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PubMed1997

Spread of measles virus through axonal pathways into limbic structures in the brain of TAP1 -/- mice.

The spread of measles virus into the brain was studied exploiting the olfactory pathway, which represents an important route of neuroinvasion by viruses. The virus was injected into the main olfactory bulb of wild-type mice and mice with disrupted TAP1 gene (TAP refers to the Transporter associated with Antigen Presentation), which codes for products essential for the cell-mediated immune response. Virus invasion was monitored for 4 weeks by immunohistochemistry. The distribution of measles virus was found to be restricted to brain areas connected with the olfactory bulbs. However, in the wild-type mice there was a marked infiltration of lymphocytes in the infected brain structures, and the virus did not pass beyond the piriform cortex. In the TAP1 -/- mice the virus spread more extensively along olfactory projections into the limbic system and monoaminergic brainstem neurons. Infected mice of both types developed seizures, which may have been focally evoked from the piriform cortex. This study provides evidence that measles virus can spread through axonal pathways in the brain. The findings obtained in the gene-manipulated mice point out that a compromised immune state of the host may potentiate targeting of virus to the limbic system through olfactory projections.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1998

Laminin-5 coating enhances epithelial cell attachment, spreading, and hemidesmosome assembly on Ti-6A1-4V implant material in vitro.

Enhancement of epithelial cell attachment to laminin-5-coated titanium alloy (Ti-6Al-4V) implant material was evaluated in vitro. Protein analysis showed that Ti-6Al-4V has a high affinity for laminin-5 and adsorbed significantly more laminin-5 than laminin-1. DNA analysis showed that laminin-5 enhanced attachment of normal human epidermal keratinocytes (NHEK) to Ti-6Al-4V significantly more than did laminin-1 or uncoated controls. The effect of passivation on laminin-5 adsorption and activity on Ti-6Al-4V also was evaluated. Passivation had no significant effect on the amount of protein adsorbed; however, AFM, ESCA, and ToF-SIMS analyses suggested that passivation affects the conformation of adsorbed laminin-5. Although laminin-5 coating significantly enhanced rapid attachment of epithelial cells to both passivated and unpassivated Ti-6Al-4V, surface area measurements showed that cells spread on laminin-5-coated passivated Ti-6Al-4V covered a significantly larger surface area than cells spread on laminin-5-coated unpassivated samples. TEM analysis showed that cells formed significantly more hemidesmosomes on the surface of laminin-5 coated passivated than on the surface of laminin-5 coated unpassivated titanium alloy. The enhancement of rapid cell attachment, spreading, and hemidesmosome assembly on laminin-5-coated passivated samples may reflect better integration between epithelial cells and titanium alloy and thus may be predictive of long-term implant stability.

Cell Adhesion↗

PubMed2000

Modulating fibroblast adhesion, spreading, and proliferation using self-assembled monolayer films of alkylthiolates on gold.

Ultrathin, highly organized functionalized alkylthiol monolayers were applied as model substrates for cell growth and protein adsorption studies. The aim of this approach was to improve the understanding of molecular surface determinants required for adhesion-dependent cell growth and proliferation using well-controlled surface chemistry. Carboxyl- and methyl-terminated alkylthiol monolayers on gold were used to monitor Swiss 3T3 fibroblast adhesion, spreading, and growth. Stress fiber and focal contact formation were determined by immunostaining of actin filaments and paxillin. Fibronectin deposition and conformation on these surface chemistries in the presence and absence of competing proteins were also determined. The relative levels of adsorbed fibronectin were assessed using radiolabeled proteins. Exposure of the 10th type III cell integrin binding domain of fibronectin was assessed using a radiolabeled monoclonal antibody. Distinct alkylthiol substrate chemistry-dependent differences were observed in fibroblast adhesion, spreading, and growth. The formation of focal contacts and stress fibers was enhanced on the carboxyl-terminated surface relative to the methyl surface. Relative deposition and conformations of adsorbed fibronectin were shown to be dependent on surface chemistry in both the presence and absence of competing proteins. The results indicated that well-controlled culture surfaces modulate differential cell adhesion, spreading, and growth through modulations of the amounts and conformations of adsorbed extracellular matrix molecules (e.g., fibronectin).

