Plant regeneration from stem-derived callus of the seed legume Lathyrus sativus L.

Callus cultures from stem explants of six Lathyrus sativus L. cultivars were tested for their morphogenic capacity. Shoot-buds were formed in calli of only one cultivar. Maximum response was observed in the medium containing 10^-8 M picloram and 10^-6 M benzylaminopurin. Supplementation with adenine sulphate was required for shoot-bud formation. The greatest frequency of shoot-bud formation was detected at the second passage and complete lack of regeneration capacity was observed after the 8th passage. Cell regenerates were diploid.     

Crystallizations and preliminary X-ray studies of calotropins DI and DII

Crystals of calotropin DI (M_r 23,400), have been prepared by microdialysis against 5% (w/v) polyethylene glycol 20,000 in water, pH 7.0. They have orthorhombic space group P2₁2₁2₁ with cell parameters $\text{a = 57.5}\text{A}\text{?}\text{, b = 86.2}\text{A}\text{?}\text{, c = 40.3}\text{A}\text{?}$. Crystals of calotropin DII (M_r 24,000), prepared by the same technique against 5% (w/v) polyethylene glycol 20,000 in phosphate buffer of low ionic strength, pH 7.0, display monoclinic space group C2 with cell parameters $\text{a = 135.8}\text{A}\text{?}\text{, b = 32.0}\text{A}\text{?}\text{, c = 47.7}\text{A}\text{?}\text{, β = 103.80 °}$. In both cases, there is only one molecule in the asymmetric unit.     

Purified vesicular stomatitis virus contains an enzyme activity that synthesizes cytidylyl (5'-3') guanosine 5'-triphosphate in vitro

An enzyme activity that synthesizes cytidylyl (5'-3') guanosine 5'-triphosphate (pppGpC) in vitro has been identified in purified vesicular stomatitis virus. The activity is discernible after a lag period which is reduced in length with increasing virus concentration. The lag is eliminated by addition of pppGpC or ppGpC which are effective primers and stimulate dinucleotide synthesis linearly. The requirements of the reaction with respect to MgCl2, NaCl, and temperature are similar to those for viral mRNA synthesis in vitro. The activity, together with the viral L and NS proteins, is removed from virions by treatment with 0.8 M NaCl. The particulate fraction from infected cells that contains the transcribing subviral ribonucleoprotein particles also contains the enzyme activity. The corresponding fraction from uninfected cells does not, indicating that the activity is mediated by virus-specific proteins. Possible functions of the dinucleotide in the life cycle of the virus are discussed.     

Freeze-fracture,ultrastructural and autoradiographic analysis of the ventral prostate glands in castrated mice

Summary Ultrastructure of the ventral prostate glands was studied in mice castrated for 1 through 60 days and for 11 and 17 months and in age-matched normals. We have described freeze-fracture and ultrastructural characteristics of acinar epithelial cells in addition to the patterns of thymidine incorporation in the cells of castrates and normal animals. Our study has shown a biphasic pattern of prostatic involution in the long-term castrated mice. In castrates the initial atrophy of prostate glands occurred by sloughing of the apical portions of columnar cells, autophagia of the cytoplasmic organelles as well as by occasional sloughing of the individual cells into the acinar lumen. Concurrent with the initial atrophy, the glands and stroma were infiltrated by neutrophils and lymphocytes. The cell loss by sloughing and leucocyte infiltration of glands became infrequent in 7- to 21-day castrates. However, the cell loss by sloughing increased secondarily in mice castrated for 21 to 37 days along with the increased leucocyte infiltration of the glands. The cell loss became minimal in castrates of 60 days and beyond. Our evidence suggests that the cell loss by sloughing was an active process in the involution of prostate glands which also showed differential sensitivity to castration stimuli in mice.This research was supported by Medical Research Service of Veterans Administration     

A new method for separating cyclic AMP from 5′-AMP with application to the assay for cyclic AMP phosphodiesterase

