R-Factor Mutant Capable of Specifying Hypersynthesis of Penicillinase

The physical characteristics of a mutant, R(M201-2), capable of conferring high and stable ampicillion resistance was analyzed. The R(M201-2) and its parent R-factor deoxyribonucleic acid (DNA) could be isolated as an extrachromosomal and covalently closed circular form. Their buoyant densities were both 1.712 g/cm(3), and their molecular weights were about 82 x 10(6) and 64 x 10(6), respectively, when measured by CsCl and sucrose density gradient analyses. The contour lengths by electron microscopy were 35.9 +/- 0.6 and 31.0 +/- 0.6 mum, respectively. By using the extracted R-factor DNA, the mutant and parent characters were transformable to another Escherichia coli strain. The mutant R factor showed an increased amount of DNA even after conjugal transfer to Proteus. An increase in the size of R-factor DNA was thus considered to be the cause of the high level of ampicillin resistance.     

Formation of Recombinants Between Nontransmissible Drug-Resistance Determinants and Transfer Factors

Noninfectious drug-resistance determinants acquired conjugal transmissibility by the formation of recombinants with transfer factors, suggesting the origin of R factors.     

Experimental Salmonellosis VIII. Postinfective Immunity and Its Significance for Conferring Cellular Immunity

In the process of live-vaccine immunization of Salmonella enteritidis infection in mice, the relation between the number of bacteria in the organs of mice and their protecting effect was studied. Treatment with antibiotics was used to control the number of immunizing bacteria in the tissues. Mice, which were infected with 10(-5) mg (1,000 mouse MLD) of virulent S. enteritidis and treated with kanamycin simultaneously, acquired high antilethal resistance against infection with the same organisms. However, the administration of large amounts of kanamycin, which caused a rapid decrease in bacterial numbers in the organs of infected mice, was incapable of conferring immunity. This indicated the necessity of persistence of live bacteria in the host for the production of immunity. A large number of microorganisms were maintained for 53 weeks in a diffusion chamber inserted into the mouse abdominal cavity. The mice implanted with diffusion chambers containing large numbers of virulent S. enteritidis did not acquire antilethal resistance against infection with the same organisms, although agglutinins against S. enteritidis were observed in these mice. Agglutinin was also found in the fluid contained in diffusion chambers inserted into mice immunized with a killed vaccine of S. enteritidis. This indicated that antibody penetrated the membrane filter of diffusion chambers from outside to inside and vice versa. From these results, it is suggested that contact of live microorganisms with the host cell is necessary for conferring postinfective immunity in salmonellosis.     

Experimental Salmonellosis X. Cellular Immunity and Its Antibody in Mouse Mononuclear Phagocytes

A cell-associated antibody was detected in the peritoneal mononuclear phagocytes (referred to as monocytes) of mice hyperimmunized with live vaccine of Salmonella enteritidis, by use of immune transfer and immune adherence hemagglutination techniques. The cellular antibody inhibited the growth of a virulent strain of S. enteritidis with the aid of complement and lysozyme on nutrient agar plates. This type of bactericidal antibody could not be detected in the monocytes of mice immunized with killed vaccine of S. enteritidis. The antibody extracted from the peritoneal monocytes of mice hyperimmunized with live vaccine was identified as a macroglobulin by ultracentrifugal analysis.     

Drug Resistance of Enteric Bacteria XIII. Distribution of R Factors in Escherichia coli Strains Isolated from Livestock

Escherichia coli strains isolated from 151 swine and 108 fowl, which were kept at the Animal Health Center, Maebashi, Japan, were surveyed for drug resistance and distribution of R factors. All of the swine and 38% of the fowl excreted E. coli strains resistant to tetracycline, chloramphenicol, streptomycin, and sulfanilamide, or certain combinations thereof. Among 278 resistant cultures isolated from swine, 13% were found to be resistant to one antibiotic, whereas 87% were resistant to more than one antibiotic. Among these resistant strains, 40% carried R factors which were transferable by the usual conjugal process. The resistance patterns of these R factors included 36% which were singly resistant and 64% which were multiply resistant. Among 54 resistant cultures isolated from fowl, 24% were singly resistant and 76% were multiply resistant. Of the resistant strains from fowl, 22% carried R factors. The resistance patterns of R factors included 50% of the singly resistant type and 50% which were multiply resistant. In spite of feeding with dairy products containing only tetracycline, a high incidence of multiple resistance was observed in the E. coli strains and the R factors isolated from these animals.     

