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Spread of MHV-JHM from nasal cavity to white matter of spinal cord. Transneuronal movement and involvement of astrocytes.

C57B1/6 mice infected intranasally with mouse hepatitis virus, strain JHM (MHV-JHM) develop hindlimb paralysis with histological evidence of demyelination several weeks after inoculation. Virus must spread from the site of inoculation, the nasal cavity, to the site of disease, the white matter of the spinal cord. It has been shown previously that after intranasal inoculation, virus enters the brain via the olfactory nerve and spreads to infect many of its neuroanatomic connections within the central nervous system (CNS). In this report, it is shown that virus infecting the spinal cord is first detected in the gray matter, with spread occurring to the white matter soon thereafter. Astrocytes are heavily infected during the process of spread from the gray to the white matter of the spinal cord. Since astrocytes are in intimate contact with neuronal synapses and are themselves connected via gap junctions, these results suggest that astrocytes may be a conduit for the spread of virus in these mice. Astrocytes provide factors for the proliferation and survival of oligodendrocytes, and widespread infection of these cells might contribute to the demyelinating process eventually observed in these mice. Additionally, since virus first appears at specific locations in the spinal cord, it should be possible to determine the source of the virus infecting the cord. While the results are not definitive, the data are most consistent with virus spreading from the ventral reticular formation to the gray matter of the cervical spinal cord.

Animals↗

Measurement of current spread from microelectrodes when stimulating within the nervous system.

Tungsten stimulating microelectrodes have been tested in monopolar, bipolar and concentric configurations for the extent to which unwanted current spread occurred. Current spread from monopolar electrodes in close conformity with the predictions of the inverse square law, both in vitro in vivo. The bipolar and concentric configurations, tested only in vitro, had current-spread characteristics which did not follow the inverse square law so closely. The bipolar configuration gave little reduction in spread, compared wpith a monopolar electrode, but the concentric configuration did reduce the extent of stimulus spread. The extent of spread depends greatly upon a number of experimental variables, including the dimensions of the microelectrode tip. For studies requiring precise localization with low stimulus currents, it is advisable to determine the inverse square law constant for each experimental situation. For more generalized stimulation purposes within the mammalian C.N.S., employing monopolar microelectrodes of moderate tip size, brief pulses of 10 muA can be expected to stimulate myelinated axons with about 0.15 mm of the tip, and 100 muA WILL Stimulate with a radius of about 0.5 mm.

Animals↗

Spreading of cells on various substrates evaluated by Fourier analysis of shape.

In order to evaluate in mathematical terms the morphological changes occurring in the course of cell spreading, Fourier analysis of shape was applied. Human urothelial Hu 961 b cells plated on type IV collagen, fibronectin, laminin, glass and bovine serum albumin (BSA) were studied. Fourier parameters describing cell shape as well as surface areas covered by the cells on the substrate were subjected to statistical analysis. Using analysis of variance and discriminant analysis it was found that parameters describing cell shape (both gross shape of cells and their fine scale contour foldings) possessed a higher power of discrimination between the cells spread on various substrates than the differences in cell surface areas. In the course of observation (75 and 150 min) the highest number of attached cells and highest degree of spreading were found when cells were plated on type IV collagen. Moderate alterations in cell shape and moderate increase of surface area were seen in the group of cells seeded on fibronectin, whereas the cells plated on laminin, glass and BSA revealed a moderate increase of surface area, but no changes in their shape were observed. The differences in attachment of cells and in the degree of their spreading might be due to the variation in expression of plasma membrane receptors for various substrates. The Fourier analysis of cell shape coupled with measurement of surface area is a good tool for quantitative evaluation of cell spreading and can be used for discrimination between cells spread on different substrates.

Cell Adhesion↗

Growth in serum-free medium of human colonic adenocarcinoma cell lines on microcarriers: a two-step method allowing optimal cell spreading and growth.