3T3 Cells↗

PubMed1998

Possible routes of spread of carcinoma of the maxillary sinus to the oral cavity.

Carcinomas arising in the maxillary sinus, occasionally present clinically in the oral cavity along the occlusal ridge of the upper alveolus. The mechanism of this spread has not been investigated but it may resemble invasion of the mandible by oral carcinomas, which tend to invade the soft tissues rather than eroding through bone. In Britain, such patients are usually edentulous. This project examines deficiencies in the bony walls of the maxillary sinus to determine possible routes of spread. Maxillae were obtained from one side of 17 dissecting room cadavers, aged 71-95 years; 15 were edentulous. The roof of the maxillary sinus was removed in all specimens. Five maxillae were each cut into six slices, 0.5 cm thick, in a vertical bucco-lingual plane. In all, soft tissues were removed by treatment in dilute bleach. Histological sections, cut in the horizontal plane, were prepared of a further three maxillae. Several possible routes were identified by which tumors could spread from the sinus to the oral cavity: 1) directly through foramina in its floor; 2) through numerous foramina in its floor to the marrow cavity of the alveolar process, which mostly contained fat amongst narrow bony trabeculae. (The marrow cavity, in turn, had numerous communications with the occlusal surface); 3) through foramina which carried branches of the superior alveolar nerves and vessels; 4) through deficiencies in the bony walls of the sinus at the neurovascular grooves and elsewhere.

Aged↗

PubMed2000

Single-pinhole confocal imaging of sub-resolution sparse objects using experimental point spread function and image restoration.

Sparse fluorescent pointlike subresolution objects have been imaged using a diffraction limited single-pinhole confocal fluorescence microscope. A Maximum likelihood image restoration algorithm has been used in conjunction with a measure of the experimental point spread function for improving the three-dimensional imaging of subresolution sparse objects. The experimental point-spread-function profiles have been improved by a factor of 1.95 in lateral direction and 3.75 in axial direction resulting in full-width half maximum (FWHM) values of 91 +/- 11 nm and 160 +/- 26 nm. This amounts to 1. 43 and 2.15 in optical units, respectively. The lateral and axial FWHM of the sparse pointlike subresolution objects is about 5 and 3 times smaller than the wavelength. This result points to the attractive possibility of utilising a compact confocal architecture for localising punctuate fluorescent objects having subresolution dimensions. The key resides in the utilisation of the measured point spread function coupled to an appropriate image restoration approach, and, of course, in the stability of the confocal system being used.

Algorithms↗

PubMed1980

The cascade theory of metastatic spread: are there generalizing sites?

The term "cascade spread," with regard to metastases, is being used ambiguously. some workers take it to mean that cancers metastasize along preferred pathways, others infer that some sites are seeded first and that further metastases proceed from these sites rather than from the primary tumor. It is our opinion that the statistical arguments used to support the second approach have serious flaws. Given our present understanding of the metastatic process, a spread along preferred pathways is supported both by physiologic as well as statistical considerations. However, the contribution "generalizing sites" to the spread of metastases remains unproven.

Humans↗

PubMed1985

Ductal spread in prostatic carcinoma.

The histologic appearances of ductal invasion were studied in 139 cases of prostatic adenocarcinoma diagnosed in the Department of Pathology, Howard University Hospital, during the period January 1980 through October 1983. Intraductal spread was found in almost half (48%) of the prostatic glands examined. Ductal spread was associated with the local extent (P less than 0.001) rather than with the grade of the tumor (P less than 0.01). Three distinct patterns of ductal penetration were recognized. The duct wall was completely destroyed in microinvasion. In foci of ductal permeation the integrity of the basement membrane was generally preserved, and the duct wall was infiltrated mainly by solitary tumor cells. When the tumor spread was by extension in continuity within the duct wall, the neoplastic cells appeared to grow between the pre-existing epithelial layers. It was concluded that prostatic carcinoma cells have the ability to penetrate the wall of benign ducts and progressively replace the normal epithelial elements. In this process the general framework of the affected duct appears to be preserved.

