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PubMed1994

Displacement and translocation of osteoblast-like cells by osteoclasts.

Rabbit osteoclasts and rabbit osteoblast-like stroma cells (OB cells) were placed onto plastic surfaces and the migration patterns of individual osteoclasts and osteoclast-OB interactions were analyzed with time-lapse recording. To induce directed migration, the cultures were exposed to an electrical field of 0.01 or 0.1 V/mm. At 0.1 V/mm, osteoclasts moved directly toward the anode in some cases, clearing OB cells from their path of migration. In other cases, osteoclasts migrated toward the anode for part of the time but then changed direction and moved toward groups of OB cells. Observations were made on osteoclasts interacting with single OB cells or small colonies and on osteoclasts interacting with OB monolayers, at both field strengths; the results were independent of field strength. There were several characteristic behaviors. With single OB cells and small OB colonies, retraction of OB cells upon contact with the osteoclast was the predominant mechanism whereby these cells begin to move out of the path of the osteoclast. A pronounced ruffling or blebbing of the OB cell membrane often followed retraction. When osteoclasts displaced OB cells that were part of a monolayer, extension of an osteoclast lamellipodium underneath the edge of the OB cell layer generally preceded partial retraction of the OB cells involved. It sometimes appeared as if the detached or partially detached OB cells were "pushed" by the osteoclast, which in some cases resulted in OB cells being moved hundreds of microns in a period of a few hours, at rates comparable to the normal speed for osteoclast migration (congruent to 100 microns/h), much faster than the normal speed for OB cells (congruent to 10 microns/h).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Chemical selectivity in micellar electrokinetic chromatography: characterization of solute-micelle interactions for classification of surfactants.

The influence of surfactant type on migration behavior and chemical selectivity in micellar electrokinetic chromatography (MEKC) is investigated through linear solvation energy relationships (LSER) and functional group selectivities. In LSER modeling, solutes' capacity factors are correlated with their structural descriptors such as size, dipolarity, and hydrogen-bonding abilities. Using the LSER methodology, useful information about the nature of solute interactions with different types of surfactant aggregates can be obtained since capacity factor in MEKC is directly related to solute distribution between the bulk aqueous solvent and micelles. High correlations were observed for different LSER models of migration behavior in MEKC for a group of 60 uncharged aromatic compounds of non-hydrogen bonding (NHB), hydrogen-bonding acceptor (HBA) bases, and hydrogen-bonding donor (HBD) acids. In two anionic, hydrocarbon micellar systems of sodium dodecyl sulfate (SDS) and sodium cholate (SC), retention is primarily influenced by the size of molecules and their hydrogen bond accepting basicity. Their dipolarity/polarizability and hydrogen bond donating acidity play minor roles. Capacity factors of solutes in SDS and SC systems increase with their size and decrease for stronger hydrogen bond acceptor bases. These results are similar to those observed for other systems where hydrophobic interactions play a major role, e.g., solute distribution in the 1-octanol-water solvent system or retention in reversed phase LC. In MEKC with an anionic fluorocarbon surfactant, lithium perfluorooctanesulfonate (LiPFOS), however, size and solute HBD acidity are the two predominant factors. The LSER results indicate that compounds find the SDS micellar environments slightly less cohesive (i.e., more apolar) than the SC micelles, while the LiPFOS micelles are the most cohesive among the three surfactant aggregates and 1-octanol provides the least cohesive environment. The fluorocarbon micelles of LiPFOS, on the other hand, are the strongest hydrogen bond donor acids, followed by SDS, SC, and 1-octanol, respectively. The SC micelles have the most hydrogen bond acceptor basic characteristics, followed by 1-octanol, SDS, and LiPFOS micelles. It can be concluded that selectivity differences between these surfactant types in MEKC is primarily due to hydrogen-bonding interactions rather than the dipolar interactions. Comparing the perfluorinated and the hydrocarbon surfactants, even solute size can play a role in selective migration patterns. In addition, information from polar and hydrophobic group selectivities confirm the LSER conclusions about the underlying interactions that control migration behavior and chemical selectivity in MEKC.(ABSTRACT TRUNCATED AT 400 WORDS)

Chromatography, Liquid↗

PubMed1997

Microgel electrophoresis: sensitivity, mechanisms, and DNA electrostretching.

