Effects of low incubation temperatures on the bactericidal activity of anti-tuberculosis drugs

OBJECTIVES: to explore the effect of low incubation temperatures and the consequent slowing of bacterial metabolism on the bactericidal action of anti-tuberculosis drugs against Mycobacterium tuberculosis. METHODS: counting of surviving bacteria during exposure of static cultures to 1 mg/L isoniazid, 2 mg/L rifampicin, 0.5 or 2 mg/L TMC207 and 40 or 160 mg/L pyrazinamide, usually for periods of 21 days at temperatures of 37, 25, 22, 19, 16 or 8°C. RESULTS: the bactericidal activities of isoniazid and rifampicin were progressively reduced at 25 and 22°C, and were minimal at lower temperatures. TMC207 was immediately bactericidal at 37°C, in contrast to the early static phase reported with log phase cultures, and showed less change in activity as incubation temperatures were reduced than did rifampicin or isoniazid. Pyrazinamide was more bactericidal when incubation temperatures were reduced below 37°C and when the static seed cultures were most dormant. CONCLUSIONS: these results can be explained by the surmise that at low temperatures bacterial energy is at a low level with only just sufficient ATP to maintain homeostasis, making the bacteria more susceptible to the blocking of ATP synthesis by TMC207. Insufficient ATP at low temperature would also hinder the export of pyrazinoic acid, the toxic product of the pro-drug pyrazinamide, from the mycobacterial cell by an inefficient efflux pump that requires energ

Novel steady state of a microtubule assembly in a confined geometry

We study the steady state of an assembly of microtubules in a confined volume, analogous to the situation inside a cell where the cell boundary forms a natural barrier to growth. We show that the dynamical equations for growing and shrinking microtubules predict the existence of two steady states, with either exponentially decaying or exponentially increasing distribution of microtubule lengths. We identify the regimes in parameter space corresponding to these steady states. In the latter case, the apparent catastrophe frequency near the boundary was found to be significantly larger than that in the interior. Both the exponential distribution of lengths and the increase in the catastrophe frequency near the cell margin is in excellent agreement with recent experimental observations.Comment: 8 pages, submitted to Phys. Rev.

Sparse Graph Codes for Quantum Error-Correction

We present sparse graph codes appropriate for use in quantum error-correction. Quantum error-correcting codes based on sparse graphs are of interest for three reasons. First, the best codes currently known for classical channels are based on sparse graphs. Second, sparse graph codes keep the number of quantum interactions associated with the quantum error correction process small: a constant number per quantum bit, independent of the blocklength. Third, sparse graph codes often offer great flexibility with respect to blocklength and rate. We believe some of the codes we present are unsurpassed by previously published quantum error-correcting codes.Comment: Version 7.3e: 42 pages. Extended version, Feb 2004. A shortened version was resubmitted to IEEE Transactions on Information Theory Jan 20, 200

Colloid osmotic parameterization and measurement of subcellular crowding

© The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Mitchison, T. J. (2019). Colloid osmotic parameterization and measurement of subcellular crowding. Molecular Biology of the Cell, 30(2), (2019): 173-180, doi:10.1091/mbc.E18-09-0549.Crowding of the subcellular environment by macromolecules is thought to promote protein aggregation and phase separation. A challenge is how to parameterize the degree of crowding of the cell interior or artificial solutions that is relevant to these reactions. Here I review colloid osmotic pressure as a crowding metric. This pressure is generated by solutions of macromolecules in contact with pores that are permeable to water and ions but not macromolecules. It generates depletion forces that push macromolecules together in crowded solutions and thus promotes aggregation and phase separation. I discuss measurements of colloid osmotic pressure inside cells using the nucleus, the cytoplasmic gel, and fluorescence resonant energy transfer (FRET) biosensors as osmometers, which return a range of values from 1 to 20 kPa. I argue for a low value, 1–2 kPa, in frog eggs and perhaps more generally. This value is close to the linear range on concentration–pressure curves and is thus not crowded from an osmotic perspective. I discuss the implications of a low crowding pressure inside cells for phase separation biology, buffer design, and proteome evolution. I also discuss a pressure–tension model for nuclear shape, where colloid osmotic pressure generated by nuclear protein import inflates the nucleus.This article was prompted by lively discussions at the Marine Biological Laboratory (MBL) Physiology Course, Woods Hole, MA. I particularly thank Annie Pipathsouk (University of California, San Franscico) and Charlotte Strandkvist (Harvard Medical School) for experimental work in frog egg extract; James Pelletier (MIT), Tony Hyman (MPI Dresden), and Rob Phillips (Cal. Tech.) for discussions; and Nikon for microscopy support at MBL. T.J.M. is supported by National Institute of General Medical Sciences 39565

How many photons are needed to distinguish two transparencies?

