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A Sievers

Publications and source records attributed to A Sievers.

At least 19 recordsLinked to original sources
PubMed2001

Comparison of diameter and perimeter methods for tumor volume calculation.

PURPOSE: Lesion volume is often used as an end point in clinical trials of oncology therapy. We sought to compare the common method of using orthogonal diameters to estimate lesion volume (the diameter method) with a computer-assisted planimetric technique (the perimeter method). METHODS: Radiologists reviewed 825 magnetic resonance imaging studies from 219 patients with glioblastoma multiforme. Each study had lesion volume independently estimated via the diameter and perimeter methods. Cystic areas were subtracted out or excluded from the outlined lesion. Inter- and intrareader variability was measured by using multiple readings on 48 cases. Where serial studies were available in noncystic cases, a mock response analysis was used. RESULTS: The perimeter method had a reduced interreader and intrareader variability compared with the diameter method (using SD of differences): intrareader, 1.76 mL v 7.38 mL (P < .001); interreader, 2.51 mL v 9.07 mL (P < .001) for perimeter and diameter results, respectively. Of the 121 noncystic cases, 23 had serial data. In six (26.1%) of those 23, a classification difference occurred when the perimeter method was used versus the diameter method. CONCLUSION: Variability of measurements was reduced with the computer-assisted perimeter method compared with the diameter method, which suggests that changes in volume can be detected more accurately with the perimeter method. The differences between these techniques seem large enough to have an impact on grading the response to therapy.

Brain↗
PubMed2000

The pattern of acropetal and basipetal cytoplasmic streaming velocities in Chara rhizoids and protonemata, and gravity effect on the pattern as measured by laser-Doppler-velocimetry.

The spatial pattern of acropetal and basipetal cytoplasmic streaming velocities has been studied by laser-Doppler-velocimetry (LDV) in the positively gravitropic (downward growing) rhizoids of Chara globularis Thuill. and for the first time in the negatively gravitropic (upward growing) protonemata. The LDV method proved to be precise and yielded reproducible results even when tiny differences in velocities were measured. In the apical parts of the streaming regions of both cell types, acropetal streaming was faster than basipetal streaming. Starting at the apical reversal point of streaming, the velocity increased basipetally with the distance from that point and became fairly constant close to the basal reversal point; subsequently, the velocity decreased slightly acropetally as the apical reversal point was again approached. There was no change in velocity at the basal reversal point. However, at the apical reversal point there was an abrupt decrease in velocity. The pattern of the ratio of acropetal to basipetal streaming velocity (VR) was a function of the relative distance of the site of measurement from the apical reversal point rather than a function of the absolute distance. Upon inversion of the rhizoids, the VR decreased on average by 3.8% (+/- 0.4%), indicating that the effect of gravity on the streaming velocity was merely physical and without a physiological amplification. Rhizoids that had developed on the slowly rotating horizontal axis of a clinostat, and had never experienced a constant gravity vector, were similar to normally grown rhizoids with respect to VR pattern. In protonemata, the VR pattern was not significantly different from that in rhizoids although the direction of growth was inverse. In rhizoids, oryzalin caused the polar organization of the cell to disappear and nullified the differences in streaming velocities, and cytochalasin D decreased the velocity of basipetal streaming slightly more than that of acropetal streaming. Cyclopiazonic acid, known as an inhibitor of the Ca2+-ATPase of the endoplasmic reticulum, also reduced the streaming velocities in rhizoids, but had slightly more effect on the acropetal stream. It is possible that the endogenous difference in streaming velocities in both rhizoids and protonemata is caused by differences in the cytoskeletal organization of the opposing streams and/or loading of inhibitors (like Ca2+) from the apical/subapical zone into the basipetally streaming endoplasm.

Cytochalasin D↗
PubMed2000

Plant cells on earth and in space.

Two quite different types of plant cells are analysed with regard to transduction of the gravity stimulus: (i) Unicellular rhizoids and protonemata of characean green algae; these are tube-like, tip-growing cells which respond to the direction of gravity. (ii) Columella cells located in the center of the root cap of higher plants; these cells (statocytes) perceive gravity. The two cell types contain heavy particles or organelles (statoliths) which sediment in the field of gravity, thereby inducing the graviresponse. Both cell types were studied under microgravity conditions (10(-4) g) in sounding rockets or spacelabs. From video microscopy of living Chara cells and different experiments with both cell types it was concluded that the position of statoliths depends on the balance of two forces, i.e. the gravitational force and the counteracting force mediated by actin microfilaments. The actomyosin system may be the missing link between the gravity-dependent movement of statoliths and the gravity receptor(s); it may also function as an amplifier.