Human colonic adenocarcinoma cells have been successfully grown on polystyrene microcarriers by modifying the culture conditions used in monolayer culture. The method can be divided into two culture phases: a) a phase of spreading, wherein cells were seeded in presence of serum-supplemented medium; b) a phase of active growth wherein spread cells on the beads were allowed to grow in a serum-free medium. Under these conditions, optimal spreading and growth of HT 29 and HRT 18 cells on the microcarriers were obtained. A differential propagation was observed between HT 29-D4 and HT 29-D9 cells (both clonal populations derived from HT 29 cells) on the microcarriers that is tentatively related to the discrepancy observed in the spreading efficiency of these clonal cells on serum-coated culture flasks. An index of spreading efficiency (IS index) has been defined to quantify the efficiency of spreading of each cell line on microcarriers. These data gave the opportunity to develop serum-free, scale-up methods to culture cells like HT 29 which release potentially useful products.

Adenocarcinoma↗

Thermodynamic aspects of cell spreading on solid substrata.

To verify the validity of thermodynamic approaches to the prediction of cellular behavior, cell spreading of three different cell types on solid substrata was determined in vitro. Solid substrata as well as cell types were selected on the basis of their surface free energies, calculated from contact angle measurements. The surface free energies of the solid substrata ranged from 18-116 erg cm-2. To measure contact angles on cells, a technique was developed in which a multilayer of cells was deposited on a filter and air dried. Cell surface free energies ranged from 60 erg cm-2 for fibroblasts, and 57 for smooth muscle cells, to 91 for HeLa epithelial cells. After adsorption of serum proteins, cell surface free energies of all three cell types converged to approx 74 erg cm-2. The spreading of these cell types from RPMI 1640 medium on the various solid substrata showed that both in the presence and in the absence of serum proteins in the medium, cells spread poorly on low energy substrata (Ys less than 50 erg cm-2), whereas good cell spreading was observed on the higher energy substrata. Calculations of the interfacial free energy of adhesion (delta Fadh) show that delta Fadh decreases with increasing Ys, and equals zero around 45 erg cm-2 for all three cell types in the presence of serum proteins and for HeLa epithelium cells in the absence of serum proteins. This explains the spreading of these cells on the various substrata upon a thermodynamic basis. The results clearly show that substratum surface free energy has a predictive value with respect to cell spreading in vitro, both in the presence and absence of serum proteins. It is noted, however, that interfacial thermodynamics fail to explain the behavior of fibroblasts and smooth muscle cells in the absence of serum proteins, most likely because of the relatively high surface charges of these two cell types.

Adsorption↗

Spread of gallbladder carcinoma: CT evaluation with pathologic correlation.

BACKGROUND: To assess the accuracy of computed tomographic (CT) imaging in the detection of spread and staging of gallbladder carcinoma. METHODS: CT findings of spread of gallbladder carcinoma in 59 Japanese patients who underwent radical surgery were correlated retrospectively with pathologic findings. RESULTS: The incidence of histologically proven nodal involvement was 54% (32 patients) and the most common spread of gallbladder carcinoma. The sensitivities in CT detection of N1 and N2 nodal involvement were 36% and 47%, respectively; positive predictive values were 94% and 92%, respectively. Direct extension to the liver, extrahepatic bile duct, and gastrointestinal tract or pancreas were histologically confirmed in 24, 18, and five patients. The sensitivities in the CT detection of direct spread to the liver of less than 2 cm, more than 2 cm, the extrahepatic bile duct, and the gastrointestinal tract or pancreas were 65%, 100%, 50%, and 57%, respectively; positive predictive values were 77%, 100%, 90%, and 100%, respectively. The incidence of liver metastases and involvement of interaortocaval nodes were 7% and 16%, respectively. The sensitivities in CT detection of liver metastases and involvement of interaortocaval nodes were 75% and 21%, respectively; positive predictive values were 100% and 86%, respectively. CT could not detect direct spread to omentum and peritoneal seedings. CONCLUSION: For detecting the spread of gallbladder carcinoma, CT imaging has low to moderate sensitivity; however, CT imaging can help in determining resectability and in planning the treatment, especially in advanced-stage gallbladder carcinoma, because of a high positive predictive value.