A new method for assay of cyclic AMP phosphodiesterase (EC 3.1.4.17) has been developed based on the observation that a mixture of cyclic AMP and AMP can be resolved on a column of florisil (activated magnesium silicate) at pH 7.0. The cyclic nucleotide is retained by the silicate and the AMP which is not adsorbed is virtually quantitatively recovered. The adsorption of cyclic AMP by florisil is greatly influenced by the pH of the buffer but independent of its ionic strength. In the actual assay cyclic[³H]AMP is incubated with the enzyme source in the presence of Mg²⁺ and the reaction is stopped by the addition of CCl₃COOH (0.3 m). The mixture is then neutralized by dilution with 10 vol of 0.5 m sodium phosphate buffer, pH 7.0, and applied on a small (0.4 × 4.0-cm) florisil column equilibrated with the same buffer. The column is eluted with 3 vol of the buffer and the radioactivity of the eluate which contains only [³H]AMP is measured. The use of cyclic[³H]AMP of high specific activity in the assay allows a high degree of sensitivity while the addition of CCl₃COOH instantaneously terminates the reaction allowing for increased precision. The assay compares favorably in simplicity and speed with those currently employed for cyclic AMP phosphodiesterase.     

Cyanobacterial biofertilizers in rice agriculture

Floodwater and the surface of soil provide the sites for aerobic phototrophic nitrogen (N) fixation by free-living cyanobacteria and theAzolla-Anabaena symbiotic N₂-fixing complex. Free-living cyanobacteria, the majority of which are heterocystous and nitrogen fixing, contribute an average of 20–30 kg N ha^-1, whereas the value is up to 600 kg ha^-1 for theAzollaAnabaena system (the most beneficial cyanobacterial symbiosis from an agronomic point of view). Synthesis and excretion of organic/growth-promoting substances by the cyanobacteria are also on record. During the last two or three decades a large number of studies have been published on the various important fundamental and applied aspects of both kinds of cyanobacterial biofertilizers (the free-living cyanobacteria and the cyanobacteriumAnabaena azollae in symbiotic association with the water fernAzolla), which include strain identification, isolation, purification, and culture; laboratory analyses of their N₂-fixing activity and related physiology, biochemistry, and energetics; and identification of the structure and regulation of nitrogenfixing (nif) genes and nitrogenase enzyme. The symbiotic biology of theAzolla-Anabaena mutualistic N₂-fixing complex has been clarified. In free-living cyanobacterial strains, improvement through mutagenesis with respect to constitutive N₂ fixation and resistance to the noncongenial agronomic factors has been achieved. By preliminary meristem mutagenesis inAzolla, reduced phosphate dependence was achieved, as were temperature tolerance and significant sporulation/spore germination under controlled conditions. Mass-production biofertilizer technology of free-living and symbiotic (Azolla-Anabaena) cyanobacteria was studied, as were the interacting and agronomic effects of both kinds of cyanobacterial biofertilizer with rice, improving the economics of rice cultivation with the cyanobacterial biofertilizers. Recent results indicate a strong potential for cyanobacterial biofertilizer technology in rice-growing countries, which opens up a vast area of more concerted basic, applied, and extension work in the future to make these self-renewable natural nitrogen resources even more promising at the field level in order to help reduce the requirement for inorganic N to the bare minimum, if not to zero.     

Mapping of movement in the isometrically contracting human soleus muscle reveals details of its structural and functional complexity.

It is becoming increasingly apparent that precise knowledge of the anatomic features of muscle, aponeurosis, and tendons is necessary for understanding how a muscle-tendon complex generates force and accomplishes length changes. This report presents both anatomic and functional data from the human soleus muscle acquired by using magnetic resonance imaging. The results show a strong relationship between the complex three-dimensional structure of the muscle-tendon system and the intramuscular distribution of tissue velocities during in vivo isometric contractions. The proximal region of the muscle is unipennate, whereas the midregion has a radially bipennate hemicylindrical structure, and the distal region is quadripennate. Tissue velocity mapping shows that the highest velocity regions overlay the aponeuroses connected to the Achilles tendon. These are located on the anterior and posterior surfaces of the muscle. The lowest velocities overlay the aponeuroses connected to the origin of the muscle and are generally located intramuscularly.