Characterization of cross-bridge elasticity and kinetics of cross-bridge cycling during force development in single smooth muscle cells

Force development in smooth muscle, as in skeletal muscle, is believed to reflect recruitment of force-generating myosin cross-bridges. However, little is known about the events underlying cross-bridge recruitment as the muscle cell approaches peak isometric force and then enters a period of tension maintenance. In the present studies on single smooth muscle cells isolated from the toad (Bufo marinus) stomach muscularis, active muscle stiffness, calculated from the force response to small sinusoidal length changes (0.5% cell length, 250 Hz), was utilized to estimate the relative number of attached cross-bridges. By comparing stiffness during initial force development to stiffness during force redevelopment immediately after a quick release imposed at peak force, we propose that the instantaneous active stiffness of the cell reflects both a linearly elastic cross-bridge element having 1.5 times the compliance of the cross-bridge in frog skeletal muscle and a series elastic component having an exponential length-force relationship. At the onset of force development, the ratio of stiffness to force was 2.5 times greater than at peak isometric force. These data suggest that, upon activation, cross-bridges attach in at least two states (i.e., low-force-producing and high-force-producing) and redistribute to a steady state distribution at peak isometric force. The possibility that the cross-bridge cycling rate was modulated with time was also investigated by analyzing the time course of tension recovery to small, rapid step length changes (0.5% cell length in 2.5 ms) imposed during initial force development, at peak force, and after 15 s of tension maintenance. The rate of tension recovery slowed continuously throughout force development following activation and slowed further as force was maintained. Our results suggest that the kinetics of force production in smooth muscle may involve a redistribution of cross-bridge populations between two attached states and that the average cycling rate of these cross-bridges becomes slower with time during contraction.     

Bacillus sporulation gene spo0H codes for sigma 30 (sigma H).

The DNA sequences of the spo0H genes from Bacillus licheniformis and B. subtilis are described, and the predicted open reading frames code for proteins of 26,097 and 25,447 daltons, respectively. The two spo0H gene products are 91% identical to one another and about 25% identical to most of the procaryotic sigma factors. The predicted proteins have a conserved 14-amino-acid sequence at their amino terminal end, typical of sigma factors. Antibodies raised against the spo0H gene product of B. licheniformis specifically react with RNA polymerase sigma factor protein, sigma 30, purified from B. subtilis. We conclude that the spo0H genes of B. licheniformis and B. subtilis code for sigma 30, now known as sigma H.     

Pancreatic islets of variable size--insulin secretion and glucose utilization

Glucose metabolism and insulin secretion were studied in isolated rat pancreatic islets of different sizes and the amount of tissue was quantitated by the measurement of DNA. It was found that larger islets (140-210 ng DNA/islet) utilized more glucose (based on the conversion of 3H-5-glucose to [3H]20) per ng of DNA than islets containing less DNA (60-120 ng/islet). However, the insulin secreted per ng of DNA in response to a given glucose concentration was the same in islets of all sizes. Also, the islet insulin and glucagon content when expressed in terms of DNA did not depend upon islet size. Thus, although glucose utilization rates expressed as a function of islet DNA content were greater in larger islets, no such relationship was found for glucose-induced insulin release or insulin and glucagon content.     

Physiological influences on carbon isotope discrimination in huon pine (Lagarostrobos franklinii)

