In-plasma analysis of plasma-surface interactions.

During deposition, modification, and etching of thin films and nanomaterials in reactive plasmas, many active species can interact with the sample simultaneously. This includes reactive neutrals formed by fragmentation of the feed gas, positive ions, and electrons generated by electron-impact ionization of the feed gas and fragments, excited states (in particular, long-lived metastable species), and photons produced by spontaneous de-excitation of excited atoms and molecules. Notably, some of these species can be transiently present during the different phases of plasma processing, such as etching of thin layer deposition. To monitor plasma-surface interactions during materials processing, a new system combining beams of neutral atoms, positive ions, UV photons, and a magnetron plasma source has been developed. This system is equipped with a unique ensemble of in-plasma surface characterization tools, including (1) a Rutherford Backscattering Spectrometer (RBS), (2) an Elastic Recoil Detector (ERD), and (3) a Raman spectroscopy system. RBS and ERD analyses are carried out using a differentially pumped 1.7 MV ion beam line Tandetron accelerator generating a beam at grazing incidence. The ERD system is equipped with an absorber and is specifically used to detect H initially bonded to the surface; higher resolution of surface H is also available through nuclear reaction analysis. In parallel, an optical port facing the substrate is used to perform Raman spectroscopy analysis of the samples during plasma processing. This system enables fast monitoring of a few Raman peaks over nine points scattered on a 1.6 × 1.6 mm2 surface without interference from the inherent light emitted by the plasma. Coupled to the various plasma and beam sources, the unique set of in-plasma surface characterization tools detailed in this study can provide unique time-resolved information on the modification induced by plasma. By using the ion beam analysis capability, the atomic concentrations of various elements in the near-surface (e.g., stoichiometry and impurity content) can be monitored in real-time during plasma deposition or etching. On the other hand, the evolution of Raman peaks as a function of plasma processing time can contribute to a better understanding of the role of low-energy ions in defect generation in irradiation-sensitive materials, such as monolayer graphene.

Thomson parabola and time-of-flight detector cross-calibration methodology on the ALLS 100 TW laser-driven ion acceleration beamline.

We report on the cross-calibration of Thomson Parabola (TP) and Time-of-Flight (TOF) detectors as particle diagnostics, implemented on the most recent setup of the ALLS 100 TW laser-driven ion acceleration beamline. The Microchannel Plate (MCP) used for particle detection in the TP spectrometer has been calibrated in intensity on the tandem linear accelerator at the Université de Montréal. The experimental data points of the scaling factor were obtained by performing a pixel cluster analysis of single proton impacts on the MCP. A semi-empirical model was extrapolated and fitted to the data to apply the calibration also to higher kinetic energies and to extend it to other ion species. Two TOF lines using diamond detectors, placed at +6° and -9° with respect to the target-normal axis, were benchmarked against the TP spectrometer measurements to determine the field integrals related to its electric and magnetic dispersions. The mean integral proton numbers obtained on the beamline were about 4.1 × 1011 protons/sr with a standard deviation of 15% in the central section of the spectrum around 3 MeV, hence witnessing the high repeatability of the proton bunch generation. The mean maximum energy was of 7.3 ± 0.5 MeV, well in agreement with similar other 100 TW-scale laser facilities, with the best shots reaching 9 MeV and nearly 1012 protons/sr. The used particle diagnostics are compatible with the development of a high-repetition rate targetry due to their fast online readout and are therefore a crucial step in the automation of any beamline.

Disentangling neighbors and extended range density oscillations in monatomic amorphous semiconductors.

