The adult fast isozyme of myosin is present in a nerve-muscle tissue culture system

Abstract. Organotypic nerve-muscle cultures were prepared from foetal mouse spinal cord and adult mouse muscle fibres. In this system, the adult fibres degenerate and new myotubes form. The regenerated muscle fibres become innervated, develop cross-striations, and survive for several months. We have investigated the isozymes of myosin present in these muscle fibres using histochemistry and immunocytochemistry with antibodies to rat embryonic, neonatal, and adult fast myosins. We demonstrate that some of the regenerated fibres contain adult fast but not embryonic or neonatal myosin. This is the first demonstration of the production of adult myosin heavy chain in tissue culture. This system therefore offers the possibility for further study of the development of adult myosin isoforms in vitro.     

Purification, characterization and antimicrobial properties of an antibiotic (wassumycin) from Streptomyces species 215

A yellow amorphous compound, wassumycin, was isolated from an unidentified strain of Streptomyces. Wassumycin inhibited the growth of several species of human and animal pathogenic bacteria especially Clostridia. Wassumycin also inhibited the growth of HeLa cells, and showed good antitumour activity against experimental mouse tumours such as L1210, P388 leukaemias, and B16 melanoma. On the basis of the chemical and antimicrobial properties, the antibiotic appeared similar to the chromomycins and olivomycins, but its ultraviolet, infrared, and proton magnetic resonance spectra differed from theirs significantly.     

Murine hybridomas secreting natural monoclonal antibodies reacting with self antigens

Spleen cells from nonimmunized BALB/c mice were fused with two nonsecreting myeloma lines. The hybrids were selected in HAT medium and screened for Ig production and for antibody activity against actin, tubulin, myosin, thyroglobulin, myoglobin, spectrin, dsDNA, fetuin, and transferrin. Among 161 hybrids secreting Ig, three were found to react with DNA, one with thyroglobulin, and one mainly with myosin. Two of these hybrids could be propagated and further characterized. On the basis of inhibition experiments, one was found to be directed against dsDNA; the other was directed mainly against myosin but at the same time reacted significantly with actin, tubulin, spectrin, and dsDNA. Reactivity with myosin seemed to be concentrated in the light meromyosin subfragment, known to be rich in alpha-helical structure. These results indicate: 1) There are reactive B cell clones directed against self antigens. 2) The antibody specificities found for these antibodies are very similar to those found for natural antibodies in normal human serum and for human monoclonal Ig. 3) The widespread reactivity found for the clone mainly reacting with myosin raises the possibility that the determinant recognized by this antibody is a conformational structure that possibly is associated with alpha-helical structures.     

Modeling the Potential Impact of Host Population Survival on the Evolution of M. tuberculosis Latency

Tuberculosis (TB) is an infectious disease with a peculiar feature: Upon infection with the causative agent, Mycobacterium Tuberculosis (MTB), most hosts enter a latent state during which no transmission of MTB to new hosts occurs. Only a fraction of latently infected hosts develop TB disease and can potentially infect new hosts. At first glance, this seems like a waste of transmission potential and therefore an evolutionary suboptimal strategy for MTB. It might be that the human immune response keeps MTB in check in most hosts, thereby preventing it from achieving its evolutionary optimum. Another possible explanation is that long latency and progression to disease in only a fraction of hosts are evolutionary beneficial to MTB by allowing it to persist better in small host populations. Given that MTB has co-evolved with human hosts for millenia or longer, it likely encountered small host populations for a large share of its evolutionary history and had to evolve strategies of persistence. Here, we use a mathematical model to show that indeed, MTB persistence is optimal for an intermediate duration of latency and level of activation. The predicted optimal level of activation is above the observed value, suggesting that human co-evolution has lead to host immunity, which keeps MTB below its evolutionary optimum.     

Cloning of the rat insulin-like growth factor binding protein-1 gene and analysis of its 5' promoter region.

To understand specific mechanisms involved in the regulation of insulin-like growth factor binding protein-1 (IGFBP-1), an important modulator of IGF bioactivity, we cloned the rat IGFBP-1 gene and sequenced a 1.5 kb Sph1-Sph1 fragment containing 1110 bases upstream from the translation start site. Computer analysis reveals the presence of ATA, CACCC, and CCAAT elements, and putative homeodomain, AP-1, insulin and glucocorticoid response elements in the 5' promoter. Primer extension and ribonuclease protection studies reveal a single cap site in RNA from rat hepatoma cells and both control and diabetic rat liver.     

Plant Life History and Residue Chemistry Influences Emissions of CO2 and N2O From Soil – Perspectives for Genetically Modified Cell Wall Mutants