Actins↗
PubMed1999

Ultrastructure and cytoskeleton of Chara rhizoids in microgravity.

Gravitropic tip growth of Chara rhizoids is dependent on the presence and functional interaction between statoliths, cytoskeleton and the tip-growth-organizing complex, the Spitzenkorper. Microtubules are essential for the polar cytoplasmic zonation but are excluded from the apex and do not play a crucial role in the primary steps of gravisensing and graviresponse. Actin filaments form a dense meshwork in the subapical zone and converge into a prominent apical actin patch which is associated with the endoplasmic reticulum (ER) aggregate representing the structural center of the Spitzenkorper. The position of the statoliths is regulated by gravity and a counteracting force mediated by actomyosin. Reducing the acceleration forces in microgravity experiments causes a basipetal displacement of the statoliths. Rhizoids grow randomly in all directions. However, they express the same cell shape and cytoplasmic zonation as ground controls. The ultrastructure of the Spitzenkorper, including the aggregation of ER, the assembly of vesicles in the apex, the polar distribution of proplastids, mitochondria, dictyosomes and ER cisternae in the subapical zone is maintained. The unaltered cytoskeletal organization, growth rates and gravitropic responsiveness indicate that microgravity has no major effect on gravitropic tip-growing Chara rhizoids. However, the threshold value of gravisensitivity might be different from ground controls due to the altered position of statoliths, a possibly reduced amount of BaSO4 in statoliths and a possible adaptation of the actin cytoskeleton to microgravity conditions.

Actins↗
PubMed1999

Electron microscopic analysis of gravisensing Chara rhizoids developed under microgravity conditions.

Tip-growing, unicellular Chara rhizoids that react gravitropically on Earth developed in microgravity. In microgravity, they grew out from the nodes of the green thallus in random orientation. Development and morphogenesis followed an endogenous program that is not affected by the gravitational field. The cell shape, the polar cytoplasmic organization, and the polar distribution of cell organelles, except for the statoliths, were not different from controls that had grown on earth (ground controls). The ultrastructure of the organelles and the microtubules were well preserved. Microtubules were excluded from the apical zone in both ground controls as well as microgravity-grown rhizoids. The statoliths (vesicles containing BaSO4 crystals in a matrix) in microgravity-grown rhizoids were spread over a larger area (up to 50 microm basal to the tip) than the statoliths of ground controls (10-30 microm). Some statoliths were even located in the subapical zone close to microtubules, which was not observed in ground controls. The crystals in statoliths from microgravity-grown rhizoids appeared more loosely arranged in the vesicle matrix compared with ground controls. The chemical composition of the crystals was identified as BaSO4 by X-ray microanalysis. There is evidence that the amount of BaSO4 in statoliths of rhizoids developed in microgravity is lower than in ground controls, indicating that the gravisensitivity of microgravity-developed rhizoids might be reduced compared with ground controls. Lack of gravity, however, does not affect the process of tip growth and does not inhibit the development of the structures needed for the gravity-sensing machinery.

Chlorophyta↗
PubMed1999

Identification and characterization of IS2404 and IS2606: two distinct repeated sequences for detection of Mycobacterium ulcerans by PCR.

Molecular analysis of Mycobacterium ulcerans has revealed two new insertion sequences (ISs), IS2404 and IS2606. IS2404 was identified by complete sequencing of a previously described repetitive DNA segment from M. ulcerans. This element is 1,274 bp long, contains 12-bp inverted repeats and a single open reading frame (ORF) potentially encoding a protein of 327 amino acids (aa), and apparently generates 7-bp direct repeats upon transposition. Amino acid similarity was found between the putative transposase and those encoded by ISs in other bacterial sequences from Aeromonas salmonicida (AsIs1), Escherichia coli (H repeat element), Vibrio cholerae (VcIS1), and Porphyromonas gingivalis (PGIS2). The second IS, IS2606, was discovered by sequence analysis of a HaeIII fragment of M. ulcerans genomic DNA containing a repetitive sequence. This element is 1,404 bp long, with 12-bp inverted repeats and a single ORF potentially encoding a protein of 445 aa. Database searches revealed a high degree of amino acid identity (70%) with the putative transposase of IS1554 from M. tuberculosis. Significant amino acid identity (40%) was also observed with transposases from several other microorganisms, including Rhizobium meliloti (ISRm3), Burkholderia cepacia (IS1356), Corynebacterium diphtheriae, and Yersinia pestis. PCR screening of DNA from 45 other species of mycobacteria with primers for IS2404 confirm that this element is found only in M. ulcerans. However, by PCR, IS2606 was also found in Mycobacterium lentiflavum, another slow-growing member of the genus Mycobacterium that is apparently genetically distinct from M. ulcerans. Testing the sensitivity of PCR based on IS2404 and IS2606 primers demonstrated the ability to detect 0.1 and 1 M. ulcerans genome equivalents, respectively. The ability to detect small numbers of cells by using two gene targets will be particularly useful for analyzing environmental samples, where there may be low concentrations of M. ulcerans among large numbers of other environmental mycobacteria.

