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At least 1,099 records · Page 61Linked to original sources
PubMed1994

Hodgkin disease: Hodgkin and Reed-Sternberg cells picked from histological sections show clonal immunoglobulin gene rearrangements and appear to be derived from B cells at various stages of development.

Hodgkin disease (HD) is characterized by a small number of putative malignant cells [Hodgkin and Reed-Sternberg (HRS) cells] among a background of lymphocytes and histiocytes. The lineage of HRS cells is still elusive and a clonal origin of these rare cells has not formally been demonstrated. We isolated HRS cells by micromanipulation from histological sections of three cases of Hodgkin lymphoma (each representing a distinct subtype of the disease) and analyzed individual cells for immunoglobulin variable (V) gene rearrangements by PCR. In each of the three cases a single heavy-chain V (VH) (and in one case, in addition, a kappa light-chain) gene rearrangement was amplified from the HRS cells, identifying these cells as members of a single clone. A potentially functional VH rearrangement was obtained from a case of nodular sclerosis HD. Somatic mutations and intraclonal diversity in the VH genes indicate a germinal center B-cell origin of the HRS cells in a case of lymphocyte-predominant HD, whereas in a case of mixed-cellularity HD the sequence analysis revealed only nonfunctional V gene rearrangements, suggesting a pre-B-cell origin. This indicates that HRS cells can originate from B-lineage cells at various stages of development.

Adult↗
PubMed1996

Magnetic resonance microscopy and spectroscopy reveal kinetics of cryoprotectant permeation in a multicompartmental biological system.

Successful cryopreservation of most multicompartmental biological systems has not been achieved. One prerequisite for success is quantitative information on cryoprotectant permeation into and amongst the compartments. This report describes direct measurements of cryoprotectant permeation into a multicompartmental system using chemical shift selective magnetic resonance (MR) microscopy and MR spectroscopy. We used the developing zebrafish embryo as a model for studying these complex systems because these embryos are composed of two membrane-limited compartments: (i) a large yolk (surrounded by the yolk syncytial layer) and (ii) differentiating blastoderm cells (each surrounded by a plasma membrane). MR images of the spatial distribution of three cryoprotectants (dimethyl sulfoxide, propylene glycol, and methanol) demonstrated that methanol permeated the entire embryo within 15 min. In contrast, the other cryoprotectants exhibited little or no permeation over 2.5 h. MR spectroscopy and microinjections of cryoprotectants into the yolk inferred that the yolk syncytial layer plays a critical role in limiting the permeation of some cryoprotectants throughout the embryo. This study demonstrates the power of MR technology combined with micromanipulation for elucidating key physiological factors in cryobiology.

Animals↗
PubMed1996

Role of cortical N-methyl-D-aspartate receptors in auditory sensory memory and mismatch negativity generation: implications for schizophrenia.

Working memory refers to the ability of the brain to store and manipulate information over brief time periods, ranging from seconds to minutes. As opposed to long-term memory, which is critically dependent upon hippocampal processing, critical substrates for working memory are distributed in a modality-specific fashion throughout cortex. N-methyl-D-aspartate (NMDA) receptors play a crucial role in the initiation of long-term memory. Neurochemical mechanisms underlying the transient memory storage required for working memory, however, remain obscure. Auditory sensory memory, which refers to the ability of the brain to retain transient representations of the physical features (e.g., pitch) of simple auditory stimuli for periods of up to approximately 30 sec, represents one of the simplest components of the brain working memory system. Functioning of the auditory sensory memory system is indexed by the generation of a well-defined event-related potential, termed mismatch negativity (MMN). MMN can thus be used as an objective index of auditory sensory memory functioning and a probe for investigating underlying neurochemical mechanisms. Monkeys generate cortical activity in response to deviant stimuli that closely resembles human MMN. This study uses a combination of intracortical recording and pharmacological micromanipulations in awake monkeys to demonstrate that both competitive and noncompetitive NMDA antagonists block the generation of MMN without affecting prior obligatory activity in primary auditory cortex. These findings suggest that, on a neurophysiological level, MMN represents selective current flow through open, unblocked NMDA channels. Furthermore, they suggest a crucial role of cortical NMDA receptors in the assessment of stimulus familiarity/unfamiliarity, which is a key process underlying working memory performance.

Animals↗
PubMed1996

Development of a bovine X chromosome linkage group and painting probes to assess cattle, sheep, and goat X chromosome segment homologies.