High energy x-ray diffraction measurements of pure amorphous Ge were made and its radial distribution function (RDF) was determined at high resolution, revealing new information on the atomic structure of amorphous semiconductors. Fine structure in the second peak in the RDF provides evidence that a fraction of third neighbors are closer than some second neighbors; taking this into account leads to a narrow distribution of tetrahedral bond angles, (8.5 ± 0.1)°. A small peak which appears near 5 Å upon thermal annealing shows that some ordering in the dihedral bond-angle distribution takes place during structural relaxation. Extended range order is detected (in both a-Ge and a-Si) which persists to beyond 20 Å, and both the periodicity and its decay length increase upon thermal annealing. Previously, the effect of structural relaxation was only detected at intermediate range, involving reduced tetrahedral bond-angle distortions. These results enhance our understanding of the atomic order in continuous random networks and place significantly more stringent requirements on computer models intending to describe these networks, or their alternatives which attempt to describe the structure in terms of an arrangement of paracrystals.

Dietary intervention in older adults with early-stage Alzheimer dementia: early lessons learned.

In older adults, an adequate diet depends on their ability to procure and prepare food and eat independently or the availability of dietary assistance when needed. Inadequate food intake or increased nutritional requirements lead to poor nutritional status, which is considered a key determinant of morbidity, increased risk of infection, and mortality in elderly individuals. Weight loss among seniors also heralds increased morbidity and mortality. Dietary behaviour disorders affecting food consumption, nutrition status and maintenance of body weight are common in older adults, and have a substantial impact on nutritional status and quality of life among older adults with Alzheimer Dementia (AD). The Nutrition Intervention Study (NIS) is ongoing. It employs a quasi-experimental pre-post intervention design in physically-well, community-dwelling early stage AD patients aged 70 y or older. To date, 34 intervention group patients and 25 control group participants have been recruited with their primary caregivers (CG) from 6 hospital-based memory and geriatric clinics in Montreal. The NIS uses clinical dietetics principles to develop and offer tailored dietary strategies to patients and their CG. This paper reports on the application of dietary intervention strategies in two intervention group participants; one was deemed successful while the other was considered unsuccessful. The report documents challenges encountered in assessing and counselling this clientele, and seeks to explain the outcome of intervention in these patients.

The role of surgical biopsy in the diagnosis of glioma in individuals with neurofibromatosis-1.

Most gliomas in neurofibromatosis type 1 (NF1) are pilocytic astrocytomas (PAs) of the optic pathway occurring in young children. However, some individuals develop gliomas that lack the typical NF1-associated clinical features or radiographic appearance. We identified 17 atypical presentations from a review of 100 patients with NF1-associated gliomas. Biopsy showed that 9 were not classic PAs. These data highlight the value of biopsy in NF1-associated gliomas with unusual clinical or radiographic presentations.

Phenotypic and genotypic characterization of esterase-producing Ureibacillus thermosphaericus isolated from an aerobic digestor of swine waste.

Eight closely related thermophilic strains were isolated from an aerobic and thermophilic treatment of swine wastes. The pleomorphic cells (short and long rods; cocci) showed peritrichous flagella, terminally swollen sporangium, and liberated spores exhibiting hairy appendages. The Gram reaction was negative for both young (4 h) and old (48 h) cultures. Several features, such as colonial morphology, growth between 35 degrees C and 65 degrees C, presence of catalase, presence of spores, and strictly aerobic metabolism (except for one strain), are similar to those found for the genus Bacillus. The inability of the strains to use sugars, except esculin, as source of carbon and energy and the whole cell fatty acid composition are similar to those found in Bacillus thermosphaericus DSM 10633. Sequence analysis of the 16S rRNA gene revealed 99.8%-99.9% identity for seven of the thermophilic strains with this species. A new genus, Ureibacillus, was recently proposed for type strain B. thermosphaericus DSM 10633 The last strain exhibits 97.8% and 97.3% identity with Ureibacillus terrenus DSM12654 and Bacillus sp. TP-84, respectively. Esterase activities were detected for all strains, and assays on p-nitrophenyl butyrate and p-nitrophenyl caprylate revealed that strains were more active on the shorter substrate.

Quantitative analysis of the transcondylar approach to the foramen magnum.