Base Sequence↗
PubMed1998

Hypergravity can reduce but not enhance the gravitropic response of Chara globularis protonemata.

The relationship between the position of the statoliths and the direction and rate of tip growth in negatively gravitropic protonemata of Chara globularis was studied with a centrifuge video microscope. Cells placed perpendicularly to the acceleration vector (stimulation angle 90 degrees) showed a gradual reduction of the gravitropic curvature with increasing accelerations from 1 g to 8 g despite complete sedimentation of all statoliths on the centrifugal cell flank. It is argued that the increased weight of the statoliths in hypergravity impairs their acropetal transport which is induced when the cell axis deviates from the normal upright orientation. When the statoliths were centrifuged deep into the apical dome at 6 g and a stimulation angle of 170 degrees the gravitropic curvature after 1 h was identical to that determined for the same cells at 1 g and the same stimulation angle. This indicates that gravitropism in Chara protonemata is either independent of the pressure exerted by the statoliths on an underlying structure or is already saturated at 1 g. When the statoliths were moved along the apical cell wall at 8 g and the stimulation angle was gradually increased from 170 degrees to 220 degrees the gravitropic curvature reverted sharply when the cluster of statoliths passed over the cell pole. This experiment supports the hypothesis that in Chara protonemata asymmetrically distributed statoliths inside the apical dome displace the Spitzenkorper and thus the centre of growth, resulting in gravitropic bending. In contrast to the positively gravitropic Chara rhizoids, no modifications either in the transport of statoliths during basipetal acceleration (6 g, stimulation angle 0 degree, 5 h) or in the subsequent gravitropic response could be detected in the protonemata. The different effects of centrifugation on the positioning of statoliths in Chara protonemata and rhizoids indicate subtle differences in the function of the cytoskeleton in both types of cells.

Centrifugation↗
PubMed1998

Statolith positioning by microfilaments in Chara rhizoids and protonemata.

The rhizoids of the green alga Chara are tip-growing cells with a precise positive gravitropism. In rhizoids growing downwards the statoliths never sediment upon the cell wall at the very tip but keep a minimal distance of approximately 10 micrometers from the cell vertex. It has been argued that this position is attained by a force acting upon the statoliths in the basal direction and that this force is generated by an interaction between actin microfilaments and myosin on the statolith membrane. This hypothesis received experimental support from (1) effects of the actin-attacking drug cytochalasin, (2) experiments under microgravity conditions, and (3) clinostat experiments. Using video-microscopy it is now shown that this basipetal force also acts on statoliths during sedimentation. As a result, many statoliths in Chara rhizoids do not simply fall along the plumb line while sedimenting during gravistimulation, but move basipetally. This statolith movement is compared to the ones occurring in the unicellular Chara protonemata during gravistimulation. Dark-grown protonemata morphologically closely resemble the rhizoids but respond negatively gravitropic. In contrast to the rhizoids a gravistimulation of the protonemata induces a transport of statoliths towards the tip. This transport is mainly along the cell axis and not parallel to the gravity vector. It is stressed that the sedimentation of statoliths in Chara rhizoids and protonemata as well as in gravity sensing cells in mosses and higher plants is accompanied by statolith movements based on interactions with the cytoskeleton. In tip-growing cells these movements direct the statoliths to a definite region of the cell where they can sediment and elicit a gravitropic curvature. In the statocytes of higher plants the interactions of the statoliths with the cytoskeleton probably do not serve primarily to move the statoliths but to transduce mechanical stresses from the sedimenting statoliths to the plasma membrane.

Actin Cytoskeleton↗
PubMed1998

Determination of the threshold acceleration for the gravitropic stimulation of cress roots and hypocotyls.

