One of the most sensitive hypoxia detection methods is based on the observation that binding of nitroimidazoles to cellular macromolecules occurs as a result of hypoxia-dependent bioreduction by cellular nitroreductases. Nitroimidazole-binding techniques provide measurements of hypoxia to virtually any degree of spatial resolution and with a multiplicity of techniques. This paper demonstrates hypoxia imaging using in vivo EF5 binding with detection by a fluorochrome-conjugated monoclonal antibody. We investigated these techniques in the 9L glioma tumour, in part because the exact nature of the hypoxia in this tumour system is controversial. Our results demonstrate that following intravenous injection of EF5, binding and detection using a monoclonal antibody in 9L gliomas is specific and oxygen dependent. Detection of binding using fluorescence microscopy can be performed on frozen tissues; tissue sections can be counterstained with haematoxylin and eosin for light microscopic analysis. Alternatively, the distribution of hypoxia in a tumour can be inferred by examining individual tumour cells using flow cytometric techniques. Based upon the results presented herein, the radiation-resistant phenotype of 9L epigastric tumours grown in our laboratories can be associated with the presence of hypoxic cells.


Accumulated knowledge in the molecular processes of tumour development combined with the availability of genetically modified viruses resemble the basis for new promising cancer therapeutics. The main advantages of employing replication-competent viruses are achievement of tumour selective killing and amplification of their oncolytic potential within the tumour mass. In this review, we describe the development of ONYX-015, one of the first and most advanced replication-competent viruses for cancer therapy. We discuss the molecular biology of this therapeutic approach and the interesting results obtained with this virus in clinical trials.British Journal of Cancer (2002) 86, 5–11. DOI: 10.1038/sj/bjc/6600006www.bjcancer.comThe Cancer Research Campaign© 2002  Infection of normal cells with ONYX-015 should evoke a p53 response: either growth arrest or apoptosis, resulting in aberrant viral replication. In contrast, tumour cells that do not possess a functional p53 gene (p53− cells) should support replication of ONYX-015. Replication of ONYX-015 should therefore be restricted to p53 deficient cells resulting in selective destruction of cancer cells.The striking similarities of tumour cells and adenovirus infected cells gave rise to the concept of using a mutant adenovirus to selectively eliminate tumour cells, which is based on the observation that in both, tumour cells and adenovirus infected cells, p53 – in its role as ‘guardian of the genome’ – is a major target for inactivation. ONYX-015 contains a 827 bp deletion in the E1B region of the viral genome and a point mutation that generates a premature stop codon preventing expression of a truncated form of the E1B55K protein . Theses In the first experiments to test this hypothesis, ONYX-015's ability to grow was tested in tumour cell lines of known p53 status  demonstrated significant anti-tumour activity of intratumourally administered ONYX-015 in tumours with mutated , and pancreatic cancer. Additional studies were performed in patients with premalignant oral dysplasia, ovarian cancer, and liver metastases from colorectal cancer  experienced objective, partial, tumour responses and an additional ten patients  showed stabilization of their disease. While detection of viral DNA using A subsequent phase-II study was conducted in patients with recurrent HNSCC. As a consequence of the animal studies described above, which had shown that sequential doses of virus are superior to single applications, two dosing regimens were examined. The therapy consisted either of intratumoural injection for five days (standard regimen) or twice daily for two consecutive weeks (hyperfractionated regimen). Main side effects were mild to moderate fever and pain at the site of injection; the latter was particularly frequent in patients receiving hyper fractionated therapy.p53 gene mutations. In an additional 12 patients (41%) the disease was stabilized. There were no significant differences in therapeutic activity between the treatment schedules. Interestingly, in the first cycle of treatment viral genomes were detected in the peripheral blood of about half the patients 24 h after the last virus administration, a finding that was interpreted as a sign of intratumoural virus replication. The number of patients with detectable virus declined significantly in subsequent cycles. In parallel, serum levels of neutralizing antibodies increased dramatically . It is most likely that such high titers of neutralizing antibodies will contribute to a rapid clearance of virus particles shed from the tumours into the peripheral blood. To what extent antibodies interfere with intratumoural virus spread is unclear at this time. Nevertheless, this study demonstrated again the favourable toxicity profile of ONYX-015 and showed unambiguously the anti-tumour activity of the virus in a subset of patients, in particular those with a mutated p53 gene.In patients receiving standard therapy, objective responses were observed in 14% (including three out of 40 patients that achieved complete responses), which were significantly correlated with the presence of As discussed above, preclinical studies suggested a potential synergistic effect of chemotherapy and ONYX-015. Based on these results, a phase-II study was initiated, aimed to evaluate the combination of ONYX-015 and standard, cisplatin-based chemotherapy