Adult↗

Survival-associated histologic spreading modes of operable intrahepatic, peripheral-type cholangiocarcinomas.

We investigated survival-associated histologic and metastatic spreading modes of intrahepatic, peripheral-type cholangiocarcinomas resected to contribute to surgical control of the tumor. Previous results have been mostly obtained from autopsies, reflecting the terminal status of patients. We clinicopathologically reviewed the resected 20 intrahepatic, peripheral-type cholangiocarcinomas and investigated the histologic findings of resected specimens and medical records to assess spreading modes along with patients' survival. The carcinoma cells superficially spread in the ductal epithelium in 75%, infiltrated along Glisson's system and migrated multidirectionally in 100%, and permeated the vascular network in 80%. The cumulative survival rate significantly related to vascular permeation, extrahepatic metastases, and lymphatic, neural, and nodal involvement but not to ductal spread, tumor size (cutoff size 5 cm), or intrahepatic metastases by the log-rank test. The patients with lymphatic, neural, or nodal involvement died early after surgery. Practically, only vascular permeation was identified as a significant independent variable for survival using multivariate analysis. Peripheral cholangiocarcinomas spread mainly in three modes: ductal spread, infiltration along Glisson's system, and vascular permeation. In the practically operable cases, vascular permeation is closely related to survival, and intrahepatic metastasis may be surgically controlled to some degree.

Aged↗

Cholesterol-lowering effect of spreads enriched with microcrystalline plant sterols in hypercholesterolemic subjects.

BACKGROUND: Plant sterols have been shown to reduce serum lipid concentrations. The effectiveness is highly dependent on the physical state of the plant sterols. By means of a new crystallizing method, plant sterols can be added into dietary fats and oils homogeneously. In this fat ingredient, plant sterols are in a microcrystalline form. AIMS OF THE STUDY: We investigated the cholesterol-lowering effect and possible side effects of vegetable oil-based spreads fortified with two different doses of microcrystalline plant sterols. METHODS: This double-blind randomized, placebo-controlled study consisted of a 6-wk run-in and a 6-month experimental period. During the run-in period, all 155 hypercholesterolemic subjects received rapeseed oil-based control spread. In the beginning of the experimental period subjects were randomly assigned into one of three experimental groups. The control group continued to use control spread, and the two test groups used spreads with added plant sterols of either 1.5 g/d or 3.0 g/d. The subjects consumed test spreads as a part of their normal diet without any restrictions in lifestyle and diet. RESULTS: Plasma total- and LDL-cholesterol concentrations were significantly reduced by 7.5-11.6% (0.46-0.62 mmol/1) in groups consuming margarine enriched with free plant sterols, compared with the control group. The effects were similar between the two groups consuming either 1.5g or 3.0 g plant sterols per day. No effect on HDL-cholesterol or triacylglycerol concentrations occurred. The test spreads did not induce any adverse effects in blood clinical chemistry, hematology or decreases in serum concentrations of lipid soluble vitamins. CONCLUSIONS: Microcrystalline plant sterols are effective in lowering serum total- and LDL-cholesterol concentrations without obvious side effects. The daily dose of 1.5 g plant sterols is enough to reach the maximum effect.

Adult↗

AP-1 (c-Fos/c-Jun) is required for corneal epithelial spreading.

