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L Vakaet

Publications and source records attributed to L Vakaet.

At least 19 recordsLinked to original sources

Execution of a single-isocenter three-field technique, using a multileaf collimator or tray-mounted cerrobend blocks: effect on treatment time.

PURPOSE: In this article, we studied the total treatment time of a single-isocenter three-field irradiation of breast and axilla, using either tray-mounted cerrobend blocks, or a multileaf collimator (MLC) for field shaping. METHODS AND MATERIALS: A total of 20 female, unselected patients were given 50 Gy (2 Gy/fraction) on breast and 46 Gy on axilla and supraclavicular region (2 Gy/fraction). Patients were randomized between two different treatment groups. The first group (n = 10) was treated on a Philips SL-75 linear accelerator (SL-75), using 5 MV photons with tray-mounted cerrobend blocks. The second group (n = 10) was treated on a Philips SL-25 linear accelerator, using 6 MV photons and a MLC (SL-25-MLC). RESULTS: Although the beam-on time on the SL-25-MLC was significantly higher (p < 0.0001) compared to the SL-75, overall treatment time was significantly shorter using a MLC instead of tray-mounted cerrobend blocks (p < 0.0001). The difference in total treatment time was in the range of 100 s per patient per day. The main difference between the two accelerators was observed when setup of the second and third field was done using the automatic setup facility of the SL-25-MLC (avoids entering the treatment room). A mean time gain of 124 s per treatment session was observed using automatic setup. Considering the yearly number of patients receiving this treatment, a total time gain equivalent to 16.15 8-h workdays was calculated. CONCLUSIONS: Compared to a technique using tray-mounted cerrobend blocks in the single-isocenter three-field irradiation of a breast and axilla, a MLC combined with automatic field setup provides a significant time advantage, by reducing the number of manipulations inside the treatment room.

Axilla

Morphotypic plasticity in vitro and in nude mice of epithelial mouse mammary cells (NMuMG) displaying an epithelioid (e) or a fibroblastic (f) morphotype in culture.

Transition from an epithelioid (e-) to a fibroblastic (f-) morphotype marks invasiveness in clinical and experimental cancer. To understand better the factors influencing such transitions, we have subcloned and manipulated mouse mammary gland (NMuMG) cell cultures and compared the invasive phenotype of multiple subclones in vitro and in vivo. Cell lines with an e-morphotype expressed E-cadherin homogeneously and were not invasive in vitro. Cells with an f-morphotype were E-cadherin-negative and became fully invasive in vitro upon expression of the ras oncogene. Invasive tumors were produced in node mice after subcutaneous injection of e-type or f-type cells. These tumors showed cystic, glandular and undifferentiated structures. Tumors from f-type cells were E-cadherin-negative whereas e-type tumors stained heterogeneously in immunohistochemical preparations. Our observations demonstrate the impact of the micro-ecosystem on the invasive phenotype, with in vivo downregulation of E-cadherin and stimulation of the e- to f-morphotype transition.

Animals

In vivo and in vitro invasiveness of a rat colon-cancer cell line maintaining E-cadherin expression: an enhancing role of tumor-associated myofibroblasts.

In various cell systems, an inverse relationship was found between expression of E-cadherin, a molecule involved in the Ca(2+)-dependent homophylic cell-to-cell attachment of epithelial cells, and the capacity to invade extracellular matrix gels or normal tissues in vitro. DHD/K12/TRb (PROb) cells, maintained as a cell line derived from a rat colon carcinoma, homogeneously expressed in vitro immunoreactive E-cadherin, which was functional as shown in cell dissociation-reassociation assays. PROb cells were found to be non-invasive in 3 different assays in vitro. However, tumors resulting from a s.c. injection of PROb cells into syngeneic BD-IX rats were invasive, although PROb cells maintained E-cadherin expression in the tumors. Cells from a freshly dissociated PROb tumor showed, not only PROb cells but also tumor-associated myofibroblasts and were able to cross a Matrigel-coated filter. PROb tumors were indeed infiltrated by numerous myofibroblasts, mainly located at the invasive edge of the tumor. Cells from an established culture of tumor-infiltrating myofibroblasts were able to confer upon PROb cells invasiveness through Matrigel-coated filter or into chick-heart fragments. PROb cells maintained their capacity to express E-cadherin after myofibroblast-enhanced Matrigel invasion. Tumor-associated myofibroblasts, but not PROb cells, secreted a 72-kDa collagenase that could play a role in tumor-cell invasion. These results strongly suggest that cells from the tumor stroma, and more specifically myofibroblasts, may be involved in the invasiveness of epithelial tumor cells in vivo, even when E-cadherin expression prevents tumor-cell invasiveness in different in vitro assays.

