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A Sievers

Publications and source records attributed to A Sievers.

At least 55 records · Page 3Linked to original sources

[Research under reduced gravity. Part II: experiments in variable gravitational fields].

Recently, the reduced gravitational field of space laboratories, rockets, or satellites in Earth orbits offers a gravitational field which is variable from 10(-4) g to 1 g by the use of centrifuges. Especially with plants, data concerning gravisensitivity are based on experiments with clinostats. First experiments in reduced gravitational fields, however, demonstrate the uncertainty of these results. Thus, the main task of gravitational biologists is to test the validity of results obtained with the aid of clinostats. On this basis it should be possible to find a common mechanism to explain the influence of gravity on organisms. Experiments under reduced gravity in sounding rockets provided new knowledge on the perception of the gravity stimulus in plant cells.

Biological Science Disciplines↗

[Research under reduced gravity. Part I: bases of gravitational biology].

The orientation of organisms in space and their morphogenesis in relation to the gravitational field of the Earth are the main topics of research in the field of gravitational biology. For more than 100 years clinostats provided the only possibility to simulate physiological weightlessness. In contrast to animals, plants are characterized by intracellular gravireceptors. Nevertheless, there are some indications, e.g., the minimal energy of approx. 10(-18) J triggering a gravity-dependent response, for similar mechanisms of gravity perception. Stretch-activated ion channels might be the common structural basis.

Biological Science Disciplines↗

Identification of a genetically distinct subspecies of Mycobacterium kansasii.

To assess the usefulness of a specific DNA probe for Mycobacterium kansasii, 105 isolates from Australia, Belgium, Japan, South Africa, and Switzerland were collected and analyzed. Twenty of these isolates were probe negative, of which 18 were from Belgium and Switzerland. Analysis of all isolates by Southern blot hybridization indicated a lack of variability among probe-positive isolates, while probe-negative isolates were clearly distinct and showed greater diversity. Sequence analysis of the 250 nucleotides at the 5' end of the 16S rRNA gene revealed that 19 of the 20 probe-negative isolates had a sequence different from that of M. kansasii. A total of five nucleotide differences were present in a cluster consisting of two nucleotide deletions and three nucleotide substitutions. These results suggest the existence of a genetic subspecies of M. kansasii.

Base Sequence↗

Isolation of a fastidious Mycobacterium species from two AIDS patients.

Two strains of fastidious mycobacteria were isolated from two patients with AIDS and clinical disease suggestive of Mycobacterium avium complex infection. Acid-fast bacilli were isolated from blood and bone marrow of both patients in BACTEC 12B and/or 13A media. The acid-fast bacilli failed to grow on subculture to routine Löwenstein-Jensen medium containing pyruvate and egg yolk agar. After several attempts, the strain from one patient was finally cultured on Middlebrook 7H9 medium with agar, charcoal, and yeast extract 13 months after the initial specimens were received in the laboratory. The second patient's strain was cultured on the same medium 6 weeks postinoculation with fresh BACTEC fluid and 5 months after specimen collection. Routine biochemical and growth tests were performed on these isolates but failed to give definitive identifications. 16S rRNA gene sequencing suggested that the organisms share at least 98.9% homology with M. simiae. Even greater homology (99.86%) was found with the recently described species "M. genavense." Recognition of the fastidious nature of some mycobacteria that infect AIDS patients is important in the treatment of infections in these patients and in understanding the epidemiology of atypical mycobacterial infections. It is suggested that a liquid culture medium such as BACTEC be employed for primary isolation of mycobacteria from AIDS patients and that subculture to the charcoal medium described here be carried out for those organisms that fail to grow on subculture to routine media.

AIDS-Related Opportunistic Infections↗

How well does the clinostat mimic the effect of microgravity on plant cells and organs?

The effect of clinostatting and microgravity on plant cells and organs is considered on the basis of distinguishing two types of gravistimulation: static and dynamic. The former is switched off both by clinostatting and microgravity, the latter is switched off by microgravity but occurs inevitably during clinostatting and may be perceived by cells if the rotation is not fast enough. Effects of clinostatting and microgravity on different examples of static gravistimulation (tonic effects, formation of compression wood, growth of "grass nodes," compensation of epinasty, stabilization of cellular polarity) are considered. The mechanism of the dynamic stimulation is presented; it is related to the displacement of the gravity sensing masses in the cell containing them, and involves disturbance of cytoskeletal tension. The low threshold for gravity perception and short minimal time of dynamic stimulation are emphasized. Only a relatively fast rotating clinostat, on which the radial distance of the cells from the rotational axis is small enough to keep the centrifugal force low, can effectively compensate gravity. However, one must take into account the extreme sensitivity of plants to mechanical stresses that may appear during clinostatting at different levels of plant organization.

