Slow cycling of unphosphorylated myosin is inhibited by calponin, thus keeping smooth muscle relaxed

A key unanswered question in smooth muscle biology is whether phosphorylation of the myosin regulatory light chain (RLC) is sufficient for regulation of contraction, or if thin-filament-based regulatory systems also contribute to this process. To address this issue, the endogenous RLC was extracted from single smooth muscle cells and replaced with either a thiophosphorylated RLC or a mutant RLC (T18A/S19A) that cannot be phosphorylated by myosin light chain kinase. The actin-binding protein calponin was also extracted. Following photolysis of caged ATP, cells without calponin that contained a nonphosphorylatable RLC shortened at 30% of the velocity and produced 65% of the isometric force of cells reconstituted with the thiophosphorylated RLC. The contraction of cells reconstituted with nonphosphorylatable RLC was, however, specifically suppressed in cells that contained calponin. These results indicate that calponin is required to maintain cells in a relaxed state, and that in the absence of this inhibition, dephosphorylated cross-bridges can slowly cycle and generate force. These findings thus provide a possible framework for understanding the development of latch contraction, a widely studied but poorly understood feature of smooth muscle.

Molecular Cloning and Characterization of a Radial Spoke Head Protein of Sea Urchin Sperm Axonemes: Involvement of the Protein in the Regulation of Sperm Motility

Monoclonal antibodies raised against axonemal proteins of sea urchin spermatozoa have been used to study regulatory mechanisms involved in flagellar motility. Here, we report that one of these antibodies, monoclonal antibody D-316, has an unusual perturbating effect on the motility of sea urchin sperm models; it does not affect the beat frequency, the amplitude of beating or the percentage of motile sperm models, but instead promotes a marked transformation of the flagellar beating pattern which changes from a two-dimensional to a three-dimensional type of movement. On immunoblots of axonemal proteins separated by SDS-PAGE, D-316 recognized a single polypeptide of 90 kDa. This protein was purified following its extraction by exposure of axonemes to a brief heat treatment at 40°C. The protein copurified and coimmunoprecipitated with proteins of 43 and 34 kDa, suggesting that it exists as a complex in its native form. Using D-316 as a probe, a full-length cDNA clone encoding the 90-kDa protein was obtained from a sea urchin cDNA library. The sequence predicts a highly acidic (pI = 4.0) protein of 552 amino acids with a mass of 62,720 Da (p63). Comparison with protein sequences in databases indicated that the protein is related to radial spoke proteins 4 and 6 (RSP4 and RSP6) of Chlamydomonas reinhardtii, which share 37% and 25% similarity, respectively, with p63. However, the sea urchin protein possesses structural features distinct from RSP4 and RSP6, such as the presence of three major acidic stretches which contains 25, 17, and 12 aspartate and glutamate residues of 34-, 22-, and 14-amino acid long stretches, respectively, that are predicted to form α-helical coiled-coil secondary structures. These results suggest a major role for p63 in the maintenance of a planar form of sperm flagellar beating and provide new tools to study the function of radial spoke heads in more evolved species.

Eine Messe des Lebens : für Soli Chor und grosses Orchester, nach Nietzsches Zarathustra /

"In die Universal Edition aufgenommen."

Also sprach Zarathustra : Tondichtung, frei nach Friedr. Nietzsche, für grosses Orchester, Op. 30 /

Originally for orchestra; arr. for 2 pianos, 4 hands.

Thus to revisit; some reminiscences of Ford Madox Hueffer.

pt. I. Before moonrise.--pt. II. Prosateurs: Credentials. Puiser dans le vide. The lordly treasure-house. Mots justes. Combien je regrette. Coda. W. H. Hudson and the simple word. Mr. Joseph Conrad and Anglo-Saxondom. Henry James, Stephen Crane and the main stream.--pt. III. The battle of the poets: "Thoughts before battle." Mr. Pound, Mr. Flint, some imagistes or cubistes, and the poetic vernacular. A descriptive interlude. "Vers libre." Second coda. Index.

