Calcineurin activity is required for the completion of cytokinesis

Successful completion of cytokinesis requires the spatio-temporal regulation of protein phosphorylation and the coordinated activity of protein kinases and phosphatases. Many mitotic protein kinases are well characterized while mitotic phosphatases are largely unknown. Here, we show that the Ca²⁺- and calmodulin-dependent phosphatase, calcineurin (CaN), is required for cytokinesis in mammalian cells, functioning specifically at the abscission stage. CaN inhibitors induce multinucleation in HeLa cells and prolong the time cells spend connected via an extended intracellular bridge. Upon Ca²⁺ influx during cytokinesis, CaN is activated, targeting a set of proteins for dephosphorylation, including dynamin II (dynII). At the intracellular bridge, phospho-dynII and CaN are co-localized to dual flanking midbody rings (FMRs) that reside on either side of the central midbody ring. CaN activity and disassembly of the FMRs coincide with abscission. Thus, CaN activity at the midbody plays a key role in regulating the completion of cytokinesis in mammalian cells.     

Induction of uncoiled chromosomes by vibration

Summary Chromatin condensation during metaphase can be removed by simple vibration of metaphase cells prior to fixation. Uncoiled chromosome arms consist of long threads with dense regions at irregular distances each from the other.Acknowledgment. This work is supported by the Hellenic Anticancer Institute.     

Routes and machinery of primary cilium biogenesis

Primary cilia are solitary, microtubule-based protrusions of the cell surface that play fundamental roles as photosensors, mechanosensors and biochemical sensors. Primary cilia dysfunction results in a long list of developmental and degenerative disorders that combine to give rise to a large spectrum of human diseases affecting almost any major body organ. Depending on the cell type, primary ciliogenesis is initiated intracellularly, as in fibroblasts, or at the cell surface, as in renal polarized epithelial cells. In this review, we have focused on the routes of primary ciliogenesis placing particular emphasis on the recently described pathway in renal polarized epithelial cells by which the midbody remnant resulting from a previous cell division event enables the centrosome for initiation of primary cilium assembly. The protein machinery implicated in primary cilium formation in epithelial cells, including the machinery best known for its involvement in establishing cell polarity and polarized membrane trafficking, is also discussed.     

Induction of uncoiled chromosomes by vibration.

Chromatin condensation during metaphase can be removed by simple vibration of metaphase cells prior to fixation. Uncoiled chromosome arms consist of long threads with dense regions at irregular distances each from the other.     

The role of motile aeromonads in the fish disease, ulcerative disease syndrome (UDS)

Ulcerative Disease Syndrome (UDS) is an epizootic fish disease characterized by the presence of severe, open dermal ulcers on the head, midbody, and dorsal regions of the fish. Aeromonas hydrophila and A. sobria were recovered more often from UDS fish than other bacteria from the genera Vibrio, Alteromonas and Plesiomonas. Representative isolates of A. hydrophila, A. sobria, V. anguillarum, V. vulnificus, Alteromonas putrefaciens, and P. shigelloides taken from UDS and healthy fish were assayed for virulence-associated factors. The aeromonads produced a wide variety of hydrolytic enzymes and expressed cell surface characteristics linked to virulence whereas the other bacterial species rarely produced the same enzymes or cell surface characteristics. The role of aeromonads in UDS is believed to be opportunistic or secondary and these bacteria are thought to play an important role in this degenerative disease.     

Syndecan-4 promotes cytokinesis in a phosphorylation-dependent manner

During mitosis, cells detach, and the cell–matrix interactions become restricted. At the completion of cytokinesis, the two daughter cells are still connected transiently by an intercellular bridge (ICB), which is subjected to abscission, as the terminal step of cytokinesis. Cell adhesion to the matrix is mediated by syndecan-4 (SDC4) transmembrane heparan sulfate proteoglycan. Our present work demonstrated that SDC4 promotes cytokinesis in a phosphorylation-dependent manner in MCF-7 breast adenocarcinoma cells. The serine179-phosphorylation and the ectodomain shedding of SDC4 changed periodically in a cell cycle-dependent way reaching the maximum at G2/M phases. On the contrary, the phospho-resistant Ser179Ala mutant abrogated the shedding. The phosphorylated full-length and shed remnants enriched along the mitotic spindles, and subsequently in the ICBs, however, proper membrane insertion was necessary for midbody localization. Expression of phosphomimicking Ser179Glu SDC4 resulted in incomplete abscission, whereas expression of the phospho-resistant SDC4 led to giant, multinucleated cells.     

