DNA-polymorphic patterns linked to the β-globin genes in German families affected with hemoglobinopathies and thalassemias: a comparison to other ethnic groups

Summary DNA haplotype constellations of the β-globin gene cluster have been analyzed in German families with hemoglobinopathies (Hb Freiburg, Hb K?ln, Hb Presbyterian) and β-thalassemias. The polymorphis patterns obtained were compared to those found in families from Greece, Italy, and Turkey affected by β-thalassemia syndromes. With the combined analysis of seven restriction site polymorphisms a DNA-diagnostic prediction for additional offspring could be made with an overall frequency of 75% in the four ethnic groups.     

Mapping of cysteine genes on the chromosome of Pseudomonas aeruginosa PAO

Summary Three loci coding for different steps in the pathway of cysteine biosynthesis have been mapped by R68.45-mediated coconjugation analysis. The cysteine auxotrophic mutants could be subdivided into sulfite and sulfide-requiring mutants. Sulfide-requiring mutants (cysIV group) were localized at a single position between pyrF and pur-67, while sulfite-requiring mutants (cysI and cysII) mapped at two different regions. The cysI group was also localized between pyrF and pur-67, although more distal to pyrF than the cysIV group. This group included the cys-54 marker, which has been mapped previously. The second group of sulfite-requiring mutants, designated as cysII, was cotransducible with hisI and localized at the end of the PAO chromosomal map. This location was also confirmed for the marker cys-59.The marker cys-59 (which was cotransducible with his1) was cotransferred by R68.45-mediated conjugations with both the late marker pur-67 and the early marker ilv-226. As the late marker hisI was positioned at about 60–65 min (Herrmann and Günther, in press) the length of the PAO chromosome was estimated to be about 70 min.     

pH dependence of the in situ formation of an organicN-chloramine water disinfectant

Summary Staphylococcus aureus was used to assess the bactericidal efficacy of aqueous solutions of the organicN-chloramine compound 3-chloro-4,4-dimethyl-2-oxazolidinone (agent I) formed in situ. The rate of in situ formation, accomplished by reacting free chlorine with the amine precursor, was a function of pH. When the reagents were combined under acidic conditions (pH5.5) and allowed to react for 22 h, sufficient residual free chlorine was present to inactivate the bacteria in less than 5 min. When combined under less acidic conditions (pH6.0), comparable bacterial inactivation required 30–60 min due to the extensive reaction of the free chlorine to form agent I. The kill rates present under less acidic and neutral conditions are equivalent to those for pre-formed agent I. In water disinfection applications for pH6.0, in situ formation of agent I would provide a combination of rapid initial and slower long-term disinfection.     

New derivatization and separation procedures for reducing oligosaccharides

4-Trifluoroacetamidoaniline was reacted with reducing oligosaccharides in the presence of sodium cyanoborohydride to give aminoalditol derivatives, useful for linkage to proteins or solid matrices. A mixture of reducing oligosaccharides, difficult to separate by HPLC, was treated in the same way. The resulting derivatives were easily separated by HPLC.Abbreviations TFAN 4-trifluoroacetamidoaniline - LcOse₄ lacto-N-tetraose - IV²Fuc-LcOse₄ lacto-N-fucopentaose l - III⁴Fuc-LcOse₄ lacto-N-fucopentaose II - III³Fuc-nLcOse₄ lacto-N-fucopentaose III - IV²Fuc, III⁴Fuc-LcOse₄ lacto-N-difucohexaose I - II⁶Galß1-4GlcNAc-LcOse₄ lacto-N-hexaose - II³NeuAc-Lac 3-sialyllactose - GlcNAcß1-4GlcNAcß1-4GlcNAc chitotriose - GalNac1-3|Fuc1-2|Galß1-4Glc A-tetrasaccharide     

Primary astroglial cultures

Conclusion Primary cultures from neonatal rat brain consist mainly of astroglial cells, immunohistochemically identified by GFAp and S-100. As other cells than astrocytes may survice in the culture, specific markers for the expected cells were used. Cells with phagocytic properties, endothelial-like cells, oligoblasts, ependymal cells and mesenchymal cells were identified. No neurons have so far been detected.The astroglial cells have a high-affinity uptake for glutamate, aspartate GABA, taurine and hypotaurine, while there is probably a non-saturable uptake of norepinephrine, dopamine and 5-HT. The enzymes MAO, COMT, GABA-T and GS have been demonstrated. It thus seems that astrocytes take part in the inactivation of neurotransmitters, although amino acids and monoamines are taken up with different mechanisms.The presence of receptors for different neurotransmitters and neuromodulators has been demonstrated on astrocytes.Astroglial-enriched cultures from various brain regions have shown that the cells express specialized functional properties concerning neurotransmitter uptake, metabolizing enzymes and receptor density.Astroglial cell differentiation in culture is shortly reviewed and one possibility to affect this maturation by co-cultivation with neuronal containing cultures is point out.     