in patients with recurrent HNSCC. The results mirrored the preclinical data, including the frequent occurrence of complete remissions. Of 30 evaluable patients, 19 (63%) experienced an objective tumour response, including eight (27%) complete responses. Most interestingly, none of the 19 tumours that had shown objective responses progressed during the observation time of this study. In patients with multiple lesions, only the largest and clinically most relevant tumour had been injected, so that the other tumours served as intra-patient controls. Despite the fact that the injected tumours were generally larger, nine of 11 responded, in contrast to three of 11 non-injected tumours. This statistically significant difference represents one of most convincing pieces of evidence for a specific anti-tumour effect of ONYX-015. Encouraged by these results, a clinical phase-III study has been initiated and is ongoing; patients with first relapse of HNSCC are receiving either combined cisplatin-based chemotherapy and intratumourally injected ONYX-015 or standard chemotherapy.13 viral particles. The main side effects included, again, fever and rigours. In contrast to the previous study, however, a transient, dose-dependent increase of serum aminotransferase was also observed. In the context of recent concerns regarding the safety of adenovirus vectors, it is important to note that these changes were mild  In one patient, intratumoural virus replication was documented although no objective tumour responses were observed. The reasons for a potential lower efficacy of systemic administration of ONYX-015, relative to tumour injection, will be discussed below.Intravenous administration of ONYX-015 has been explored in a pilot study of advanced lung tumours . To overcome the need to infect all tumour cells to achieve complete remission, new versions of ONYX-015 have been created that deliver pro-drug converting enzymes to tumour cells, which create high local concentrations of cytotoxic compounds following systemic administration of non-toxic substrates. For example, an E1B55K-deleted adenovirus has been devised, expressing herpes simplex virus-thymidine kinase (HCV-TK), which catalyzes phosphorylation of gancyclovir resulting in highly cytotoxic metabolites (The results from the clinical phase-III study in HNSCC will be available in the first half of the year 2002 and will have a crucial impact on the further development of this therapeutic approach. In addition, future studies of ONYX-015 might include new strategies to overcome the limitations described above. These include the design of replication-restricted viruses with altered receptor specificity, capable of circumventing the problem of loss of expression of adenovirus receptors on target cells, in particular CAR, by utilizing other, cancer specific proteins as alternate receptors. We are currently investigating the possibility of pharmacological restoration of CAR expression at the surface of cancer cells (Anders abolites .ONYX-015 is an interesting new agent for the treatment of solid tumours that shows unambiguous evidence of antitumour activity in a broader range of tumours than initially anticipated. However, the biological mechanisms defining the interaction of this virus with its human host need further exploration. Based on this knowledge, strategies can be developed to overcome problems of bio-distribution and resistance, and, ultimately, to increase the efficacy of systemically administered ONYX-015.


P = 0.0155). The degree of bcl-2 expression was significantly related with tumour size (P = 0.0155). The expression of bcl-2 and p53 protein was not associated with the cure rate due to PDT. Tumour length and T status in TNM staging were significantly related to the cure by univariate analysis. T status was the only predictor of the cure according to mutivariate analysis. Of 42 CR lesions, the expression of neither bcl-2 nor p53 protein was associated with local recurrence; only T status was significantly associated (P = 0.008). The relationship between the expression of oncoprotein and local recurrence after PDT was not documented in this study. The success of PDT may depend on the exact assessment of tumour size under optimized PDT illumination. © 2000 Cancer Research CampaignPhotodynamic therapy (PDT) in early squamous cell carcinoma of the bronchus has been shown to result in complete response (CR) and cure. However, local recurrence after PDT develops frequently even after complete remission. Because the effect of PDT had been reported to depend on apoptosis, and apoptosis is inhibited by bcl-2 protein, the relationship between the expression of bcl-2 protein and local recurrence after PDT was examined immunohistochemically. From 1983 to 1997, 50 patients with 59 early squamous cell carcinoma of the bronchus received PDT, and a CR was obtained in 43 lesions (72.8%). As there was no recurrence among tumours that were disease-free for more than 2 years, in this study the tumours were defined as cured when recurrence did not occur 2 years subsequent to the receiving of PDT. Of these CR lesions, 31 carcinomas (53.4%) resulted in a cure. Bcl-2 immunoreactivity was detected in 23 tumours (46.9%) and p53 immunoreactivity was detected in 22 tumours (44.9%). When all tumours were divided into either a large tumour with a longitudinal tumour length of 10 mm or more, or a small tumour with a length of less than 10 mm, the large tumour expressed more bcl-2 protein than the small tumour (


In the Funding section, the second grant number from the NIH Models of Infectious Disease Agent Study program was incorrect. The correct grant number is: 1U54GM088558.