To determine the range of the threshold acceleration (a-threshold) for the gravitropic stimulation of Lepidium sativum L. roots and hypocotyls, experiments were performed on a centrifuge-clinostat with two-orthogonal axes. The rotation rate of the clinostat was 4 rpm (< or = 1.8 x 10(-4) g), while that of the centrifuge was from 3 to 17 rpm (3 x 10(-3) to 10(-1) g). The gravitropic response was determined: (i) after growth of roots and hypocotyls in their normal vertical position and subsequent gravitropic stimulation for 3 h by accelerations of 4 x 10(-3) to 10(-1) g, and (ii) after continuous stimulation in the lateral direction by centripetal accelerations of 4 x 10(-3) to 10(-1) g. The a-threshold was defined by an extrapolation of the regression line of R = p + rx, where x was either ln a or l/a for 3 h or a continuous stimulation, respectively. The a-threshold estimated after 3 h stimulation was equal to 2.6 x 10(-3) g for roots and 3.1 x 10(-3) g for hypocotyls. The threshold accelerations that were unable to evoke a gravitropic response even with continuous stimulation of cress roots and hypocotyls were approximately 3.1 x 10(-3) g and 3.6 x 10(-3) g, respectively. Increasing the stimulation acceleration up to 4.1 x 10(-3) g led to a statistically confirmed gravitropic response of a definite proportion of both the root and hypocotyl populations. In the experiments where acceleration and stimulation time were variable, the threshold dose (D-threshold) for roots was determined to be about 14 to 22 g x s, depending on the stimulation duration and the range of accelerations. The kinetics of gravitropic response at a near-threshold acceleration (4 x 10(-3) to 1.9 x 10(-2) g) differed from that at 1 g (horizontal stimulation). At low forces, the maximal response dependent on the magnitude of acceleration could not be enhanced by increasing the stimulation time up to at least 210 min.

Acceleration↗
PubMed1998

Temporal course of graviperception in intermittently stimulated cress roots.

Gravitropic bending of Lepidium roots caused by intermittent stimulation lasting approximately 1 h was the same for a particular sum of stimulation intervals and was independent of (i) the length of a single stimulation interval (from 1 to 12.2 s) during which the roots were exposed unilaterally and horizontally, and (ii) rest intervals (from 60 to 300 s) during which roots were horizontally rotated at two revolutions per minute on a clinostat. The same effectiveness of equal sums of short stimulations separated by relatively long rest intervals indicates that the signals into which the stimuli are transduced are: (i) additive; (ii) proportional to the duration of a single stimulation; and (iii) stable for at least 5 min. The perception time is shorter than 1 s, the presentation time is approximately 10 s. The effects of intermittent stimulation fit the hypothesis that the gravity-induced movement of statoliths changes asymmetrically the stress in cytoskeletal actin filaments, thereby inducing gravitropic bending.

Brassicaceae↗
PubMed1997

Central root cap cells are depleted of endoplasmic microtubules and actin microfilament bundles: implications for their role as gravity-sensing statocytes.

Indirect immunofluorescence, using monoclonal antibodies to actin and tubulin, applied to sections of root tips of Lepidium, Lycopersicon, Phleum, and Zea, revealed features of the cytoskeleton that were unique to the statocytes of their root caps. Although the cortical microtubules (CMTs) lay in dense arrays against the periphery of the statocytes, these same cells showed depleted complements of endoplasmic microtubules (EMTs) and of actin microfilament (AMF) bundles, both of which are characteristic of the cytoskeleton of other post-mitotic cells in the proximal portion of the root apex. The scarcity of the usual cytosketetal components within the statocytes is considered responsible for the exclusion of the larger organelles (e.g., nucleus, plastids, ER elements) from the interior of the cell and for the absence of cytoplasmic streaming. Furthermore, the depletion of dense EMT networks and AMF bundles in statocyte cytoplasm is suggested as being closely related to the elevated cytoplasmic calcium content of these cells which, in turn, may also favour the formation of the large sedimentable amyloplasts by not permitting plastid divisions. These latter organelles are proposed to act as statoliths due to their dynamic interactions with very fine and highly unstable AMFs which enmesh the statoliths and merge into peripheral AMFs-CMTs-ER-plasma membrane complexes. Rather indirect evidence for these interactions was provided by showing enhanced rates of statolith sedimentation after chemically-induced disintegration of CMTs. All these unique properties of the root cap statocytes are supposed to effectively enhance the gravity-perceptive function of these highly specialized cells.

Actin Cytoskeleton↗
PubMed1997

Detection of Mycobacterium ulcerans in environmental samples during an outbreak of ulcerative disease.

Mycobacterium ulcerans is an environmental bacterium which causes chronic skin ulcers. Despite significant epidemiological evidence to suggest that water is the source of infection, the organism has never been identified in the environment. Environmental water samples were collected from a small town in which an outbreak of 29 cases had occurred in a 3-year period. These were examined by mycobacterial culture and PCR amplification. Similar to previous studies, M. ulcerans was not cultured from the water samples. However, five samples were positive for M. ulcerans by PCR. These samples were collected from a swamp and a golf course irrigation system within the outbreak area. This is the first time that M. ulcerans has been demonstrated to be present in the environment and supports the postulated epidemiology of disease due to this organism.