Improvements in and Environmental Applications of Double-Vial Radiorespirometry for the Study of Microbial Mineralization

Several variations in the scintillation mixture and the filter paper arrangements for double-vial radiorespirometry were compared. Improved efficiencies (44%) and shorter response times were found by adding wetting agents and methanolic NaOH to the scintillation mixture in the filter paper. The scintillation chemicals used did not contain dioxane and were found to be nontoxic to the test microbiota in this system. Covering the inner reaction vial with aluminum foil minimized the reduction in counting efficiency when testing colored or dense environmental samples. Mineralization rates were determined with ¹⁴C-labeled glucose, acetate, and glutamate and [¹⁴C]cellulose- and [¹⁴C]lignin-labeled lignocellulose for composting cow manure, forest soil, and arctic lake sediment microbiota. This improved method can be used in a variety of procedures involving the measurement of microbial mineralizations of organic compounds. Since no liquid scintillation cocktail is used for counting, the radioactive wastes are aqueous or can be incinerated, making disposal easy.     

Sugar ring isomerization in C-arabinosylflavones

6-C-α-l-Arabinopyranosyl- and furanosylacacetins have been synthesized. They are isomerized by short acid treatment to give a mixture of the four anomer/ring size combinations without any Wessely-Moser isomerization. In the same conditions molludistin (8-C-α-l-arabinopyranosylgenkwanin) led only to a mixture of molludistin and 8-C-α-l-arabinofuranosylgenkwanin. This is the first demonstration of ring sugar isomerization in C-glycosylflavones. In usual solvent systems, α-anomers are easily separated from β-anomers, whereas corresponding pyranosyl and furanosyl anomers are not. However, they are easily separated after permethylation and characteristic features are found in the mass spectra of PM 6-C-arabinofuranosyl isomers.     

Immunocytochemistry of brain-reactive monoclonal antibodies in peripheral tissues

Among a total of 135 tissue-reactive monoclonal antibodies previously prepared, 81 were brain-selective and were classified into neuronal and non-neuronal categories. The neuronal antibodies were again subdivided into antineurofibrillar, antiperikaryonal-neurofibrillar, and antisynapse-associated groups. On the basis of morphologic, developmental, biochemical, and pathologic criteria, the antibodies in at least two of these groups were found to detect heterogeneous antigens (called "neurotypes") rather than different antigenic determinants in single antigens. On examining the distribution in peripheral organs of staining patterns of 11 antineuronal brain-reactive antibodies, we now confirm that these antibodies are, indeed, largely brain-specific. In general, non-neuronal elements in liver, lung, heart, thymus, intestine, adrenal, and spleen remained unstained. However, most of the antibodies stained peripheral neural elements. Occasional antibodies did stain selected, non-neuronal structures. Four out of five antineurofibrillar antibodies stained nerve fibers in adrenal medulla, intestine and thymus. All of three antiperikaryonal-neurofibrillar antibodies also stained nerve fibers in the adrenal medulla, but not in other organs. Two out of three antisynapse-associated antibodies stained what appear to be nerve contacts on adrenal medullary cells, but not on any other peripheral cells examined. The non-neuronal peripheral staining patterns were restricted to selective nuclear staining exhibited by two out of five antineurofibrillar antibodies and the staining of macrophage and selected cardiac muscle nuclei by two of three antisynapse-associated antibodies. However, one antineurofibrillar antibody also stained the cytoplasm of selected liver cells. Among non-neuronally reacting antibodies, two antibodies stained nuclei of all cells except neurons in brain as well as peripheral organs. An antibody staining the ciliary epithelium of choroid plexus also stained basal bodies of ciliated bronchial epithelium. The overall data suggest that the specificity of brain-reactive antibodies is high and that their cross-reactivity with epitopes in non-nervous tissue is rare. In these cases, the antibodies seem to provide specific reagents for these additional structures as well as for their specific brain antigens.     

