Image-guided stereotactic ablative radiotherapy for the liver: a safe and effective treatment.

AIMS: Stereotactic ablative body radiotherapy (SABR) is a non-invasive treatment option for inoperable patients or patients with irresectable liver tumors. Outcome and toxicity were evaluated retrospectively in this single-institution patient cohort. PATIENTS AND METHODS: Between 2010 and 2014, 39 lesions were irradiated in 33 consecutive patients (18 male, 15 female, median age of 68 years). All the lesions were liver metastases (n = 34) or primary hepatocellular carcinomas (n = 5). The patients had undergone four-dimensional respiration-correlated PET-CT for treatment simulation to capture tumor motion. We analyzed local control with a focus on CT-based response at three months, one year and two years after treatment, looking at overall survival and the progression pattern. RESULTS: All patients were treated with hypofractionated image-guided stereotactic radiotherapy. The equivalent dose in 2 Gy fractions varied from 62.5 Gy to 150 Gy, delivered in 3-10 fractions (median dose 93.8 Gy, alpha/beta = 10). The CT-based regression pattern three months after radiotherapy revealed partial regression in 72.7% of patients with a complete remission in 27.3% of the cases. The site of first progression was predominantly distant. One- and two-year overall survival rates were 85.4% and 68.8%, respectively. No toxicity of grade 2 or higher according to the NCI Common Terminology Criteria for Adverse Events v4.0 was observed. CONCLUSION: SABR is a safe and efficient treatment for selected inoperable patients or irresectable tumors of the liver. Future studies should combine SABR with systemic treatment acting in synergy with radiation, such as immunological interventions or hypoxic cell radiosensitizers to prevent distant relapse.

Cell-lineage and clonal-contribution map of the trochophore larva of Patella vulgata (mollusca).

Molluscan development is characterised by its extremely regular cleavage pattern. In numerous molluscs the fate of various early-cleavage stage blastomeres has been determined and fate maps have been constructed. On the basis of similarities between these fate maps, a generalised molluscan cell-lineage map has been constructed. Recently, the validity of this map has been challenged. In this study, the cell-lineage of the first-, second-, and third-quartet micromeres and third-generation macromeres of the equally-cleaving gastropod mollusc Patella vulgata was studied by fluorescent cell-lineage tracer injection followed by epifluorescence microscopy and confocal laser scanning microscopy. For the first time, a complete cell-lineage map, in the form of a clonal-contribution map of the trochophore, has been constructed with the use of fluorescent cell-lineage tracers. This map both agrees and differs in a number of respects with the generalised cell-lineage map of molluscs. The most important deviation is that the micromere 2d, formerly referred to as the first somatoblast, is not the only cell that forms the foot and shell gland in Patella.

Organiser role of the stem cell of the mesoderm in prototroch patterning in Patella vulgata (Mollusca, Gastropoda).

During early development of the gastropod mollusc Patella vulgata, the stem cell of the mesoderm (3D-macromere) is induced. As a result of this induction, the embryo becomes dorsoventrally organised. At about the same time in development, ciliated cells, so-called trochoblasts, are formed. Later in development, some trochoblasts deciliate and, together with the ciliated trochoblast, form the dorsoventrally organised prototroch, the locomotory organ of the larva. In order to study the role of the 3D-macromere in the specification of trochoblasts and in the induction of the dorsoventral organisation of the prototroch, induction of 3D has been prevented in various ways. it is shown that preventing 3D-induction results in the formation of a radially symmetrical prototroch. The trochoblasts of all four quadrants developed like corresponding trochoblasts of the A-quadrant. Somewhere between 30 and 120 min after fifth cleavage the 3D-macromere induces the formation of specific trochoblasts and organises the dorsoventral pattern of the prototroch. Besides a role of the 3D-macromere, a role of other cells has been demonstrated in the conditionally specified deciliation of trochoblasts.

Spatial and temporal coincidence of induction processes and gap-junctional communication in Patella vulgata (Mollusca, Gastropoda).

