Rat kinesin light chain 3 associates with spermatid mitochondria

AbstractWe recently discovered that in rat spermatids, kinesin light chain KLC3 can associate with outer dense fibers, major sperm tail components, and accumulates in the sperm midpiece. Here, we show that mitochondria isolated from rat-elongating spermatids have bound KLC3. Immunoelectron microscopy indicates that the association of KLC3 with mitochondria coincides with the stage in spermatogenesis when mitochondria move from the plasma membrane to the developing midpiece. KLC3 is able to bind in vitro to mitochondria from spermatids as well as somatic cells employing a conserved kinesin light chain motif, the tetratrico-peptide repeats. Expression of KLC3 in fibroblasts results in formation of large KLC3 clusters close to the nucleus, which also contain mitochondria: no other organelles were present in these clusters. Mitochondria are not present in KLC3 clusters after deletion of KLC3's tetratrico-peptide repeats. Our results indicate that the rat spermatid kinesin light chain KLC3 can associate with mitochondria

Interruption of visually perceived forward motion in depth evokes a cortical activation shift from spatial to intentional motor regions

Forward locomotion generates a radially expanding flow of visual motion which supports goal-directed walking. In stationary mode, wide-field visual presentation of optic flow stimuli evokes the illusion of forward self-motion. These effects illustrate an intimate relation between visual and motor processing. In the present fMRI study, we applied optic flow to identify distinct interfaces between circuitries implicated in vision and movement. The dorsal premotor cortex (PMd) was expected to contribute to wide-field forward motion flow (FFw), reflecting a pathway for externally triggered motor control. Medial prefrontal activation was expected to follow interrupted optic flow urging internally generated action. Data of 15 healthy subjects were analyzed with Statistical Parametric Mapping and confirmed this hypothesis. Right PMd activation was seen in FFw, together with activations of posterior parietal cortex, ventral V5, and the right fusiform gyms. Conjunction analysis of the transition from wide to narrow forward flow and reversed wide-field flow revealed selective dorsal medial prefrontal activation. These findings point at equivalent visuomotor transformations in locomotion and goal-directed hand movement, in which parietal-premotor circuitry is crucially implicated. Possible implications of an activation shift from spatial to intentional motor regions for understanding freezing of gait in Parkinson's disease are discussed: impaired medial prefrontal function in Parkinson's disease may reflect an insufficient internal motor drive when visual support from optic flow is reduced at the entrance of a narrow corridor. (C) 2010 Elsevier B.V. All rights reserved

Developmental Expression of the 84-kDa ODF Sperm Protein: Localization to both the Cortex and Medulla of Outer Dense Fibers and to the Connecting Piece

AbstractOuter dense fibers (ODF) are specialized cytoskeletal elements of the mammalian sperm tail which are composed of several prominent proteins. We previously reported the isolation of a cDNA (111-450) encoding a putative 84-kDa ODF protein. Here we demonstrate by independent cDNA isolations and by translational/immunoprecipitation of testicular mRNAs using anti-ODF 84 antibodies that 111-450 cDNA encodes the 84-kDa protein. We then analyzed the testicular expression of the ODF 84 mRNA and protein. Riboprobes generated from the clones recognized four testicular-specific transcripts of 1.6, 2.2, 2.4, and 2.8 kb in both rat and bull of which the immunoprecipitable product of the 2.4-kb mRNA comigrates with ODF 84 protein. Developmental Northerns indicated that the 2.2- and 2.4-kb mRNAs are first transcribed during meiotic prophase while the other two species are first expressed in round spermatids. The levels of all the transcripts steadily increased up to elongated spermatids. Immunocytochemistry revealed that the anti-84 reactive ODF proteins were synthesized and assembled in the cytoplasm of elongated spermatids (steps 9–18) with peak activity occurring in step 16 of spermiogenesis. Immunogold labeling was selective to the assembling ODF and connecting piece of the tail and to granulated bodies of the cytoplasmic lobe. Both the striated collar and capitulum of the connecting piece were immunolabeled as well as the basal plate of the implantation fossa. A combination of pre- and postembedding immunogold labeling provided evidence that the 84-kDa ODF protein is localized to both the cortex and medulla of the ODF in contrast to the sole medullary localization of the major 27-kDa ODF protein. Thus the 84-kDa ODF protein, encoded by the 2.4 transcript, is translationally regulated, packaged after synthesis into granulated bodies, assembled in a proximal to distal direction along the axoneme and may interact by means of leucine zippers specifically with the 27-kDa ODF protein during assembly. Its localization to both the cortex and medulla of the ODF, as opposed to exclusive medullary localization of the 27-kDa ODF protein, and the presence of two leucine zippers, only one of which interacts with the 27-kDa ODF, suggests that it could act as a link between proteins of the two regions of the ODF

A critical evaluation of spectrum-based fault localization techniques on a large-scale software system