Animals

Behaviour of dissociated hypoblast cells on the basal lamina and on extracellular fibrils in the gastrulating chicken embryo.

The spreading behaviour of dissociated hypoblast cells on and besides a band of aligned fibrils associated with the basal lamina of the epiblast was investigated by the use of scanning electron microscopy. A horse-shaped band of aligned fibrils, first demonstrated by Wakely and England (1979), is present during the gastrulation stages of chicken embryos on the ventral side of the epiblast at the cranial and lateral borders of the area pellucida. The basal lamina of the area pellucida situated inside the fibrillar band enables the spreading and probably the locomotion of dissociated cells, which appeared as polarized cells. Numerous cells were also found on the fibrillar band, and these cells lacked distinct lamellae and a polarized shape. Extensions of the cells contacted the extracellular fibrils and, at these sites of contact, the pattern of the fibrils was frequently deformed. From these observations and from previous results emerged the concept that spreading and locomotion of dissociated hypoblast cells, as well as single mesoblast cells and healing hypoblast epithelium, are inhibited by the band of extracellular fibrils, which acts as a physical barrier. The cell biological basis of the mechanism by which extracellular fibrils associated with the basal lamina arrest the migration of hypoblast and mesoblast cells, but guide the migration of primordial germ cells, is discussed.

Animals

Loss of epithelial differentiation and gain of invasiveness correlates with tyrosine phosphorylation of the E-cadherin/beta-catenin complex in cells transformed with a temperature-sensitive v-SRC gene.

Loss of histotypic organization of epithelial cells is a common feature in normal development as well as in the invasion of carcinomas. Here we show that the v-src oncogene is a potent effector of epithelial differentiation and invasiveness. MDCK epithelial cells transformed with a temperature-sensitive mutant of v-src exhibit a strictly epithelial phenotype at the nonpermissive temperature for pp60v-src activity (40.5 degrees C) but rapidly loose cell-to-cell contacts and acquire a fibroblast-like morphology after culture at the permissive temperature (35 degrees C). Furthermore, the invasiveness of the cells into collagen gels or into chick heart fragments was increased at the permissive temperature. The profound effects of v-src on intercellular adhesion were not linked to changes in the levels of expression of the epithelial cell adhesion molecule E-cadherin. Rather, we observed an increase in tyrosine phosphorylation of E-cadherin and, in particular, of the associated protein beta-catenin. These results suggest a mechanism by which v-src counteracts junctional assembly and thereby promotes invasiveness and dedifferentiation of epithelial cells through phosphorylation of the E-cadherin/catenin complex.

Animals

Testis tumors.

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Adolescent

Expression of E-cadherin in embryogenetic ingression and cancer invasion.

Homophilic interactions of E-cadherin serve the organization of embryonic and adult epithelia and counteract cancer invasion. The role of E-cadherin as an invasion-suppressor molecule has been demonstrated cancer. Regulation of embryonic ingression and cancer invasion via E-cadherin occurs at transcriptional, translational and post-translation levels.

Animals

Study of yolk precursor transport in the avian ovary with the use of horseradish peroxidase.

Ovaries of adult Japanese quails were exposed in vivo to the exogenous protein horseradish peroxidase (HRP) for varying lengths of time to investigate ultrastructurally the permeability of the wall of these follicles, the protein uptake capacity of granulosa and oocyte and the kinetics of protein uptake in different stages. There is a sudden increase in permeability of the follicle wall from previtellogenesis to vitellogenesis. This is not due to a loss of sealing (tight) junctions in the granulosa cell layer, but is probably related to a permeability change in the basement membrane. The transition from the slow growth phase to the rapid growth during vitellogenesis is accompanied by a limited widening of the intercellular channels and the concomitant development of a complex endocytotic apparatus in the ooplasm. The slowing down of yolk deposition during the last day before ovulation is accompanied by a narrowing of the intercellular channel width. The granulosa cells show a high intracellular HRP uptake during intermediary yolk formation. Transcytosis through the granulosa cannot be excluded but is probably a minor pathway at certain stages. The light microscopically detectable uptake of HRP by the oocyte coincides with the start of exogenous vitellogenesis. After 90 sec of exposure to HRP (intravenous injection) the tracer can be found in the intercellular channels of the granulosa and in superficially located yolk spheres. On the other hand it takes 10 min for the tracer to traverse the cortex of the oocyte.

Animals

The role of chemotherapy in the management of advanced laryngeal cancer.