The X chromosome linkage group is conserved in placental mammals. However, X chromosome morphological differences, due to internal chromosome rearrangements, exist among mammalian species. We have developed bovine chromosome painting probes for Xp and Xq to assess segment homologies between the submetacentric bovine X chromosome and the acrocentric sheep and goat X chromosomes. These painting probes and their corresponding DNA libraries were developed by chromosome micromanipulation, DNA micropurification, microcloning, and PCR amplification. The bovine Xp painting probe identified an interstitially located homologous segment in the sheep and goat Xq region, most probably resulting from chromosome inversion. Ten type II (microsatellite) markers obtained from the bovine Xq library and five other X chromosome assigned, but unlinked, markers were used to generate a linkage map for Xq spanning 89.4 centimorgans. The chromosome painting probes and molecular markers generated in this study would be useful for comparative mapping and tracing of internal X chromosome rearrangements in all ruminant species and would contribute to the understanding of mammalian sex chromosome evolution.

Animals↗
PubMed1997

Hodgkin and Reed-Sternberg cells in lymphocyte predominant Hodgkin disease represent clonal populations of germinal center-derived tumor B cells.

Among the four subtypes of Hodgkin disease (HD), lymphocyte-predominant (LP) HD is now generally considered as a separate entity. The B cell nature of the typical Hodgkin and Reed-Sternberg (HRS) cells and their variants (L and H, lymphocytic and histiocytic cells) in LP HD has long been suspected, but the question of whether these cells represent a true tumor clone is unclear. We previously demonstrated clonal Ig gene rearrangements in one case of LP HD. In the present study, five cases of LP HD were analyzed by micromanipulation of single HRS cells from frozen tissue sections and DNA amplification of rearranged Ig heavy chain genes from those cells. Clonal V gene rearrangements harboring somatic mutations were detected in each case. In three cases ongoing somatic mutation was evident. This shows that HRS cells in LP HD are a clonal tumor population derived from germinal center B cells. The pattern of somatic mutation indicates that HRS cells in LP HD are selected for antibody expression. This, and the presence of ongoing mutation discriminates LP from classical HD.

Adolescent↗
PubMed1997

Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure.

We report here that living cells and nuclei are hard-wired such that a mechanical tug on cell surface receptors can immediately change the organization of molecular assemblies in the cytoplasm and nucleus. When integrins were pulled by micromanipulating bound microbeads or micropipettes, cytoskeletal filaments reoriented, nuclei distorted, and nucleoli redistributed along the axis of the applied tension field. These effects were specific for integrins, independent of cortical membrane distortion, and were mediated by direct linkages between the cytoskeleton and nucleus. Actin microfilaments mediated force transfer to the nucleus at low strain; however, tearing of the actin gel resulted with greater distortion. In contrast, intermediate filaments effectively mediated force transfer to the nucleus under both conditions. These filament systems also acted as molecular guy wires to mechanically stiffen the nucleus and anchor it in place, whereas microtubules acted to hold open the intermediate filament lattice and to stabilize the nucleus against lateral compression. Molecular connections between integrins, cytoskeletal filaments, and nuclear scaffolds may therefore provide a discrete path for mechanical signal transfer through cells as well as a mechanism for producing integrated changes in cell and nuclear structure in response to changes in extracellular matrix adhesivity or mechanics.

Acrylamide↗
PubMed1998

RecA binding to a single double-stranded DNA molecule: a possible role of DNA conformational fluctuations.

Most genetic regulatory mechanisms involve protein-DNA interactions. In these processes, the classical Watson-Crick DNA structure sometimes is distorted severely, which in turn enables the precise recognition of the specific sites by the protein. Despite its key importance, very little is known about such deformation processes. To address this general question, we have studied a model system, namely, RecA binding to double-stranded DNA. Results from micromanipulation experiments indicate that RecA binds strongly to stretched DNA; based on this observation, we propose that spontaneous thermal stretching fluctuations may play a role in the binding of RecA to DNA. This has fundamental implications for the protein-DNA binding mechanism, which must therefore rely in part on a combination of flexibility and thermal fluctuations of the DNA structure. We also show that this mechanism is sequence sensitive. Theoretical simulations support this interpretation of our experimental results, and it is argued that this is of broad relevance to DNA-protein interactions.

Bacteriophage lambda↗
PubMed2000

Manipulating the genetic identity and biochemical surface properties of individual cells with electric-field-induced fusion.