Based on the treatment of microgels to remove proteins, we speculate that proteins may be bound to DNA in the microgels even after electrophoresis. We speculate that some DNA single-strand breaks may be a reflection of these protein-DNA complexes. We suggest methods to limit such artifacts, and present data demonstrating a lymphocyte DNA double-strand break sensitivity of 12.5 rads and day-to-day reproducibility of microgel electrophoresis using these principles. Extending these principles, we describe DNA behavior during alkaline and neutral microgel electrophoresis based on observations of the stained DNA and its migration patterns. During microgel electrophoresis, individual DNA molecules behave as if anchored at one end while the other end is free to migrate in response to the electric field. We capitalize on this behavior by developing a neutral microgel method to stretch chromosomes.

DNA Damage↗

PubMed1994

Cell viability and migration in nerve isografts and allografts.

Even though autogenous nerve grafts are used frequently, there is little information concerning cell survival rates and migration patterns, following peripheral nerve grafting. Labeling techniques with a vital fluorescent stain (PKH-26, Zynaxis Cell Science, Malvern, PA) allow cell migrations from both the nerve graft and host nerve to be tracked for up to 45 days from the time of nerve transplantation. With this labeling technique, two phases of nerve graft incorporation were identified, early and late, in an animal model using inbred Lewis and Brown-Norway rats. In genetically identical Lewis rats, isografts were performed as a means of modeling the autografts used clinically. At approximately 3 days after isogeneic transplantation, with the proximal host nerve end labeled, there was an early migration of host cells from the proximal nerve end into the epineural tissue of the nerve graft. At 25 days, a late phase was evident, with fluorescent labeling of host cells into the perineural and endoneural tissues. When the nerve grafts were labeled, the label persisted for up to 45 days, indicating viability of the graft. Cells migrated from the labeled nerve graft into the distal host nerve segment. Cellular migration from peripheral nerve tissue, following allograft transplantation, was initially similar to the isograft studies. But after 25 days, with the proximal host nerve end labeled, a significant decrease in the labeled host cells migrating into the graft was noted (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1999

Gene genealogies in geographically structured populations.

Population genetics theory has dealt only with the spatial or geographic pattern of degrees of relatedness or genetic similarity separately for each point in time. However, a frequent goal of experimental studies is to infer migration patterns that occurred in the past or over extended periods of time. To fully understand how a present geographic pattern of genetic variation reflects one in the past, it is necessary to build genealogy models that directly relate the two. For the first time, space-time probabilities of identity by descent and coalescence probabilities are formulated and characterized in this article. Formulations for general migration processes are developed and applied to specific types of systems. The results can be used to determine the level of certainty that genes found in present populations are descended from ancient genes in the same population or nearby populations vs. geographically distant populations. Some parameter combinations result in past populations that are quite distant geographically being essentially as likely to contain ancestors of genes at a given population as the past population located at the same place. This has implications for the geographic point of origin of ancestral, "Eve," genes. The results also form the first model for emerging "space-time" molecular genetic data.

Genealogy and Heraldry↗

PubMed1980

Re-evaluation of the lymphocyte migration through the high-endothelial venules. Light and electron microscopic studies on the opossum's lymph node.

We have described two types of lymphocyte migration through the HEV. In the first case the lymphocytes do not show constriction ring during the diapedesis. 97% of lymphocytes migrate in this manner. In this migration pattern the lymphocyte gets through the endothelium by means of flexibility and pliability of the endothelium. It is suggested that these lymphocytes migrate into the blood from the lymph node. In the second case the lymphocytes undergo morphological changes, they form constriction ring and they migrate from the blood into the lymph node.