In contrast, the ganglioside content of the cultured VM tumour cells was high  and consisted almost exclusively of N-acetylneuraminic acid. The ganglioside pattern of the tumour grown subcutaneously was complex, while GM3, GM2, GM1, and GD1a were the major gangliosides in the cultured tumour cells. This tumour will be a useful natural model for evaluating the role of gangliosides and other glycolipids in tumour cell invasion and metastasis. © 2001 Cancer Research Campaign http://www.bjcancer.comGlycosphingolipid abnormalities have long been implicated in tumour malignancy and metastasis. Gangliosides are a family of sialic acid-containing glycosphingolipids that modulate cell–cell and cell–matrix interactions. Histology and ganglioside composition were examined in a natural brain tumour of the VM mouse strain. The tumour is distinguished from other metastatic tumour models because it arose spontaneously and metastasizes to several organs including brain and spinal cord after subcutaneous inoculation of tumour tissue in the flank. By electron microscopy, the tumour consisted of cells (15 to 20 μm in diameter) that had slightly indented nuclei and scant cytoplasm. The presence of smooth membranes with an absence of junctional complexes was a characteristic ultrastructural feature. No positive immunostaining was found for glial or neuronal markers. The total ganglioside sialic acid content of the subcutaneously grown tumour was low (12.6 ± 0.9 μg per 100 mg dry wt,


P=0.034) and poor metastasis-free survival. In multivariate analysis, after allowing for stage, Bid expression was a significant prognostic factor for both disease-specific and metastasis-free survival (P=0.026). It is concluded that strong tumour Bid expression is associated with poor outcome following radiotherapy regardless of intrinsic tumour cell radiosensitivity, and is adverse prognostic for disease-specific and metastasis-free survival in younger patients.The Bcl-2 family of apoptotic regulators is thought to play an essential role in cancer development and influence the sensitivity of tumour cells to radiotherapy. Bid is an abundantly expressed Bcl-2 family protein playing a central role in various pathways of apoptosis by integrating and converging signals at the mitochondria. The relevance of apoptotic modulation by Bcl-2 and related proteins in tumour development and radiation response for human tumours remains undefined. Therefore, a study was made regarding the expression of Bid in patients with locally advanced cervix carcinoma who received radiotherapy. Bid expression was assessed using immunohistochemistry in pretreatment archival biopsies from 98 patients. The data were correlated with clinicopathologic characteristics and treatment outcome. Pretreatment tumour radiosensitivity data were available for 60 patients. Strong Bid expression was associated with a patient age less than the median of 52 years ( Radiotherapy is the main treatment modality for locally advanced carcinoma of the cervix. In spite of the reduction in incidence and mortality in Europe and the USA, this disease remains one of the major causes of female cancer-related deaths worldwide. Defective apoptotic mechanisms may contribute to both malignant progression and radiation resistance of this tumour .L. They are related by homology domains, known as BH domains 1–4, which are crucial for protein function and the intermolecular association of family members. Apoptotic signals, from intrinsic or extrinsic stimuli, are regulated via the relative actions and interactions of prosurvival and prodeath Bcl-2 proteins (C from mitochondria), proteolytic caspase cascade activation and ultimately cell death  and were registered at the Department of Oncology, Christie Hospital between 1987 and 1993. They were a consecutive series who received radiation therapy with curative intent and for whom a biopsy was taken for research. Patients were treated according to previous standard techniques of the Manchester School as described elsewhere  before cin vitro clonogenic assay to obtain surviving fraction at 2 Gy radiation (SF2).For some of the patients included in the study, data were available from previous published work on tumour radiosensitivity . RadioseM citrate buffer (pH 6.0) and endogenous peroxidase was blocked with 3% methanolic hydrogen peroxide. A standard DAB-based immunostaining procedure employing an avidin–biotin complex reagent (DAKO) was used to detect Bid protein after application of specific goat anti-Bid antibody  at 1 μg μl−1 overnight at 4°C. Sections prepared from cell pellets of wild-type and Bid knock-out mouse embryonic fibroblasts . Bid stain intensity was weighted as either absent (0), very weak (1), weak (2), moderate (3) or strong (4) and, for each biopsy section, the estimated percentage of tumour cells (to the nearest 10%) that expressed Bid at each intensity was assessed and multiplied by the corresponding weighted intensity value. These values were added to obtain a final score for each biopsy with a theoretical range of 0–400. The frequency distribution of the final scores was normal and quartiles were used to categorise scores (I–IV) for statistical purposes. Examples from each of these categories are illustrated in Tumour sections were assessed for the extent and intensity of Bid expression specifically within tumour cells without the observer's knowledge of clinical data. Necrotic areas, stroma and non-neoplastic epithelial cells were ignored. For the majority of tumours, 90–100% of the tumour cells were Bid positive. However, intensity of Bid stain varied from biopsy to biopsy and within tumour cells of individual biopsy sections. A comparative semiquantitative scoring index recommended for immunohistochemical biomarker validation was used to reflect the intensity of staining of individual cells and the percentage of cells that stained at different intensities .The reproducibility and concordance of scores were tested by Spearman's rank correlation and Altmann–Bland plot, respectively. The distribution of Bid score in relation to patient and tumour characteristics was investigated using either the Mann–Whitney Bid immunostaining demonstrated a granular, cytoplasmic pattern consistent with the cytoplasmic/mitochondrial/intercellular membrane association of Bid . Weak tor=0.92, P<0.001) with an Altman–Bland plot equation of y=−0.0029x−2.058 showing