Tubulin Polyglycylation: Differential Posttranslational Modification of Dynamic Cytoplasmic and Stable Axonemal Microtubules in Paramecium

Polyglycylation, a posttranslational modification of tubulin, was discovered in the highly stable axonemal microtubules of Paramecium cilia where it involves the lateral linkage of up to 34 glycine units per tubulin subunit. The observation of this type of posttranslational modification mainly in axonemes raises the question as to its relationship with axonemal organization and with microtubule stability. This led us to investigate the glycylation status of cytoplasmic microtubules that correspond to the dynamic microtubules in Paramecium. Two anti-glycylated tubulin monoclonal antibodies (mAbs), TAP 952 and AXO 49, are shown here to exhibit different affinities toward mono- and polyglycylated synthetic tubulin peptides. Using immunoblotting and mass spectrometry, we show that cytoplasmic tubulin is glycylated. In contrast to the highly glycylated axonemal tubulin, which is recognized by the two mAbs, cytoplasmic tubulin reacts exclusively with TAP 952, and the α- and β- tubulin subunits are modified by only 1–5 and 2–9 glycine units, respectively. Our analyses suggest that most of the cytoplasmic tubulin contains side chain lengths of 1 or 2 glycine units distributed on several glycylation sites. The subcellular partition of distinct polyglycylated tubulin isoforms between cytoplasmic and axonemal compartments implies the existence of regulatory mechanisms for glycylation. By following axonemal tubulin immunoreactivity with anti-glycylated tubulin mAbs upon incubation with a Paramecium cellular extract, the presence of a deglycylation enzyme is revealed in the cytoplasm of this organism. These observations establish that polyglycylation is reversible and indicate that, in vivo, an equilibrium between glycylating and deglycylating enzymes might be responsible for the length of the oligoglycine side chains of tubulin.

Nitric oxide partitioning into mitochondrial membranes and the control of respiration at cytochrome c oxidase

An emerging and important site of action for nitric oxide (NO) within cells is the mitochondrial inner membrane, where NO binds to and inhibits members of the electron transport chain, complex III and cytochrome c oxidase. Although it is known that inhibition of cytochrome c oxidase by NO is competitive with O2, the mechanisms that underlie this phenomenon remain unclear, and the impact of both NO and O2 partitioning into biological membranes has not been considered. These properties are particularly interesting because physiological O2 tensions can vary widely, with NO having a greater inhibitory effect at low O2 tensions (<20 μM). In this study, we present evidence for a consumption of NO in mitochondrial membranes in the absence of substrate, in a nonsaturable process that is O2 dependent. This consumption modulates inhibition of cytochrome c oxidase by NO and is enhanced by the addition of exogenous membranes. From these data, it is evident that the partition of NO into mitochondrial membranes has a major impact on the ability of NO to control mitochondrial respiration. The implications of this conclusion are discussed in the context of mitochondrial lipid:protein ratios and the importance of NO as a regulator of respiration in pathophysiology.

Geographical structuring in the mtDNA of Italians.

Geographical patterns of mtDNA variation were studied in 12 Italian samples (1072 individuals) by two different spatial autocorrelation methods. Separate analyses of the frequencies of 12 restriction morphs show North-South clines, differences between Sardinia and the mainland populations, and the effects of isolation by distance. A recently developed autocorrelation statistic summarizing molecular similarity at all sites (AIDA; autocorrelation index for DNA analysis) confirms the presence of a clinical pattern; differences between random pairs of haplotypes tend to increase with their geographical distance. The partition of gene diversity, however, reveals that most variability occurs within populations, whereas differences between populations are minor (GST = 0.057). When the data from the 12 samples are pooled, two descriptors of genetic variability (number of polymorphic sites and average sequence difference between pairs of individuals) do not behave as expected under neutrality. The presence of clinal patterns, Tajima's tests, and a simulation experiment agree in suggesting that population sizes increased rapidly in Italy and Sicily but not necessarily so in Sardinia. The distribution of pairwise sequence differences in the Italian peninsula (excluding Sardinia) permits a tentative location of the demographic increase between 8000 and 20,500 years ago. These dates are consistent with archaeological estimates of two distinct expansion processes, occurring, respectively, in the Neolithic and after the last glacial maximum in the Paleolithic. Conversely, there is no genetic evidence that such processes have had a major impact on the Sardinian population.

