1,971 research outputs found
Role of the Kinesin-like Protein KipB in Aspergillus nidulans
Molecular motors are protein machines, which power
almost all forms of movement in the living world. Among the
best known are the motors that hydrolyze ATP and use the
derived energy to generate force. They are involved in a
variety of diverse cellular functions as vesicle and organelle
transport, cytoskeleton dynamics, morphogenesis, polarized
growth, cell movements, spindle formation, chromosome movement,
nuclear fusion, and signal transduction. Three superfamilies of
molecular motors, kinesins, dyneins, and myosins, have so far
been well characterized. These motors use microtubules (in the
case of kinesines and dyneins) or actin filaments (in the case
of myosins) as tracks to transport cargo materials within a
cell. Analysis of fungal genomes revealed at least 10 distinct
kinesins in filamentous fungi, some of which are not found in
yeasts. We used the motor domain of conventional kinesin (KinA)
from Aspergillus nidulans to perfom BLAST searches at the
public A. nidulans genome database, at the Whitehead Center for
Genome Research (Cambridge USA), and identified eleven putative
kinesin motors. They grouped into nine of the eleven families,
two kinesins being found in the Unc104 familiy and
interestingly, one did not fall into any of the known families.
The present work analyses the function of a kinesin-like
protein in A. nidulans, KipB, which is a member of the Kip3
kinesin family. This family includes one representative in
Saccharomyces cerevisiae (Kip3, the family founding member),
two in Schizosaccharomyces pombe, Klp5 and Klp6 and one in
Drosophila, Klp67A, the single one reported so far for higher
eukaryotes in this family. Kip3 kinesins are implicated in
microtubule disassembly and are required for chromosome
segregation in mitosis and meiosis. To assess the function of
KipB kinesin in A. nidulans, a kipB disruption strain was
constructed. Analysis of the DkipB mutant revealed new features
concerning the cellular functions of Kip3 proteins, but also
some conserved ones. kipB is not essential for vegetative
growth, and meiosis and ascospore formation were not affected
in the DkipB mutant. The KipB protein was shown to be involved
in the turnover of interphase cytoplasmic, mitotic and astral
microtubules. DkipB mutants are less sensitive to the
microtubule-destabilizing drug benomyl, and the microtubule
cytoskeleton of interphase cells in DkipB mutants appears
altered. Interestingly, spindle morphology and positioning were
severely affected. Spindles were highly mobile, could overpass
each other, moved over long distances through the cytoplasm,
and displayed in 64% of the cases an extremely bent shape,
latter feature being the first time reported for Kip3 kinesins.
Mitotic progression was delayed in the DkipB mutant and a
higher number of cytoplasmic microtubules remained intact
during mitosis. DkipB heterozygous strains showed an increased
instability of diploid nuclei, which proved once more KipB
involvement in mitosis, along with DkipB clear genetic
interaction with a mutation in another mitotic kinesin in A.
nidulans, bimC4. An N-terminal GFP-KipB construct localized to
cytoplasmic microtubules in interphase cells and to spindle and
astral microtubules during mitosis, in a discontinuous pattern.
Speckles of GFP-KipB appeared to be aligned in the cell.
Time-lapse video microscopy indicated that the spots were
moving independently towards the microtubule plus ends. This
advanced the hypothesis that KipB could display processivity
and intrinsic motility along microtubules, or that other
kinesins involved in organelle motility are able to target the
KipB protein to the microtubule plus ends. In the case of
C-terminally truncated GFP-KipB protein versions, a stronger
GFP signal was obtained and colocalization with a-tubulin-GFP
revealed that they uniformly stain cytoplasmic, mitotic and
astral microtubules. This suggests that the C-terminus is
important for the correct localization and the movement of KipB
protein along microtubules
Immunological characterization of chromogranins A and B and secretogranin II in the bovine pancreatic islet
Antisera against chromogranin A and B and secretogranin II were used for analysing the bovine pancreas by immunoblotting and immunohistochemistry. All three antigens were found in extracts of fetal pancreas by one dimensional immunoblotting. A comparison with the soluble proteins of chromaffin granules revealed that in adrenal medulla and in pancreas antigens which migrated identically in electrophoresis were present. In immunohistochemistry, chromogranin A was found in all pancreatic endocrine cell types with the exception of most pancreatic polypeptide-(PP-) producing cells. For chromogranin B, only a faint immunostaining was obtained. For secretorgranin II, A-and B-cells were faintly positive, whereas the majority of PP-cells exhibited a strong immunostaining for this antigen. These results establish that chromogranins A and B and secretogranin II are present in the endocrine pancreas, but that they exhibit a distinct cellular localization
Helminths and HIV infection: epidemiological observations on immunological hypotheses
Parasitic helminths have co-evolved with the mammalian immune system. Current hypotheses suggest that immunological stimulation in the presence of helminths is balanced by immuno-regulation and by the broad spectrum of mechanisms possessed by helminths for countering the host immune response. The degree to which this balance is perfected, and the mechanisms by which this is achieved, vary between helminth species; we suggest that this is reflected not only in the degree of pathology induced by helminths but also in a variety of relationships with HIV infection and HIV disease. Available epidemiological data regarding interactions between helminths and HIV are largely observational; results are variable and generally inconclusive. Well designed, controlled intervention studies are required to provide definitive information on the species-specific nature of these interactions and on the advantages, disadvantages and optimal timing of de-worming in relation to HIV infection
Backbone rigidity and static presentation of guanidinium groups increases cellular uptake of arginine-rich cell-penetrating peptides