Cytoskeleton↗

Effects of submergence on development and gravitropism in the coleoptile of Oryza sativa L.

Caryopses of rice (Oryza sativa L. cv. Sasanishiki) were germinated in air or under water. In submerged seedlings a twofold increase in coleoptile growth rate and an inhibition of root growth was observed. The amount of starch in the amyloplasts of submerged coleoptiles was substantially reduced compared to the air-grown control plants and plastids had a proplastidic character. During the rapid elongation of coleoptiles under water, the osmotic concentration of the press sap remained constant, whereas in air-grown coleoptiles a decrease was measured. Determination of curvature of gravistimulated air-grown and submerged shoots was carried out by placing the coleoptiles horizontally in air of 98% relative humidity. Air-grown coleoptiles reached a vertical orientation within 5 h after onset of gravistimulation. In coleoptiles germinated under water the first signs of consistent negative gravitropic bending occurred after 4-5 h and curvature was complete after 24 h. During the first 5 h of gravistimulation the water-grown coleoptiles grew at an average rate of 0.39 mm h-1, whereas in air-grown coleoptiles a rate of 0.27 mm h-1 was measured. Concomitant with the delayed onset of gravitropic bending of the water-grown coleoptiles, a change in plastid ultrastructure and an increase in starch content was observed. We conclude that the gravitropic responsiveness of the rice coleoptile depends on the presence of starch-filled amyloplasts.

Cotyledon↗

Graviresponse and the localization of its initiating cells in roots of Phleum pratense L.

Roots of Phleum pratense L. were photographed during both vertical growth and gravitropic bending, and positions of anticlinal rhizodermal cell walls were digitized on the physically upper and lower flanks of the root in the curvature plane. By using B-splines, arc lengths of these positions, i.e. distances along the root surface, values of curvature, and relative elemental rates of elongation were estimated. The whole graviresponse can be divided into phases according to growth-rate values: (i) an increase of rates on the upper side of the root and a decrease on the lower side during the first 1-1 1/2 h after the root has been moved from the vertical to a horizontal position, (ii) a transient equality of the rates on both sides, (iii) 2-3 h after the beginning of graviresponse, the growth gradient is inverted, and (iv) finally, after about 4 h, the growth rates of both flanks are approximately equal again. Curvature begins 15-20 min after horizontal placement of the root. During the first 2 h of graviresponse, plots of curvature versus arc length show one maximum value. After 2-2 1/2 h, two maximum values can be observed, the apical one near the root tip always keeping the same distance from the tip, the other one drifting basipetally relative to the growing tip. By evaluating photographs of high magnification, a group of six rhizodermal cells on each side of the root was identified which are the first cells showing gravitropic bending. These cells are located at the beginning of the elongation zone, enclosing the region 480-680 micrometers from the root tip. These cells might be target cells for a signal which the statenchyma, the site of graviperception, sends to the reacting zone of gravicurvature.

Gravitropism↗

Cytoplasmic streaming in Chara rhizoids: studies in a reduced gravitational field during parabolic flights of rockets.

In-vivo videomicroscopy of Chara rhizoids under 10(-4)g demonstrated that gravity affected the velocities of cytoplasmic streaming. Both, the acropetal and basipetal streaming velocities increased on the change to microgravity. The endogenous difference in the velocities of the oppositely directed cytoplasmic streams was maintained under microgravity, yet the difference was diminished as the basipetal streaming velocity increased more than the acropetal streaming velocity. Direction and structure of microfilaments labeled by rhodamine-phalloidin had not changed after 6 min of microgravity.

Acceleration↗

Differentiation of Mycobacterium tuberculosis strains by use of a nonradioactive Southern blot hybridization method.

The only means of dividing strains of Mycobacterium tuberculosis is by phage typing. Attempts at developing a convenient method based on differences in chromosomal DNA by detecting restriction fragment length polymorphisms (RFLP) have had limited success. This report describes the development of a nonradioactive RFLP technique that differs from most methods by using enzymes that have four base recognition sites rather than six. The restriction enzymes AluI, DdeI, HinfI, NdeII, RsaI, and TaqI were used to digest genomic DNA, and high-molecular-weight fragments were visualized after Southern blotting with digoxigenin-labeled M. tuberculosis DNA. Digestion with AluI resulted in different banding patterns among all eight clinical isolates of M. tuberculosis and three type strains from the tuberculosis complex. NdeII distinguished all but two strains, while HinfI and RsaI were unable to distinguish two pairs of strains. Digestion with the enzymes DdeI and TaqI failed to result in clearly discernible bands. These preliminary results suggest that this method provides a fine differentiation of M. tuberculosis strains and may be useful as an epidemiologic tool.

Blotting, Southern↗

The polar organization of the growing Chara rhizoid and the transport of statoliths are actin-dependent.