Grouping genetic operators for the delineation of functional areas based on spatial interaction

The delineation of functional economic areas, or market areas, is a problem of high practical relevance, since the delineation of functional sets such as economic areas in the US, Travel-to-Work Areas in the United Kingdom, and their counterparts in other OECD countries are the basis of many statistical operations and policy making decisions at local level. This is a combinatorial optimisation problem defined as the partition of a given set of indivisible spatial units (covering a territory) into regions characterised by being (a) self-contained and (b) cohesive, in terms of spatial interaction data (flows, relationships). Usually, each region must reach a minimum size and self-containment level, and must be continuous. Although these optimisation problems have been typically solved through greedy methods, a recent strand of the literature in this field has been concerned with the use of evolutionary algorithms with ad hoc operators. Although these algorithms have proved to be successful in improving the results of some of the more widely applied official procedures, they are so time consuming that cannot be applied directly to solve real-world problems. In this paper we propose a new set of group-based mutation operators, featuring general operations over disjoint groups, tailored to ensure that all the constraints are respected during the operation to improve efficiency. A comparative analysis of our results with those from previous approaches shows that the proposed algorithm systematically improves them in terms of both quality and processing time, something of crucial relevance since it allows dealing with most large, real-world problems in reasonable time.

The Critical Strips of the Sums 1 + 2z + ... + nz

We give a partition of the critical strip, associated with each partial sum 1 + 2z + ... + nz of the Riemann zeta function for Re z < −1, formed by infinitely many rectangles for which a formula allows us to count the number of its zeros inside each of them with an error, at most, of two zeros. A generalization of this formula is also given to a large class of almost-periodic functions with bounded spectrum.

Europe’s reactive and protective muddling through: the results of a summit in fire fighting mode. EPC Post-Summit Analysis, 19 October 2015

The EU’s October summit was dominated by one issue; the migration and refugee crisis, with EU leaders intent on putting on a public display of unity after weeks of bitter arguments, and concentrating on fire-fighting and immediate measures to tackle the most pressing reasons for, and impacts of, the crisis. Longer-term measures to address some of the root causes of increased migratory flows, support for the integration of newly arrived refugees or the introduction of new channels of legal migration, were not discussed. The Summit also spent little time on two issues that had originally been expected to be a key part of the agenda: the forthcoming British referendum on EU membership, where irritation with the slow pace of talks and British vagueness about its demands were in evidence; and the governance of Economic and Monetary Union (EMU), where EU leaders missed another opportunity for a thorough debate about future perspectives on the basis of the ‘Five Presidents’ Report’.

Optimization of the Modular Educational Program Structure

Optimized structure of the educational program consisting of a set of the interconnected educational objects is offered by means of problem solution of optimum partition of the acyclic weighed graph. The condition of acyclicity preservation for subgraphs is formulated and the quantitative assessment of decision options is executed. The original algorithm of search of quasioptimum partition using the genetic algorithm scheme with coding chromosomes by permutation is offered. Object-oriented realization of algorithm in language C++ is described and results of numerical experiments are presented.

Posthumous works of Frederic II, King of Prussia.

"Translated from the French by Thomas Holcroft."

