Effect of tubulozole, a new synthetic microtubule inhibitor, on the induction of casein gene expression by prolactin

Colchicine and related drugs are known to inhibit milk secretion. They are also able to prevent stimulation of casein and DNA synthesis by prolactin in the mammary gland. The present report reports data obtained with tubulozole, a new antimitotic drug. Tubulozole C added to culture medium of isolated rabbit epithelial mammary cells strongly inhibited their multiplication. Simultaneously, at a concentration of 1 microM, it prevented almost completely the induction of beta-casein mRNA. Induced cells were rapidly deinduced by addition of the drug to the medium. A similar inhibition was observed when the induction was obtained with prolactin alone or with its two stimulators insulin and glucocorticoids. Tubulozole T, an isomer of tubulozole C which is known to be ineffective in disrupting microtubules, did not alter prolactin actions. These data and those obtained with other tubulin-binding drugs strongly suggest that the integrity of microtubules is required for prolactin to deliver its message to the mammary cell.     

Lipid peroxidation in plumbagin administered rats

Plumbagin was administered to rats at a concentration of 1,2,4,8 and 16 mg per kg body weight. After 24 h lipid peroxide levels were found to decrease in subcellular fractions of liver. Plumbagin inhibited ascorbate and nicotinafde adenine dinucleotide phosphate (reduced) dependent lipid peroxidation but was without any effect on cumene hydroperoxide dependent lipid peroxidation. Injection of 16 mg of plumbagin per kg body weight was found to decrease liver total reduced glutathione and also fcrosomal glucose-6-phosphatase. The results are discussed with reference to the anti- and prooxidant properties of plumbagin.     

The use of genetic markers in the analysis of competitive interactions in Drosophila melanogaster

A range of long established inbred lines derived from the TEXAS population of Drosophila melanogaster have been used to elucidate the nature of the competitive interactions which occur in genetically heterogeneous mixtures. A prerequisite for this type of investigation is the ability to distinguish the genotypes which compete in mixed culture. Specific marker alleles are generally used to achieve this distinction although in the past little attention has been given to the possibility of competitive bias introduced by the marker alleles themselves. For the experiments reported in this paper two specific marker alleles (y ² and w ^a) have been introduced independently into the TEXAS inbred lines. In this way the original wild type inbred lines could be compared with similar series of genotypes marked with either y ² or w ^a and the effects of the marker alleles determined.The results indicated that the body colour mutation (y ²) was neutral in its effect on the competitive interaction of recipient strains. The introduction of the white apricot eye colour mutation (w ^a) however, had a pronounced and deleterious effect on competitive ability. This effect was to render genotypes less able to compete effectively in mixed culture by depressing inter-genotypic competitive ability. These effects were found to be consistent over a range of genotypes and for each of two characters measuring competitive success.     

Multiple incompatibilities within populations of Culex pipiens L. in southern France

Strains of Culex pipiens derived from natural populations collected in southern France were crossed to determine their ability to give fertile offspring. Uni- and bi-directional incompatibilities occurred between these strains and each of them had its own crossing properties. Compatibility or incompatibility of a cross seemed to be related to the geographic distance separating the parental colonies, but not to their ecological origin (hypogeous or epigeous). Our results showed evidence for the variation in time of crossing properties of a strain.     

Macrozoobenthos of three Pennsylvania lakes: responses to acidification

The littoral macrozoobenthos (MZB) of three northeastern Pennsylvania lakes was sampled seasonally from summer 1981 until summer 1983, to determine if any changes were occurring in response to acid deposition. In the acidified lake (total alkalinity 0.0 eq L^–1) the mean pH decreased from 5.5 in 1981 to 4.2 in 1983. Chironomidae comprised 71.30% of the MZB numbers and 19.6% of the wet weight. Over the study period the wet weight of Chironomidae increased (p < 0.04) as did the total numbers of Chironomidae in general (p < 0.01) and Tanytarsini (p < 0.01) in particular. Total numbers of MZB also increased (p < 0.02) in the acidified lake, but there was no significant change in the number of taxa, diversity or total wet weight. In the moderately sensitive lake (total alkalinity 47.4 eq L^–1, mean pH 6.1) Chironomidae were numerically (43%) dominant but Odonata (18.6%) and Mollusca (12.7%) dominated wet weight. There were no significant changes in the MZB of the moderately sensitive lake over the study period. In the least sensitive lake (total alkalinity 190 eq L^–1, mean pH 6.6) the Amphipoda (31.3%) and Chironomidae (27.3%) together provided 58.6% of the MZB numbers, and the Mollusca formed 55.1% of wet weight. Wet weight at the least sensitive lake was higher (p < 0.01) and there were more Ephemeroptera, Pelecypoda and Gastropoda than at the other two lakes. There were no differences in total numbers, diversity or number of taxa among the three lakes.     

