Recent advances in plant cell cultures in bioreactors

Two key issues in the application of plant-cell-culture technology to the production of valuable secondary metabolites are reviewed: the selection of cell lines with suitable genetic, biochemical and physiological characteristics; and the optimization of bioreactor environments. Although great progress has been made in recent years in the design, selection and optimization of bioreactor hardware, optimization of environmental factors such as medium components, light irradiation and O₂ supply needs detailed investigations for each case. With a better understanding of plant cell metabolism and physiology, further developments in cultivation processes, such as process integration and on-line monitoring and control, can be expected in the near future.J.-J. Zhong and J.-T. Yu are with the Research Institute of Biochemical Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China T. Yoshida is with the International Center of Cooperative Research in Biotechnology (ICBiotech), Faculty of Engineering, Osaka University, Suita, Osaka 565, Japan.     

Effects of substance P on Na and K transport in perfused main submandibular duct of rat

Substance P (2 and 4 micrograms/kg . min, iv) caused an inhibition of net efflux of Na and net influx of K in perfused main excretory duct of rat submandibular gland. These effects could not be blocked by atropine sulfate. The data suggest that substance P receptors are present in the duct cells and play a role in the regulation of transductal electrolyte transport.     

Replication of chromosomal DNA in cultured abnormal human cells

Summary The replication of chromosomal DNA in a series of abnormal human cell cultures has been studied by means of DNA-fiber autoradiography. In lymphocytes with trisomy 21, in fibroblasts of 45,X; 47,XXX; 49,XXXXY; and 49,XXXXX chromosomal constitution, and in fibroblasts from a patient with xeroderma pigmentosum (De Sanctis-Cacchione syndrome), the rate of DNA replication does not differ from that in normal cells, varying in a single fork from 0.2 to 1.0 m/min with a mean of about 0.6 m/min. In fibroblasts with trisomy 7 the rate of DNA replication is greater, varying from 0.3 to 1.2 m/min with a mean of about 0.8 m/min. The sizes of replication units in all cells examined are from 80 to 500 m with a mean of about 200–300 m.     

Respiratory system of Fusidium coccineum and regulation of biosynthesis of fusidic acid and sporogenesis

Summary The respiratory system, sporulation, and dynamics of alkaline protease formation were studied in three strains of the fungus Fusidium coccineum, differing in their ability to make antibiotics.Oxidative phosphorylation provided most of the energy in high and low activity strains and their respiratory activity was exclusively related to mitochondria functioning.In inactive and low activity strains, the terminal oxidation of reduced equivalents proceeds mainly by the respiratory chain with cytochrome oxidase as the terminal component. In the high activity strain there is a cyanide-resistant alternative pathway which is parallel to the classical cytochrome chain. The complete transition to the use of this pathway coincides with the stage of maximum antibiotic biosynthesis. The induction of the alternative pathway in the high activity strain was not concerned with the inhibition of the cytochrome site of the respiratory chain by fusidic acid. It was shown that the quantity of the antibiotic synthesized and the character of cellular differentiation can be altered by changing the oxidation pathwats used with inhibitors such as chloramphenicol and salicyl hydroxamate.We suggest that there must be common regulation of antibiotic formation, sporulation and induction of the alternative oxidation pathway.     

Functional properties of locust locomotor muscles

The ultrastructure of the muscle fibers and the electrical constants and responses of the membrane to microapplication of L-glutamate and acetylcholine were investigated in the longitudinal flight muscle and the flexor tibiae ofLocusta migratoria migratorioides. The twitch flight muscle differs from the slower leg muscle in the smaller size of its sarcomeres and the lower values of the space attenuation factor of the electrotonic potential, time constant, and resistance of the membrane. Microapplication of sodium L-glutamate at strictly definite points of the fibers of both muscles evoked depolarization responses of the membrane. In experiments on normal and denervated muscle, during microapplication of acetylcholine, changes in the level of the membrane potential were never observed. It is concluded that L-glutamic acid is the excitatory mediator of the twitch and slow muscle systems of insects.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 9, No. 5, pp. 532–538, September–October, 1977.     