Comparison of grinding media - Cylpebs versus balls

Cylpebs are slightly tapered cylindrical grinding media with a ratio of length to diameter of unity. The manufactures have made conflicting claims regarding the milling performance of Cylpebs in comparison with balls. One major point of interest is which one grinds finer at the same operating conditions. The difficulty in comparison is due to the shape difference. The two grinding media have different surface area, bulk density and contact mechanisms in grinding action. Comparative tests were conducted using the two types of grinding media in a laboratory Bond ball mill at various conditions of equality such as media mass, size distribution, surface area and input specific energy. The laboratory results indicate that at the same specific energy input level the Cylpebs produce a product with slightly less oversize due to their greater surface area, but essentially the same sizing at the fine end as that produced with the balls. The reason may be that the advantage of greater surface area is balanced by the line contact and area contact grinding actions with the Cylpebs. A new ball mill scale-up procedure [Man, Y.T., 2001. Model-based procedure for scale-up of wet, overflow ball mills, Part 1: outline of the methodology. Minerals Engineering 14 (10), 1237-1246] was employed to predict grinding performance of an industrial mill from the laboratory test results. The predicted full scale operation was compared with the plant survey data. Some problems in the original scale-up procedures were identified. The scale-up procedure was therefore modified to allow the predicted ball mill performance to match the observed one. The calibrated scale-up procedure was used to predict the Cylpebs performance in the full scale industrial mill using the laboratory tests results. (C) 2004 Elsevier Ltd. All rights reserved.

Extending the theory of Owicki and Gries with a logic of progress

This paper describes a logic of progress for concurrent programs. The logic is based on that of UNITY, molded to fit a sequential programming model. Integration of the two is achieved by using auxiliary variables in a systematic way that incorporates program counters into the program text. The rules for progress in UNITY are then modified to suit this new system. This modification is however subtle enough to allow the theory of Owicki and Gries to be used without change.

Typical kernel size and number of sparse random matrices over Galois fields: a statistical physics approach

Using methods of statistical physics, we study the average number and kernel size of general sparse random matrices over GF(q), with a given connectivity profile, in the thermodynamical limit of large matrices. We introduce a mapping of GF(q) matrices onto spin systems using the representation of the cyclic group of order q as the q-th complex roots of unity. This representation facilitates the derivation of the average kernel size of random matrices using the replica approach, under the replica symmetric ansatz, resulting in saddle point equations for general connectivity distributions. Numerical solutions are then obtained for particular cases by population dynamics. Similar techniques also allow us to obtain an expression for the exact and average number of random matrices for any general connectivity profile. We present numerical results for particular distributions.

The percutaneous absorption of ionisable compounds

The effects of ionisation on transdermal drug delivery using excised human epidermis (HS) and silastic rubber (SR) as model permeation barriers were investigated in vitro using Franz-type absorption cells. Suspensions and solutions of salicylic acid (SA), the model ionogenic permeant, were used as donors and the variables studied were vehicle pH and trans-membrane pH-gradients. For solutions, the pH effect was related to the level of ionisation of the drug and the degree of saturation of the solution. With suspensions, the observed permeation rate was unaffected by pH. The penetration profiles through HS and SR were similar, although the overall flux through HS was about 70% of that observed through SR. Pretreatment of the membranes with various enhancer regimens, including oleic acid, Azone and N, N-dimethylamides in propylene glycol (PG) and isopropyl myristate (IPM) promoted the penetration of SA. SR was not a suitable model for enhancer pretreatment using IPM as a vehicle as the membrane was significantly disrupted by this vehicle. The results from comparable experiments with and without a trans-membrane pH-gradient did not have a significant effect upon flux or flux enhancement after pretreatment with the above enhancers. A theoretical model for the extraction coefficients of weak acids was derived using the partition coefficients of the ionised and unionised species, pH and pKa. This model was shown to account for the variation in overall partition of salicylic acid dependent upon pH and pKa. This model was shown to account for the variation in overall partition of salicylic acid dependent upon pH and pKa. The distribution of this solute between aqueous and oily phases, with and without added enhancer, was measured as a function of pH. The extraction coefficients determined were consistent with the model and showed that the behaviour of the system can be explained without referral to ion-pair mechanisms. Phosphonoacetate is an effective antiviral agent. However, as it is charged at physiological pH, its permeation across cell membranes is limited. To assess the improvement of the transport properties of this molecule, mono-, di- and tri-ester prodrugs were examined. These were assessed for stability and subsequent breakdown with respect to pH by HPLC. In vitro percutaneous absorption was observed using the triester, but not the ionic mono- or di-esters. The triester absorption could be potentiated using a range of enhancers with oleic acid being the most effective. Cyclodextrins (CD) have a role as absorption enhancers for peptide compounds across nasal epithelium. One potential mode of action is that CDs include these compounds, protect them from enzymic attack and thereby increase their residence time in the nasal epithelium. This study investigated the potential of CDs to protect ester prodrugs from enzymatic breakdown and prevent production of poorly transportable ionic species. Using a range of CD to ester molar ratios (10:1 to 2500:1) a small, but measurable, protection for the model esters (parabens) against esterase attack was observed. Possible mechanisms for this phenomenon are that CDs include the ester, making it unavailable for hydrolysis, the CDs may also affect the esterase in some way preventing access for the ester into the active site.