OBJECTIVE: Condylar resection with suboccipital craniotomy increases foramen magnum exposure, but guidelines for when this is necessary are not defined. Cadaveric and computed tomography evaluations were completed to guide decision-making regarding the use and extent of condylar resection. METHODS: Quantitative analysis of foramen magnum surgical exposures was performed on 32 skulls (64 sides) and 6 cadaveric dissections (12 sides). Computed tomographic (CT) scans were performed on cadaveric heads before and after condylar resections. Digitized images of dry skulls and CT images of cadaver heads were quantitatively analyzed. Predissection CT measurements of cadaveric heads guided extent of condylar resections, and resection accuracy was assessed with postdissection CT scans. RESULTS: Skull measurements (means in parentheses) included the foramen magnum area (7.8 cm(2)), length (3.6 cm), width (3.1 cm), anteroposterior condylar length (2.3 cm), and axial condylar length (2.5 cm). Mean widths of potential surgical exposures for skulls were obtained for A) suboccipital craniotomy (2.3 cm), B) with 25% (2.6 cm), and C) 50% condylar resection (3.0 cm). Mean angles of exposure were as follows: A, 38.4 degrees; B, 49.1 degrees; and C, 54.3 degrees. CT scans of cadaveric heads before and after dissections yielded measurements of exposure equivalent to measurements found on the dry skulls. CONCLUSION: On average, lateral exposure increases by 3 mm (13%) and 7 mm (30%) for 25 and 50% condylar resection, respectively, compared with suboccipital craniotomy alone. Angles of exposure increase by 10.7 degrees (28%) and 15.9 degrees (41%). Measurements of CT images can be used preoperatively to help analyze the need for condylar resection and intraoperatively to guide the extent of condylar resection.

Clinicopathologic assessment and grading of embolized meningiomas: a correlative study of 64 patients.

BACKGROUND: Preoperative embolization of meningiomas is commonly performed to minimize intraoperative bleeding, thereby facilitating surgery and reducing the necessity for transfusion. However, the resulting necrosis and compensatory proliferation reportedly have hampered subsequent histologic grading. METHODS: The clinicopathologic features of 64 meningiomas embolized between 1989 and 1997 were assessed. Tumors were graded according to recently published criteria. RESULTS: A good embolization result (> 75% reduction in angiographic blush) was achieved in 52%. Histologically, embolized meningiomas showed higher frequencies of necrosis (89%), nuclear atypia (72%), macronucleoli (58%), sheeting (31%), high mitotic index (30%), and brain invasion (14%) when compared with nonembolized counterparts. Median mitotic and MIB-1 indices were slightly elevated (1.5 of 10 high-power fields and 1.6%, respectively). A significant degree of necrosis (> 10%) was found in 43% and was only roughly correlated with extent of angiographic blush reduction or embolization particle size. Histologic grade was benign in 57.8%, atypical in 40.6%, and anaplastic in 1.6%. At last follow-up, there were 13 recurrences, 11 in the atypical/anaplastic (41%) versus 2 in the benign (5%) subsets (P = 0.001). CONCLUSIONS: The authors conclude that 1) their grading scheme accurately stratifies embolized meningiomas, 2) extent of necrosis is difficult to predict using standard clinical parameters, and 3) their high incidence of atypical meningioma more likely reflects patient selection biases rather than artifacts induced by the embolization procedure.

Intratumoral injection of lipopolysaccharide causes regression of subcutaneously implanted mouse glioblastoma multiforme.