A method for cell-cell and cell-liposome fusion at the single-cell level is described. Individual cells or liposomes were first selected and manipulated either by optical trapping or by adhesion to a micromanipulator-controlled ultramicroelectrode. Spatially selective fusion of the cell-cell or cell-liposome pair was achieved by the application of a highly focused electric field through a pair of 5-micrometer o.d. carbon-fiber ultramicroelectrodes. The ability to fuse together single cells opens new possibilities in the manipulation of the genetic and cellular makeup of individual cells in a controlled manner. In the study of cellular networks, for example, the alteration of the biochemical identity of a selected cell can have a profound effect on the behavior of the entire network. Fusion of a single liposome with a target cell allows the introduction of the liposomal content into the cell interior as well as the addition of lipids and membrane proteins onto the cell surface. This cell-liposome fusion represents an approach to the manipulation of the cytoplasmic contents and surface properties of single cells. As an example, we have introduced a membrane protein (gamma-glutamyltransferase) reconstituted in liposomes into the cell plasma membrane.

Animals↗
PubMed2000

The mechanical properties of Saccharomyces cerevisiae.

Cell-wall mechanical properties play an integral part in the growth and form of Saccharomyces cerevisiae. In contrast to the tremendous knowledge on the genetics of S. cerevisiae, almost nothing is known about its mechanical properties. We have developed a micromanipulation technique to measure the force required to burst single cells and have recently established a mathematical model to extract the mechanical properties of the cell wall from such data. Here we determine the average surface modulus of the S. cerevisiae cell wall to be 11.1 +/- 0.6 N/m and 12.9 +/- 0.7 N/m in exponential and stationary phases, respectively, giving corresponding Young's moduli of 112 +/- 6 MPa and 107 +/- 6 MPa. This result demonstrates that yeast cell populations strengthen as they enter stationary phase by increasing wall thickness and hence the surface modulus, without altering the average elastic properties of the cell-wall material. We also determined the average breaking strain of the cell wall to be 82% +/- 3% in exponential phase and 80% +/- 3% in stationary phase. This finding provides a failure criterion that can be used to predict when applied stresses (e.g., because of fluid flow) will lead to wall rupture. This work analyzes yeast compression experiments in different growth phases by using engineering methodology.

Cell Wall↗
PubMed2005

P2Y6 nucleotide receptors activate NF-kappaB and increase survival of osteoclasts.

Nucleotides, released from cells during inflammation and by mechanical stimulation, act through the P2 family of nucleotide receptors. Previous studies have demonstrated the expression of P2Y1 and P2Y2 receptors in osteoclasts. The aim of this study was to determine whether osteoclast P2Y receptors signal through NF-kappaB, a key transcription factor regulating osteoclastogenesis. Immunofluorescence was used to detect the p65 subunit of NF-kappaB, which upon activation translocates from the cytosol to nuclei. Low levels of NF-kappaB activation were observed in untreated rabbit osteoclasts and in those exposed to 2-methylthio ADP (P2Y1 agonist) or ATP or UTP (P2Y2 agonists). In contrast, UDP or INS48823 (P2Y6 agonists) induced a significant increase in the number of cells exhibiting NF-kappaB activation, a process sensitive to the proteasome inhibitor lactacystin. In osteoclasts purified by micromanipulation, reverse transcription-PCR revealed the presence of P2Y1, P2Y2, and P2Y6 receptor transcripts, and application of agonists for these receptors induced the transient rise of cytosolic calcium. Treatment of rat osteoclasts with UDP or INS48823, but not 2-methylthio ADP or UTP, increased osteoclast survival. Osteoprotegerin (a decoy receptor for RANK ligand) did not significantly alter the effects of UDP on NF-kappaB localization or osteoclast survival, consistent with a direct action. Moreover, SN50 (cell-permeable peptide inhibitor of NF-kappaB) suppressed the enhancement of cell survival induced by UDP and INS48823. Our findings demonstrate the presence of functional P2Y6 receptors in osteoclasts. Thus, nucleotides, following their release at sites of inflammation and mechanical stimulation, can act through P2Y6 receptors to initiate NF-kappaB signaling and enhance osteoclast survival.

Active Transport, Cell Nucleus↗
PubMed2002

Arcus as a tensegrity structure in the arolium of wasps (Hymenoptera: Vespidae).