Horizontally positioned Chara rhizoids continue growth without gravitropic bending when the statoliths are removed from the apex by basipetal centrifugation. The transport of statoliths in centrifuged rhizoids is bidirectional: 50-60% of the statoliths are retransported on a straight course to the apex at velocities from 1 to 14 micrometers min-1, increasing towards the rhizoid tip The centrifuged statoliths which are located closest to the nucleus are basipetally transported and caught up in the cytoplasmic streaming of the cell. Those statoliths which are located near the apical side of the nucleus are transported either apically or basally. A de-novo-formation of statoliths was not observed. After retransport to the apex some statoliths transiently sediment, a process which can induce a local inhibition of cell wall growth. The rhizoid bends again gravitropically only if a few statoliths finally sediment in the apex; the more statoliths that sediment in the apex the shorter the radius of bending becomes. The transport of statoliths is mediated by actin filaments which form a network of thin filaments in the apical and subapical zone of the rhizoid, and thicker parallel bundles in the basal zone where cytoplasmic streaming occurs. Both subpopulations of actin filaments overlap in the nucleus zone.

Actin Cytoskeleton↗

Effect of agitation of BACTEC 13A blood cultures on recovery of Mycobacterium avium complex.

The effect of agitation of BACTEC 13A bottles (Becton Dickinson) on the recovery of Mycobacterium avium complex (MAC) from blood was compared with that of static incubation. A total of 265 blood specimens was inoculated in duplicate into BACTEC 13A bottles. One specimen was statically incubated at 35 degrees C, and the other was incubated with agitation on a Gyrotory shaker at 35 degrees C for the first 2 weeks and thereafter without shaking for up to 12 weeks. Of the 265 specimens, 77 (29.1%) were positive in either one or both of the paired bottles. The average detection times for the shaken and nonshaken bottles were 12.7 and 15.9 days, respectively. A total of 10.4% of the specimens in the shaken bottles became positive 1 week before those in the nonshaken bottles, and 16.9% of the shaken cultures were positive more than 2 weeks before their counterparts. A further 46.8% of the agitated specimens became positive while the corresponding nonagitated cultures remained negative. When both specimens became positive at the same time, 88% of the shaken cultures had higher growth indices than their nonshaken counterparts. A further 11 paired blood cultures were taken from patients known to be infected with MAC to assess the effect of agitation of bottles on the utility of making twice-weekly readings during the first 2 weeks of incubation. Ten of the 11 sets of specimens in the shaken bottles were positive 1 or more weeks before those in the corresponding nonshaken bottles. In the remaining set, both specimens became positive on the same day; however, the growth index of the agitated culture was higher.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteremia↗

Tuberculosis in Australia: an analysis of cases identified in reference laboratories in 1986-88.

The Special Interest Group in Mycobacteria within the Australian Society for Microbiology has carried out a collaborative study of cases of tuberculosis diagnosed in Australian reference laboratories in the years 1986, 1987 and 1988. Annual totals of 574, 584 and 613 respectively, suggest that the incidence of bacteriologically-positive tuberculosis is continuing at 3-4 cases per 100,000 population. The highest rates were detected in males over 50 and females over 65 years of age. Three-quarters of the total cases relate to pulmonary disease. Resistance to at least 1 anti-tuberculosis drug was detected in 78 (12.7%) of isolates tested in 1988. The negligible decline in incidence of tuberculosis in Australia, the high prevalence in S.E. Asian countries, and the fact that HIV-infection is an important risk factor, make it imperative that Australia's diagnostic and management programmes be maintained.

Adolescent↗

Gravity sensing mechanisms in plant cells.

Sensing of gravity is essential for the survival of plant seedlings. Therefore it is understandable that gravistimulation of only 0.5 sec-duration causes a graviresponse. The earliest graviresponses could be measured within seconds as alterations in membrane potentials of the statocytes in the root cap. Root statocytes are polarly organized. From a 6-day microgravity (10(-3) - 10(-4) g) experiment in the Spacelab D1 Mission it has been concluded that the observed polar differentiation is a result of a genetically prepatterned developmental program. Statoliths, the sedimentable organelles of statocytes, are surrounded by actin filaments which partly keep them in position. Under 6 min of microgravity during parabolic flights of rockets it could be demonstrated that the statoliths moved in the opposite direction to the initial gravity vector. It is concluded that shearing forces are exerted by microfilaments. It is supposed that the change of the position of statoliths is transmitted to gravisensitive structures of the statocytes (ER, plasma membrane) via microfilaments. As graviperception is influenced by calcium ions, it is suggested that these interactions regulate the activity of ion channels and/or pumps in the membranes thus initiating the graviresponse chain. In the case of cytoplasmic streaming in Chara rhizoids, the endogenous difference between the opposing streaming directions is diminished under microgravity during the flights of rockets. Possibly, shear stresses are affected by gravity, thus inducing gravity-related differences in the streaming velocities via actin filaments.

