerve fibers belong to the fiber group B. Their myelin sheath begins or ends in the ventricular cavity. Peculiarities of the ependyma of this region are discussed.

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Die Untersuchung wurde mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Über die Blutkapillaren und perivaskulären Strukturen der Area postrema des Kaninchens und über ihr Verhalten im Pentamethylentetrazol-(„Cardiazol“-)Krampf

Zusammenfassung Die sinusförmigen Kapillaren der Area postrema werden von zwei Basalmembranen umgeben, die einen Raum einschließen, in dem Makrophagen liegen. Verästelte Ausläufer der äußeren Basalmembran dringen tief zwischen die Gliafortsätze ein und bilden ein Membranlabyrinth. Das Kapillarendothel ist fenestriert. Nach Cardiazolkrampf entsteht ein perivaskuläres Oedem, das das Membranlabyrinth öffnet und den perivaskulären Raum vergrößert. Das Endothel zeigt hierbei starke Cytopempsis. Die funktionelle Bedeutung der Strukturen wird diskutiert.

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Summary The sinusoid capillaries of the area postrema are surrounded by two basement membranes enclosing a perivascular space, in which macrophages are found. Ramified extensions of the outer basement membrane penetrate deeply between the glial processes forming a labyrinth of basement membranes. The capillary endothelium is fenestrated. After a Cardiazolinduced convulsion a perivascular edema opens the basement membrane labyrinth and the perivascular space is greatly enlarged. The endothelium shows pinocytotic activity. The functional significance of these structures is discussed.

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Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Zur Frage einer intraventrikulären Neurosekretion

Zusammenfassung In der vorderen Wand des IV. Ventrikels und des Canalis centralis gegenüber der Area postrema liegen beim Kaninchen Nervenzellen, deren Fortsätze intraventrikulär mit 6 großen Protoplasmakugeln endigen, die mit Mitochondrien angefüllt sind. Das Perikaryon ist reich an Ergastoplasma, alle Teile der Zelle enthalten leere, 600 bis 1500 Å messende, und ein dichtes Granulum enthaltende, 650–1000 Å große Bläschen. Vermutlich werden Stoffe in den Liquor cerebrospinalis abgegeben. Dem Plasmalemm der Nervenzellen liegen Synapsen an.

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Summary In the anterior wall of the IVth ventricle, opposite to the area postrema, nerve cells are found. Their processes end with spheroid swellings extending into the ventricle. These bulbs contain a large number of mitochondria, while ergastoplasm is seen in the perikaryon. The cytoplasm of the whole cell shows vesicles (600–1500 Å) and dense-core vesicles (650–1000 Å) which might be extruded into the liquor cerebrospinalis. The plasmalemma of the perikaryon is in contact with synapses.

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Die Untersuchung wurde mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Subcellular distribution of carbonic anhydrase in Solanum tuberosum L. leaves

The intracellular compartmentation of carbonic anhydrase (CA; EC 4.2.1.1), an enzyme that catalyses the reversible hydration of CO₂ to bicarbonate, has been investigated in potato (Solanum tuberosum L.) leaves. Although enzyme activity was mainly located in chloroplasts (87% of total cellular activity), significant activity (13%) was also found in the cytosol. The corresponding CA isoforms were purified either from chloroplasts or crude leaf extracts, respectively. The cytosolic isoenzyme has a molecular mass of 255 000 and is composed of eight identical subunits with an estimated M _r of 30000. The chloroplastic isoenzyme (M _r 220000) is also an octamer composed of two different subunits with M _r estimated at 27 000 and 27 500, respectively. The N-terminal amino acid sequences of both chloroplastic CA subunits demonstrated that they were identical except that the M _r-27 000 subunit was three amino acids shorter than that of the M _r-27 500 subunit. Cytosolic and chloroplastic CA isoenzymes were found to be similarly inhibited by monovalent anions (Cl^–, I^–, N ₃ ^- and NO ₃ ^- ) and by sulfonamides (ethoxyzolamide and acetozolamide). Both CA isoforms were found to be dependent on a reducing agent such as cysteine or dithiothreitol in order to retain the catalytic activity, but 2-mercaptoethanol was found to be a potent inhibitor. A polyclonal antibody directed against a synthetic peptide corresponding to the N-terminal amino acid sequence of the chloroplastic CA monomers also recognized the cytosolic CA isoform. This antibody was used for immunocytolocalization experiments which confirmed the intracellular compartmentation of CA: within chloroplasts, CA is restricted to the stroma and appears randomly distributed in the cytosol.Abbreviations BSA bovine serum albumin - CA carbonic anhydrase - PMSF phenylmethylsulphonyl fluoride - BAM benzamidine - DTT dithiothreitol - 2-ME 2-mercaptoethanol - PVDF polyvinylidene difluoride The authors thanks P. Carrier and Dr. B. Dimon for technical assistance with the mass-spectrometry measurements.