Immunocytochemical study of the intracellular localization of M protein of vesicular stomatitis virus

Summary The purpose of this paper is to describe the immunocytochemical localization of M protein of vesicular stomatitis virus (VSV) in infected cells. Vero cells, MDBK cells, Swiss 3T3 cells, and BHK cells were examined at various times after infection. For immunofluorescent staining, the cells were fixed with PLP fixative and then treated with 0.05% Triton X-100 before incubation with antibodies. Three hours after infection, M protein exhibited diffuse immunostaining throughout the cytoplasm and later accumulated along the cell membrane. The localization of M protein differed from the granular localization of the nucleocapsid N protein of VSV in the cytoplasm. For electron microscopy, the cells were fixed first in a mixture of 2% paraformaldehyde and 0.05% glutaraldehyde and then with PLP fixative, this being followed by treatment with 0.05% saponin. They were then immunostained using the immunoperoxidase method. The M protein was found to be distributed throughout the cytoplasm and later under the cell membrane, especially at virus budding sites. We also used postembedding immunostaining and freeze-fracture immunostaining to avoid the translocation of M protein caused by the detergent treatment. These techniques confirmed our previous results. Our findings are consistent with the view that the M protein of VSV is synthesized on free ribosomes and is then associated with the cell membrane where viral assembly may occur.S. Ohno was a visiting fellow from the Fogarty International Center at the National Institutes of Health, USA, from September 1981 to August 1983, while some parts of this work were in progress.     

Current approaches to the synthesis and reconstruction of genetic material

Advanced approaches to the synthesis and reconstruction of genetic material developed in the Institutes of Molecular Biology and Genetics during the past years are summarized. The evolution of methods for oligonucleotide synthesis and scopes for their use in gene production are discussed. The principles of localised mutagenesis methods developed in the Institute are described, such as: a) mutagenesis directed to the regulatory gene regions; b) segment-localized mutagenesis; c) mutagenesis directed by phosphotriester analogues of oligonucleotides. Examples of employing these methods for induction of regulatory mutants of phage lambda, production of fused genes, mutant interferon genes, construction of new DNA vectors, construction of hybrid H1-H3 subtype haemagglutinine gene of influenza virus etc. are presented. The approach to in vivo site-directed mutagenesis is experimentally substantiated.     

Cytogenetic study of the effect of gossypol on mouse germ cells

The clastogenic and mutagenic activities of a new antifertility and antitumor agent gossypol were studied in the mouse male germ cells. Results of the present work indicate that at the doses 125 and 250 mg/kg the drug does not significantly increase frequencies of the micronuclei in the early spermatids and sperm head abnormalities. Hence, genotoxic influence can not be proposed as responsible for the antifertility effect of gossypol.     

Intratumoural light distribution in an experimental mouse tumour irradiated by a diffuse-light irradiator compared with unilateral helium-neon light for photodynamic therapy

Investigations on dosimetry in photodynamic therapy (PDT) of experimental mouse tumours transplanted into the right hind leg revealed a significant variability in the fluence rate reaching tumour cells in different parts of the tumour when irradiated by a 50 mW collimated He-Ne laser. Based on intratumoural fibre-optical light measurements, a new irradiation source was constructed, in which variability in the radiant energy fluence rate between different parts of the solid tumour was reduced. The new diffuse-light irradiator was constructed basically from two concentric water chambers surrounded by three linear 3000 W xenon flash lamps. The outer chamber was an optical band filter, and the inner chamber contained a light-dispersing solution of lipid droplets which created an isotropic light field in which the tumours were submerged for PDT. Compared with unilateral He-Ne laser irradiation, an enhancement factor of 7.3 in radiance was obtained for the diffuse-light irradiator measured in the tumour. The new apparatus provides a nearly isotropic light field for in vivo experimental PDT.