OBJECTIVE: Anecdotal reports documented extended survival times for patients who developed infections at the site of resection of malignant gliomas. Hypothesized mechanisms for this phenomenon include immune responses triggered by lipopolysaccharide (LPS). This investigation assessed whether LPS could produce tumor regression in an in vivo model of malignant glioma. METHODS: Delayed brain tumor cells (2 x 10(6)) were injected subcutaneously into female BALB/c mice. LPS (300-500 microg) was injected intratumorally or subcutaneously at a contralateral site on Days 10, 17, and 24. Control animals received phosphate-buffered saline intratumorally or subcutaneously. Mice were killed on Day 28, and tumors were removed. Mean tumor masses for control animals and the two LPS-treated groups (intratumoral or contralateral subcutaneous treatment) were compared. Histological assessments of treated and control tumors were performed. RESULTS: Complete or nearly total tumor regression was achieved in all 20 mice with subcutaneous delayed brain tumor cell tumors treated intratumorally with 400 microg of LPS (mean tumor mass of 0.09 +/- 0.38 g versus 2.42 +/- 2.46 g for control animals, P < 0.0001). Intratumoral administration of 300 microg of LPS or subcutaneous injection of 300 or 400 microg of LPS at a contralateral site resulted in less consistent regression of subcutaneous tumors. Administration of 500 microg of LPS resulted in tumor regression similar to that observed with lower doses but was limited by treatment-related deaths in 40% of animals. Histological assessment revealed lymphocytic infiltration of LPS-treated tumors. CONCLUSION: Intratumoral injections of LPS caused dramatic regression of subcutaneously implanted delayed brain tumor cell mouse gliomas. Investigation of this antitumoral effect may improve treatment responses for patients with malignant gliomas.

Gliomas in rodent whisker barrel cortex: a new tumor model.

OBJECT: Surgical treatment of gliomas is difficult because they are invasive. Invasion of essential cortex often limits or precludes surgical resection. A tumor model was developed in which the rodent whisker barrel cortex was used to examine how gliomas affect cortical function and structure. METHODS: Both DBT (mouse) and C6 (rat) glioma cell lines were grown in culture and labeled with the fluorescent marker Dil in vitro. Labeled tumor cells were then injected into the whisker barrel cortex of adult mice and rats. Neurological assessments were made daily and magnetic resonance (MR) images were obtained. Animals were killed by perfusion 6 to 14 days after injection, and histological sections were prepared and studied. Tumors were found in all 20 rats and 10 mice that had been injected with the C6 and DBT cell lines, respectively. The animal cells had been labeled with Dil in vitro, and all in vivo tumors proved to be Dil positive. The MR images revealed the tumor locations and serial MR images demonstrated tumor growth. Histological evaluation confirmed the location of the tumor and the disruption of barrel cortex architecture. CONCLUSIONS: Both DBT and C6 glioma cell lines can be used to generate malignant glial tumors reproducibly in the whisker barrel cortex. Fluorescent labeling and cytochrome oxidase staining permit visualization of tumor growth patterns, which disrupt the barrel cortex by microscopic invasion and by gross tissue deformation. Magnetic resonance imaging demonstrates the anatomical extension of these tumors in live rodents. Using this model for further studies on the effects of malignant glioma growth on functional cerebral cortex should advance our understanding of the neurological issues and management of patients with these tumors.

Isolation and characterization of human malignant glioma cells from histologically normal brain.

Brain invasion prevents complete surgical extirpation of malignant gliomas; however, invasive cells from distant, histologically normal brain previously have not been isolated, cultured, and characterized. To evaluate invasive human malignant glioma cells, the authors established cultures from gross tumor and histologically normal brain. Three men and one woman, with a mean age of 67 years, underwent two frontal and two temporal lobectomies for tumors, which yielded specimens of both gross tumor and histologically normal brain. Each specimen was acquired a minimum of 4 cm from the gross tumor. The specimens were split: a portion was sent for neuropathological evaluation (three glioblastomas multiforme and one oligodendroglioma) and a portion was used to establish cell lines. Morphologically, the specimens of gross tumor and histologically normal brain were identical in three of the four cell culture pairs. Histochemical staining characteristics were consistent both within each pair and when compared with the specimens sent for neuropathological evaluation. Cultures demonstrated anchorage-independent growth in soft agarose and neoplastic karyotypes. Growth rates in culture were greater for histologically normal brain than for gross tumor in three of the four culture pairs. Although the observed increases in growth rates of histologically normal brain cultures do not correlate with in vivo behavior, these findings corroborate the previously reported stem cell potential of invasive glioma cells. Using the radial dish assay, no significant differences in motility between cultures of gross tumor and histologically normal brain were found. In summary, tumor cells were cultured from histologically normal brain acquired from a distance greater than 4 cm from the gross tumor, indicating the relative insensitivity of standard histopathological identification of invasive glioma cells (and hence the inadequacy of frozen-section evaluation of resection margins). Cell lines derived from gross tumor and histologically normal brain were usually histologically identical and demonstrated equivalent motility, but had different growth rates.