At the time of diagnosis about half the patients with laryngeal cancer have locally advanced disease. In our series of 720 larynx carcinoma patients, treated in the department of radiotherapy at the University hospital in Ghent, in a period between 1975 and 1991, 45.2% of the patients had stage III or IV disease. The excellent results from locoregional treatment of limited disease are given elsewhere in this issue (De Schryver). Results for treatment of stage III and IV disease remain however poor. Locoregional failure rates of 50% and overall survival rates of 30% after 5 years indicate the need for better initial local and systemic control in advanced stage disease.

Antineoplastic Agents

The arrest of cell migration in the chicken blastoderm: experimental evidence for the involvement of a band of extracellular fibrils associated with the basal lamina.

This article overviews our current knowledge of the occurrence and distribution of oriented extracellular fibrils associated with the basal lamina, and their presumptive role in contact guidance of cells in early embryos. To investigate the role of the band of extracellular fibrils situated at the basal side of the epiblast at the cranial edge of the area pellucida of the chicken blastoderm, we determined the precise location and morphology of the fibrils using TEM and SEM, described the relationship between migrating mesoblast cells and the fibrillar band, and, finally, tested experimentally the behavior of homologous and heterologous tissues in the vicinity of the fibrillar band. The descriptive analysis demonstrated that a horseshoe-shaped, 170 microns-wide band of fibrils occurs at the cranial and lateral edges of the area pellucida and area opaca, the highest density being found in the area pellucida. Migrating mesoblast cells presented a surface morphology that was different from the morphology of cells reaching the fibrils at the lateral edge of the area pellucida. Mesoblast cells never crossed the fibrils, an observation that may explain why during gastrulation, mesoblast cells invade the area opaca only in the caudal region, where no fibrillar band is present. The experimental analysis, which involved transplantation and healing experiments, demonstrated that the arrest of cell migration, that occurred in all cases in the vicinity of the fibrillar band, was correlated with changes in surface morphology suggesting a decreased cell adhesion to the fibrils. From these observations emerged the view that the horseshoe-shaped fibrillar band functions as a barrier inhibiting migration of individual mesoblast cells and expansion of tissue sheets, rather than as an extracellular substrate mediating the oriented guidance of cells. In addition to its inhibitory role in cell migration, the extracellular band may also be regarded as a factor that stabilizes the polarity of the early embryo by determining the cranial and lateral limits between embryonic and extraembryonic tissues.

Animals

Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role.

A cDNA encoding the cell-cell adhesion molecule E-cadherin was transfected into highly invasive epithelial tumor cell lines of dog kidney or mouse mammary gland origin. Transfectants with a homogeneously high expression of E-cadherin showed a reproducible loss of activity in two types of in vitro invasion assays. Invasiveness of these transfectants could be reinduced specifically by treatment with anti-E-cadherin antibodies. In vivo, they formed partly differentiated tumors, instead of fully undifferentiated tumors. Alternatively, a plasmid encoding E-cadherin-specific anti-sense RNA was introduced into noninvasive ras-transformed cells with high endogenous E-cadherin expression. The resulting down-regulation, albeit partial, rendered the cells invasive. These data provide direct evidence that E-cadherin acts as an invasion suppressor molecule.

Animals

Features of polyingression and primitive streak ingression through the basal lamina in the chicken blastoderm.

The de-epithelialization of cells of the upper layer during the phenomena of polyingression and primitive streak ingression was studied by analyzing, from the time of laying to the end of gastrulation, the ultrastructure of the basal lamina underlying the upper layer. The electron density of the basal lamina and associated extracellular materials was enhanced by addition of tannic acid to the fixative. Special attention was also paid to the spatial and temporal distribution of blebs at the basal surface of the upper layer, and to the contribution of the de-epithelialized cells to the formation of the deep layer. The results indicate that a nascent basal lamina is already present at the time of laying, especially beneath regions of the area pellucida where polyingression is not apparent. From the onset of incubation, the basal lamina rapidly develops, and it is interrupted by a large number of blebs. However, during the first 6-8 h of incubation, i.e., stages 1-2 of Vakaet (Arch. Biol. (Liège) 81:387-426, 1970), a downward movement of de-epithelialized cells that insert into the deep layer and form the endophyll persists cranially. This phenomenon of polyingression, which starts during the intrauterine period, probably extends from caudal to cranial and comes to an end by stage 3. During these first three stages, the number of blebs progressively decreases, especially in the cranial part of the area pellucida, and a thicker, continuous basal lamina associated with numerous interstitial bodies is laid down. The caudal part of the upper layer is still actively blebbing at that time. Due to the convergence of this area toward the axis of the blastoderm, which leads to ingression at and elongation of the primitive streak up to and including stage 6, the number of blebs at the basal surface of the upper layer progressively decreases. From stage 7 on, blebs are virtually absent; shortening of the primitive streak and formation of the head process begin. At the level of the head process, primitive streak ingression has ceased and a novel basal lamina is progressively deposited beneath the upper layer. By stage 9, a thick, smooth basal lamina physically separates the upper layer from the head mesenchyme. Summarizing, at the time of gastrulation, the presence of blebs that perforate the basal lamina is correlated with the de-epithelialization of cells. Before incubation, however, de-epithelialization of upper-layer cells occurs before the assembly of the basal lamina.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Esophageal cancer.