The unfolding of the hymenopteran attachment pad (arolium) may be achieved in two ways, hydraulic and mechanical. The first was confirmed in experiments with pressure applied to more proximal leg parts and on immersion in hypotonic solutions. Presumably, this way of unfolding does not play an important role for a living hornet. Mechanical unfolding was studied experimentally with the aid of a micromanipulator pulling the tendon of the musculus retractor unguis. Ablation experiments on different parts of the arolium indicated that the arcus is the most crucial element for mechanical unfolding. The shape of the arcus in closed and open conditions was measured using a 3D measurement microscope and reconstructed by means of 3D computer graphics. The arcus coils up upon being freed from the arolium tissues, and coils up even more after immersion into a 10% aqueous solution of NaOH. Geometrical models of the arcus are proposed, from which the rotational moment of elasticity is derived. Conformations and deformations of the arcus are quantified in order to explain its role in the folding and unfolding processes of the arolium. The diversity of approaches supports the idea that the arcus is a prestressed (tensegrity) structure providing immediate, soft, and graded transmission of forces during folding and unfolding action of the arolium.

Journal Article↗
PubMed1999

Advantages and dangers of erbium laser application in stapedotomy.

Among different types of lasers, the erbium laser exhibits particularly favourable characteristics for ear surgery. Experiments with application of erbium laser pulses to the isolated stapes connected to an inner ear model confirmed that there was virtually no thermal effect to the inner ear liquid and that the border damage zone on the stapes footplate perforation did not exceed 5-10 microm. Erbium laser pulses, however, produce pressure waves due to the explosive ablation of tissue. Pulses of 10 to 17 J/cm2 producing pressure waves between 140 and 160 dB appear to be a limit for clinical application. With these criteria, an in-house built erbium YAG laser with a fiberoptic delivery device was used in 15 patients for stapedotomy. A special microhandpiece, where a zirconium fluoride fiber was connected to a quartz tip, was developed. In addition, three patients had stapedotomy with a commercially available Zeiss (Opmi TwinER) microscope equipped with a micromanipulator-operated erbium laser beam. One year after surgery, the air-bone gap was closed in all patients to within 20 dB between 0.5 and 3 kHz with only minor permanent bone conduction threshold losses (< 20 dB). However, we observed an immediate postoperative middle and high frequency loss of up to 75 dB on bone conduction threshold measurements 2 h after surgery, suggesting an acoustic traumatization by the erbium laser. This threshold shift recovered close to preoperative values within 6 h. These observations prompted us to discontinue the clinical use of erbium laser for stapedotomy until the problem of temporary acoustic traumatization is resolved.

Adult↗
PubMed1989

Evidence for selection of virulent sub-populations of Entamoeba histolytica by cholesterol.

Quantitatively much higher Concanavalin A (Con. A) agglutinability, haemolytic potency, and activities of acid hydrolases, namely phosphatase (EC 3.1.3.2), ribonuclease (EC 2.7.7.16), deoxyribonuclease (EC 3.1.4.5) and proteinase--were observed in a virulent strain of Entamoeba histolytica (IP-106), as compared to attenuated and avirulent strains (200-NIH) and DKB respectively. In addition, significant differences in these parameters were observed among clonal cultures derived from the latter two cultures by cultivation of single amoebic cells picked out by micromanipulation. Repeated sub-culturing of parent cultures of both these strains in cholesterol-enriched medium resulted in marked enhancement of all the above activities, but no such change occurred in the derived clonal cultures following similar cholesterol treatment. The implication of these findings in relation to enhancement of the virulence of E. histolytica by cholesterol is discussed.

Acid Phosphatase↗
PubMedDate not supplied

Mechanical properties of melamine-formaldehyde microcapsules.

The mechanical properties of melamine-formaldehyde (M-F) microcapsules were studied using a micromanipulation technique. Single microcapsules with diameters of 1-12 microm were compressed and held between two parallel planes, compressed and released, and compressed to burst at different speeds, whilst the force being imposed on the microcapsules and their deformation were measured simultaneously. This force increased as single microcapsules were compressed and then relaxed slightly as they were held. When the microcapsules were repeatedly compressed and released, a pseudo yield point was found for each microcapsule. Before the microcapsules were compressed to this point, the deformed microcapsules recovered to their original shape once the force was removed. However, when the deformation was beyond the 'yield point' there was profound hysteresis and the microcapsules showed plastic behaviour. As the microcapsules were compressed to burst at different speeds, ranging from 0.5-6.0 microm/s, it was found that their mean bursting forces did not change significantly. The deformations at the pseudo yield point and at bursting were also independent of the compression speed. On average, these melamine-formaldehyde microcapsules reached their 'yield point' at a deformation of about 19 +/- 1%, and burst at a deformation of 70 +/- 1%.