good concordance. A significant correlation was obtained  when a series of 35 randomly selected tumour sections were scored by a second observer (GH).Tumour sections were scored twice by the first observer (MG) and ranged from 100 to 380. A significant correlation was obtained between the two scores (P=0.034). This finding was supported by a weak but significant inverse correlation between patient age and Bid expression . There was no relationship between the level of Bid expression and disease stage, differentiation status and tumour radiosensitivity (SF2).Bid expression was examined in relation to disease-specific, local recurrence-free and metastasis-free survival by Kaplan–Meier and log-rank analysis. A preliminary analysis revealed a trend for a worse survival for patients with strong Bid expression  and the availability of complete long-term followup survival data.Bid expression was widespread in normal and neoplastic epithelial cells of the cervix tissues, although heterogeneity was observed within and between individual tumour biopsies. The extent and intensity of Bid expression between tumour biopsies was compared and found to range widely, indicating substantially altered regulation of Bid protein levels within the carcinomas examined. Tumour Bid expression was higher in younger patients, and was related to a more aggressive tumour phenotype which was more likely to metastasise, but was not related to intrinsic tumour cell radiosensitivity. Notably, strong tumour Bid expression was prognostic for poor outcome, particularly for younger patients, and could provide additional outcome prediction information given tumour stage.Variable expression of Bid was previously found in several types of human cancers other than cervix carcinoma, including neural tumours, colorectal adenocarcinomas, ovarian and prostate cancers , and an in vitro studies  and caspases 3 and 8 (that directly affect Bid function) have also been reported to be differentially regulated in cervix carcinoma , indicating a link between expression of the two proteins. Recently, Bid was shown to be negatively regulated by the hypoxia-inducible transcription factor HIF-1 (A typical regulation or aberrant expression of other Bcl-2 family members may influence the level of Bid protein detectable within the cell and/or its functional ability. For example, Bcl-2 is commonly overexpressed in a variety of tumour types including cervix carcinoma (Tumour cells with other compensatory defects in apoptotic mechanisms may tolerate elevated levels of Bid and/or prevent the normal proapoptotic function of Bid. Elevated expression of apoptosis inhibitor proteins, such as XIAP, cIAP or survivin, might also allow downstream accumulation of Bid  and may If indeed Bid proapoptotic function is compromised, an attractive theory to explain increased metastatic potential may be the increased ability of tumour cells with reduced Bid function to avoid Fas/TNFR/Granzyme B-mediated apoptosis, induced by immune cells circulating in the body, thus enabling malignant cells to invade surrounding tissue early in tumour development. Similarly, defective Bid function could reduce the overall apoptotic potential in response to radiation and may partly explain increased rates of tumour recurrence following radiotherapy.in vitro measurement of intrinsic tumour cell radiosensitivity, but was associated with a more metastatic and aggressively malignant tumour phenotype in younger patients. Further study is required to examine the mechanism of Bid overexpression and its subsequent impact on apoptotic potential and disease phenotype. However, the results link Bid apoptotic signalling to tumour progression and treatment response, and may aid the development of tailored combined treatment and disease monitoring schedules for individual patients.In conclusion, altered regulation of Bid expression within cervical carcinoma was shown and overexpression of Bid was an adverse prognostic factor for radiotherapy outcome. The relationship between strong Bid expression and poor outcome following radiotherapy was unrelated to


Adriamycin (10 mg/kg) administered s.c. to male ACI rats bearing Hepatoma H-4-II-E caused a 9-day delay in tumour growth but no changes in the clonogenic fraction of the tumour were detectable by in vitro assay at any time after treatment. There is no significant decrease in the yield of cells on enzymatic dissociation of the tumour nor a reduction in mg DNA/g tumour that might indicate a decrease in tumour cellularity. Mitotic and labelling indices and [3H]dT uptake into DNA remain essentially unchanged, relative to age-equivalent controls, but are slightly lower than in controls of equal weight. The reliability of the clonogenic assay and possible mechanisms by which Adriamycin delays tumour growth are discussed.


We investigated the role of platelets in human melanoma cell (line 397) interaction with vascular endothelial cells (ECs) under flow conditions. The ability of the tumour cells to adhere to the EC monolayer was significantly reduced by application of flow at a shear rate of 250 s(-1). A 2.2-fold increase in tumour cell adhesion to ECs under flow was observed upon addition of thrombin receptor agonist peptide (TRAP)-activated platelets but not resting platelets. A similar increase (2.5-fold) in tumour cell adhesion to ECs under flow was observed when the tumour cells were incubated with resting platelets on thrombin-treated ECs. However, thrombin treatment of the ECs alone had no effect on tumour cell adhesion in the absence of platelets. The enhancement of tumour cell adhesion to ECs by TRAP-activated platelets was virtually abolished by blockade of the platelet glycoproteins P-selectin and GPIIb-IIIa by monoclonal antibodies. Blockade of P-selectin also inhibited the direct adhesion of TRAP-activated platelets to ECs, but did not affect the interaction of the tumour cells with platelets immobilized on subendothelial extracellular matrix (ECM). Blockade of GPIIb-IIIa inhibited both platelet-EC and platelet-tumor cell interactions. Our results indicate that tumour cell adhesion to the endothelium under flow is enhanced by platelets under conditions that allow platelet adhesion to ECs. Inhibition studies suggest that activated platelet adhesion to ECs is mediated by P-selectin and GPIIb-IIIA, and tumour cell adhesion to EC-bound platelets--mainly by GPIIb-IIIa.