Summary Measurements of the light environment and stomatal and photosynthetic behaviour are reported for Huon Pine (Lagarostrobos franklinii, family Podocarpaceae) in western Tasmanian rainforest. For a variety of microenvironments, these are used in an analysis of stable carbon isotope measurements in the air, and in branch and leaf material, using a model for carbon isotope fractionation in leaves (Farquhar et al. 1982).The major features of ¹³C variations with respect to branch position can be explained in terms of the direct influence of light level acting via the rate of CO₂ assimilation. In addition a relatively constant ¹³C gradient of about 2.6 between leaf tip and branch wood is observed.Alternative explanations are advanced for the tip-towood gradient in ¹³C. If the ¹³C of leaf tips is taken to represent the value for photosynthate, maintenance respiration is proposed as a mechanism for the further fractionation; a significant ¹³C depletion in respired CO₂ is implied which is not supported by indirect measurements of atmospheric isotope ratio. Furthermore, an assumption of significant sampling errors (e.g. related to humidity effects on assimilation) is required to obtain good quantitative prediction of the light influence.If the branch wood ¹³C is taken to represent that of the photosynthate, the tip-to-wood gradient may find an explanation, via the model, in terms of tip tissue comprising carbon from immature cells. Translocation of photosynthate from exposed to shaded branches is then proposed as a means of obtaining quantitative agreement with the predicted light influence.The support provided for the applicability of the Farqunar et al. (1982) model in the field is discussed in the context of the problem of obtaining past global atmospheric CO₂ levels from ¹³C in tree-rings.     

DNA-binding proteins present in varicella-zoster virus-infected cells.

DNA-binding proteins present in varicella-zoster virus-infected cells were identified by DNA-cellulose chromatography of radioactively labeled cell extracts. Seven virus-specific proteins, ranging in molecular weight from approximately 175,000 to 21,000, showed affinity for single- or double-stranded DNA or both. These proteins include the varicella-zoster virus major capsid protein, a phosphorylated tegument protein, and a 125,000-molecular-weight species which may be analogous to the major DNA-binding protein of herpes simplex virus. We also identified a number of DNA-binding phosphoproteins by these procedures. Finally, protein blot studies were carried out to determine whether these proteins bind preferentially to virus rather than to host cell DNA.     

Multiline varieties and disease control

Summary A general model for the evolution of pathogen populations on mixtures or multilines is developed. This model is used to extend previous analyses of the effects of the widespread cultivation of multilines on the evolution of virulence in obligate parasites to mixtures of lines carrying different numbers of resistance genes. It is concluded that the composition of an equilibrium pathogen population growing on a multiline may vary within wide limits and the prinicipal determinant of its composition is the number of components in the multiline and the resistance genes they carry. Other factors of importance are (i) the relative contribution made by each host class (with different numbers of resistance genes) to the pathogen spore pool each generation; (ii) the levels of stabilizing selection against unnecessary virulence genes; and (iii) the way in which unnecessary genes for virulence combine to reduce pathogen fitness. Conditions for the fixation of avirulent biotypes in the pathogen population and the evolution of a pathogen superrace are given for multilines of various compositions.Paper No. 9246 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina. This investigation was supported in part by NIH Research Grant No. GM 11546 from the National Institute of General Medical Sciences     

Temporal Patterns of Reported Single-Vehicle Car and Truck Accidents in Texas, U.S.A. During 1980-1983

Reported single car and truck accidents due specifically to driver 'fatigue' for the 4-year span of 1980-1983 were analyzed for 24-hr, 7-d and 1-yr patterns. Three types of data were examined: (1) accidents in rural Texas, (2) accidents only in San Antonio, the fourth largest city in Texas and (3) accident ratio (number of accidents per time interval divided by traffic volume for the interval) in this city.

A statistically significant seasonal variation of relatively low amplitude with peaks in May and July was detected only for accidents in rural areas. High-amplitude periodicities of 7 d were evident in all the data sets, with a peak on the weekend. A very prominent 24-hr pattern was detected as well, with the acrophase determined by single cosinor around 0300. A slight secondary peak in accidents, around 1500, was evident in the data from the rural areas. This secondary peak represented a 12-hr rhythm in addition to the 24-hr. Overall, features of the temporal patterns in accidents over 24 hr and over the year in urban and rural areas differed only slightly. Accident ratios yielded results similar to accident frequencies. However, with regard to the rural data, the shape of the 24-hr temporal pattern varied with driver age; older drivers exhibited a flatter curve with a more pronounced afternoon rather than early morning (around 0300) peak.