Cooperative attachment of cross bridges predicts regulation of smooth muscle force by myosin phosphorylation

Serine 19 phosphorylation of the myosin regulatory light chain (MRLC) appears to be the primary determinant of smooth muscle force development. The relationship between MRLC phosphorylation and force is nonlinear, showing that phosphorylation is not a simple switch regulating the number of cycling cross bridges. We reexamined the MRLC phosphorylation-force relationship in slow, tonic swine carotid media; fast, phasic rabbit urinary bladder detrusor; and very fast, tonic rat anococcygeus. We found a sigmoidal dependence of force on MRLC phosphorylation in all three tissues with a threshold for force development of approximately 0.15 mol P(i)/mol MRLC. This behavior suggests that force is regulated in a highly cooperative manner. We then determined whether a model that employs both the latch-bridge hypothesis and cooperative activation could reproduce the relationship between Ser(19)-MRLC phosphorylation and force without the need for a second regulatory system. We based this model on skeletal muscle in which attached cross bridges cooperatively activate thin filaments to facilitate cross-bridge attachment. We found that such a model describes both the steady-state and time-course relationship between Ser(19)-MRLC phosphorylation and force. The model required both cooperative activation and latch-bridge formation to predict force. The best fit of the model occurred when binding of a cross bridge cooperatively activated seven myosin binding sites on the thin filament. This result suggests cooperative mechanisms analogous to skeletal muscle that will require testing.     

Molecular characterization of birnaviruses isolated from wild marine fishes at the Flemish Cap (Newfoundland)

Several isolates of aquatic birnaviruses were recovered from different species of wild fish caught in the Flemish Cap, a Newfoundland fishery close to the Atlantic coast of Canada. The nucleotide sequence of a region of the NS gene was identical among the isolates and was most similar to the Dry Mills and West Buxton reference strains of infectious pancreatic necrosis virus (IPNV). Phylogenetic analysis of the sequence of a region of the VP2 gene demonstrated that the isolates were most closely aligned with the American strains of IPNV serotype A1. Electron microscopy of virus structures clarified and concentrated from cultures of infected chinook salmon embryo (CHSE-214) cells revealed a majority of typical IPNV-like icosahedral particles, as well as a low proportion of type I tubules having a diameter of approximately 55 nm and a variable length of up to 2 microm. The tubules could be propagated in cell cultures, but always in the presence of low proportions of icosahedral particles. Cloning of selected isolates by serial dilution yielded preparations with a high proportion of the tubular structures with a density in CsCl gradients of approximately 1.30 g cm(-3). Polyacrylamide gel electrophoresis revealed the material in the band was composed of the IPNV pVP2 and VP2 proteins.     

香菇的学名及其在当代真菌分类中的位置

本文综述了香菇(Lentinula edodes)的分类历史,确认其在蘑菇目(Agaricales)Tricholomataceae科下的分类地位,并证实了它与多孔菌目(Poriales)Lentinaceae科的Lentinus属没有联系。根据《真菌、地衣汉语学名命名法规》,作者讨论了译为“香菇属”的Lentinus和“小香菇属”的Lentinellus两属的汉语学名问题,提出Lentinus的汉语学名应订正为“韧伞属”,Lentinellus为“螺壳菌属”。香菇所在的Lentinula属的汉语学名建议为“木菇属”。     

Root systems and root:mass ratio-carbon allocation under current and projected atmospheric conditions in arable crops

Roots of annual crop plants are a major sink for carbon particularly during early, vegetative growth when up to one-half of all assimilated carbon may be translocated belowground. Flowering marks a particularly important change in resource allocation, especially in determinate species, with considerably less allocation to roots and, depending on environmental conditions, there may be insufficient for maintenance. Studies with ¹⁴C indicate the rapid transfer belowground of assimilates with typically 50% translocated in young cereal plants of which 50% is respired; exudation/rhizodeposition is generally <5% of the fixed carbon. Root: total plant mass decreases through the season and is affected by soil and atmospheric conditions. Limited water availability increased the allocation of ¹³C to roots of wheat grown in columns so that at booting 0.38 of shoot C (ignoring shoot respiration) was belowground compared to 0.31 in well-watered plants. Elevated CO₂ (700 mol CO₂ mol^–1 air) increased the proportion of root:total mass by 55% compared with normal concentration, while increasing the air temperature by a mean of 3 °C decreased the proportion from 0.093 in the cool treatment to 0.055 in the warm treatment.     

The duration and rate of grain growth, and harvest index, of wheat (Triticum aestivum L.) in response to temperature and CO2

Winter wheat (Triticum aestivum L. cv. Hereward) was grown inthe field inside polyethylene-covered tunnels at a range oftemperatures at either 380 or 684 µmol mol^–1 CO₂.Serial harvests were taken from anthesis until harvest maturity.Grain yield was reduced by warmer temperatures, but increasedby CO₂ enrichment at all temperatures. During grain-filling,individual grain dry weight was a linear function of time fromanthesis until mass maturity (attainment of maximum grain dryweight) within each plot. The rate of progress to mass maturity(the reciprocal of time to mass maturity) was a positive linearfunction of mean temperature, but was not affected by CO₂ concentration.The rate of increase in grain dry weight per ear was 2.0 mgd^–1 greater per 1 C rise, and was 8.0 mg d^–1 greaterat 684 compared with 380 µmol mol^–1 CO₂ at a giventemperature. The rate of increase in harvest index was 1.0%d^–1 in most plots at 380 µmol mol^–1 CO₂ andin open field plots, compared with 1.18% d^–1 in all plotsat 684 µmol mol^–1 CO₂. Thus, the increased rateof grain growth observed at an elevated CO₂ concentration couldbe attributed partly to a change in the partitioning of assimilatesto the grain. In contrast, the primary effect of warmer temperatureswas to shorten the duration of grain-filling. The rate of graingrowth at a given temperature and the rate of increase in harvestindex were only independent of the number of grains per earabove a critical grain number of 23–24 grains per ear({small tilde}20 000 grains m^–2). Key words: Winter wheat, grain growth, temperature, CO₂, harvest index, critical grain number     

The effect of temperature and CO2 on seed quality development in wheat (Triticum aestivum L.)