Adenocarcinoma↗

PubMed2000

Monoclonal antibody MS13 identifies a plasmatocyte membrane protein and inhibits encapsulation and spreading reactions of Manduca sexta hemocytes.

Lepidopterans generally can successfully defend themselves against a variety of parasites or parasitoids. One mechanism they use is to encapsulate the invader in many layers of hemocytes. For encapsulation to occur, the hemocytes must attach to the foreign material, spread, and adhere to each other. The molecules that mediate these processes are not known. One method to identify proteins potentially necessary for adhesion, spreading, and, thus, encapsulation is to use monoclonal antibodies that interfere with these functions. In this paper, we report that a monoclonal antibody against Manduca sexta plasmatocytes effectively inhibited encapsulation of synthetic beads in vitro and in vivo. Furthermore, it inhibited plasmatocyte spreading in vitro. Other anti-hemocyte antibodies did not have these effects. The plasmatocyte-specific monoclonal antibody, mAb MS13, recognized a protein of approximately 90,000 daltons as indicated by Western blot analysis of hemocyte lysate proteins. The epitope recognized by mAb MS13 was present on the exterior surface of plasmatocytes. Using indirect immunohistochemistry with hemocyte-specific antibodies, we also determined that during encapsulation plasmatocytes were the first cells bound to latex beads and later layers consisted of both plasmatocytes and granular cells. Arch.

Animals↗

PubMed1993

TNF-alpha induces spreading of B-CLL via the CD11c/CD18 molecule.

Activation of malignant B cells can lead to extensive morphological changes of these cells. A combination of PMA and TNF-alpha can induce adherence of purified B-CLL cells, which acquire a dendritic cell-like appearance. This phenomenon that we have termed spreading, is accompanied by upregulation of expression of both beta 1 and beta 2 integrin molecules. Spreading was inhibited by the addition of antibodies against CD18 or CD11c. In other B cell malignancies (HCL and NHL), morphological changes could be induced by PMA in the absence of TNF-alpha. Culturing in the presence of prednisolone resulted in an inhibition of spreading, most likely mediated via a down regulation of CD18 and CD11c expression. These data indicate that the CD11c/CD18 complex might be important for the adhesive properties of B cells.

Antigens, CD↗

PubMed1988

Spread and maintenance of a disease in a structured population.

The patterns of interactions among different groups of individuals in a structured population is an important factor affecting the ease of establishment of a disease and the rate of spread of the disease in a population. A model for the spread of a disease in such a population is described. The model considers two levels of interaction: interactions between individuals within a subpopulation because of geographic proximity, and interactions between individuals of the same or different sub-populations because of attendance at common social functions. Analysis of the model leads to sufficient conditions which determine maximum subpopulation sizes which allow the disease to be maintained in the population. These conditions are inequalities relating the removal rate of infectives to the infection rate of susceptibles. Results are discussed for both idealized movement patterns and for the spread of hepatitis A among day care centers in Albuquerque, New Mexico. In addition, the basic structures of the contact matrices necessary to extend the model to deal with vector-borne diseases and with sexually transmitted diseases are discussed.

Hepatitis A↗

PubMed1979

Subarachnoid spread of ependymoma: diagnosis and evaluation of therapy by computerized tomography.

The potential of ependymomas to spread throughout the intracranial subarachnoid space is well documented, though the incidence is disputed. This relates in part to the time required for such metastases to produce symptoms. The diagnosis of such spread, however, is important for therapeutic management. This report demonstrates the effectiveness of computerized tomography in diagnosing spread of ependymoma throughout the basal cisterns and cerebral fissures.

Adult↗

PubMed2004

Active sites in the carboxyl-terminal region of the laminin alpha chain in Drosophila neuronal cell spreading.