In addition to endocytosis-mediated cellular uptake, hydrophilic cell-penetrating peptides are able to traverse biological membranes in a non-endocytic mode termed transduction, resulting in immediate bioavailability. Here we analysed structural requirements for the non-endocytic uptake mode of arginine-rich cell-penetrating peptides, by a combination of live-cell microscopy, molecular dynamics simulations and analytical ultracentrifugation. We demonstrate that the transduction efficiency of arginine-rich peptides increases with higher peptide structural rigidity. Consequently, cyclic arginine-rich cell-penetrating peptides showed enhanced cellular uptake kinetics relative to their linear and more flexible counterpart. We propose that guanidinium groups are forced into maximally distant positions by cyclization. This orientation increases membrane contacts leading to enhanced cell penetration
A Condensation-Ordering Mechanism in Nanoparticle-Catalyzed Peptide Aggregation
Nanoparticles introduced in living cells are capable of strongly promoting
the aggregation of peptides and proteins. We use here molecular dynamics
simulations to characterise in detail the process by which nanoparticle
surfaces catalyse the self- assembly of peptides into fibrillar structures. The
simulation of a system of hundreds of peptides over the millisecond timescale
enables us to show that the mechanism of aggregation involves a first phase in
which small structurally disordered oligomers assemble onto the nanoparticle
and a second phase in which they evolve into highly ordered beta-sheets as
their size increases
Natural variation in sensory-motor white matter organization influences manifestations of Huntington's disease
While the HTT CAG-repeat expansion mutation causing Huntington's disease (HD) is highly correlated with the rate of pathogenesis leading to disease onset, considerable variance in age-at-onset remains unexplained. Therefore, other factors must influence the pathogenic process. We asked whether these factors were related to natural biological variation in the sensory-motor system. In 243 participants (96 premanifest and 35 manifest HD; 112 controls), sensory-motor structural MRI, tractography, resting-state fMRI, electrophysiology (including SEP amplitudes), motor score ratings, and grip force as sensory-motor performance were measured. Following individual modality analyses, we used principal component analysis (PCA) to identify patterns associated with sensory-motor performance, and manifest versus premanifest HD discrimination. We did not detect longitudinal differences over 12 months. PCA showed a pattern of loss of caudate, grey and white matter volume, cortical thickness in premotor and sensory cortex, and disturbed diffusivity in sensory-motor white matter tracts that was connected to CAG repeat length. Two further major principal components appeared in controls and HD individuals indicating that they represent natural biological variation unconnected to the HD mutation. One of these components did not influence HD while the other non-CAG-driven component of axial versus radial diffusivity contrast in white matter tracts were associated with sensory-motor performance and manifest HD. The first component reflects the expected CAG expansion effects on HD pathogenesis. One non-CAG-driven component reveals an independent influence on pathogenesis of biological variation in white matter tracts and merits further investigation to delineate the underlying mechanism and the potential it offers for disease modification
Scabies Mite Peritrophins Are Potential Targets of Human Host Innate Immunity
The gut of most invertebrates is lined by a protective layer of chitin and glycoproteins, often designated as a peritrophic matrix. Previous research suggests that it forms a barrier that may protect the midgut epithelium from abrasive food particles and pathogens. Parasitic invertebrates ingesting vertebrate plasma have evolved additional strategies to protect themselves from hazardous host molecules consumed during feeding. An important part of the immediate defense in vertebrate plasma is complement-mediated killing. The Complement system is a complex network of more than 35 proteins present in human plasma that results in killing of foreign cells including the gut epithelial cells of a feeding parasite. Recently we found that scabies mites, who feed on skin containing plasma, produce several proteins that inhibit human complement within the mite gut. The mites excrete these molecules into the upper epidermis where they presumably also inhibit complement activity. Mite gut antigens that initially trigger the complement cascade have not been identified previously. Obvious possible targets of complement attack within the mite gut could be peritrophins. Our study describes the first peritrophin identified in scabies mites and indicates a possible role in complement activation
Regional movements of the tiger shark, Galeocerdo cuvier, off northeastern Brazil: inferences regarding shark attack hazard
An abnormally high shark attack rate verified off Recife could be related to migratory behavior of tiger sharks. This situation started after the construction of the Suape port to the south of Recife. A previous study suggested that attacking sharks could be following northward currents and that they were being attracted shoreward by approaching vessels. In this scenario, such northward movement pattern could imply a higher probability of sharks accessing the littoral area of Recife after leaving Suape. Pop-up satellite archival taus were deployed on five tiger sharks caught off Recife to assess their movement patterns off northeastern Brazil. All tags transmitted from northward latitudes after 7-74 days of freedom. The shorter, soak distance between deployment and pop-up locations ranged between 33-209 km and implied minimum average speeds of 0.02-0.98 km.h(-1). Both pop-up locations and depth data suggest that tiger shark movements were conducted mostly over the continental shelf. The smaller sharks moved to deeper waters within 24 hours after releasing, but they assumed a shallower (< 50 m) vertical distribution for most of the monitoring period. While presenting the first data on tiger shark movements in the South Atlantic, this study also adds new information for the reasoning of the high shark attack rate verified in this region,State Government of Pernambuco and Petrobras (Brazil); Fundacao para a Ciencia e Tecnologia (Portugal) [MCTES/FCT/SFRH/BD/37065/2007
Prototypes for Content-Based Image Retrieval in Clinical Practice
Content-based image retrieval (CBIR) has been proposed as key technology for computer-aided diagnostics (CAD). This paper reviews the state of the art and future challenges in CBIR for CAD applied to clinical practice
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