Nepenthes, widely distributed in the Asian tropics, rely mostly on nutrients derived from arthropods trapped in their pitcher-shaped leaves and digested by their enzymatic fluid. The genus exhibits a great diversity of prey and pitcher forms and its mechanism of trapping has long intrigued scientists. The slippery inner surfaces of the pitchers, which can be waxy or highly wettable, have so far been considered as the key trapping devices. However, the occurrence of species lacking such epidermal specializations but still effective at trapping insects suggests the possible implication of other mechanisms.The carnivorous plants of the genus Nepenthes rafflesiana is highly viscoelastic and that this physical property is crucial for the retention of insects in its traps. Trapping efficiency is shown to remain strong even when the fluid is highly diluted by water, as long as the elastic relaxation time of the fluid is higher than the typical time scale of insect movements.Using a combination of insect bioassays, high-speed video and rheological measurements, we show that the digestive fluid of Nepenthes pitchers as simple passive traps and is of great adaptive significance for these tropical plants, which are often submitted to high rainfalls and variations in fluid concentration. The viscoelastic trap constitutes a cryptic but potentially widespread adaptation of Nepenthes species and could be a homologous trait shared through common ancestry with the sundew (Drosera) flypaper plants. Such large production of a highly viscoelastic biopolymer fluid in permanent pools is nevertheless unique in the plant kingdom and suggests novel applications for pest control.This finding challenges the common classification of  Sarracenia, Cephalotus and Nepenthes, flypaper-traps in sundews (Drosera) and butterworts (Pinguicula) or even be very sophisticated devices such as the snap traps of Dionaea or the suction bladders of Utricularia. All these carnivorous plants secrete a digestive fluid involved in the process of prey digestion Carnivorous plants live in nutrient-poor soils and have circumvented this shortage of resources by deriving most of their nutrients from the digestion of arthropods captured through a variety of trapping mechanisms Nepenthes pitcher plants, prey capture and retention is mainly thought to be fulfilled by the slippery waxy layer which covers the upper inner part of the pitcher in most species N. bicalcarata for instance) Nepenthes species lack such specialized surfaces Nepenthes pitcher plants is more complex than commonly acknowledged. Moreover, reports of secretion of wetting agents In N. rafflesiana, one of the most widespread species of the genus in northern Borneo Sarracenia pitcher plants Nepenthes by Juniper and co-authors Nepenthes pitcher plants. Moreover, up to now, the rheological properties of the fluid, which govern how a fluid moves under forces, and their possible role in insect capture have never been investigated. We thus focused our study on the digestive fluid of N. rafflesiana and first tested whether the fluid alone was able to retain insects by comparing retention of insects thrown into glass vials filled with water or pure digestive fluid. Then, to determinate which physical properties of the digestive fluid was implicated in retention, we compared surface properties  and rheology  of pure fluid, water and intermediate dilutions. Our study unveils the peculiar viscoelastic properties of the digestive fluid of N. rafflesiana and its crucial role in prey capture.Here we focus on N. rafflesiana. The insects (ant workers and flies) were chosen because they represent non-flying and flying insects and are part of the two insect orders  most frequently captured by the pitcher plants Drosophila melanogaster, Calliphora vomitoria) and ants (Linepithema humile) escaped easily from water . Moreover, while observations on moving insects showed that the pure digestive fluid wet flies and ants once they began to move in the fluid, observations on paralyzed insects showed that the pure digestive fluid did not spontaneously wet insects. This suggests that wetting of insects occurs dynamically rather than statically, probably because of fluid viscosity. More quantitative data were obtained by measuring the quasi-static advancing contact angles of the fluid (same mixing as in the retention experiment) over model surfaces using the sessile drop method Since insects greatly rely on their hydrophobic body surfaces and on the surface tension of water to sustain their weight and move at liquid interfaces η of the digestive fluid , the shear viscosity of the pure fluid was rather low, about 15 times the viscosity of water . However, when insects move, the digestive fluid is not only sheared but also stretched. For simple liquids such as water, resistance to both shearing and stretching are equivalent and given by the same value of the viscosity  and apparent extensional viscosity Εη  of the digestive fluid in a controlled capillary break-up extensional geometry 4 times larger than the shear viscosity at typical shear rates and strains imposed by insect motions  as the ratio of the fluid elastic relaxation time λ . As shown by the results of a logistic regression . The important result is that the abrupt transition in capture rate occurred when the Deborah number became inferior to 1, i.e. when the elastic relaxation time became inferior to the typical half-period of the swimming stroke of insects . The elastic forces are likely to be the key force of retention of insects in the fluid of N. rafflesiana. First, the fluid did not spontaneously wet insects and its surface tension was shown to be similar to that of water from which insects can escape easily. Therefore, surface properties of the fluid are unlikely to be involved in trapping in N. rafflesiana. Moreover, the shear viscosity of the fluid at typical shear rates imposed by insect movements was rather low, and became similar to that of water at dilutions for which the capture rate was still maximal. We therefore suggest that the shear viscosity of the digestive fluid is unlikely to explain the retention of insects, although further investigation using standardized simple (non elastic) fluids would be helpful to precisely quantify a possible role of shear viscosity in trapping. By contrast, the digestive fluid exhibits a strong extensional viscosity, which appears