Vascular plants have lignified tissues that transport water, minerals, and photosynthetic products throughout the plant. They are the dominant primary producers in terrestrial ecosystems and capture significant quantities of atmospheric carbon dioxide (CO₂) through photosynthesis. Some of the fixed CO₂ is respired by the plant directly, with additional CO₂ lost from rhizodeposits metabolized by root-associated soil microorganisms. Microbially-mediated mineralization of organic nitrogen (N) from plant byproducts (rhizodeposits, dead plant residues) followed by nitrification generates another greenhouse gas, nitrous oxide (N₂O). In anaerobic soils, reduction of nitrate by microbial denitrifiers also produces N₂O. The plant-microbial interactions that result in CO₂ and N₂O emissions from soil could be affected by genetic modification. Down-regulation of genes controlling lignin biosynthesis to achieve lower lignin concentration or a lower guaiacyl:syringyl (G:S) ratio in above-ground biomass is anticipated to produce forage crops with greater digestibility, improve short rotation woody crops for the wood-pulping industry and create second generation biofuel crops with low ligno-cellulosic content, but unharvested residues from such crops are expected to decompose quickly, potentially increasing CO₂ and N₂O emissions from soil. The objective of this review are the following: 1) to describe how plants influence CO₂ and N₂O emissions from soil during their life cycle; 2) to explain how plant residue chemistry affects its mineralization, contributing to CO₂ and N₂O emissions from soil; and 3) to show how modification of plant lignin biosynthesis could influence CO₂ and N₂O emissions from soil, based on experimental data from genetically modified cell wall mutants of Arabidopsis thaliana. Conceptual models of plants with modified lignin biosynthesis show how changes in phenology, morphology and biomass production alter the allocation of photosynthetic products and carbon (C) losses through rhizodeposition and respiration during their life cycle, and the chemical composition of plant residues. Feedbacks on the soil environment (mineral N concentration, soil moisture, microbial communities, aggregation) affecting CO₂ and N₂O emissions are described. Down-regulation of the Cinnamoyl CoA Reductase 1 (CCR1) gene is an excellent target for highly digestable forages and biofuel crops, but A. thaliana with this mutation has lower plant biomass and fertility, prolonged vegetative growth and plant residues that are more susceptible to biodegradation, leading to greater CO₂ and N₂O emissions from soil in the short term. The challenge in future crop breeding efforts will be to select tissue-specific genes for lignin biosynthesis that meet commercial demands without compromising soil CO₂ and N₂O emission goals.     

Wind turbine noise limits propagation of greater prairie‐chicken boom chorus,but does it matter?

Acoustic signals are often critical elements of mating displays, and lekking male greater prairie‐chickens (Tympanuchus cupido pinnatus) use their boom vocalization for this purpose. We quantified the acoustic characteristics of the boom chorus created by multiple male greater prairie‐chickens vocalizing simultaneously at leks in Brown County, NE, USA, in 2013 and 2014. We used these data to evaluate (a) the role of the boom chorus in prairie‐chicken breeding dynamics and (b) the impact of a wind energy facility on the acoustic signal of the boom chorus. We sampled the chorus using audio recorders placed in transects extending from leks; the chorus exhibited an average peak frequency of 297 ± 13 Hz. The mean chorus signal‐to‐noise ratio declined from 15.7 dB (50 m) to 2.6 dB (800 m), and wind speed and direction, topography, and relative humidity caused variation in signal‐to‐noise ratio at a given distance and location. Chorus recordings from leks within 1,000 m of a wind turbine had lower signal‐to‐noise ratio (β_turbine = ?5.659, SE = 1.289) than leks farther from turbines. The chorus signal‐to‐noise ratio increased slightly with more males present on the lek (~0.1 dB for each additional male; β_males = 0.177; SE = 0.037) and considerably more as more females visited the lek (~1.4 dB for each additional female; β_females = 2.498, SE = 0.235; β_females² = ?0.309, SE = 0.039). Our results provide support for the signal enhancement hypothesis that proposes the boom chorus is influenced notably by male–male competition for females on the lek, rather than functioning solely to advertise the presence of the lek to recruit females. Our results also suggest the choruses emanating from small leks have the greatest potential to be masked by anthropogenic (wind turbine) noise, which may affect the breeding success of male and female prairie‐chickens.     

A comparative analysis of intermediate filament proteins in bovine heart Purkinje fibres and gastric smooth muscle

The intermediate filament (IF) composition of muscle cells of various sources is still a controversial issue. In the present study, the IF composition of bovine Purkinje fibres (PFs), atrial and ventricular myocardium, and gastric smooth muscle (SM) has been compared using biochemical and immunocytochemical methods. The Mr of the major IF subunit protein in all four tissues was 55,000. In two-dimensional (2-D) electrophoresis gels of Triton-treated ordinary atrial and ventricular myocardium and the gastric muscular wall, two or three isoelectric isoforms were seen, whereas in PFs up to seven isoforms caused by phosphorylation were observed. In immunofluorescence studies antibodies against the Mr 55,000 subunit of PFs and gastric SM, respectively, both showed identical reactivity with PFs, atrial and ventricular myocytes, gastric SM cells and some SM cells in intramyocardial and gastric muscular wall blood vessels. A small amount of vimentin (Mr 57,000) was also detected in 2-D gel electrophoresis in all four tissues as well as in immunoblotting of PFs with antibodies to vimentin. Immunofluorescence studies using both polyclonal and monoclonal antibodies to vimentin showed that vimentin was present in the endothelium and SM cells of both intramyocardial and gastric muscular wall vessels, sometimes together with desmin in the vascular SM cells, but was never seen in PF, atrial, ventricular or gastric SM cells proper. As expected, vimentin was present in interstitial tissue, i.e., fibroblasts and capillaries. However, interestingly, the monoclonal antibodies, which recognized different antigenic determinants of vimentin, did not give identical staining patterns. Especially the staining of the vascular SM cells differed. Since this staining pattern did not change upon denaturation and unmasking experiments, it seems that the organization of vimentin in different mesenchymal cell types varies. Vimentin was also detected in isolated PFs but here it was located solely in the contaminating interstitial tissue. Thus, desmin is the sole IF protein expressed in PFs, in atrial and ventricular myocytes and in gastric SM cells proper; vimentin alone being present in the interstitial tissue cells, whilst in vascular SM cells desmin and vimentin are coexpressed in various proportions. The variation in number of isoforms of desmin and the heterogeneity in staining of mesenchymal tissues with monoclonal vimentin antibodies probably indicates that the IF cytoskeletons are differently organized in various cell types, even though they contain IFs of the same class.