DNA, Bacterial↗
PubMed1997

Displacement of statoliths in Chara rhizoids during horizontal rotation on clinostats.

The basipetal movement of statoliths in Chara rhizoids, similar to that during parabolic flights of TEXUS rockets occurs also during rotation on clinostats. Within 15 min on fast-rotating clinostat, the distance between the center of the statolith complex and the cell vertex increases for 60% of that in positively gravitropic downward growing rhizoids. Cytochalasin D experiments confirm that the movement of statoliths is actin-dependent and the actin filaments exert basipetal forces on statoliths in gravity field. The clinostat and/or cytochalasin experiments confirm the suggestion that on earth the position of statoliths depends on the balance of the gravitational force and the counteracting force mediated by actin filaments. The statolith center keeps a stable position during about 30 min on a fast-rotating clinostat, i.e. it is then in a new dynamically stable state. This new state is achieved 15 min after the basipetal acting filament-mediated force has been disturbed by clinostatting. Further experiments on the fast-rotating clinostat show that this new position brings about a reorganization of actin filaments which makes the process of acropetal transport of statoliths possible. The amplitude of particle oscillatory movement decreases as the rotational speed of the clinostat increases. This explains the differences of the results obtained from the experiments on fast-rotating and slow-rotating clinostats. It should be kept in mind that rhizoids are unicellular. The fast-rotating clinostat is suitable for simulation of conditions without gravity when a rhizoid is on the axis of rotation. The interaction of statoliths and actin filaments at zero gravity can be studied by means of such a clinostat.

Actin Cytoskeleton↗
PubMed1996

Fixation procedure for transmission electron microscopy of Chara rhizoids under microgravity in a Spacelab (IML-2).

A special fixation device and fixation procedure have been developed to investigate for the first time the ultrastructure of gravity-sensing, unicellular Chara rhizoids grown for 30 h under microgravity (MG) conditions during the IML-2 mission. The fixation unit allowed culture, fixation and storage of Chara rhizoids in the same chamber without transferring the samples. The procedure was easy and safe to perform and required a minimum of crew time. Rhizoids fixated with glutaraldehyde in space and further processed for electron microscopy on ground showed that the fixation was of high quality and corresponded to the fixation quality of rhizoids in the ground controls. Thus, the equipment accomplished the manifold problems related to the physical effects of MG. The polarity of the rhizoids was maintained in MG. Well-preserved organelles and microtubules showed no obvious difference in ultrastructure or distribution after 30-h growth in MG compared to ground controls. The statoliths were more randomly distributed, however, only up to 50 microns basal to the tip. Thus, changing the gravity conditions does to disturb the cellular organisation of the rhizoids enabling the tip-growing cells to follow their genetic program in development and growth also under MG.

Cell Culture Techniques↗
PubMed1996

Gravitropism of the primary root of maize: a complex pattern of differential cellular growth in the cortex independent of the microtubular cytoskeleton.

The spatio-temporal sequence of cellular growth within the post-mitotic inner and outer cortical tissue of the apex of the primary root of maize (Zea mays L.) was investigated during its orthogravitropic response. In the early phase (0-30 min) of the graviresponse there was a strong inhibition of cell lengthening in the outer cortex at the lower side of the root, whereas lengthening was only slightly impaired in the outer cortex at the upper side. Initially, inhibition of differential cell lengthening was less pronounced in the inner cortex indicating that tissue tensions which, in these circumstances, inevitably develop at the outer-inner cortex interface, might help to drive the onset of the root bending. At later stages of the graviresponse (60 min), when a root curvature had already developed, cells of the inner cortex then exhibited a prominent cell length differential between upper and lower sides, whereas the outer cortex cells had re-established similar lengths. Again, tissue tensions associated with the different patterns of cellular behaviour in the inner and outer cortex tissues, could be of relevance in terminating the root bending. The perception of gravity and the complex tissue-specific growth responses both proceeded normally in roots which were rendered devoid of microtubules by colchicine and oryzalin treatments. The lack of involvement of microtubules in the graviresponse was supported by several other lines of evidence. For instance, although taxol stabilized the cortical microtubules and prevented their re-orientation in post-mitotic cortical cells located at the lower side of gravistimulated roots, root bending developed normally. In contrast, when gravistimulated roots were physically prevented from bending, re-oriented arrays of cortical microtubules were seen in all post-mitotic cortical cells, irrespective of their position within the root.

Cell Size↗