Epithelial impedance analysis in experimentally induced colon cancer.

Epithelial impedance analysis was used to measure the alterations in resistance of the large bowel in a murine model of large bowel cancer. The technique was able to resolve the epithelial resistance from the total resistance of the bowel wall. A progressive decrease in resistance of the bowel epithelium occurs during carcinogenesis induced with dimethyhydrazine. About a 21% decrease in epithelial resistance from 22.0 +/- 1.3 omega.cm-2 to 17.5 +/- 1.1 omega cm-2 (p less than 0.025) was observed after 20 wk of carcinogen administration. The sensitivity of the technique in detecting altered epithelial resistance in premalignant bowel mucosa was improved by examining the impedance profile in a sodium-free Ringer's solution where the epithelium of control colons had a resistance of 24.4 +/- 1.8 omega.cm-2 compared with 19.0 +/- 1.1 omega.cm-2 (p less than 0.02) in colons from animals treated for only 4 wk with the carcinogen. Epithelial impedance analysis would seem to be a sensitive technique capable of identifying changes in the electrical properties or the large bowel early in disease states.     

The Putative Oncogene Pim-1 in the Mouse: Its Linkage and Variation among t Haplotypes

Pim-1, a putative oncogene involved in T-cell lymphomagenesis, was mapped between the pseudo-alpha globin gene Hba-4ps and the alpha-crystallin gene Crya-1 on mouse chromosome 17 and therefore within the t complex. Pim-1 restriction fragment variants were identified among t haplotypes. Analysis of restriction fragment sizes obtained with 12 endonucleases demonstrated that the Pim-1 genes in some t haplotypes were indistinguishable from the sizes for the Pim-1b allele in BALB/c inbred mice. There are now three genes, Pim-1, Crya-1 and H-2 I-E, that vary among independently derived t haplotypes and that have indistinguishable alleles in t haplotypes and inbred strains. These genes are closely linked within the distal inversion of the t complex. Because it is unlikely that these variants arose independently in t haplotypes and their wild-type homologues, we propose that an exchange of chromosomal segments, probably through double crossingover, was responsible for indistinguishable Pim-1 genes shared by certain t haplotypes and their wild-type homologues. There was, however, no apparent association between variant alleles of these three genes among t haplotypes as would be expected if a single exchange introduced these alleles into t haplotypes. If these variant alleles can be shown to be identical to the wild-type allele, then lack of association suggests that multiple exchanges have occurred during the evolution of the t complex.     

The Influence of Nonautonomous P Elements on Hybrid Dysgenesis in Drosophila melanogaster

An inbred line of the M' strain Muller-5 Birmingham was studied for its abilities to affect P-M hybrid dysgenesis. This strain possesses 57 P elements, all of which are apparently defective in the production of the P transposase. In combination with transposase-producing elements, these nonautonomous elements can enhance or diminish the incidence of hybrid dysgenesis, depending on the trait that is studied. Dysgenic flies that have one or more paternally-derived chromosomes with these elements partially repress the instability of the P element insertion mutation, snw; however, such flies have elevated frequencies of another dysgenic trait, GD sterility, and also show distorted segregation ratios. An explanation is presented in which all of these phenomena are unified as manifestations of the kinetics of P element activation in the germ line. The progeny of Muller-5 Birmingham females exhibit partial repression of both snw instability and GD sterility. This repression appears to involve a factor that can be transmitted maternally through at least two generations. This mode of repression therefore conforms to the pattern of inheritance of the P cytotype, the condition that brings about nearly total repression of P element activity in some strains. Models in which this repression could arise from the nonautonomous P elements of Muller-5 Birmingham are discussed.     

A New Epistasis Group for the Repair of DNA Damage in Bacteriophage T4: Replication Repair

The gene 32 mutation amA453 sensitizes bacteriophage T4 to the lethal effects of ultraviolet (UV) irradiation, methyl methanesulfonate and angelicin-mediated photodynamic irradiation when treated particles are plated on amber-suppressing host cells. The increased UV sensitivity caused by amA453 is additive to that caused by mutations in both the T4 excision repair (denV) and recombination repair (uvsWXY) systems, suggesting the operation of a third kind of repair system. The mutation uvs79, with many similarities to amA453 but mapping in gene 41, is largely epistatic to amA453. The mutation mms1, also with many similarities to amA453, maps close to amA453 within gene 32 and is largely epistatic to uvs79. Neither amA453 nor uvs79 affect the ratio of UV-induced mutational to lethal hits, nor does amA453 affect spontaneous or UV-enhanced recombination frequencies. Gene 32 encodes the major T4 ssDNA-binding protein (the scaffolding of DNA replication) and gene 41 encodes a DNA helicase, both being required for T4 DNA replication. We conclude that a third repair process operates in phage T4 and suggest that it acts during rather than before or after DNA replication.     

The white gene as a marker in a new P-element vector for gene transfer in Drosophila.

We describe new vectors suitable for P-element mediated germ line transformation of Drosophila melanogaster using passenger genes whose expression does not result in a readily detectable phenotypic change of the transformed flies. The P-element vectors contain the white gene fused to the heat shock protein 70 (hsp70) gene promoter. Expression of the white gene rescues the white phenotype of recipient flies partly or completely even without heat treatment. Transformed descendents of most founder animals (GO) fall into two classes which are distinguishable by their orange and red eye colours. The different levels of white expression are presumably due to position effects associated with different chromosomal sites of insertion. Doubling of the gene dose in orange eyed fly stocks results in an easily visible darkening of the eye colour. Consequently, the generation of homozygous transformants is easily possible by simple inbreeding due to the phenotypic distinction of homo- and heterozygous transformants. Cloning into these P-element vectors is facilitated by the presence of polylinkers with 8 and 12 unique restriction sites.     