In the past, spectrum-based fault localization (SBFL) techniques have been developed to pinpoint a fault location in a program given a set of failing and successful test executions. Most of the algorithms use similarity coefficients and have only been evaluated on established but small benchmark programs from the Software-artifact Infrastructure Repository (SIR). In this paper, we evaluate the feasibility of applying 33 state-of-the-art SBFL techniques to a large real-world project, namely ASPECTJ. From an initial set of 350 faulty version from the iBugs repository of ASPECTJ we manually classified 88 bugs where SBFL techniques are suitable. Notably, only 11 bugs of these bugs can be found after examining the 1000 most suspicious lines and on average 250 source code files need to be inspected per bug. Based on these results, the study showcases the limitations of current SBFL techniques on a larger program

Performance engineering for microservices and serverless applications: the RADON approach

Microservices and serverless are becoming integral parts of mod-ern cloud-based applications. Tailored performance engineering isneeded for assuring that the applications meet their requirementsfor quality attributes such as timeliness, resource efficiency, andelasticity. A novel DevOps-based framework for developing mi-croservices and serverless applications is being developed in theRADON project. RADON contributes to performance engineeringby including novel approaches for modeling, deployment optimiza-tion, testing, and runtime management. This paper summarizes thecontents of our tutorial presented at the 11th ACM/SPEC Interna-tional Conference on Performance Engineering (ICPE)

Nearest-neighbour directed random hyperbolic graphs

Undirected hyperbolic graph models have been extensively used as models of scale-free small-world networks with high clustering coefficient. Here we presented a simple directed hyperbolic model, where nodes randomly distributed on a hyperbolic disk are connected to a fixed number m of their nearest spatial neighbours. We introduce also a canonical version of this network (which we call "network with varied connection radius"), where maximal length of outgoing bond is space-dependent and is determined by fixing the average out-degree to m. We study local bond length, in-degree and reciprocity in these networks as a function of spatial coordinates of the nodes, and show that the network has a distinct core-periphery structure. We show that for small densities of nodes the overall in-degree has a truncated power law distribution. We demonstrate that reciprocity of the network can be regulated by adjusting an additional temperature-like parameter without changing other global properties of the network.Comment: 26 papers, 12 figure

Hyponatraemia in imported malaria is common and associated with disease severity

<p>Abstract</p> <p>Background</p> <p>Hyponatraemia (serum sodium < 135 mmol/L) has long been recognized as a complication of malaria. However, few studies have been done in non-immune adult populations. It has not been investigated previously how hyponatraemia is distributed among the various <it>Plasmodium </it>species, and its association with malaria severity is unknown.</p> <p>The aim of this retrospective cohort study was to determine the prevalence of hyponatraemia and its association with malaria severity in a large cohort of patients with imported malaria caused by various <it>Plasmodium </it>species.</p> <p>Methods</p> <p>All patients that were diagnosed with malaria in the Harbour Hospital and Institute for Tropical Diseases in Rotterdam in the period 1999-2009 and who had available serum sodium on admission were included. Severe malaria was defined according to the modified WHO criteria. Prevalence of hyponatraemia and its association with malaria severity were investigated by univariate comparison, ROC analysis and multivariate logistic regression analysis.</p> <p>Results</p> <p>A total of 446 patients with malaria (severe falciparum malaria n = 35, non-severe falciparum malaria n = 280, non-falciparum malaria n = 131) was included. Hyponatraemia was present in 207 patients (46%). Prevalence and severity of hyponatraemia were greatest in severe falciparum malaria (77%, median serum sodium 129 mmol/L), followed by non-severe falciparum malaria (48%, median serum sodium 131 mmol/L), and non-falciparum malaria (34%, median serum sodium 132 mmol/L). Admission serum sodium < 133 mmol/L had a sensitivity of 0.69 and a specificity of 0.76 for predicting severe malaria. Multivariate logistic regression showed that serum sodium < 131 mmol/L was independently associated with severe falciparum malaria (odds ratio 10.4, 95% confidence interval 3.1-34.9). In patients with hyponatraemia, hypovolaemia did not appear to play a significant role in the development of hyponatraemia when prerenal azotaemia and haematocrit were considered as surrogate markers for hypovolaemia.</p> <p>Conclusions</p> <p>Hyponatraemia is common in imported malaria and is associated with severe falciparum malaria. From a clinical point of view, the predictive power of hyponatraemia for severe malaria is limited. The precise pathophysiological mechanisms of hyponatraemia in malaria require further study.</p

On the evolution of decoys in plant immune systems

The Guard-Guardee model for plant immunity describes how resistance proteins (guards) in host cells monitor host target proteins (guardees) that are manipulated by pathogen effector proteins. A recently suggested extension of this model includes decoys, which are duplicated copies of guardee proteins, and which have the sole function to attract the effector and, when modified by the effector, trigger the plant immune response. Here we present a proof-of-principle model for the functioning of decoys in plant immunity, quantitatively developing this experimentally-derived concept. Our model links the basic cellular chemistry to the outcomes of pathogen infection and resulting fitness costs for the host. In particular, the model allows identification of conditions under which it is optimal for decoys to act as triggers for the plant immune response, and of conditions under which it is optimal for decoys to act as sinks that bind the pathogen effectors but do not trigger an immune response.Comment: 15 pages, 6 figure