PURPOSE: We previously demonstrated that the AP-1 components c-fos/c-jun are up-regulated in healing rat corneal epithelium in a relatively early phase following epithelial débridement, implicating the AP-1 function in the initiation of cell movement. To explore this hypothesis, we examined the effect of lack of c-Fos and c-Jun protein expressions on the spreading of corneal epithelium and in situ in organ culture. Antisense-oligonucleotide (AS) c-fos-null mice were used for this purpose. METHODS: A rectangular piece of corneal tissue (2 x2 mm) was obtained from each eye of recently killed adult C57BL/6 mice and was incubated for 11 h in culture medium with 8 microM c-fos AS or c-jun AS probe. Sense probes were used for negative control. A rectangular section of corneal tissue was also obtained from each eye of c-fos(-/-), c-fos(+/-) and c-fos(+/+) mice and was organ-cultured for 11 h. The length of the path of epithelial spreading on stromal cut surface (both sides) was measured in hematoxylin-eosin-stained specimens. Data were analyzed by unpaired Student's t-test. RESULTS: Addition of c-fos AS to the medium decreased the length of epithelial spread to 40.36% of that in the control with the S probe. Addition of c-jun AS decreased the length of epithelial spreading rate to 42.71% of control with S probe. Lacking c-Fos decreased the epithelial spreading to 17.73% of control data from c-fos(+/-) and c-fos(+/+) mice. CONCLUSION: AP-1 (c-Fos/c-Jun) is required for the corneal epithelial spreading.

Animals↗

Mode of tumor spread and surgical strategy in gallbladder carcinoma.

BACKGROUND AND AIMS: The mode and degree of tumor spread in gallbladder carcinoma is poorly documented. The present study classifies the patterns of dissemination of this tumor with a focus on surgical strategy. PATIENTS AND METHODS: Surgical specimens from 112 patients who underwent curative resection were reviewed. There were stage I, II, III, and IV in 9, 11, 14, and 78 patients, respectively. RESULTS: Six types of spread were identified. In the hepatic bed type ( n=20) a large mass in the fundus and body penetrated into the liver through the gallbladder bed with or without contiguous spread to the gastrointestinal tract. The extent of hepatectomy was individualized from wedge resection to extended right hepatectomy based on the clinical findings. In the hepatic hilum type ( n=26) a relatively small tumor in the neck infiltrated the hepatic hilum causing obstructive jaundice. Extended right hepatectomy plus bile duct resection with or without portal vein resection was necessary for curative resection because the tumor had extended into the right portal pedicle, and postoperative hepatic failure was common. In the bed and hilum type ( n=18) a huge mass occupying the entire gallbladder involved both the gallbladder bed and the hepatic hilum. Extended right hepatectomy with combined resection of contiguous spread was necessary for curative resection. In the lymph node type ( n=15) enlarged metastatic lymph nodes were the most prominent feature, and the primary tumor remains limited to the gallbladder in most cases. Extended lymphadenectomy with combined individualized resection was performed. In the cystic duct type ( n=9) a small mass arising from the cystic duct involved the common bile duct. This type presented at an earlier stage than the first four types. In the localized type ( n=24) tumor spread is localized to the gallbladder and presented at the earliest stage of any type. Simple cholecystectomy with or without wedge hepatic resection and regional lymphadenectomy resulted in a satisfactory outcome. Prognosis depends on the stage rather than on the mode of tumor spread. Even in the advanced types favorable results may be obtained in selected patients undergoing radical resection for M0 tumors without portal vein invasion. Success also was achieved in the rare patients with para-aortic lymph node metastases that were not infiltrative. CONCLUSIONS: These six types of gallbladder cancer can be diagnosed preoperatively by clinical and radiological examination. This information should assist the surgeon in the choice of operation and predict outcome.

Adult↗

Specific amino acid restriction inhibits attachment and spreading of human melanoma via modulation of the integrin/focal adhesion kinase pathway and actin cytoskeleton remodeling.

We had previously found that selective restriction of amino acids inhibits invasion of human A375 melanoma. Integrins, cell surface receptors for the components of extracellular matrix (ECM), are activated during cell adhesion and spreading, and initiate signaling pathways that control growth and invasion of tumor cells. We examined the effect of tyrosine (Tyr) and phenylalanine (Phe), methionine (Met) or glutamine (Gln) restriction on attachment and spreading of A375 and MeWo melanoma cell lines on fibronectin and laminin. In A375 cells, restriction of Tyr/Phe or Met inhibited attachment to and spreading on laminin and fibronectin, inhibited alpha3 and alpha4 integrin expression, and inhibited accumulation of FAK-Tyr397 and F-actin at leading edges of cell protrusions. Tyr/Phe restriction also inhibited attachment-induced autophosporylation of FAK-Tyr397. In MeWo cells, the order of inhibition by amino acid restriction on cell attachment and spreading was as follows: Gln > Tyr/Phe > Met. Restriction of Gln reduced alpha5 integrin expression. All amino acid restrictions similarly inhibited phosphorylation of FAK-Tyr397, FAK-Tyr577, FAK-Tyr861 and paxillin-Tyr31. Gln restriction exhibited the strongest inhibition of actin cytoskeleton remodeling during the cell spreading. The present study reveals that specific amino acid restriction inhibits attachment and spreading of melanoma via inhibition of specific integrin expression, inhibition of integrin-mediated FAK phosphorylation, and modulation of actin cytoskeleton remodeling. These data provide additional understanding of the mechanism by which specific amino acid restriction controls invasion and migration of melanoma.