Nucleotide sequence of the genome of the filamentous bacteriophage I2-2: Module evolution of the filamentous phage genome

Summary The nucleotide sequence of the circular single-stranded genome of the filamentous Escherichia coli phage I2-2 has been determined and compared with those of the filamentous E. coli phages Ff(M13, fl, or fd) and IKe. The I2-2 DNA sequence comprises 6744 nucleotides; 139 nucleotides less than that of the N- and I2-plasmid-specific phage IKe, and 337 (336) nucleotides more than that of the F-plasmid-specific phage Ff. Nucleotide sequence comparisons have indicated that I2-2, IKe, and Ff have a similar genetic organization, and that the genomes of I2-2 and IKe are evolutionarily more closely related than those of I2-2 and Ff. The studies have further demonstrated that the I2-2 genome is a composite replicon, composed of only two-thirds of the ancestral genome of IKe. Only a contiguous I2-2 DNA sequence of 4615 nucleotides encompassing not only the coat protein and phage assembly genes, but also the signal required for efficient phage morphogenesis, was found to be significantly homologous to sequences in the genomes of IKe and Ff. No homology was observed between the consecutive DNA sequence that contains the origins for viral and complementary strand replication and the replication genes. Although other explanations cannot be ruled out, our data strongly suggest that the ancestor filamentous phage genome of phages I2-2 and IKe has exchanged its replication module during evolution with that of another replicon, e.g., a plasmid that also replicates via the so-called rolling circle mechanism. Offprint requests to: R.N.H. Konings     

Conformational properties of oxytocin in dimethyl sulfoxide solution: NMR and restrained molecular dynamics studies.

The conformation of oxytocin, the neurohypophyseal nonapeptide hormone, in solution in deuterated dimethyl sulfoxide has been determined by 1H-nmr. The structural determination is based on the experimental data set of nuclear Overhauser effect restraints. Obtained after the restrained molecular dynamics simulation on an initial structure of extended conformation, five resultant structures satisfy the experimental restraints well. These structures resemble that of the crystal structure of deamino-oxytocin, an analogue of oxytocin, in terms of a close correlation observed both at two beta-turn regions of the 20-membered tocin ring and at the tripeptide tail end. Based on this comparison and analysis of restrained molecular dynamics trajectories, we found that, although the turns are stabilized by the formation of hydrogen bonds, the oxytocin molecule possesses a slight twist in DMSO solution relative to the orientation of deamino-oxytocin in the crystalline state. Analyses of oxytocin conformation indicate that the tripeptide tail is more flexible than the tocin ring.     

Early stages of formation and dispersal of the temperate flora in the Boreal Region

Budantsev, L. Yu. (Komarov Botanical Institute, Prof. Popov Str. 2, 197376 St. Petersburg, Russia). Early stages of formation and dispersal of the temperate flora in the Boreal Region. Bot. Rev.58(1): 1–48, 1992.—The thesis of this review is that, as stated as early as 1908 by V. L. Komarov, the composition of a flora can be understood only as a process, or separate stage, in the context of migration in time and space of various floristic assemblages and their isolation, as induced by transformation of continental and ocean shapes, changes in climate, and the environment as a whole. Thus the formation of geofloras of the past was influenced by gradually changing environments that determined the spread, patterning, and spatial differentiation of floras and their evolution. Parallel to the more commonly-seen names of eras—Paleozoic, Mesozoic, and Cenozoic—we can speak of the Paleophytic, Mesophytic, and Cenophytic eras. Eras defined in these two ways (by faunistic or by floristic criteria) do not completely coincide. Generally, changes in the flora have, necessarily, preceded changes in the fauna. It is the Cenophytic with which this review is mostly concerned, the era of Angiosperm dominance. The movement of early subtropical and warm temperate floras in the Early Cenophytic, followed by temperate or even boreal floras, as the climate changes, is traced in detail. The regions discussed most fully are the Boreal-Atlantic and Boreal-Pacific, with emphasis on the Angaro-Beringian flora. The disappearance of archaic forms (e.g., cycadophytes) and the gradual predominance of angiosperms is documented. The movements of the floral assemblages in response to environmental changes are mapped and described. The early development and diversification of the boreal temperate flora is considered to have taken place mainly in Angaro-Beringia, associated with the invasive migration of tropical angiosperms from southeastern Asia.