Cardiac arrest following an iatrogenic 3,4-diaminopyridine intoxication in a patient with Lambert-Eaton myasthenic syndrome.

Syndromes with impaired neuromuscular transmission are frequently treated with pyridine derivates. 3,4-diaminopyridine is thought to have fewer side effects than the commonly used, but less potent, 4-aminopyridine. We describe a patient with an initially unrecognized iatrogenic intoxication with 3,4-diaminopyridine. Except for a life threatening arrhythmia, symptoms were similar to a 4-aminopyridine intoxication. The patient made a full recovery with symptomatic treatment and withdrawal of the drug.

Complexity and abundance of ribonucleic acid transcribed from restriction endonuclease fragments of Euglena chloroplast deoxyribonucleic acid during chloroplast development.

Chloroplast DNA from Euglena gracilis was used to construct a partial library of recombinant plasmids representing 45% of the DNA. Each plasmid was radioactively labeled in vitro by nick translation and hybridized in liquid to a vast excess of total cellular RNA isolated either from cells grown continually in the dark or from cells containing developing chloroplasts. The complexity and abundance of the RNA that hybridized to the different chloroplast restriction endonuclease DNA fragments were calculated from the RNA-DNA hybridization saturation values and the pseudo-first-order hybridization rate constants, respectively. The complexity of these transcripts showed little change during chloroplast development. In several cases, the complexity of the RNA was greater than expected for asymmetrical transcription, suggesting the possibility that transcription may be symmetrical in some regions of chloroplast DNA. The abundance of the transcripts ranged from 0.0001% to nearly 10% of the total cellular RNA, and in some cases changed by as much as 5-10-fold during chloroplast development.

DNA sequence organization in the alga Euglena gracilis.

The sequence organization of nuclear DNA in the single-celled alga Euglena gracilis has been studied by a combination of techniques: (1) the comparison of the reassociation kinetics of DNA fragments 300, 2000 and 8100 nucleotides long; (2) the reassociation of 32P-labeled DNA fragments of various lengths with driver fragments 300 nucleotides long; (3) the hyperchromicity of DNA structures formed by the reassociation of repetitive sequences; (4) and the direct measurement of the size of the duplex regions of reassociated repetitive DNA resistant to S1 nuclease. The single copy DNA sequences are approximately 1500 nucleotide pairs long and are interspersed with repetitive DNA sequences. The repetitive DNA, consisting of both highly repetitive and middle repetitive sequences, consists of one fraction of nucleotide sequences (0.67) with an average size of 4900 nucleotide pairs and a second fraction (0.33) with an average size of 1000 nucleotide pairs, 34% of the DNA consists of foldback sequences which are present on 45% of the DNA 4000 nucleotides long.

Influence of growth conditions upon the number of chloroplast DNA molecules in Euglena gracilis.

The number of chloroplast DNA molecules in Euglena gracilis cells was measured by determining the shift in the observed second-order rate constant for the reassociation of (125)I-labeled chloroplast DNA in the presence of unlabeled total cell DNA. Cells grown to stationary phase in the dark contained 217 molecules of chloroplast DNA. Cells grown to stationary phase in the light in either heterotrophic or autotrophic medium contained 590 and 1014 chloroplast DNA molecules, respectively. The observed second-order rate constant for the reassociation of (125)I-labeled chloroplast DNA was not significantly altered in the presence of total cell DNA from a heat-bleached mutant, ZHB, which lacks chloroplast DNA. This evidence suggests that there is less than 0.3 of a chloroplast DNA molecule present in the nucleus of Euglena.

A measurement of the fraction of chloroplast DNA transcribed during chloroplast development in Euglena gracilis.

The fraction of chloroplast DNA transcribed at different stages of chloroplast development in Euglena gracilis was measured by RNA-DNA hybridization. Euglena cells were grown in the dark in a heterotrophic medium to stationary phase and then transferred to the light. Chloroplast development was monitored by the increase in the cellular chlorophyll content in the absence of cell division. Total cell RNA was isolated at various stages of chloroplast development, and hybridized in a vast excess to [125I]chloroplast DNA. The fraction of [125I]chloroplast DNA in the form of a duplex was monitored by chromatography on hydroxylapatite columns. The amount of RNA-DNA hybrid in the duplex mixture was determined by correcting for the contribution of DNA-DNA renaturation under the same conditions. The fraction of chloroplast DNA transcribed was calculated by multiplying by two the amount of single-stranded DNA in the form of an RNA-DNA hybrid. Prior to the initiation of chloroplast development (i.e., in dark grown cells) the fraction of chloroplast DNA represented as RNA transcripts in the cell is 0.53. As chloroplast development proceeds, the fraction of the chloroplast DNA transcribed decreases to 0.47. Experiments in which mixtures of various RNA samples were hybridized to the chloroplast DNA indicate that there is a small portion of chloroplast DNA transcribed at later stages of chloroplast development which is not represented as transcripts at the onset of chloroplast development. Melting properties of the RNA-DNA hybrids show that the RNA-DNA duplexes are slightly less stable than renatured [125I]chloroplast DNA.