The relative facts interpretation and Everett's note added in proof

In the published version of Hugh Everett III's doctoral dissertation, he inserted what has become a famous footnote, the “note added in proof”. This footnote is often the strongest evidence given for any of various interpretations of Everett (the many worlds, many minds, many histories and many threads interpretations). In this paper I will propose a new interpretation of the footnote. One that is supported by evidence found in letters written to and by Everett; one that is suggested by a new interpretation of Everett, an interpretation that takes seriously the central position of relative states in Everett's pure wave mechanics: the relative facts interpretation. Of central interest in this paper is how to make sense of Everett's claim in the “note added in proof” that “all elements of a superposition (all “branches”) are “actual,” none any more “real” than the rest.”     

TTDN1 is a Plk1-interacting protein involved in maintenance of cell cycle integrity

Polo-like kinase 1 (Plk1) is a highly conserved serine/threonine kinase that plays critical roles in many cell cycle events, especially in mitosis. In the present study, we identified TTDN1 as a potential interacting partner of Plk1 in yeast two-hybrid screens. Sequence analysis indicates that TTDN1 contains a consensus Plk1-binding motif at its C terminus. TTDN1 colocalizes with Plk1 at the centrosome in mitosis and the midbody during cytokinesis. TTDN1 is phosphorylated by Cdk1 in mitosis, and this is required for its interaction with Plk1. Site-directed mutagenesis indicates that TTDN1 is phosphorylated at multiple residues, including Ser93 and Ser104. Mutation of Thr120 of TTDN1 abolishes its interaction with Plk1, suggesting phosphorylation of Thr120 in the consensus Plk1-binding motif is required for its interaction with Plk1. Overexpression of TTDN1 or its knockdown by siRNA causes multi-polar spindles and multiple nuclei, suggesting that TTDN1 plays a role in regulating mitosis and cytokinesis. Received 27 November 2006; received after revision 4 January 2007; accepted 25 January 2007 Y. Zhang, Y. Tian: These authors contribute equally to this work.     

Localization of sound producing animals using the arrival time differences of their signals at an array of microphones

Summary This method provides a reliable means to determine distance between vocalizing individuals and their locations through a prolonged period of time without disturbance to the communication process by the presence of an observer. The accuracy of this method was tested by varying the arrival time differences on the microphone grid for known locations in a random fashion. This test shows that a vocalizing animal within an area of 4 ha surrounding the microphone array can be localized with an accuracy of ±1 m. The accuracy decreases with the distance from the central area and as a function of the geometry of the array. The location of an individual can be determined based on 5 vocalizations given in sequence.Acknowledgment. This research was supported by funds granted to Irene Magyar from the Chapman Fund, The American Museum of Natural History and Sigma Xi, Grants in Aid of Research.     

Numerical simulations of self-propelled swimming of 3D bionic fish school

Numerical simulations of self-propelled swimming of a three dimensional bionic fish and fish school in a viscous fluid are carried out.This is done with the assistance of a parallel software package produced for 3D moving boundary problems.This computational fluid dynamics package combines the adaptive multi-grid finite volume method,the immersed boundary method and VOF (volume of fluid) method.By using the package results of the self-propelled swimming of a 3D bionic fish and fish school in a viscous fluid...     

Effect of parathyroidectomy on the fasting-refeeding response in the rat colon

Following fasting and refeeding, the colonic epithelium of the rat exhibits a marked hyperplasia. This response is of a similar magnitude but of a longer duration to that observed in mice. This response is not affected by reducing serum calcium levels to those reported to alter normal tissue proliferation in vivo.     

Problems in the determination of isoelectric points of beta lactamases]

Analytical isoelectric focusing in polyacrylamide gels and preparative electrofocusing in density gradients may give very different pI determination for the same beta lactamase. This difference seems due to a lack of enzyme migration in some parts of the cross-linked polyacrylamide gel. This migration appears easier in density gradient isoelectricfocusing, and thus this technique yields more significant absolute pI values. Analytical isoelectric focusing should only be used for comparison studies.     