An established Drosophila neuronal cell line (BG2-c6) proved to be useful to analyze laminin-mediated cell spreading and signal transduction [Takagi et al. (2000) Biochem Biophys Res Commun 270:482-487]. Here, we report, in addition to the whole molecule, the truncated alpha chain of Drosophila laminin (containing the entire carboxyl-terminal globular domain) and two dodecapeptides corresponding to the cell-binding sites identified in the alpha1 chain of mouse laminin were also active to stimulate BG2-c6 cell spreading. Our previous study [Takagi et al. (1996) J Biol Chem 271:18074-18081] revealed that these recombinant protein and synthetic peptides promoted neurite outgrowth in the primary cell culture system prepared from Drosophila embryo. Therefore, the similar effects by these proteins and peptides suggest the presence of a common mechanism of laminin and neuronal cell interaction working in both primary and established cells. One of the two active peptides contains the sequence SIKVGV. Its murine counterpart carries the sequence SIKVAV by which the interaction of laminin and cells is mediated. Furthermore, laminin-dependent BG2-c6 cell spreading was inhibited by heparin. This observation suggests that cell surface glycoproteins participate in the interaction of laminin and BG2-c6 cells.

Amino Acid Sequence↗

PubMed1990

Use of a spread sheet to calculate the current-density distribution produced in human and rat models by low-frequency electric fields.

The current-density distribution produced inside irregularly shaped, homogeneous human and rat models by low-frequency electric fields is obtained by a two-stage finite-difference procedure. In the first stage the model is assumed to be equipotential. Laplace's equation is solved by iteration in the external region to obtain the capacitive-current densities at the model's surface elements. These values then provide the boundary conditions for the second-stage relaxation solution, which yields the internal current-density distribution. Calculations were performed with the Excel spread-sheet program on a Macintosh-II microcomputer. A spread sheet is a two-dimensional array of cells. Each cell of the sheet can represent a square element of space. Equations relating the values of the cells can represent the relationships between the potentials in the corresponding spatial elements. Extension to three dimensions is readily made. Good agreement was obtained with current densities measured on human models with both, one, or no legs grounded and on rat models in four different grounding configurations. The results also compared well with predictions of more sophisticated numerical analyses. Spread sheets can provide an inexpensive and relatively simple means to perform good, approximate dosimetric calculations on irregularly shaped objects.

Animals↗

PubMed1995

Total mesorectal excision and local recurrence: a study of tumour spread in the mesorectum distal to rectal cancer.

Total mesorectal excision (TME) appears to be associated with a reduced local recurrence rate following surgery for rectal cancer. Of 20 patients with rectal cancer in whom TME was performed, adenocarcinoma was found in the distal mesorectum in four. Distal mesorectal spread often extended further than intramural spread. Patients with tumour in the distal mesorectum had a worse outcome at 4-year follow-up, a greater risk of local recurrence and an increased frequency of distant metastasis. Distal tumour spread is, therefore, a marker of poor prognosis in rectal cancer. This study provides further evidence that incomplete excision of the mesorectum contributes to local recurrence in a proportion of patients with rectal cancer, particularly in those with tumours in the middle and lower third of the rectum.

Adenocarcinoma↗

PubMed1989

A computer model for unconscious spread of anxiety-linked inhibition in cognitive networks.

Unconscious inhibitory processes, triggered by a potential anxiety reaction, are reviewed in the context of an emerging rapprochement between psychodynamic and cognitive approaches in experimental psychology. Conditions underlying spread of inhibitory action to other cognitive networks are first explored in three tachistoscopic experiments utilizing words posthypnotically tied to a potential anxiety, pleasure, or neutral reaction. Response times of subjects, instructed to ignore those words while naming pictures or solving anagrams as quickly as possible, reveal a highly differentiated pattern of circumstances governing likelihood of inhibitory spread from anxiety-linked words to target stimuli. Next a computer model is constructed to simulate cognitive processes from onset of display to eventual response, and the model is then tested for its fit to the empirical data. Finally, an illustrative study shows that a subset of computer-generated predictions for spread of inhibitory action is verifiable experimentally.

Adult↗

PubMed1983

Fibronectin is not a serum spreading factor for HeLa cells.