Animals↗

PubMed1998

Immunological characterization of adenosine A2A receptors in human and porcine cardiovascular tissues.

Antipeptide antibody was raised in rabbit against the sequence (361-390) of RDC-8, the presumed adenosine A2A receptor cDNA from canine. The antibody titer was estimated by solid phase radioimmunoassay. Western blot analysis under reducing conditions identified a major 45 +/- 1 kDa protein in bovine striatal membranes. This immunoreactive band was competed in the presence of excess peptide. Furthermore, the antibody recognized a single 45-kDa immunoreactive band in membranes from cells transfected with the recombinant human adenosine A2A receptors, whereas, fail to cross-react with membranes from cells transfected with recombinant rat A1 and human A3 receptors. Membranes from human and porcine coronary artery, ventricle, atria and platelets (human only) showed a major immunoreactive band at 45 +/- 1 kDa size. Under nonreducing conditions, the migration patterns of the immunoreactive bands were not altered indicating the absence of interchain disulfide bond. The 45-kDa immunoreactive band co-migrated with 2-[4-(2-¿2-[(4-aminophenyl)methylcarbonylamino]ethyl-aminocarbo nyl¿et hyl)phenyl]ethylamino-5'-Nethylcarboxamidoadenosine photoaffinity labeled A2A adenosine receptor using SANPAH as the photoaffinity cross-linker. We provide immunological evidence for the presence of A2A adenosine receptor in human cardiovascular tissues that exists as a 45-kDa monomeric protein. This study also presents evidence for the presence of A2A adenosine receptor in ventricle and atria in both human and porcine.

Adolescent↗

PubMed2001

Results of hip revision using the Exeter stem, impacted allograft bone, and cement.

The Exeter stem and impacted, morselized allograft bone and cement were used in the revisions of 18 consecutive femoral components (17 patients). The primary arthroplasty had been done because of osteoarthritis. All of the femoral components were revised for the first time because of aseptic loosening. The migration pattern of the Exeter stem after revision was studied using roentgen stereophotogrammetric analysis. At 2 years after surgery, all 18 femoral stems had migrated in the distal direction (average, 2.5 mm). In addition, seven of the stems had migrated in the medial direction (average, 1.3 mm), and two stems had migrated in the lateral direction (0.5 mm and 1 mm, respectively). Sixteen of the femoral stems also had migrated in the posterior direction (average, 2.9 mm), but none migrated in the anterior direction. The migration rate decreased gradually with time during the followup. Six femoral stems continued to migrate between 1.5 and 2 years after surgery. In patients with major femoral bone deficiency at the time of hip revision, the use of impacted morselized allograft bone and cement yielded an initial fixation similar to that obtained in conventionally cemented revisions. Pain had improved in all patients at the 2-year followup.

Aged↗

PubMed1981

Inverse correlation between neutrophil microtubule numbers and enhanced random migration.