Amino Acids↗

Perineural spread of malignant mesothelioma resulting in an intradural spinal cord mass: case report.

Spinal cord involvement by perineural spread of malignant mesothelioma is rare. We report a case of malignant mesothelioma that spread locally to invade the bony spine with both extradural and intradural perineural spread into the spinal canal that resulted in spinal cord compression. A 61-year-old man with a history of malignant mesothelioma presented with progressive leg weakness and right-sided arm weakness. Magnetic resonance imaging showed an enhancing lesion in the apex of the right lung with extension through the C7-T1 foramina with right hemicord enhancement. The patient underwent a C7-T1 laminectomy and right-sided C7-T1 and T1-T2 foraminotomies for neural decompression and biopsy of the lesion. Intraoperatively, tumor extended epidurally, and intradural perineural tumor spread along the C8 and T1 nerve roots into the spinal cord. Because it adhered to the spinal cord, no dissectible plane could be identified that would allow for safe total removal of the tumor. The epidural portion of the tumor, the adjacent involved bone, and the T1 nerve root were resected. Pathologic examination revealed malignant mesothelioma with bony invasion and perineural spread along the T1 nerve root. After decompression of the spinal cord, the patient had moderate improvement of his hand and leg function. Perineural spread of malignant mesothelioma resulting in spinal cord compression is an unusual clinical presentation. Intimate involvement of the spinal cord may prohibit aggressive tumor resection.

Humans↗

Mixing behavior of binary insoluble phospholipid monolayers. Analysis of the mixing properties of binary lecithin and cephalin systems by application of several surface and spreading techniques.

The detailed miscibility analysis of binary phospholipid monolayers requires the application of a variety of spreading and surface techniques which often yield complementary results. Testing the equilibrium state of the binary monolayer by long-time experiments is also of great importance. Studies of the compression and spreading behavior of binary monomolecular systems form a basis for the determination of binary monomolecular phase diagrams. Within these plots different phase regions occur which permit clear statements regarding the miscibility state. Additional knowledge of the miscibility properties (phase diagrams) of the binary bulk systems is required. From the analogy of the properties of the bulk systems, the miscibility state of the monolayers is also determined by the temperature, and we can classify the monolayers of binary lecithin and cephalin systems into systems of complete miscibility, partial miscibility and complete immiscibility. In addition to the differences in the chemical structure of the mixing components, the film states in the monolayer and the miscibility behavior of the bulk systems are also influencing factors. If one of the components does not produce a spreading pressure, miscibility gaps occur in the phase diagram of the phospholipid monolayer. The miscibility gap, expressed by a constant spreading pressure, indicates complete immiscibility within this concentration range. If both components produce spreading pressures, and condensed and liquid-expanded film states within the considered temperature range, partial miscibility of the components becomes probable. The most effective parameter is then the difference in the chemical structure of the components. When both components produce spreading pressures and condensed films, the chemical structure of the mixing phospholipid compounds within their hydrophilic and hydrophobic parts is of essential importance. Depending on the differences in the chemical structures of their chains and their head groups in the case of binary phospholipid monolayers, the following possibilities result: complete miscibility, partial miscibility and complete immiscibility of the lecithins and the cephalins. Complete miscibility within the binary phospholipid monolayer takes place in the case of identical head-group structure and where there are only small differences in the chain length of the fatty acid groups. With increasing hydrocarbon chain length differences, partial miscibility or even complete immiscibility can occur within the monolayer. Chemical differences in the head-group structure of the mixing components have a similar influence. In the case of binary lecithin/cephalin mixtures, the differences in the head-group structure affect the miscibility behavior more than the chain length differences do in the case of lecithin/lecithin and cephalin/cephalin mixtures.(ABSTRACT TRUNCATED AT 400 WORDS)