to be several orders of magnitude larger than the classical (shear) viscosity. This is a clear signature of elasticity in complex fluids. Unlike the shear viscosity, the extensional viscosity of the digestive fluid remains high even when the fluid was highly diluted by water. Such a large extensional viscosity means that the digestive fluid offers a large resistance to stretching or squeezing flows, hence penalizing insects when they try to escape from the fluid or to climb on the pitcher wall. The strong correlation we found between the capture rate of insects and the Deborah number supports the hypothesis that trapping occurs when the elastic forces created by insect movements have no time to relax. It is also compatible with recent models of locomotion in viscoelastic fluids showing that propulsion is much less efficient at high Deborah numbers Misumenops nepenthicola, which lives and reproduces in the trap of N. rafflesianaOutside of any effect of the pitcher walls, the pitcher fluid alone is able to retain insects (flies and ants) in the trap of NepenthesN. rafflesiana, the slippery waxy surface of the pitchers was shown to play a minor role in the trapping function of the plant, being even a variable character of weak adaptive significance Nepenthes rafflesiana, may also apply to other species and be more frequent than commonly acknowledged within the Nepenthes genus. Already, casual observations of insects attached to the inner pitcher wall, as if they were glued by plant secretions, were made in N. inermisN. inermis lack a waxy zone. It is also the case of N. eymae, N. aristolochioides, N. talagensis, N. dubia and N. jacquelinae, whose trapping fluid has been reported to be viscous in touching Nepenthes inermis was reported to be (under the name of N. bongso) specialised in trapping midges N. aristolochioides, while N. jacquelinae was observed to trap essentially flying preys of bigger sizes Our result undermines the claim that the pitcher surfaces are the main component of the trapping mechanism in Nepenthes could exhibit a composition close to the acidic polysaccharide mucilage The exact composition of the viscoelastic fluid remains to be studied. However, we can suppose that the fluid is composed of polysaccharides, as these macromolecules are the main component of mucilages in plants All measurements were performed in the laboratory on pitchers of plants grown in greenhouses in Montpellier (CEFE-CNRS) at 25–30°C and 90% humidity. Complementary measurements of surface tension and viscosity were carried out in the fields  and gave similar results.Calliphora vomitoria larvae were bred at 27°C until adult emergence. The flies Drosophila melanogaster were bred on a nutritive substrate and the ants Linepithema humile were collected on the grounds of the campus of the University in Marseille . Pure digestive fluid of seven newly opened pitchers from seven plants was collected and mixed in a glass vial in the laboratory at 25°C. Nine vials (each filled with 50 ml) were prepared from this mixture using different solutions, respectively 100% pure fluid, 37%, 17%, 7%, 4%, 2%, 1%, 0.5% fluid and distilled water. The experiment consisted in drawing a fly into a soft tube, blowing it onto the solution in a given vial and observing it during five minutes. Ten trials, each using a different fly, were carried out for each of the nine fluid solutions. The capture rate was defined as the percentage of flies that were still retained within the fluid solution after 5 minutes. All the flies that did not escape within five minutes eventually died within the fluid. Similar experiments were conducted with the ants.For the retention experiment, about 100 http://rsb.info.nih.gov/ij/) and Matlab® softwares in order to compute each drop's equatorial diameter D and diameter d at the distance D from the bottom of the drop. The interfacial surface tension was then computed from 2/Hσ = ρgD, where ρ is the density of the digestive fluids , g = 9.81 m.s−2 is the gravity and H is a shape parameter depending on the ratio d/Dσ = 0.0720 N.m−1) we obtained was equal to the reference value given at 25°C (σ = 0.07197 N.m−1) Interfacial surface tension between air and pure digestive fluids (12 fluids from 12 opening pitchers of 6 different plants) was measured using the pendant drop method δt = 0.056 s, final separation 12.5 mm) was applied to the fluid by rapidly vertically removing a thin rod (radius 0R = 1.5 mm) from a small sample of fluid, thus creating an elongated liquid filament. The subsequent capillary thinning and break-up dynamics of the filament were recorded at a high spatial and temporal resolution  using a high-speed camera (IDT Monochrome) mounted on a stereomicroscope (Leica MZ16). Videos (n = 5 for each tested fluid) were then post-processed using Image J and Matlab® software in order to compute the time evolution of the midpoint filament's radius R(t). In all experiments, gravity was small compared to capillary forces (Bond number 02/σBo = ρgR ∼0.3) and the filament's relaxation time scale was large compared to the inertial time scale λ is the longest relaxation time scale of the internal elastic forces R(t) curve, we computed the elastic relaxation time Fluid elasticity of pure and diluted fluids  was investigated using capillary break-up extensional rheometry Video S1Calliphora vomitoria) thrown into water on its ventral surface .This high-speed video  shows the fall and escape of a fly ((2.89 MB MOV)Click here for additional data file.Video S2Calliphora vomitoria) thrown into water on its dorsal surface .This high-speed video  shows the fall and escape of a fly ((3.53 MB MOV)Click here for additional data file.Video S3Calliphora vomitoria) thrown into the digestive fluid on its ventral surface. The motion of the fly involves both shearing and stretching of the fluid at large Deborah numbers (10<De<100), thereby inducing high retentive elastic forces  .This high-speed video  shows the fall and retention of a fly ((3.92 MB MOV)Click here for additional data file.Video S4Calliphora vomitoria) thrown into the digestive fluid on its dorsal surface. The fly is wetted by the digestive fluid and is unable to move its wings and extract its legs, which are retained by sticky filaments typical of complex fluids such as mucus or saliva .This high-speed video  shows the fall and retention of a fly ((9.16 MB MOV)Click here for additional data file.