The random migration of neutrophils under agarose as measured by the number of cells leaving the well, is enhanced when the pH or the osmolality of the medium is reduced or when microtubule agents are used. Concentrations of colchicine above 5 x 10-7 M increased the number of cells migrating and decreased the mean number of centriolar microtubules in a dose-dependent fashion from 16 to 4 per 4 micron 2 at 10-5 M. The distance that colchicine-treated neutrophils migrated from the well was not different from the control. Lowering the pH from 7.4 to 6.0 also increased random migration and decreased pericentriolar microtubules from a mean of 16 to a mean of 10. At pH 6.0, both the number of cells that migrated and the distance the cells forming the leading edge travelled from the well were increased. Since peripheral microtubules may play a greater role in cell migration than centiolar ones, we examined the numerical density of microtubules in the peripheral cytoplasm. Lowering the medium pH reduced the mean number of microtubules per 10 micron 2 from 6 to 2. Colchicine reduced micro-tubules in the same area to I. At the low pH, colchicine reduced even further the numbers of both centriole-associated and peripheral microtubules but the migration pattern was the same as that seen at pH 6.0 without colchicine. Lowering medium osmolality from 280 to 230 m-osmol increased random migration but did not affect microtubule numbers. The addition of colchicine to this system decreased microtubule numbers and increased migration even further. Conditions that enhanced neutrophil migration also affected cell shape. Whereas cells at pH 7.4 were generally fan-shaped with a broad, smooth leading edge, cells at pH 6.0 with or without colchicine were long and narrow. Neutrophils at pH 7.4 but 230 m-osmol had a scalloped edge, which often appeared thickened. This too was not altered by colchicine. The morphology of cells treated with colchicine was similar to controls except for the more frequent presence of long retraction fibres. Each of these treatments thus appears to act on a different aspect of the cell's locomotory apparatus. The mechanisms by which colchicine and lowered ph enhance migration may partially overlap since both significantly decrease peripheral microtubules. The data suggest that microtubules play a constraining role within the cell, limiting the ability of the cell to move and change direction.

Adult↗

PubMed1992

Analysis of genetic variation of epizootic hemorrhagic disease virus and bluetongue virus field isolates by coelectrophoresis of their double-stranded RNA.

Thirty-two bovine field isolates of bluetongue virus (BTV), 6 field isolates of epizootic hemorrhagic disease virus (EHDV) from deer, 4 BTV prototype serotypes (10, 11, 13, and 17), and 2 EHDV prototype serotypes (1 and 2) were coelectrophoresed, using polyacrylamide gels. Field isolates were obtained from various regions of the United States. Analysis of polyacrylamide gels and scattered plots generated for comparison of migration patterns for different isolates within each serotype of BTV revealed wide variation among the individual segments. The BTV serotypes 10 and 11 had more variation, compared with BTV serotypes 13 and 17, especially for migration of genome segment 5. A definitive correlation was not seen between the double-stranded RNA migration profiles on polyacrylamide gel electrophoresis, geographic origin, herd of origin, or year of collection. One BTV field isolate contained more than 1 electropherotype, with 2 bands at the segment-7 position, and it was further characterized as BTV serotype 11. Segments 2 and 5 of EHDV isolates were more variable in their migration than were the other gene segments. Generally, migration profiles for EHDV double-stranded RNA were more variable, compared with those of BTV isolates. Although a correlation was found between migration profiles and serotype of 2 isolates of EHDV, a study of additional EHDV isolates is required before the diversity of electrophoretic patterns of EHDV can be determined.

Animals↗

PubMed1999

The role of CCR7 in TH1 and TH2 cell localization and delivery of B cell help in vivo.

Subsets of murine CD4+ T cells localize to different areas of the spleen after adoptive transfer. Naïve and T helper 1 (TH1) cells, which express the chemokine receptor CCR7, are home to the periarteriolar lymphoid sheath, whereas activated TH2 cells, which lack CCR7, form rings at the periphery of the T cell zones near B cell follicles. Retroviral transduction of TH2 cells with CCR7 forces them to localize in a TH1-like pattern and inhibits their participation in B cell help in vivo but not in vitro. Thus, differential expression of chemokine receptors results in unique cellular migration patterns that are important for effective immune responses.

Adoptive Transfer↗

PubMed1991

Generation of avian cells resembling osteoclasts from mononuclear phagocytes.