Formamidines interact withDrosophila octopamine receptors,alter the flies' behavior and reduce their learning ability

Summary We have studied the effect of formamidines onDrosophila melanogaster. Low concentrations of formamidines are toxic to adultDrosophila. A mutant with reduced cAMP synthesis displays increased resistance to the toxin. Formamidines also reduce viability ofDrosophila eggs and retard imago eclosion. At sublethal concentrations, formamidines markedly affect the flies' behavior. Upon injection, the compounds increase muscle activity. Upon feeding, formamidines induce motor excitation, reduce phototaxis and impair olfactory learning without affecting the ability to recognize an olfactory cue. In vitro, two formamidines were found to inhibit octopamine-stimulated adenylate cyclase without affecting the basal activity of the enzyme, while a third one was found to stimulate adenylate cyclase; this stimulation was blocked by phentolamine and to a lesser degree by propranolol, thus resembling the effect of octopamine. The binding of [³H]octopamine toDrosophila head membranes was also inhibited. Taken together, our results indicate that formamidines interact with octopaminergic systems inDrosophila, exert both peripheral and central effects in the fly, and could be used to dissect the roles of octopamine in development and behavior, including behavioral plasticity. The results also suggest that formamidines could be used to select mutants in aminergic transmission and in the cAMP cascade.Abbreviations CDMF chlordimeform - DMPF N,N-dimethyl-N2-(2,4-dimethylphenyl) formamidine     

Electrolocation in the presence of jamming signals: behavior

Electrolocation behavior of Apteronotus leptorhynchus was studied by monitoring the animal's ability to maintain orientation to a variety of moving electrolocation targets. The primary goal of this study was to determine the relative effectiveness of various types of electrical 'jamming signals' in disrupting electrolocation performance. 1. Two measures of electrolocation performance were used: The latency between the electrolocation target motion and the fish's following response, and the minimum distance separating the fish from the target during the target movement sequence. Latency increased and minimum fish-target distance decreased as target size was decreased, and when large diameter ceramic targets were used as control stimuli the fish were less able to avoid, and frequently collided with, these 'electrically transparent' objects. 2. Four types of jamming signals were used in attempts to mask the electrosensory input used for electrolocation. Broad-band noise and sinusoidal signals, different in frequency by a few Hz from the animal's personal electric organ discharge (DF stimuli), were used to jam the tuberous electroreceptors. Five Hz and 50 Hz sinusoidal signals were used to jam the low-frequency or ampullary receptor system. Both the noise and the DF stimuli were effective in reducing electrolocation performance, and the threshold intensity for behavior deterioration was about three-fold lower for DF stimuli as compared to the noise. The rate of change of response deterioration as a function of increasing jamming intensity was, however, not different for these two types of stimuli. Neither the 50 Hz nor the 5 Hz jamming signals caused electrolocation deterioration when presented alone. However, 5 Hz jamming, when added to either the noise or DF jamming, did result in significant increments in response deterioration. This suggests that the ampullary receptors can provide supplementary information for electrolocation. 3. Electrolocation performance deterioration was also studied with various difference frequencies between an animal's EOD and the sinusoidal jamming stimulus. Increasing DF results in decreased electrolocation deterioration, but even at the highest DF frequencies used (128 Hz) significant response degradation was observed. 4. The apparent differences in the effectiveness of noise and DF stimulation in reducing electrolocation performance are largely accounted for by the differential effects of the tuberous electroreceptor filter characteristics on these two types of signals.     

Isolation ofAcholeplasma oculi from human amniotic fluid in early pregnancy

Acholeplasmas have been isolated from a variety of animals, insects, and plants, but onlyAcholeplasma laidlawii has previously been found in humans. We have isolatedAcholeplasma oculi in pure culture from the amniotic fluid of a woman at 19 weeks of gestation. The organism was positively identified by growth inhibition, epi-immunofluorescence, and arbutin hydrolysis. Demonstration of organisms directly in amniotic fluid by DNA fluorochrome and immunofluorescence staining provided additional evidence that the isolate was genuine and not a medium contaminant. The remainder of the pregnancy was unremarkable, and a full-term male infant was delivered without complications. Even though there is some evidence possibly associatingA. oculi with various diseases in livestock, the prevalence and significance ofA. oculi in humans has not been determined.     

Mutational analysis of the Bradyrhizobium japonicum common nod genes and further nod box-linked genomic DNA regions

Summary By insertional and deletional marker replacement mutagenesis the common nod region of Bradyrhizobium japonicum was examined for the presence of additional, essential nodulation genes. An open reading frame located in the 800 bp large intergenic region between nodD1 and nodA did not appear to be essential for nodulation of soybean. Furthermore, a strain with a deletion of the nodI- and nodJ-like genes downstream of nodC had a Nod⁺ phenotype. A mutant with a 1.7 kb deletion immediately downstream of nodD1 considerably delayed the onset of nodulation. This region carried a second copy of nodD (nodD2). A nodD1-nodD2 double mutant had a similar phenotype to the nodD2 mutant. Using a 22-mer oligonucleotide probe partially identical to the nod box sequence, a total of six hybridizing regions were identified in B. japonicum genomic DNA and isolated from a cosmid library. Sequencing of the hybridizing regions revealed that at least three of them represented true nod box sequences whereas the others showed considerable deviations from the consensus sequence. One of the three nod box sequences was the one known to be associated with nodA, whereas the other two were located 60 to 70 kb away from nif cluster I. A deletion of one of these two sequences plus adjacent DNA material mmutant 308) led to a reduced nodulation on Vigna radiata but not on soybean. Thus, this region is probably involved in the determination of host specificity.Dedicated to Prof. Giorgio Semenza on the occasion of his 60th birthday