Juvenile open angle glaucoma: fine mapping of the TIGR gene to 1q24.3–q25.2 and mutation analysis

Autosomal dominant juvenile open angle glaucoma (JOAG) is an early-onset form of primary open angle glaucoma (POAG), which has been linked to chromosome 1q21–q31. Recently, mutations in the trabecular meshwork inducible glucocorticoid response gene (TIGR), one of the candidate genes mapped in this region, were identified in glaucoma patients of several families. We screened for mutations of the TIGR gene in two German families with JOAG and in 100 unselected sporadic cases of POAG. In the first family we identified a Pro370Leu mutation and in the second family a Gly367Arg mutation cosegregating with the glaucoma phenotype. No pathogenic mutation was found in 100 sporadic cases but a Tyr347Tyr polymorphism was found in two patients. Furthermore, fluorescence in situ hybridization (FISH) analysis was used to map a TIGR-specific yeast artificial chromosome to 1q24.3–q25.2. Received: 19 June 1997 / Accepted: 12 August 1997     

Physiological and Molecular Biological Characterization of Ammonia Oxidation of the Heterotrophic Nitrifier Pseudomonas putida

The heterotrophic nitrifier Pseudomonas putida aerobically oxidized ammonia to hydroxylamine, nitrite, and nitrate. Product formation was accompanied by a small but significant release of NO, whereas N₂O evolution could not be detected under the assay conditions employed. The isolate reduced nitrate to nitrite and partially further to NO under anaerobic conditions. Aerobically grown cells utilized γ-aminobutyrate as a carbon source and as a N-source by ammonification. The physiological experiments, in particular the inhibition pattern by C₂H₂, indicated that P. putida expressed an ammonia monooxigenase. DNA-hybridization with an amoA gene probe coding for the smaller subunit of the ammonia monooxigenase of Nitrosomonas europaea allowed us to identify, to clone, and to sequence a region with an open reading frame showing distinct sequence similarities to the amoA gene of autotrophic ammonia oxidizers. Received: 9 April 1998 / Accepted: 15 May 1998     

Studies on the reaction of fluorescamine with primary amines

Fluorescamine is a useful reagent for the fluorometric assay of primary amines. The extent of the reaction between fluorescamine and primary amines, as well as the fluorescence intensities of the resulting fluorophors depend on pH, solvent composition and reagent concentration. Optimum values for these variables further depend on the amine under study. The influence of these parameters on the fluorogenic reaction of representative amines, and on their fluorophoric derivatives has been investigated, and the results are reported here.     

Cryptogenic Cerebral Embolism in Women Taking Oral Contraceptives

Fourteen women taking oral contraceptives were admitted during a five-year period because of acute cerebrovascular lesions. A diagnosis of major cerebral embolism was established in four of them. No source of embolism was found, and thorough investigation failed to reveal any predisposing illness. Cerebral embolism was a probable diagnosis in several of the remaining 10 patients. A comparison was made with the strokes occurring in women not taking contraceptive pills in corresponding age groups.     

Chromatography of Microbial Lipids by Centrifugation Through Microparticulate Gel

An improved apparatus and a procedure are described by which the migration of sample components in column chromatography is accelerated by centrifugal force, thereby making it possible to use beds of densely packed gel prepared with ultrafine silica. This technique was used to resolve components of certain lipid mixtures where other methods have failed, and it has been found generally useful as an adjunct to other methods for the fractionation of lipids. Biologically active phosphoglycolipids from Mycobacterium tuberculosis and a phosphatidylglycerol-like substance from Mycoplasma pneumoniae which formed single spots on thin-layer chromatographic plates were each found to contain a major and several minor components by centrifugal chromatography. The method enabled us to isolate individual components of Wax D from M. tuberculosis rather than a spectrum of components. Minor components were resolved which, although present in insufficient quantity to influence results of chemical analyses, may be responsible for biological activity. The apparatus provides an essentially closed system which reduces highly volatile solvents to minimal evaporation during the chromatographic process. Samples are applied in solution and are not allowed to dry on the columns until after separation has been achieved. Consequently, polar, labile microbial lipids can be resolved without the use of harsh reagents which destroy some of their properties. Single components may be harvested by cutting and removing appropriate segments of the larger chromatograms or by eluting them from the columns.     