Winter wheat (Triticum aestivum L.) cv. Hereward was grown inthe field in two double-walled polyethylene-covered tunnelswithin each of which a temperature gradient was superimposedon diurnal and seasonal fluctuations in temperature. The meantemperature between anthesis and harvest maturity varied from14.3 to 18.4C among plots within these tunnels. The CO₂ concentrationwas controlled at different values in each tunnel; seasonalmean concentrations were 380 and 684 µmol CO₂ mol^–1air. Crops were also grown outside the tunnels at ambient temperaturesand CO₂. Samples of seeds were harvested sequentially from eachplot between anthesis and harvest maturity. Seed germinationand seed survival during subsequent air-dry storage were determinedfor each sample. The onset of both ability to germinate anddesiccation tolerance (ability to germinate after rapid desiccationto 10–15% moisture content and subsequent rehydration)coincided in all environments. Full germination capacity (>97%, determined at 10C) was reached 4–18 d before theend of the seed-filling phase (mass maturity) in most cases.There was little or no decline in germination capacity duringsubsequent seed development and maturation. Differences in seedquality were evident, however, throughout seed development andmaturation when seed survival curves during subsequent storagewere compared. Potential longevity in air-dry storage (assessedby the value K₁ of the seed viability equation) improved consistentlyboth before and after mass maturity. There was a significantpositive relation between the rate of increase in potentiallongevity (dK₁Idt) and temperature (the minimum temperaturefor seed quality development was 4.8 C), but neither CO₂ concentrationnor production within the polyethylene tunnels affected thisrelation. Key words: Wheat, Triticum aestivum L., seed development, seed longevity, carbon dioxide, temperature     

Multiple forms of tubulin in the cytoskeletal and flagellar microtubules of polytomella

The alga polytomella contains several organelles composed of microtubules, including four flagella and hundreds of cytoskeletal microtubules. Brown and co-workers have shown (1976. J. Cell Biol. 69:6-125; 1978, Exp. Cell Res. 117: 313-324) that the flagella could be removed and the cytoskeletans dissociated, and that both structures could partially regenerate in the absence of protein synthesis. Because of this, and because both the flagella and the cytoskeletons can be isolated intact, this organism is particularly suitable for studying tubulin heterogeneity and the incorporation of specific tubulins into different microtubule-containing organelles in the same cell. In order to define the different species of tubulin in polytonella cytoplasm, a (35)S- labeled cytoplasmic fraction was subjected to two cycles of assembly and disassembly in the presence of unlabeled brain tubulin. Comparison of the labeled polytomella cytoplasmic tubulin obtained by this procedure with the tubulin of isolated polytomella flagella by two-dimensional gel electrophoresis showed that, whereas the β-tubulin from both cytoplasmic and flagellar tubulin samples comigrated, the two α-tubulins had distinctly different isoelectic points. As a second method of isolating tubulin from the cytoplasm, cells were gently lysed with detergent and intact cytoskeletons obtained. When these cytoskeletons were exposed to cold temperature, the proteins that were released were found to be highly enriched in tubulin; this tubulin, by itself, could be assembled into microtubules in vitro. The predominant α-tubulin of this in vitro- assembled cytoskeletal tubulin corresponded to the major cytoplasmic α-tubulin obtained by coassembly of labeled polytomella cytoplasmic extract with brain tubulin and was quite distinct from the α-tubulin of purified flagella. These results clearly show that two different microtubule-containing organelles from the same cell are composed of distinct tubulins.     

Identification of restriction-fragment-length polymorphisms in genomic DNA of the lesser snow goose (Anser caerulescens caerulescens)

A genomic library of partially EcoRI-digested DNA from the lesser snow goose, Anser caerulescens caerulescens, was constructed in the phage vector Charon 4. Phage containing only unique sequences were identified by screening plaques with 32P-labeled genomic DNA. Restriction-fragment- length polymorphisms (RFLPs) were identified by probing DNA from 11-13 male birds from the breeding colony at La Perouse Bay. Of the 17 probes examined, all detected RFLPs with at least one of EcoRi, HindIII, Msp1, and Taq1. Several of them identified highly variable regions with multiple alleles. These RFLPs are valuable DNA markers that can be used for (1) the examination of DNA variation, relatedness, and genetic distance and (2) assessing paternity and maternity. These data suggest that there are higher levels of variation of DNA sequence in birds than had previously been thought to exist.      

Discovery of diphenyl amine based sodium channel blockers, effective against hNav1.2

The development of new therapies for chronic pain is an area of unmet medical need. Central to pathways of chronic pain is the upregulation of voltage-gated sodium channels. The use of tricyclic antidepressants, which also have sodium channel activity, in chronic pain therapy prompted us to develop novel compounds from this scaffold. Herein, we show that the tricyclic moiety is not needed for effective inhibition of the [³H]-BTX binding site and sodium currents of hNa_v1.2. Our lead compound 6, containing a diphenyl amine motif, demonstrated a 53% inhibitory block of Na_v1.2 currents at 10 μM, which is greater than 50% increase in current block in comparison to the amitriptyline standard. Altogether our study establishes that the tricyclic motif is unnecessary for hNa_v1.2 activity and modification of the amine portion is detrimental to sodium channel block.