High tumour interstitial fluid pressure (IFP) has been adversely linked to poor drug uptake in patients, and to treatment response following radiotherapy in cervix cancer patients. In this study we measured IFP values in a selection of murine and xenograft models, spontaneously arising or transplanted either intramuscularly (i/m) or orthotopically and analysed their relationship to tumour vascularity and metastatic spread.KHT-C murine fibrosarcoma, ME180 and SiHa human cervix carcinoma were grown either intramuscularly (i/m), sub-cutaneously (s/c) or orthotopically. Polyoma middle-T (MMTV-PyMT) transgenic spontaneous mammary tumours were studied either as spontaneous tumours or following orthotopic or i/m transplantation. IFP was measured in all tumours using the wick-in-needle method. Spontaneous metastasis formation in the lungs or lymph nodes was assessed in all models. An immunohistochemical analysis of tumour hypoxia, vascular density, lymphatic vascular density and proliferation was carried out in ME180 tumours grown both i/m and orthotopically. Blood flow was also assessed in the ME180 model using high-frequency micro-ultrasound functional imaging.KHT-C i/m: 2–42 mmHg, s/c: 1–14 mmHg, ME180: i/m 5–68 mmHg, cervix 4–21 mmHg, SiHa: i/m 20–56 mmHg, cervix 2–26 mmHg, MMTV-PyMT: i/m: 13–45 mmHg, spontaneous 2–20 mmHg and transplanted 2–22 mmHg. Additionally, there was significant variation between individual tumours growing in the same mouse, and there was no correlation between donor and recipient tumour IFP values. Metastatic dissemination to the lungs or lymph nodes demonstrated no correlation with tumour IFP. Tumour hypoxia, proliferation, and lymphatic or blood vessel density also showed no relationship with tumour IFP. Speckle variance analysis of ultrasound images showed no differences in vascular perfusion between ME180 tumours grown i/m versus orthotopically despite differences in IFP.Tumour IFP was heterogeneous in all the models irrespective of growth site: Our studies across a range of different tumour models showed substantial heterogeneity in tumour IFP, suggesting differences in the vascular development and interstitial fluid dynamics in the individual tumours. The results demonstrate a strong stochastic aspect to tumour IFP development, notably the variation apparent between different tumours within the same animal and the lack of correlation between donor and recipient tumours. Solid tumours show interstitial fluid pressures (IFP) that are elevated above that of normal tissues. Tumour growth and development is supported by both the pre-existing host vasculature and by neovasculature generated through the process of angiogenesis. Tumour angiogenesis generates abnormal vessels -3 that dElevated tumour IFP plays a role in the pathophysiological microenvironment that characterises solid tumours contributing to disease progression and therapeutic resistance ,6-9. TheThese data, coupled with the breadth of data demonstrating elevated IFP in a wide range of human tumours ,19,21-25in vitro/in vivo growth cycle. In vitro cells were maintained as monolayers in plastic tissue culture flasks using α-MEM medium  supplemented with 10% fetal bovine serum . The cervical carcinoma cell lines were maintained under G-418 selection (400 μg/ml). Cells between their 2nd and 5th in vitro passage were removed from the flasks during exponential growth using 0.05% trypsin for transplantation into mice. KHT-C cells were transplanted into syngeneic 8–12 week old C3H/HeJ male mice . ME180 and SiHa cells were transplanted into 8–12 week old female CB-17/SCID mice obtained from an in-house breeding program. PyMT cells were transplanted into female FVB  or SCID mice. Tumours were initiated either intramuscularly (i/m) in the left gastrocnemius muscle, or sub-cutaneously (s/c) on the flank by injection of 2.5 × 105 or 5 × 105 cells respectively in a 50 μl volume of α-MEM media. Tumours growing i/m were monitored by measuring the external leg diameter of the mouse. Tumours growing s/c were measured directly. Orthotopic cervical and mammary gland tumours were initiated from donor tumours using protocols described below. Animals were housed at the Ontario Cancer Institute animal facility and had access to food and water ad libitum. All experiments were performed under protocols approved according to the regulations of the Canadian Council on Animal Care.Experiments were performed using MMTV-polyoma middle-T transgenic mice  bred inth mammary gland. A small incision was made in the fat pad of the 4th mammary glands and a tumour fragment was sutured in place using a single 8-0 silk suture. The skin was closed using stainless steel wound clips. The left and right 4th mammary glands were implanted with a donor tumour fragment from either the same or a different donor tumour . Pressure levels used were based on the average IFP values apparent when grown i/m. Pressure was controlled through adjustments of a dual scale low pressure gauge  on the outlet port. Control cells were gassed using the same system, but without the addition of a pressure gauge on the outlet port, allowing the gas to flush through freely at atmospheric pressure. The metastatic potential of pressure treated tumour cells was assessed using an experimental lung metastases assay ; obtained from Dr. Cameron Koch, University of Pennsylvania, at 10 mg/kg 2.5 h prior to tumour excision. Once excised, half of the tumour