Superfocusing of high-M2 semiconductor laser beams:experimental demonstration

The focusing of multimode laser diode beams is probably the most significant problem that hinders the expansion of the high-power semiconductor lasers in many spatially-demanding applications. Generally, the 'quality' of laser beams is characterized by so-called 'beam propagation parameter' M2, which is defined as the ratio of the divergence of the laser beam to that of a diffraction-limited counterpart. Therefore, M2 determines the ratio of the beam focal-spot size to that of the 'ideal' Gaussian beam focused by the same optical system. Typically, M2 takes the value of 20-50 for high-power broad-stripe laser diodes thus making the focal-spot 1-2 orders of magnitude larger than the diffraction limit. The idea of 'superfocusing' for high-M2 beams relies on a technique developed for the generation of Bessel beams from laser diodes using a cone-shaped lens (axicon). With traditional focusing of multimode radiation, different curvatures of the wavefronts of the various constituent modes lead to a shift of their focal points along the optical axis that in turn implies larger focal-spot sizes with correspondingly increased values of M2. In contrast, the generation of a Bessel-type beam with an axicon relies on 'self-interference' of each mode thus eliminating the underlying reason for an increase in the focal-spot size. For an experimental demonstration of the proposed technique, we used a fiber-coupled laser diode with M2 below 20 and an emission wavelength in ~1μm range. Utilization of the axicons with apex angle of 140deg, made by direct laser writing on a fiber tip, enabled the demonstration of an order of magnitude decrease of the focal-spot size compared to that achievable using an 'ideal' lens of unity numerical aperture. © 2014 SPIE.

Large Distinct Part Sizes in a Random Integer Partition

A partition of a positive integer n is a way of writing it as the sum of positive integers without regard to order; the summands are called parts. The number of partitions of n, usually denoted by p(n), is determined asymptotically by the famous partition formula of Hardy and Ramanujan [5]. We shall introduce the uniform probability measure P on the set of all partitions of n assuming that the probability 1/p(n) is assigned to each n-partition. The symbols E and V ar will be further used to denote the expectation and variance with respect to the measure P . Thus, each conceivable numerical characteristic of the parts in a partition can be regarded as a random variable.

The Multidirectional Memory of Charlie Hebdo

This article will discuss notions and concepts of remembering in the aftermath of the Charlie Hebdo attacks. Much has been written about the immediate response to the attacks, both commending the collective spirit of unity that defined the ‘marche républicaine’ of 11 January 2015, and criticising the alleged hypocrisy and cynicism of, most notably, the political figures that took to the streets that day, hand in hand. I will consider a selection of the memory practices that have emerged since then, notably on the anniversary of the event. This demonstration of memory provides key insights into the form and manner of remembering within a particular cultural group, but also reflects how the present moment is integral to our understanding of memory. The purpose of this article is to consider how official and non-official remembering of Charlie Hebdo can intertwine as well as pull in separate directions. A focus on the politics, the language, the aesthetics and the geography of commemorative activities in this article will enable an appreciation of the multidirectional character of remembering Charlie Hebdo.