We give a bound on the minimum number of photons that must be absorbed by any quantum protocol to distinguish between two transparencies. We show how a quantum Zeno method in which the angle of rotation is varied at each iteration can attain this bound in certain situations.Comment: 5 pages, 4 figure

Transport enhancement from incoherent coupling between one-dimensional quantum conductors

We study the non-equilibrium transport properties of a highly anisotropic two-dimensional lattice of spin-1/2 particles governed by a Heisenberg XXZ Hamiltonian. The anisotropy of the lattice allows us to approximate the system at finite temperature as an array of incoherently coupled one-dimensional chains. We show that in the regime of strong intrachain interactions, the weak interchain coupling considerably boosts spin transport in the driven system. Interestingly, we show that this enhancement increases with the length of the chains, which is related to superdiffusive spin transport. We describe the mechanism behind this effect, compare it to a similar phenomenon in single chains induced by dephasing, and explain why the former is much stronger

Meiotic Spindle: Sculpted by Severing

Katanin is a conserved AAA ATPase with the ability to sever microtubules, but its biological function in animal cells has been obscure. A recent study using electron tomography has found that katanin stimulates the production of microtubules in the meiotic spindles of Caenorhabditis elegans oocytes

Small-molecule and mutational analysis of allosteric Eg5 inhibition by monastrol

BACKGROUND: A recent crystal structure of monastrol in a ternary complex with the kinesin Eg5 motor domain highlights a novel, induced-fit drug binding site at atomic resolution. Mutational obliteration of the monastrol binding site results in a monastrol-resistant, but otherwise catalytically active Eg5 motor domain. However, considering the conformational changes at this site, it is unclear what specific interactions stabilize the interaction between monastrol and the Eg5 motor domain. RESULTS: To study the molecular complementarity of the monastrol-Eg5 interaction, we used a combination of synthetic chemistry and targeted mutations in Eg5 to measure the contribution of specific contacts to inhibition of Eg5 in vitro and in cultured cells. Structure-activity data on chemical derivatives, sequence analysis of Eg5 homologs from different species, and the effect of mutations near the drug binding site were consistent with the crystal structure. CONCLUSION: The mechanism of monastrol revealed by our data rationalizes its specificity for Eg5 over other kinesins and highlights a potential mechanism of drug resistance for anti-cancer therapy targeting this site in Eg5

Force and length in the mitotic spindle

Author Posting. © The Author(s), 2009. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Current Biology 19 (2009): R749-R761, doi:10.1016/j.cub.2009.07.028.The mitotic spindle assembles to a steady-state length at metaphase through the integrated action of molecular mechanisms that generate and respond to mechanical forces. While molecular mechanisms that produce force have been described, our understanding of how they integrate with each other, and with the assembly-disassembly mechanisms that regulate length, is poor. We review current understanding of the basic architecture and dynamics of the metaphase spindle, and some of the elementary force producing mechanisms. We then discuss models for force integration, and spindle length determination. We also emphasize key missing data that notably includes absolute values of forces, and how they vary as a function of position, within the spindle.S.D. received support from a Milton Fund (Harvard University) and T.J.M. was supported by NIH grants GM039565 and P50 GM068763

Mitotic spindle assembly by two different pathways in vitro

Abstract. We have used Xenopus egg extracts to study spindle morphogenesis in a cell-free system and have identified two pathways of spindle assembly in vitro using methods of fluorescent analogue cytochemistry. When demembranated sperm nuclei are added to egg extracts arrested in a mitotic state, individual nuclei direct the assembly of polarized microtubule arrays, which we term half-spindles; half-spindles then fuse pairwise to form bipolar spindles. In contrast, when sperm nuclei are added to extracts that are induced to enter interphase and arrested in the following mitosis, a single sperm nucleus can direct the assembly of a complete spindle. We find that microtubule arrays in vitro are strongly biased towards chromatin, but this does not depend on specific kinetochore-microtubul