Bacteriochlorophyllgehalt und Proteinmuster der Thylakoide von Rhodospirillum rubrum während der Morphogenese des Photosynthese-Apparates

Zusammenfassung Der Zusammenhang zwischen dem spezifischen Bacteriochlorophyllgehalt der Zellen und der Thylakoidmorphogenese wird an Rhodospirillum rubrum untersucht. Bei der Induktion des Photosynthese-Apparates wird zunächst Bacteriochlorophyll synthetisiert, obgleich noch keine Thylakoide gebildet werden (1. Phase). Werden Thylakoide ausgebildet, so steigt der spezifische Bacteriochlorophyllgehalt der Thylakoide in Abhängigkeit vom spezifischen Bacteriochlorophyllgehalt der Zellen an, bis ein Wert von 12–13 g Bacteriochlorophyll je mg Zellprotein erreicht ist (2. Phase). Während der Wert für die Zellen darüber hinaus weiter erhöht werden kann, bleibt der Wert der Thylakoide konstant bei 100 g Bacteriochlorophyll je mg Thylakoid-Protein (3. Phase). Isolierte Thylakoide aus Zellen mit niedrigem Bacteriochlorophyllgehalt besitzen geringere Durchmesser als Thylakoide aus Zellen mit höheren Werten. Auch hinsichtlich der Zusammensetzung der Thylakoide in Abhängigkeit von den steigenden Bacteriochlorophyllgehalten bei induzierten Zellen konnten Unterschiede festgestellt werden. Ähnlich wie die spezifischen Bacteriochlorophyllgehalte der Thylakoide, nähern sich die verschiedenen Proteinbausteine der Thylakoide einem festen Verhältnis zueinander, das sich oberhalb von 10–14 g Bacteriochlorophyll je mg Zellprotein nicht mehr ändert. Mit Zunahme des Bacteriochlorophyllgehaltes der Zellen steigt der Gehalt an thylakoidspezifischen Proteinen in den Membranen an und der Anteil der für die Cytoplasmamembran spezifischen Komponenten nimmt ab.

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The bacteriochlorophyll content and protein composition of chromatophores (=thylakoids) of Rhodospirillum rubrum during morphogenesis of the photosynthetic apparatus

Summary The correlation between the specific bacteriochlorophyll content of whole cells and the morphogenesis of chromatophores was investigated in Rhodospirillum rubrum. During the first phase after induction of the photosynthetic apparatus bacteriochlorophyll is synthesized without formation of chromatophores. In a second phase chromatophores increases in a linear correlation to the specific bacterichlorophyll content of the cells. In a third phase, when the specific bacteriochlorophyll content of the cells has reached 12–13 g/mg protein, the specific bacteriochlorophyll content of the chromatophores remains constant (100 g/mg protein). Isolated chromatophores from the second phase have smaller diameters, than chromatophores from the third phase. The composition of the protein compounds of isolated chromatophores changes during the development of the chromatophores in a similar fashion as the specific bacteriochlorophyll content of chromatophores does change. With increasing bacteriochlorophyll content of the cells the chromatophore specific proteins in the membranes increase whereas proteins specific for the cytoplasmic membrane decrease until both reach a constant level.

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Verwendete Abkürzungen BChl. Bacteriochlorophyll     

Quantitative DNS-Bestimmungen von Ameisenhirnzellen

Zusammenfassung Gehirnzellen von Myrmica laevinodis-Arbeiterinnen wurden Feulgengefärbt und dabei 1–6 oder 15 Std im Schiffschen Reagens behalten. Die Farbintensität wurde mikrospektrophotometrisch bestimmt. Die Tiere des einen Nestes zeigten ein Extinktionsmaximum nach 1 Std Aufenthalt im Schiffschen Reagens, die eines anderen Nestes nach 5 Std. Die Färbedauer ist also ein weiterer Faktor, der bei quantitativen DNS-Bestimmungen zu berücksichtigen ist.

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Quantitative determination of DNA in feulgen stained brain cells of antsI. The effect of the staining duration on the extinction

Summary Brains of adult Myrmica laevinodis (Hymenoptera, Formicidae) workers were Feulgen stained. They were kept in Schiff's reagens for 1 to 6 hours. In specimens from one nest the colour maximum was reached after 1 hour, in others from a different nest after 5 hours. The colour intensity measured by microspectrophotometry depended on the time the brains stayed in Schiff's reagens. This result means that the time of staying in Schiffs reagens is another factor influencing the amount of measurable DNA.

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Die Untersuchung wurde mit Unterstützung durch den Schweizerischen Nationalfond durchgeführt.

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Die Laborantin Frl. Elisabeth Räber trug mit tatkräftiger, selbständiger Arbeit zum Fortgang der Untersuchung bei. Prof. Dr. F. Ruch stellte uns großzügigerweise sein Mikrospektrophotometer zur Verfügung, wofür ich ihm herzlich danke.