Selective dorsal rhizotomy and rates of orthopedic surgery in children with spastic cerebral palsy.

If the spasticity of cerebral palsy (CP) is reduced in children at a young age by selective dorsal rhizotomy, the incidence of lower-extremity deformities requiring orthopedic surgery may be reduced; however, this has never been investigated in detail. The authors examined the effects of selective dorsal rhizotomy on rates of lower-extremity orthopedic surgery in 178 children with CP. Age at selective dorsal rhizotomy ranged from 2 to 19.3 years (mean 5.5 years) with follow-up intervals ranging from 24 to 70 months (mean 44 months). Spastic CP was classified as quadriplegia (33%), diplegia (65%), and hemiplegia (2%). To assess the effects of early versus late rhizotomy on rates of orthopedic surgery, patients were grouped as follows: Group I underwent rhizotomy between 2 and 4 years of age (54 patients), and Group II underwent rhizotomy between 5 and 19 years of age (124 patients). Comparison of Kaplan-Meier plots of lifetime orthopedic surgery rates revealed that Group II underwent orthopedic surgery at a higher rate than Group I (p = 0.037). Analysis by procedure type revealed higher orthopedic surgery rates in Group II than Group I for heel cord releases (p = 0.0025), adductor releases (p = 0.018), and hamstring releases (p = 0.02). Orthopedic surgery rates were no higher for Group II compared to Group I for ankle/foot operations (p = 0.023), femoral osteotomy (p = 0.25), iliopsoas releases (p = 0.35), and "other" operations (p = 0.013). The data indicate that early rhizotomy reduces the need for orthopedic surgery for heel cord, hamstring, and adductor releases.

Mitogens as motogens.

Numerous in vivo methodologies have documented the invasive behavior of glioma cells through normal brain parenchyma. Glioma cell locomotion has also been assessed with a number of in vitro assays including the Boyden chamber and other chemotaxis assays, colloidal gold cell tracking, analysis of migration of cells tumor cells from spheroids, confrontation cultures of glioma cells with aggregates of non-neoplastic tissue, time-lapse video microscopy, electron microscopic examination of the cytomorphologic correlates of cell motility, the radial dish assay, and quantitative enzyme immunoassay of proteins associated with invasion (e.g. laminin). Several of these techniques have been specifically modified to assess the effects of cytokines on glioma cell motility in vitro. Cytokines studied utilizing these methods include: epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), the bb dimer of platelet-derived growth factor (PDGFbb), nerve growth factor (NGF), interleukin 2 (IL-2), transforming growth factors alpha and beta 1 (TGF alpha and TGFstraat1), and tumor necrosis factor alpha (TNF alpha). This review summarizes the investigational methods used to evaluate random and directional glioma cell motility and invasion in vivo and in vitro. The roles of specific mitogens as motogens, as evaluated with these methods are then presented.

Predictors of ability to walk after selective dorsal rhizotomy in children with cerebral palsy.

Serial evaluations were completed after selective dorsal rhizotomy on 90 children with spastic cerebral palsy to analyze whether age, the preoperative gait score, voluntary dorsiflexion at the ankle, the diagnosis (quadriplegia or diplegia), or the length of follow-up correlated with the ability to walk after rhizotomy. The preoperative gait score (P < 0.0001), the diagnosis (diplegia versus quadriplegia, P < 0.0001), unilateral dorsiflexion (P = 0.0029), and bilateral dorsiflexion (P < 0.0001) were significant predictors of the maximal postoperative gait score in the univariate regression analysis, but only the preoperative gait score (P < 0.0001) and the diagnosis (P = 0.0015) retained significant predictive power in the multivariate analysis. These data suggest that the preoperative gait score and the diagnosis are the strongest predictors of ability to walk after selective dorsal rhizotomy. Dorsiflexion demonstrated predictive power only in the univariate model, suggesting that it might have some prognostic value but less than the preoperative gait score or the diagnosis.