Liposomes↗

Halothane, but not alpha-chloralose, blocks potassium-evoked cortical spreading depression in cats.

The effects of two anesthetics, halothane and alpha-chloralose, on induction of spreading depression and on extracellular glutamate elevation after intracortical potassium administration were investigated in artificially ventilated (30% oxygen/70% nitrous oxide) cats. High potassium concentrations were achieved using either direct KCl injections (7 microliters, 150 mM via a micropipette) or microdialysis by supplementing 100, 300 or 500 mM KCl, respectively, for 10 min to the perfusion solution (Ringer's). Changes of the cortical DC potential were recorded adjacent (1-2 mm: electrode DC1) and distant (6-7 mm: electrode DC2) to the injection site. Either under halothane (0.75% in the respiratory gas mixture) or under alpha-chloralose (60 mg/kg i.v.) anesthesia, prolonged negative shifts of the DC potential reflecting the elevated potassium levels after KCl injection were measured near the injection site (electrode DC1). In contrast, spreading depressions (transient short DC deflections) were almost exclusively observed under alpha-chloralose. Spreading depressions recorded with electrode DC1 were superimposed on the prolonged negative DC shifts and they propagated frequently to the more distant site (DC2). Upon KCl administration, dose dependent elevations of extracellular glutamate were measured. These elevations were not significantly altered by the type of anesthesia. Our results suggest that in cats, spreading depression induction is affected by anesthesia, i.e., spreading depression induction is inhibited by halothane as compared to alpha-chloralose. Furthermore, factors other than glutamate or high potassium seem to contribute to spreading depression induction.

Animals↗

Human monocyte spreading induced by activated factor B of the complement alternative pathway: differential effects of Fab' and F(ab')2 antibody fragments directed to C5, C6, and C7.

Human peripheral blood mononuclear phagocytes are induced by activated Factor B (Bb) of the complement alternative pathway to undergo morphological shape changes in vitro which have been described as "spreading." The spreading reaction induced by Bb has previously been shown to depend upon the enzymatic activity of Bb and to be inhibited by Fab' antibody fragments directed to C5 (but not anti-C3 Fab'). The possibility that Bb may exert its effect on monocytes by initiating assembly of terminal complement complexes comprised of C5b, 6, 7, C5b-8, or C5b-9 was addressed in the present study. The effects were tested of Fab' and F(ab')2 antibody fragments directed to C5, C6, C7, and C8 and to neoantigens expressed in the assembling terminal complement complexes on the monocyte spreading reaction induced by Bb. Differential effects of monovalent Fab' and divalent F(ab')2 antibody fragments were observed. Anti-C5, C6, and C7 Fab' were found to inhibit the spreading reaction induced by Bb in an immunologically specific manner. Divalent F(ab')2 fragments directed to these same proteins (but not to C3, C4, C8, or C9) induced monocyte spreading in the complete absence of Bb or other recognized inducing agents. Monocyte spreading induced by hybridoma immunoglobulin (Ig) directed to C5 and C7 was found to be correlated with the binding of 10(6) molecules Ig per cell. These findings support the notion that C5, C6, and C7 (or an analogous system of cellular proteins) are associated with the surface of human peripheral blood monocytes and that these proteins may play a role in certain reactions by which mononuclear phagocytes are induced to altered states of cellular physiology.

Antibodies, Anti-Idiotypic↗

Unique morphology of HeLa cell attachment, spreading and detachment from microcarrier beads covalently coated with a specific and non-specific substratum.