The spreading of HeLa cells on plastic substratum is mediated by fibronectin-depleted foetal calf serum but not by fibronectin isolated by gelatin-Sepharose affinity chromatography. The same is true for freshly explanted chick embryonic chondrocytes. In contrast, BHK cell spreading exceeds 67% after 120 min at 37 degrees C in fibronectin-supplemented (10 micrograms ml-1) serum-free medium. Long-term cultivation of HeLa cells in Eagle's MEM supplemented with fibronectin-free serum is associated with the accumulation of cells in mitosis or before cytokinesis; many cells die and the remaining living cells, characterized by marked changes in morphology, multiply very slowly. It can be concluded therefore that fibronectin does not produce spreading in HeLa cells but forces them into mitosis.

Animals↗

PubMed2001

Over-expression of betaI tubulin in MDCK cells and incorporation of exogenous betaI tubulin into microtubules interferes with adhesion and spreading.

Little is known about the presence and distribution of tubulin isotypes in MDCK cells although essential epithelial functions in these monolayers are regulated by dynamic changes in the microtubule architecture. Using specific antibodies, we show here that the betaI, betaII, and betaIV isotypes are differentially distributed in the microtubules of these cells. Microtubules in subconfluent cells radiating from the perinuclear region contain betaI and betaII tubulins, while those extending to the cell edges are enriched in betaII. Confluent cells contain similar proportions of betaI and betaII along the entire microtubule length. betaIV is the less abundant isotype and shows a similar distribution to betaII. The effect of modifying tubulin isotype ratios in the microtubules that could affect their dynamics and function was analyzed by stably expressing in MDCK cells betaI tubulin from CHO cells. Three recombinant clones expressing different levels of the exogenous betaI tubulin were selected and subcloned. Clone 17-2 showed the highest expression of CHO beta1 tubulin. Total betaI tubulin levels (MDCK+CHO) in the clones were approximately 1.8 to 1.1-fold higher than in mock-transfected cells only expressing MDCK beta1 tubulin. In all the cells, betaII tubulin levels remained unchanged. The cells expressing CHO beta1 tubulin showed defective attachment, spreading, and delayed formation of adhesion sites at short times after plating, whereas mock-transfected cells attached and spread normally. Analysis of cytoskeletal fractions from clone 17-2 showed a MDCK betaI/CHO betaI ratio of 1.89 at 2 h that gradually decreased to 1.0 by 24 h. The ratio of the two isotypes in the soluble fraction remained unchanged, although with higher values than those found for the polymerized betaI tubulin. By 24 h, the transfected cells had regained normal spreading and formed a confluent monolayer. Our results show that excess levels of total betaI tubulin, resulting from the expression of the exogenous beta1 isotype, and incorporation of it into microtubules affect their stability and some cellular functions. As the levels return to normal, the cells recover their normal phenotype. Regulation of betaI tubulin levels implies the release of the MDCK betaI isotype from the microtubules into the soluble fraction where it would be degraded.

Animals↗

PubMed2006

Myoblast attachment and spreading are regulated by different patterns by ubiquitous calpains.

The calcium-dependent proteolytic system is a large family of well-conserved ubiquitous and tissue-specific proteases, known as calpains, and an endogenous inhibitor, calpastatin. Ubiquitous calpains are involved in many physiological phenomena, such as the cell cycle, muscle cell differentiation, and cell migration. This study investigates the regulation of crucial steps of cell motility, myoblast adhesion and spreading, by calpains. Inhibition of each ubiquitous calpain isoform by antisense strategy pinpointed the involvement of each of these proteases in myoblast adhesion and spreading. Moreover, the actin cytoskeleton and microtubules were observed in transfected cells, demonstrating that each ubiquitous calpain could be involved in the actin fiber organization. C2C12 cells with reduced mu- or m-calpain levels have a rounded morphology and disorganized stress fibers, but no modification in the microtubule cytoskeleton. Antisense strategy directed against MARCKS, a calpain substrate during C2C12 migration, showed that this protein could play a role in stress fiber polymerization. A complementary proteomic analysis using C2C12 cells over-expressing calpastatin indicated that two proteins were under-expressed, while six, which are involved in the studied phenomena, were overexpressed after calpain inhibition. The possible role of these proteins in adhesion, spreading, and migration was discussed.

Actins↗