Biomechanical Phenomena↗
PubMed2006

Synergy between visual and auditory signals and its influence on the follow-up regulation quality.

The study aimed at verifying a hypothesis that supporting a visual signal of regulation deviation with an auditory one could improve the quality of regulation; the operator would have better information on machine operation. A special simulator was applied to follow-up tracking with a manual lever which controlled vertical movements of a cursor on a monitor screen. Simultaneously with visual information on screen, the operator was provided with an auditory deviation signal of pre-determined characteristics. 33 young males underwent the test. It was found that supporting a visual signal with an auditory one resulted in an improvement in the regulation quality by 5-6%, which proved synergy between those signals. The results may be used in designing tele- and servo-mechanisms, especially for remote control machinery, e.g., inspection robots or micromanipulators controlled by operators in a follow-up system.

Acoustic Stimulation↗
PubMed2000

Intracytoplasmic sperm injection in assisted reproductive technology: an evaluation.

Since the first reports of successful pregnancies in humans after treatment with intracytoplasmic sperm injection (ICSI), intensive investigations have focused on several important aspects of this form of assisted reproductive technology. In addition to the technical development of ICSI and increasing understanding of the biochemical and biophysical processes involved during fertilization after injection of an immobilized sperm, studies have aimed to define the indications for patients for a first-line ICSI treatment. One of the major concerns is of course the safety of the technique in terms of the health and reproductive life of the babies born after ICSI. The rhesus monkey is an excellent model to investigate all aspects of this micromanipulation technique. This article provides an evaluation of ICSI.

Journal Article↗
PubMed2004

Mechanical characterization of biocompatible microspheres and microcapsules by direct compression.

The mechanical properties of biocompatible microparticles including alginate microspheres and alginate-chitosan microcapsules with different wall thickness were determined using a micromanipulation technique. Single microparticles with diameters of 20-60 microm were compressed to a given deformation and held, and compressed to rupture at different speeds. The corresponding force imposed on them was measured simultaneously by a force transducer. Results showed that the force imposed on these particles increased when they were compressed, but relaxed significantly when they were held. For alginate microspheres, the faster the compression speed was, the greater the force being imposed on them at a given deformation. Alginate-chitosan microcapsules showed less force relaxation when they were held, compared with alginate microspheres. The thicker their wall was, the less significant force relaxation the microcapsules exhibited. The mean rupture force of alginate microspheres increased with the compression speed, but this effect in general became less for alginate-chitosan microcapsules, which depended on their wall thickness. However, the deformation at rupture for all three samples was independent of the compression speed. On average, the alginate-chitosan microcapsules were bigger than alginate microspheres and had a greater rupture force.

Alginates↗
PubMed1985

The total length of spindle microtubules depends on the number of chromosomes present.

We extracted chromosomes by micromanipulation from Melanoplus differentialis spermatocytes, producing metaphase spindles with only one or a few chromosomes instead of the usual complement of 23. Cells with various numbers of chromosomes were prepared for electron microscopy, and spindle microtubule length was measured. A constant increment of microtubule length was lost upon the removal of each chromosome; we estimate that only approximately 40% of the original length would remain in the total absence of chromosomes. Unexpectedly, kinetochore microtubules were not the only ones affected when chromosomes were removed: nonkinetochore microtubules accounted for a substantial fraction of the total length lost. No compensatory increase in microtubule length outside the spindle was found. Studies by others show that the kinetochore microtubules of extracted chromosomes are left behind in the cell and dissassemble. The resulting increase in subunit concentration would be expected from in vitro studies to drive microtubule assembly until the original total microtubule length was restored, but that did not happen in these living cells. We conclude that the assembly of a certain, large fraction of microtubule subunits into stable microtubules is dependent on the presence of chromosomes. Possible explanations include (a) limits on microtubule length that prevent any net assembly of the subunits released after chromosomes are removed or (b) a promotion of microtubule assembly by chromosomes, which therefore is reduced in their absence. Chromosome-dependent regulation of microtubule length may account for some features of normal mitosis.

Animals↗