The authors invited eminent Belgian specialists to report on their experience in esophageal cancer. A summary is given of the most striking features of their reports.

Combined Modality Therapy

Numerical evaluation of the invasion of closely related cell lines into collagen type I gels.

Cells were seeded on top of a reconstituted collagen gel layer, and their migration into the gel was evaluated as an assay for invasive behavior. The method was standardized by measuring the depth of migration of each cell in a defined volume of the gel. We developed a microscope stage, controlled by a computer program. This semiautomatic counting method allowed precise vertical localization of each cell in a collagen gel with an error of less than 0.1 micron. To test the discriminative power of the assay, we used cell lines which were known to be invasive or noninvasive in other assays. Closely related variants of 2 cell families were chosen: (1) one family derived from a mouse mammary gland (NMuMG), and (2) one derived from a mouse T cell lymphoma (BW5147). The assay could discriminate between invasive and noninvasive variants of related cell lines within the same family. The profile of the number of cells in each layer of the gel provided additional discrimination between the different cell lines. Furthermore, the assay allowed direct microscopic observation of cells migrating in the collagen gel. The present standardization makes the collagen assay suitable for semiautomatic testing of the invasive phenotypes in cell populations from the same as well as from different cell families.

Animals

Fate mapping the avian neural plate with quail/chick chimeras: origin of prospective median wedge cells.

The origin of prospective M cells, which are median neuroepithelial cells that become wedge-shaped during bending of the neural plate and eventually form the midline floor of the neural tube, was determined by constructing quail/chick chimeras and using the quail nucleolar marker to identify quail donor cells in chick host blastoderms. Two possible sites of prospective M-cell origin in the epiblast were examined: a single, midline rudiment located just rostral to Hensen's node and paired rudiments flanking the cranial part of the primitive streak. Our results suggest that M cells arise exclusively from the midline, prenodal rudiment. From this rudiment, M cells extend caudally throughout the entire length of the neuroepithelium. This new information on the origin of prospective M cells will aid in the analysis of their role in neurulation.

Animals

Neural plate- and neural tube-forming potential of isolated epiblast areas in avian embryos.

Formation, shaping, and bending of the neural plate and closure of the neural groove are complex processes resulting in formation of the neural tube. Two experiments were performed using avian embryos as model systems to examine these events. First, we transected blastoderms near the level of Hensen's node to determine the potential of prenodal neural plate to form neural tube in isolation from primitive streak regression. Our results demonstrate that shaping and bending of the prenodal neural plate occur under these conditions, but neural groove closure is inhibited. Second, we isolated various areas of postnodal epiblasts to determine their potential to form neural plate. Our results suggest that the area of the postnodal epiblast that can form neural plate consists of paired tracts lying adjacent to the definitive primitive streak and extending caudally at least 1 mm from its cranial end.

Animals

Immunohistochemistry of laminin in early chicken and quail blastoderms.

We have used immunohistochemical techniques to study laminin in quail blastoderms milked from the oviduct and the distribution of laminin in laid chicken and quail blastoderms. Laminin is a constituent of the basement membrane in both chicken and quail blastoderms. It is found at the ventral side of the upper layer cells. Laminin is first observed under individual upper layer cells in prelaid quail blastoderms 15 h post-ovulation, but is absent at the ingression site of endophyll cells. The presence of a continuous laminin layer coincides with the epithelialization of the epiblast after 5-10 h incubation. The laminin layer is discontinuous at the primitive streak and at Hensen's node. It is thinner and partly discontinuous at the median part of the neural plate. By induction, either of an ectopic primitive streak or a neural plate, we have demonstrated, using the chicken-quail nucleolar marker technique, that at these sites the laminin layer is interrupted. A laminin layer might confer rigidity onto the epiblast, whereas disruption of a laminin layer seems to be correlated with ingression of cells or bending of the neural plate.

Animals