In the early development of the gastropod mollusc Patella vulgata, two inductive processes take place between the animal and vegetal hemispheres. One is the induction of the vegetally-located stem cell of the mesoderm (the 3D macromere) by the first-quartet micromeres (which are located in the animal hemisphere). The other is the induction of dorsoventral symmetry in the first-quartet micromeres by the 3D macromere. As a consequence of the latter induction process, a dorsoventrally-organised prototroch is formed. In this paper, the moment of 3D induction is determined by deleting the inducing first-quartet micromeres at successive stages of development. Furthermore, the role of gap junctions in the two above-mentioned induction processes is investigated. This was done by studying lucifer yellow dye transfer between the 3D macromere and the first-quartet micromeres, in relation to these two induction processes. Analysis of the pattern of dye-transfer in vivo, in sections and with confocal laser scanning microscopy demonstrated that the moment of dye transfer coincides with the moment of 3D induction and with the moment of induction of the dorsoventral pattern of the prototroch. This indicates that gap-junctional communication may be involved in these two induction processes.

Cell lineage of the prototroch of Patella vulgata (Gastropoda, Mollusca).

The trochophore larva of the archaeogastropod mollusc Patella vulgata has a well-developed locomotory organ, the ciliated prototroch. This structure is formed from specific founder cells, the trochoblasts. Two methods were employed to determine the composition and cell lineage of the prototroch. Fluorescent cell-lineage tracer injection in trochoblasts and trochoblast founder cells was used to show how the various trochoblasts became incorporated into the prototroch. Scanning electron microscopy was used to study both differentiation, more specifically ciliation, of trochoblasts and localization of trochoblasts in the prototroch. The results obtained with both methods are in accordance with each other. During early development all trochoblasts involved in prototroch formation become cell cycle-arrested and develop cilia. Subsequently, the trochoblasts shift in position to form a circular prototroch and a number of trochoblasts deciliate. As a result of these processes the mature prototroch consists of a number of heavily ciliated cells as well as a number of deciliated cells. Comparison of prototrochs from a number of spiralian species shows that this structure is very conserved during evolution. The significance of this is discussed.

Hexose monophosphate shunt activity in erythrocytes related to cell age.

Erythrocytes were separated by age using a combination of density centrifugation and counterflow centrifugation and tested for basal activity of the hexose monophosphate shunt (HMP-shunt) as well as the methylene blue-stimulated maximal capacity by measuring CO2 production. No significant differences were found in basal HMP-shunt activity, but the maximal methylene blue-stimulated activity of old erythrocytes reached only half of the activity of the total cell population. The maximal HMP-shunt activity showed a significant correlation with hexokinase activity, but not with glucose-6-phosphate dehydrogenase activity in all but the youngest cells. The sensitivity to oxidative stress was tested by measuring the kinetics of pyruvate kinase isolated from erythrocytes incubated in presence and absence of methylene blue. Pyruvate kinase kinetics were affected more in the old cell population than in the total cell population: the K0.5 for phosphoenol-pyruvate increased four times in the unseparated cells and eight times in old cells.

Communication compartments in the ectoderm of embryos of Patella vulgata.

Patterns of gap junctional communication in the ectoderm of embryos of Patella vulgata have been studied by intracellular injection of the fluorescent dye Lucifer Yellow, and by analysis of its subsequent spread to adjacent cells (dye-coupling). We found that dye-coupling became progressively restricted to different domains of the ectoderm, forming communication compartments. These communication compartments are characterized by their high coupling abilities within the compartment, and reduction of coupling across their boundaries. During development, the pretrochal (anterior) ectoderm becomes subdivided into two communication compartments, the apical organ and the anlage of the head ectoderm. The posttrochal (posterior) ectoderm becomes subdivided into different communication compartments in two successive phases. Firstly, in the 15-h embryo the dorsal and ventral domains of the ectoderm form separate communication compartments. A dorso-ventral communication boundary restricts the passage of dye between the two domains. Secondly, in the 24-h embryo dye-coupling becomes further compartmentalized in both the dorsal and ventral domains. These compartments correspond to the anlagen of different ectodermal structures. In order to study whether any level of coupling persists between the ectodermal compartments we injected currents through a microelectrode inserted into one cell of one compartment and monitored its spread by means of a second microelectrode inserted into one cell of another compartment (electrical coupling). Despite the absence of dye-coupling, electrical coupling between the ectodermal dye-coupling compartments was detected, which suggests that some level of communication is maintained between compartments. Our results demonstrate that within the ectoderm layer of Patella vulgata the transfer of dyes becomes progressively restricted to communication compartments and, concomitantly with the specification of the different ectodermal anlagen, these compartments become subdivided into smaller communication compartments.