Comparative cytological study of Phasianus colchicus,Meleagris gallopavo,and Gallus domesticus

Summary Since viable intergeneric hybrids between the chicken (Gallus domesticus) and the pheasant (Phasianus colchicus) have been reported, as well as interfamilial hybrids between the chicken and the turkey (Meleagris gallopavo), the chromosome complements of the pheasant and the turkey were compared with that of the chicken. In these three species belonging to the order Galli, the Z-chromosomes appeared to be identical, while the autosomal complements of the pheasant and the turkey differed radically from that of the chicken. It was noted with some surprise that the pheasant of the family Phasianidae and the turkey of the family Meleagridae have very similar chromosome complements, at least so far as gross morphology of somatic metaphase chromosomes is concerned.This work was supported in part by grant C-5138 from the National Cancer Institute, U.S. Public Health Service, and grant C-17601 from the National Science Foundation.The authors gratefully acknowledge the generosity of Rea's Game Birds, Paramount, California, who supplied the pheasant chicks, and the McPherin Hatcheries, Sunnymead, California, who furnished the turkey chicks. The authors also appreciate the editorial assistance of'Patricia A. Ray.     

The Effects of Various Ions on Resting and Spike Potentials of Barnacle Muscle Fibers

Effects of monovalent cations and some anions on the electrical properties of the barnacle muscle fiber membrane were studied when the intra- or extracellular concentrations of those ions were altered by longitudinal intra-cellular injection. The resting potential of the normal fiber decreases linearly with increase of logarithm of [K⁺]_out and the decrement for a tenfold increase in [K⁺]_out is 58 mv when the product, [K⁺]_out ·[Cl^-]_out, is kept constant. It also decreases with decreasing [K⁺]_in but is always less than expected theoretically. The deviation becomes larger as [K⁺]_in increases and the resting potential finally starts to decrease with increasing [K⁺]_in for [K⁺]_in > 250 mM. When the internal K⁺ concentration is decreased the overshoot of the spike potential increases and the time course of the spike potential becomes more prolonged. In substituting for the internal K⁺, Na⁺ and sucrose affect the resting and spike potentials similarly. Some organic cations (guanidine, choline, tris, and TMA) behave like sucrose while some other organic cations (TEA, TPA, and TBA) have a specific effect and prolong the spike potential if they are applied intracellularly or extracellularly. In all cases the active membrane potential increases linearly with the logarithm of [Ca⁺⁺]_out/[K⁺]_in and the increment is about 29 mv for tenfold increase in this ratio. The fiber membrane is permeable to Cl^- and other smaller anions (Br^- and I^-) but not to acetate^- and larger anions (citrate^-, sulfate^-, and methanesulfonate^-).     

Close karyological kinship between the reptilian suborder serpentes and the class aves

Summary In contrast to the situation found in two classes of warm-blooded vertebrates, mammals and birds, the class Reptilia is not uniform with regard to total genetic content; rather, it contains two distinct categories. The close cytological kinship between snakes and birds was revealed. Both are almost identical in total genetic content, which is about 50 per cent that of placental mammals. Both have microchromosomes, as well as Z-chromosomes very similar in absolute size, comprising nearly 10 per cent of the homogametic haploid (AZ) set. This leads to the implication that snakes and birds originated from the same lineage, and that their Z-chromosomes have not changed substantially since the Jurassic period of the Mesozoic era, about 180 million years ago.Within the reptilian suborder Serpentes, the step-by-step differentiation from the primitive ZW pair to the grossly heteromorphic ZW pair could be observed. In the ancient family Boidae, the sex chromosomes were still homomorphic to each other. In the family Colubridae, the beginning of heteromorphism was manifested in two ways. In some species, a pericentric inversion on the W caused it to differ from the Z; in others, duplication of the W occurred. In the family Crotalidae, the W had apparently achieved its very specialized status; it was a distinctly smaller element.In Säo Paulo, this work was supported by Fundacão de Amparo a Pesquisa do Estado de São Paulo e Fundo de Pesquisas do Instituto Butantan. In Duarte, this work was supported in part by grant CA-05138-05, National Cancer Institute, U. S. Public Health Service. Contribution No. 36-64, Department of Biology, City of Hope Medical Center.