Evolution of the genetic code, protein synthesis and nucleic acid replication

A better definition of the structural and thermodynamic determinants of the interaction of nucleic acids with proteins is shedding light on the origin of the genetic code, protein synthesis, and nucleic acid replication. This is also allowing to show a consistent biochemical framework for the appearance of these fundamental synthetic mechanisms. This article reviews recent significant developments in the field, and discusses an integrated model for a biochemically plausible evolution of these fundamental mechanisms of synthesis. This model is based on sequence-specific interactions between abiotically synthesized polynucleotides and polypeptides, and can account for a coordinate evolution of the genetic code, protein synthesis, and nucleic acid replication in living cells.     

Papaverine enhances the negative inotropic effect of acetylcholine in rat auricles

Summary The negative inotropic effect of acetylcholine in rat left auricles is enhanced in the presence of the phosphodiesterase inhibitor papaverine. This result favours the idea of a cyclic GMP-mediated action of acetylcholine in the heart.I thank Lydia Paragnik for help. This research was supported by a grant from the Deutsche Forschungsgemeinschaft.     

A technical improvement in the radioimmunoassay of cyclic-AMP.

An improvement in the technique for the radioimmunoassay of cyclic-AMP, wherein ammonium sulfate precipitation is replaced with zirconyl phosphate gel, is presented. This substitution produces a more stable pellet than that obtained with ammonium sulfate. This greatly reduces a potential source of error due to pellet instability.     

Scientific patronage in the age of Darwin: The curious case of William Boyd Dawkins

This essay examines the curious relationship between Charles Darwin and the palaeontologist William Boyd Dawkins (1837–1929). Dawkins was a beneficiary of Darwin's patronage and styled himself as a Darwinian to Darwin and the public, yet viciously attacked Darwin and his theory in anonymous reviews. This has confused historians who have misunderstood the exact nature of Dawkins's attitude towards evolution and his relationship to Darwin. The present study explains both the reasons for Dawkins's contradictory statements and his relationship with Darwin. I introduce Batesian mimicry as a conceptual framework to make sense of Dawkins's actions, suggesting that Dawkins mimicked a Darwinian persona in order to secure advancement in the world of Victorian science. Dawkins's pro-Darwinian stance, therefore, was a façade, an act of mimicry. I argue that Dawkins exploited Darwin for his patronage – which took the form of advice, support from Darwin's well-placed friends, and monetary assistance – while safely expressing his dissent from Darwinian orthodoxy in the form of anonymous reviews. This is, therefore, a case study in how scientific authority and power could be gained and maintained in Victorian science by professing allegiance to Darwin and Darwinism.     

HCG-insensitivity of the postnatal rat ovary is due to the lack of a specific receptor

Summary Rat ovary is insensitive to luteinizing hormone and human chorionic gonadotrophin till to the 8th to 10th day of postnatal development. This insensitivity is due to the lack of a specific hormone receptor.This work was supported in part by the Deutsche Forschungsgemeinschaft (SFB 46). Purified HCG was a gift of Schering AG, Berlin-West. We thank Dr F. Peters for performing the iodination of HCG.     

Effect of insulin on lateral diffusion of pyrene in rat liver plasma membrane

Summary The yield of excimer formation by pyrene molecules inserted in rat liver plasma membranes is sensibly decreased in the presence of 1 nM insulin. This effect can be interpreted as indicating a decrease of the value of the translational diffusion coefficient of the dye within the membrane.This investigation was partially supported by Consiglio Nazionale delle Ricerche, Rome, Italy.     

Specific inhibition of a calcium dependent activation of brain cyclic AMP phosphodiesterase activity by vinblastine.

Vinblastine selectively inhibits the activation of brain cyclic AMP phosphodiesterase activity by Ca++-protein activator (50% inhibition by 2 x 10(-5) M). This inhibitory effect was reversed by excessive amounts of the activator, whereas large quantities of Ca++ caused only a slight suppression of the vinblastine effect. This result of vinblastine suggests a new site of its action and also suggests the possible role of protein activator, phosphodiesterase proteins or cyclic nucleotides in the previously known effects of vinblastine in vivo and in vitro.     

Intraluminal bile salt increases rate of firing in afferent fibers from the small intestine of the rat

Summary Perfusion of a rat intestinal segment with a solution containing sodium deoxycholate (8 mM) increases the rate of firing in periarterial afferent nerves from the gut. This observation indirectly supports our earlier proposal that bile salt evokes a net fluid secretion in the small intestine via an activation of the enteric nervous system.This study was supported by grants from the Swedish Medical Research Council (2855), The Medical Council of Swedish Life Insurance Companies and the Faculty of Medicine, University of Göteborg.