A SEM and TEM evaluation of adhesion of HeLa-S3 cells to suspensions of culture microcarriers coated with various substrata revealed two unique cell morphologies. One is similar to that for cells attaching to culture dishes and the other one only appeared with microcarriers stirred under high shear conditions. The usual appearance of a spreading cell is to change from a sphere to the shape of a 'fried egg'. This proceeded in HeLa cells by a radial extension of the filopodia in between which the cytoplasm subsequently filled. Fluorescent antibody staining of actin suggested that more actin was present at the periphery of the spreading edges of the cell than inwards. The above morphology was characteristic of HeLa cell attachment to gelatin-coated microcarriers. However, the morphology of the attachment to microcarriers coated with non-biological substances such as negatively charged sulfonate groups or positively charged polyethyleneimine or even with the attachment protein laminin was quite different. Here the cells attached and began to spread as with gelatin-microcarriers, however, the spreading was not radial but occurred from one or two major regions of the cell periphery. The cell then appeared to constrict with the formation of a substratum attached pedestal upon which the cell body was perched. With time the cell pinched-off from pedestal. Evidence indicated that the pedestal was quite fragile. Furthermore, fluorescent antiactin staining indicated that the initial spreading region contained abundant actin which was depleted upon pedestal formation and detachment. The above in addition to previous kinetic measurements provided the information to classify cell substrate attachment materials into two distinct types. One is specific substrata which promote normal attachment and spreading and appear to interact with specific cell surface proteins. The other is non-specific substrata which in high shear conditions induces pedestal formation followed by pinching-off of the cells. Had previous attachment assays been done under high shear as done with the microcarriers and HeLa cells it is likely that substrata classified as specific might be reclassified into non-specific.

Actins↗

The sigma 1 protein determines the extent of spread of reovirus from the gastrointestinal tract of mice.

After intragastric inoculation of adult mice, type 1 reovirus was initially concentrated in Peyer's patches over the first 4 hr after inoculation, then spread sequentially to the mesenteric lymph nodes and spleen. For type 3 reovirus, however, initial entry into Peyer's patches in adult mice was followed by loss of viral infectivity so that by 4 hr after inoculation virtually no infectious virus was detected in the intestine, and spread to extraintestinal tissues did not occur. In 10-day-old mice, type 3 was capable of spread to the mesenteric lymph nodes but not the spleen. Thus, as animals aged there was a greater restriction of the spread of type 3 from the intestine. Studies using a field isolate of type 3 reovirus that is resistant to intestinal proteases, and genetic studies utilizing type 1 x type 3 viral reassortants, revealed that the viral sigma 1 protein determined the capacity of reovirus to spread from the intestine in both adult and 10-day-old mice. Thus, the interaction of reovirus with host defense mechanisms, and the age-dependent restriction of spread of type 3 reovirus from the intestine are mediated by the viral sigma 1 protein.

Aging↗

Variation in cytoskeletal assembly during spreading of progressively subcultivated human embryo fibroblasts (IMR-90).

The cytoskeletons of early and late passage IMR-90 human diploid fibroblasts have been directly imaged in replicas of Triton X-100 extracted cells during spreading following reseeding. All cells from both young and sensescent cultures exhibit a cytoskeletal network of actin microfilaments, intermediate (10 nm) filaments, microtubules, and interconnecting thin filaments (6-8 nm in diameter) which do not interact with heavy meromyosin. Early passage cells assemble linear aggregates of actin filaments within 1 h of spreading. By 4 h of incubation, these bundles establish a structural bond with the cell membrane which results in resistance by the plasmalemma to detergent extraction at these sites. Furthermore, these membrane regions are associated with developing stress fibers of well-spread cells. In contrast, late passage cells exhibit slower spreading which correlates with a retarded assembly of actin bundles. In addition, by 8 h of spreading, cells of older cultures do not exhibit the regions of membrane-actin interaction which impart detergent resistance to the plasmalemma. We conclude that the ability to reassemble actin-actin and actin-membrane association during cell spreading is reduced with increased serial subcultivation of cells.

Aging↗