I. An exposition of the symbolic terms of the second part of Faust. II. How this part thus proves itself to be a dramatic treatment of the modern history of Germany worthy of the genius of Goethe and the life he devoted to the task. (The prologues and first act completely explained; the other acts generally).

Mode of access: Internet.

Also sprach Zarathustra; ein buch für alle und keinen.

Mode of access: Internet.

Speech of Mr. Holmes, of Maine, in the Senate of the United States, on his resolutions calling upon the President of the United States for the reasons of his removing from office and filling the vacancies thus created, in the recess of the Senate.

Mode of access: Internet.

A history of the fight at Concord, on the 19th of April, 1775, with a particular account of the military operations and interesting events of that ever memorable day; showing that then and there the first regular and forcible resistance was made to the British soldiery, and the first British blood was shed by armed Americans, and the Revolutionary War thus commenced.

Mode of access: Internet.

Sixth annual report of the Commissioner of lands and immigration of the state of Florida, for the year ending December 31, 1874, together with reports from the several counties ... descriptive of the soil, products and general characteristics, thus forming a complete manual of information for immigrants.

Cover-title reads The Florida settler; or, Immigrants' guide 2d ed.

Also sprach Zarathustra : ein Büch für Alle und Keinen. /

Advertisements on last 12 unnumbered pages.

Four who spoke out: Burke, Fox, Sheridan, Pitt,

Bibliography: p. 184-196.

Zarathustra's evolutionary epistemology

Friedrich Nietzsche was the first great philosopher to be influenced at the core by Darwinian ideas. He regarded Also Sprach Zarathustra as his masterpiece and most subsequent commentators have agreed. There have been many interpretations of the Zarathustra, and like all great works it has many levels of meaning. An exposition in terms of evolutionary epistemology, however, has not yet been attempted. This article rectifies this omission and shows how Nietzsche's work carries Darwinian ideas into the domain of philosophical anthropology. It shows through the prism of Nietzsche's mature thought some of the consequences of an evolutionary epistemology both in opening up alternative visions of the world and in permitting a profound criticism of our commonsense metaphysics and ontology. © 1992.

An examination of the development of LTL hub-and-spoke freight distribution systems

Over recent years, hub-and-spoke distribution techniques have attracted widespread research attention. Despite there being a growing body of literature in this area there is less focus on the spoke-terminal element of the hub-and-spoke system as being a key component in the overall service received by the end-user. Current literature is highly geared towards discussing bulk optimization of freight units rather than to the more discrete and individualistic profile characteristics of shared-user Less-than-truckload (LTL) freight. In this paper, a literature review is presented to review the role hub-and-spoke systems play in meeting multi-profile customer demands, particularly in developing sectors with more sophisticated needs, such as retail. The paper also looks at the use of simulation technology as a suitable tool for analyzing spoke-terminal operations within developing hub-and spoke systems.

Improving “last mile” delivery performance to retailers in hub and spoke distribution systems

Purpose – The purpose of this paper is to investigate the “last mile” delivery link between a hub and spoke distribution system and its customers. The proportion of retail, as opposed to non-retail (trade) customers using this type of distribution system has been growing in the UK. The paper shows the applicability of simulation to demonstrate changes in overall delivery policy to these customers. Design/methodology/approach – A case-based research method was chosen with the aim to provide an exemplar of practice and test the proposition that simulation can be used as a tool to investigate changes in delivery policy. Findings – The results indicate the potential improvement in delivery performance, specifically in meeting timed delivery performance, that could be made by having separate retail and non-retail delivery runs from the spoke terminal to the customer. Research limitations/implications – The simulation study does not attempt to generate a vehicle routing schedule but demonstrates the effects of a change on delivery performance when comparing delivery policies. Practical implications – Scheduling and spreadsheet software are widely used and provide useful assistance in the design of delivery runs and the allocation of staff to those delivery runs. This paper demonstrates to managers the usefulness of investigating the efficacy of current design rules and presents simulation as a suitable tool for this analysis. Originality/value – A simulation model is used in a novel application to test a change in delivery policy in response to a changing delivery profile of increased retail deliveries.