Several lines of indirect evidence suggest that a monocyte family precursor gives rise to the osteoclast, although this hypothesis is controversial. Starting with a uniform population of nonspecific esterase positive, tartrate-sensitive, acid phosphatase-producing, mannose receptor-bearing mononuclear cells, prepared from dispersed marrow of calcium-deprived laying hens by cell density separation and selective cellular adherence, we generated multinucleated cells in vitro. When cultured with devitalized bone, these cells show, by electron microscopy, the characteristic osteoclast morphology in that they are mitochondria-rich, multinucleated, and, most importantly, develop characteristic ruffled membranes at the matrix attachment site. Moreover, as documented by scanning electron microscopy, these cells pit bone slices in a manner identical to freshly isolated osteoclasts. In addition, isoenzymes of acid phosphatase from generated osteoclasts, separated by 7.5% polyacrylamide gel electrophoresis at pH 4, are identical to those of mature osteoclasts in migration pattern and tartrate resistance, although the precursor cells from which the osteoclasts are generated produce an entirely different isoenzyme, which is tartrate-sensitive and migrates less rapidly at pH 4. The fused cells also exhibit a cAMP response to prostaglandin E2. Therefore, osteoclast-like cells can be derived by in vitro culture of a marrow-derived monocyte cell population.

Animals↗

PubMed1990

In vivo migration and tissue localization of highly purified lymphokine-activated killer cells (A-LAK cells) in tumor-bearing rats.

Our laboratory has previously reported that the adoptive transfer of highly purified lymphokine-activated killer cells (adherent-LAK, A-LAK) into Fischer 344 (F344) rats bearing established lung or liver micrometastases effectively reduced the resultant tumor growth more than 90%, leading to significant increases in animal survival (Cancer Res. 49, 1441, 1989). To begin to investigate the mechanism(s) by which A-LAK cells mediate this anti-tumor effect, we studied their migration patterns in F344 rats bearing experimentally induced lung and liver metastases as well as subcutaneous tumors. A-LAK cells which were phenotypically 95 to 100% natural killer cells/large granular lymphocytes were labeled with either 51Chromium or fluorescein diacetate (so as to be visualized microscopically). Intravenous injection of such labeled A-LAK cells did not show significant differences in their tissue distribution patterns in tumor-bearing versus normal rats, even when high levels of exogenous recombinant interleukin-2 (rIL-2) was administered. A-LAK cells first migrated to the lungs and then subsequently migrated to the liver and spleen as early as 2 to 6 hr following iv injection. The kinetics of exit of A-LAK cells from the pulmonary capillary beds was not significantly different in rats bearing 3-day micrometastases or 14-day macrometastases compared to normal rats. Moreover, the presence of metastases in the liver did not alter the extent or kinetics of entry of A-LAK cells into the liver even in the presence of exogenously administered rIL-2. Finally, in rats bearing subcutaneous tumors, no evidence could be obtained that A-LAK cells were selectively localized to the tumor site. Tissue sections of livers from metastases-bearing animals injected with fluorescein diacetate labeled A-LAK cells did not demonstrate significant numbers of A-LAK cells infiltrating tumor nests with or without the administration of exogenous IL-2. These data suggest that A-LAK cells may mediate tumor regression in vivo by direct and indirect mechanisms, possibly through the secretion of cytokines and/or the recruitment of secondary effector cells.

Adenocarcinoma↗

PubMed1980

Invited review: guidance cue patterns and cell migration in multicellular organisms.

In multicellular organisms, guidance cues are either diffusible molecules or cellular or extracellular surfaces that are found in reproducible locations and that orient migrating cells and cell processes. The pattern of the guidance cues usually determines the complex in vivo migration routes of motile cells and cell processes. Within organisms, guidance cues are found to be organized in two general patterns: a) broad gradients--such as diffuse chemotactic gradients; b) discrete routes (substrate pathways)--such as chemotactic gradients confined to long channels, and such as the axon surface which represents a long specific highway for migrating Schwann cells.

Amphibians↗

PubMed1989

[Transplantation of embryonic cerebella into adult rat cerebella].