Parent-to-Progeny Transfer and Recombination of T4rII Bacteriophage

Transfer of parental, light (not substituted with 5-bromodeoxyuridine) (32)P-deoxyribonucleic acid (DNA) from rII(-) mutants of T4 bacteriophage to heavy (5-bromodeoxyuridine-substituted) progeny in Escherichia coli B was less homogeneous than in wild phages. The net transfer was 5 to 20% of the value for wild T4 phage, and the parental contribution per progeny DNA molecule amounted to 7 to 100% of the genome. Three classes could be distinguished, based on the density distribution of parental label in CsCl analysis of the progeny phages. "Far recombined" phages contain parental material only in semiconservatively replicated subunits covalently attached to progeny DNA, amounting to 5 to 10% parental contribution per genome. "Intermediate recombinants" contain, aside from conventional recombinant DNA, parental DNA banding at the original, light density. This DNA may be unattached to heavy progeny DNA or attached by weak bonds which are very sensitive to shearing during the extraction procedure. The parental contribution is 10 to 50% per progeny DNA molecule in this class. "Conservative" phages band close to the parental, light density in CsCl; their DNA is purely light. When the parental phage is labeled with both (3)H-leucine (capsid) and (32)P (DNA), the specific activity of (3)H/(32)P in the "conservative progeny" is 10 to 40% of that in the parental, showing that at least some of the (32)P in this area belongs to phages with parental DNA as the sole DNA component inside an unlabeled capsid, i.e., parental DNA which has been injected into the host and matured in a new capsid without replication or recombination. This phenomenon occurs to about the same extent in both single and multiple infection.     

Multiple and Specific Initiation of T4 DNA Replication

Partially replicated T4 DNA molecules (PRM) whose parental or progeny DNA was labeled with bromodeoxyuridine BUdR was analyzed by gradual shearing followed by CsCl banding of the sheared product. Analysis of PRM containing 18-mum replicated DNA showed that each replicated region was 3- to 6-mum long, indicating three to 6 replicative sites per molecule. Analysis of PRM containing 9-mum replicated DNA similarly indicated two to three replicated regions per molecule. DNA from the replicated regions of PRM containing 10-mum replicated DNA ("donor") was hybridized to DNA from mature phage ("recipient"), and the resulting hybrid was subjected to digestion with exonuclease I. The extent of protection of the recipient and more efficient self-annealing of progeny fragments from PRM indicated that the replicated regions represented 8 to 10 nonrandom locations of the genome. Possible significance of multiple sites for initiation of DNA replication is discussed.     

Die jahresperiodische Entwicklung des Wurzelund Sprossystems von Symphytum officinale L. und ihre Beziehung zu Speicherung und Verbrauch der Kohlenhydrate

Summary The development of the root and shoot system of Symphytum officinale always begins with the formation of a rape with 6–8 leaves on its epicotyl. After this stage development is determined by the length of the day. If the day is shorter than 15 hours, the subterraneous organs grow thicker. Flower formation needs a day length of at least 12 hours, shoot growth a day length of 14 hours. The shorter the day length the more leaves are formed.Starch is always stored until 6–8 leaves are formed by the young plant. After this stage it is only stored when the days are longer than 14 hours. When the days are shorter the amount of starch is reduced, partly during the thickening of the subterraneous organs. Fructosans are stored and reduced independent of day length. They are consumed during shoot elongation and flowering.A hydrolysis of starch and fructosans is also caused by low temperatures during the winter.Starch and fructosans are stored to a different degree in the various subterraneous organs: the most is stored in the shoot-born roots and the least in the subterraneous shoot parts. In the latter organs the carbohydrate content is influenced the most by the growth processes.