Invading C6 glioma cells maintaining tumorigenicity.

To characterize rat glioma cell invasion, 2 x 10(6) fluorophore-labeled or transfection-labeled C6 rat glioma cells were implanted in the rat frontal lobe. Eighty percent of the rats implanted formed bulk tumors (3-4 mm in diameter). Two weeks after implantation, fluorescence microscopy revealed single tumor cells in sites over 16 mm from the bulk brain tumor. Tumor cells distant from the bulk tumor remained single without mass formation and invaded primarily along white matter tracts. Two weeks after tumor implantation, three cell lines were created from each brain by disaggregation and initiation in culture of 1) bulk tumor, 2) contralateral hemisphere, and 3) cerebellum; all disaggregated specimens generated viable cultures. Cells cultured from the contralateral hemisphere were morphologically indistinguishable from cells from the bulk tumor and from the original C6 cell line. Cells cultured from the cerebellum were morphologically quite distinct from the C6 cell line. Cells from disaggregated specimens obtained from the tumor, contralateral hemisphere, and cerebellum were implanted in the frontal lobe of naive rats to test tumorgenicity. Bulk tumor formed in 58% of the rats implanted with specimens from tumor, in 75% of the rats implanted with specimens from contralateral hemisphere, and in only 12.5% of the rats implanted with specimens from the cerebellar hemispheres. Experiments using C6 cells labeled by transfection with the p3' ss DNA vector prior to implantation confirmed that the cells cultured from the contralateral hemisphere were derived from the implanted C6 cells. Experiments with C6 cells anchored in agar served to verify that movement to the contralateral hemisphere was secondary to parenchymal invasion rather than dispersion in the cerebrospinal fluid.

Modification of human glioma locomotion in vitro by cytokines EGF, bFGF, PDGFbb, NGF, and TNF alpha.

Cytokines exert receptor-mediated control over glia. Up-regulation of receptor expression of cytokine production corresponds with the acquisition of a neoplastic phenotype. A modified radial dish assay was used to determine whether in vitro locomotion of glioma cells is modified by the epidermal growth factor, the basic fibroblast growth factor, the bb dimer of platelet-derived growth factor, the nerve growth factor, or the tumor necrosis factor alpha. Human glioma cells were plated in the center of a petri dish with one of these cytokines in 0.5 ml agar (50 ng/ml if the cytokine was distributed evenly throughout the dish) at one edge, and 0.5 ml plain agar at the opposite edge. After 24 hours, a central zone of cells was established; the agar was gelatinized. Feeding medium was added to the dish, and slow elution from the agar established a cytokine gradient. Cell counts were performed daily over 6 to 10 days at predetermined distances on both sides of the central zone to assess directional cellular movement with respect to the cytokine gradient and the plain agar. The epidermal growth factor caused continuous chemoattraction, whereas the tumor necrosis factor alpha caused slight chemorepulsion for 24 to 48 hours, followed by strong chemoattraction. The bb dimer of platelet-derived growth factor, the basic fibroblast growth factor, and the nerve growth factor all maintained chemorepulsion over the entire 6 to 10 days. Therefore, the cytokines did affect glioma cell motility in vitro, and the modified radial dish assay used in this study provided a useful in vitro model for assessing the impact of the cytokines on glioma cell locomotion.(ABSTRACT TRUNCATED AT 250 WORDS)

The in vitro motility of human gliomas increases with increasing grade of malignancy.