Cerebellar primordia of rat embryos (embryonic days of 14-20) were transplanted into mature cerebella of normal adult rats (Fischer 344). After 1-12 months, animals were sacrificed, and the development of grafts and the interaction between host and graft were examined by light and electron microscopy. Grafts from E14 rat cerebella showed the highest survival rate (85%), which decreased with increasing donor age. In some experiments, the donor tissue developed and differentiated to form folia with the trilaminar organization of the cerebellar cortex. Synaptic connections between neuronal elements in the graft showed basically the normal pattern. Thus, mossy terminals formed synaptic contacts with dendrites of granule cells, and axons of basket cells made synaptic contacts with somata of Purkinje cells. Many spines of Purkinje dendrites were contacted with parallel fibers, while others were surrounded by processes of astroglia. Migration of donor Purkinje and granule cells in the host molecular layer was frequently observed. Some donor granule cells, implanted in the superficial aspect of the host molecular layer, migrated down along processes of Bergmann glia, which resembled the migration pattern of the normal development. Other granule cells attached to neuroglial processes which ensheathed capillary walls. While the migration of granule cells was found to be glial-dependent, the glial role in the migration of Purkinje cells was not clear. Purkinje cells probably migrated into the host molecular layer under favorable conditions where the loss of host Purkinje cells had been occurred to some extent and consequently followed by the increase of free axon endings of parallel fibers and interneurons of the host.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1984

Observations on epidermal exsorption in mice following injections of procion dyes and ethidium bromide and topically applied dimethyl sulfoxide.

Back skin of hairless mice and external ears of CD-1 white mice were used to study the details of epidermal exsorption . Ethidium bromide, a DNA ligand, and two dichlorotriazinyl (procion) dyes were injected, i.v. or i.p. Migration patterns from the skin vasculature into the epidermis were observed by fluorescence microscopy and microspectrofluorometry . Topically applied DMSO greatly enhanced the exsorption process and produced intensely labeled epidermis. Ethidium bromide reacted primarily with nuclear DNA of living cells while the procion dyes tended to migrate intercellularly to label the stratum corneum. External ears of white mice treated topically with DMSO showed a pattern of labeling which included intense fluorescence of the cartilage and perichondrium as well as the ear epidermis.

Administration, Topical↗

PubMed1977

Decay of genetic variability in geographically structured populations.

The ultimate rate and pattern of approach to equilibrium of a diploid, monoecious population subdivided into a finite number of equal, large, panmictic colonies are calculated. The analysis is restricted to a single locus in the absence of selection, and every mutant is assumed to be new to the population. It is supposed that either the time-independent backward migration pattern is symmetric in the sense that the probability that an individual at position x migrated from y equals the probability that one at y migrated from x, or it depends only on displacements and not on initial and final positions. Generations are discrete and nonoverlapping. Asymptotically, the rate of convergence is approximately (I-u)2t[I-(2NT)-1]t, where u, NT, and t denote the mutation rate, total population size, and time in generations, respectively; the transient part of the probability that two homologous genes are the same allele is approximately independent of their spatial separation. Thus, in this respect the population behaves as if it were panmictic.

Genetic Variation↗

PubMed1985

Spatial distribution of immature Culicoides variipennis (Coq.).

Circadian fluctuation in the spatial distribution of immature Culicoides variipennis (Coq.) was studied in Saltville, Virginia, USA during the summer of 1983. Pupae and early instar larvae (1st and 2nd) were localized above the shoreline in the top cm of mud and displayed little migration from this region during the 24 hr sampling period, however, later instar larvae (3rd and 4th) showed marked migration patterns. Most larval movement was horizontal rather than vertical and was confined to the top cm of mud. Late instar larvae moved towards the water during the day but migrated up above shoreline during the night. Seasonal fluctuation in larval spatial distribution was also studied. Larvae appeared to move progressively downward into the mud during the autumn. However, when the site became frozen the majority of larvae occupied a narrow liquid interface between the ice cover and the frozen mud beneath.

Animals↗