Actin Cytoskeleton↗

Oriented movement of statoliths studied in a reduced gravitational field during parabolic flights of rockets.

During five rocket flights (TEXUS 18, 19, 21, 23 and 25), experiments were performed to investigate the behaviour of statoliths in rhizoids of the green alga Charo globularia Thuill. and in statocytes of cress (Lepidium sativum L.) roots, when the gravitational field changed to approx. l0(-4) g (i.e. microgravity) during the parabolic flight (lasting for 301-390 s) of the rockets. The position of statoliths was only slightly influenced by the conditions during launch, e.g. vibration, acceleration and rotation of the rocket. Within approx. 6 min of microgravity conditions the shape of the statolith complex in the rhizoids changed from a transversely oriented lens into a longitudinally oriented spindle. The center of the statolith complex moved approx. 14 micrometers and 3.6 micrometers in rhizoids and root statocytes, respectively, in the opposite direction to the originally acting gravity vector. The kinetics of statolith displacement in rhizoids demonstrate that the velocity was nearly constant under microgravity whereas it decreased remarkably after inversion of rhizoids on Earth. It can be concluded that on Earth the position of statoliths in both rhizoids and root statocytes depends on the balance of two forces, i.e. the gravitational force and the counteracting force mediated by microfilaments.

Actin Cytoskeleton↗

Hormone treatment of roots causes not only a reversible loss of starch but also of structural polarity in statocytes.

Treatment of cress (Lepidium sativum L.) roots with phytohormones (4.3 x 10(-5) M gibberellic acid plus 4.3 x 10(-5) M kinetin, 30 h; T.H. Iversen, 1969, Physiol. Plant. 22, 1251-1262) caused not only complete destarching of amyloplasts but also destruction of the polar arrangement of cell organelles in statocytes. The nucleus was not positioned exclusively near the proximal cell pole as in the controls but was also found near the distal cell pole. The endoplasmic reticulum (ER) was no longer organized in parallel sheets at the distal cell pole but instead the ER-cisternae were randomly distributed. Additionally, the statocytes from hormone-treated roots contained a large central vacuole instead of numerous small ones as in the controls. The starch-free plastids had a reduced volume and an amoeboid shape. They did not sediment but were randomly distributed in the statocytes. The loss of structural polarity was accompanied by loss of graviresponsiveness although root growth still occurred. Twenty-two hours after removal of the hormones, structural polarity was restored and starch was resynthesized. The newly formed starch grains were smaller and more numerous per amyloplast compared to the controls. It is concluded that loss of gravisensitivity of roots after hormone treatment cannot be solely attributed to the loss of amyloplastic starch because there is a concomitant loss in the polar organisation of the statocyte.

Adenine↗

Statoliths and microfilaments in plant cells.

Microfilaments have been demonstrated in rhizoids of Chara fragilis Desvaux by labelling of actin with rhodamine-conjugated phalloidin. Each rhizoid contains thick microfilament-bundles arranged longitudinally in the basal region. In the subapical and apical regions, much thinner bundles exist which contact the statoliths and encircle them in the form of a dense envelope. In root statocytes from Lepidium sativum L. the presence of an actin network is indicated by the fact that application of cytochalasin B (25 micrograms ml-1 for 4 h) results in an approximately threefold increase in the rate of statolith (amyloplast) sedimentation relative to controls. It is concluded that in gravity-perceiving plant cells statoliths may trigger the transduction mechanism via actin filaments.

Actin Cytoskeleton↗

The polarity of statocytes and the gravisensitivity of roots are dependent on the concentration of calcium in statocytes.

In order to examine a possible role of calcium in graviperception, the calcium ionophore A23187 was used to elevate the concentration of free cytoplasmic calcium in statocytes of the roots of Lepidium sativum L. After a brief incubation (30 min) in a medium that contained 10 micromoles A23187 and 5.5 micromoles CaCl2, 50% of the roots bent gravitropically during a subsequent 2 h of horizontal exposure, with an angle of curvature that varied from 5 degrees to 70 degrees. The corresponding statocytes exhibited a polar arrangement of cell organelles as did the controls. However, in statocytes from 50% of the roots which were not curved after gravistimulation a portion of the distal endoplasmic reticulum (ER) complex was displaced in the direction of gravity within 30 min of horizontal exposure. After washing of the briefly treated roots for 24 h with 1% dimethylsulfoxide the percentage of gravitropically bending roots increased to approximately 80%, but the angle of curvature amounted to only 5 degrees-10 degrees. Longer treatment (2 h) with A23187 caused a complete loss of graviresponsiveness which was accompanied by disintegration of statocyte polarity. We concluded from these results that i) calcium is involved in graviperception and ii) gravisensitivity depends on the integrity of statocytes.

Brassicaceae↗