BACKGROUND: Malignant gliomas are a highly invasive disease with a dismal prognosis despite aggressive therapeutic interventions. METHODS: The authors tested the hypothesis that increasing in vitro motility of human glial tumors correlates with increasing grade of malignancy. A radial dish assay was used to quantitatively assess the brain tumor cell motility of 14 low passage cell lines derived from human glial tumors of varied malignancy, and 3 cell lines derived from human glia. The egress of cells from a central region of high tumor cell density to a region of lower tumor cell density was determined at various time points. A motility coefficient (MC), the slope of distance traveled against time, was generated by simple linear regression analysis. RESULTS: The MC increased with the increased histologic grade of malignancy. Generation of a t-statistic was used to determine the significance of differences in motility among 5 histopathologic groups: 10 glioblastoma (World Health Organization Grade IV) cell lines (mean MC, 0.00396), 2 mixed anaplastic oligoastrocytoma (Grade III) cell lines (mean MC, 0.00382), 1 anaplastic astrocytoma (Grade III) cell line (mean MC, 0.00295), 1 Grade I glioma cell line (mean MC, 0.00206), and 3 human glial cell lines (mean MC, 0.00110). CONCLUSIONS: The radial dish assay provided a reproducible method for quantitatively assessing brain tumor cell motility. The higher MC observed with the malignant human glioma cell lines correlated with the tendency of these tumors to invade into adjacent brain tissue. The potential to inhibit glial tumor motility may provide an important therapeutic avenue in the future.

Assessment of brain tumor cell motility in vivo and in vitro.

Brain tumor dispersal far from bulk tumor contributes to and, in some instances, dominates disease progression. Three methods were used to characterize brain tumor cell motility in vivo and in vitro: 1) 2 weeks after implantation in rat cerebral cortex, single C6 cells labeled with a fluorescent tag had migrated to brain sites greater than 16 mm distant from bulk tumor; 2) time-lapse videomicroscopy of human brain tumor cells revealed motility of 12.5 microns/hr. Ruffling leading edges and pseudopod formation were most elaborate in more malignant cells; 3) an in vitro assay was devised to quantitatively evaluate motility from a region of high cell density to one of lower cell density. Human brain tumor cells were plated in the center of a petri dish, washed, and refed, establishing a 2-cm circular zone of cells in the dish center. Motility was determined by counting cells daily at predetermined distances from the central zone perimeter. Cells were found 1 cm from the perimeter by 24 hours and 3 cm from the perimeter by 4 days. Increasing serum concentration increased motility; however, neither fibronectin nor arrest of cells in the G0 phase by hydroxyurea altered motility. The addition of cytochalasin B to block cytoskeletal assembly prevented cell motility. Motility increased with increased malignancy. Subpopulations of cells were created by clonal amplification of cells that had migrated most rapidly to the dish periphery. Although morphologically indistinguishable when compared to the original cell line from which they were derived, these subpopulations demonstrated significantly increased motility.

In vitro assessment of Taxol for human glioblastoma: chemosensitivity and cellular locomotion.

Although the antineoplastic efficacy of Taxol against a variety of tumors has been established, it has only recently been used for malignant brain tumors. We evaluated in vitro chemosensitivity of glioblastoma to Taxol and the affect of Taxol on glioblastoma cell locomotion. The clonogenic assay was used to evaluate the chemosensitivity of five human glioblastomas and the C6 rat glioma. Cells exposed to Taxol (0-250 nM) were suspended in agar in capillary tubes. Following incubation, colonies were counted to determine percent survival. All six cell lines demonstrated sensitivity to Taxol (LD50 1 nM to > 250 nM). However, even at concentrations exceeding those achievable clinically, all cell lines had surviving cells, indicating a saturation threshold for Taxol cytotoxicity. Cell locomotion was evaluated using the radial dish assay to determine the rate of egress of cells from a region of high cell density to the periphery. Increasing Taxol concentration caused increased locomotion in all six cell lines (p < 0.0001). Although Taxol has significant cytocidal impact, it increases in vitro locomotion of glioblastoma cells. These findings suggest that the clinical use of Taxol for glioblastoma may slow the growth of bulk disease, but may also lead to increased tumor invasion.