was fixed in 10% neutral buffered formalin, and the remaining half placed in optimal cutting temperature (OCT) embedding medium , and snap frozen in liquid nitrogen. Paraffin-embedded tissue was used for all markers with the exception of CD31.Analyses were carried out using immunohistochemistry for tumour hypoxia (EF5), vascular density (CD31), lymphatic vessel density (LYVE-1), and proliferation (Ki67). Tumour bearing animals were injected with the hypoxia marker EF5  for all fields analysed. The data are presented as median for all tumours and error bars show the range. The median is shown for each marker according to whether the tumour was grown in the cervix (n = 10) or i/m (n = 10). B-I) The median % positive staining and range for all 20 (or 10 for CD31) frames analysed is shown for each tumour (y-axis) plotted against tumour IFP ; B) EF-5 cervix, C) EF5 i/m, D) Ki67 cervix, E) Ki67 i/m, F) CD31 cervix, G) CD31 i/m, H) LYVE-1 cervix and I) LYVE-1 i/m.Click here for fileImmunohistochemical analyses of tumour hypoxia, proliferation and vascular area. Tumour sections from the PyMT tumour model growing in different sites were stained for EF5 as a marker of hypoxia, Ki67 as a marker of proliferation, CD31 as a marker of tumour vasculature and LYVE-1 as a marker of lymphatic vasculature. The median percentage of positive staining [) × 100] and the range is shown for each tumour model and site of growth; A) EF5 , B) Ki67  CD31 , D) LYVE-1 . An asterix indicates significance (p ≤ 0.05).Click here for file


The chromatin-bound enzyme adenosine diphosphate ribosyl transferase is activated by DNA-damaging agents. Substances that inhibit the enzyme, such as benzamide analogues, are known to increase the cytotoxicity of ionising radiation and cytotoxic drugs. The purpose of the present study was to investigate whether the anti-emetic drug metoclopramide, a benzamide derivative (4-amino-N-2-(diethylaminoethyl)-5-chloro-2-methoxybenzamide; MCA), potentiates the effect of cisplatin  on squamous cell carcinoma (SCC). For that purpose human SCC of the head and neck (i.e. tumour line AB and EH) xenografted to nude mice were used. Two administration schedules were tested: (a) MCA (2.0 mg kg-1 i.p.) one hour before CDDP (7.5 mg kg-1 i.p.); and (b) MCA (3 x 2.0 mg kg-1) given concomitant to, 24 and 48 hours after CDDP (7.5 mg kg-1) administration. Treatment efficacies were compared using the area under the growth curves (AUC), tumour volumes and specific growth delay (SGD). There was no mortality and no weight loss of significance in any treatment group. MCA alone did not induce any significant reduction in AUC, tumour volume or SGD with either treatment schedule. CDDP alone gave a significant reduction of tumour growth in tumour line AB but not in tumour line EH. In schedule (a) the addition of MCA did not give any additive effect. However, in schedule (b), for both tumour lines, MCA enhanced the effect of CDDP by significantly reducing the AUC  and increasing SGD  when compared to the tumours given CDDP alone. These effects were observed at a MCA dose currently being administered to humans.


Smoke condensate from two types of cigarette, dissolved in two solvents, has been applied regularly to the backs of mice at each of seven different dose levels. Treatment was continued 3 times weekly for up to 110 weeks, by which time 509 of the 1428 treated mice had developed skin tumours. The dependence of tumour incidence on age was adequately described by the Weibull distribution. The relation - ship between dose of smoke condensate and tumour incidence rate was, however, erratic. It was less regular than the simple relationship which has in previous work been found to obtain when the pure carcinogen benzo(a)pyrene is applied to mouse skin.


Clonogenic growth  of i.v. injected allogeneic W256 tumour cells in the lungs was markedly enhanced by treatment of rats with alpha-naphthyl thiourea (ANTU) injected i.p. from 2 h before to 2 h after the tumour cells. ANTU specifically increases pulmonary vascular permeability in adult rats and causes acute pulmonary oedema and pleural effusion. Inhibition of drug toxicity to the lungs by tachyphylaxis, specific antimetabolites or iodides did not abolish the effect of ANTU on CFE. CFE was not increased when cells were seeded by i.v. injection the lungs affected by advanced pulmonary oedema at 6 to 24 h after treatment with drug. ANTU did not enhance growth of intratracheally injected cells. Although ANTU has no cytotoxic or immunosuppressive action, treatment of tumour-immunized rats with ANTU caused apparent "breakdown" of tumour immunity in 50% of rats, by causing growth of tumour colonies in the lungs. Possible mechanisms for the ANTU-induced decrease in innate resistance to growth of tumour in the lungs are discussed.


A retrospective analysis of 321 gastric cancer patients was made to assess the prognostic value of TNM classification, tumour differentiation, Laurén classification, proliferative rate, inflammatory reaction and tumour invasion in vascular or neural structures of the gastric wall. The TNM classification showed the strongest correlation with survival in univariate and multivariate analyses (P < 0.0001). The invasion in lymphatic or vascular system and Laurén classification were also independent prognosticators in multivariate analysis (P < 0.05). In univariate analysis, the WHO-grade, the size and the location of the tumour and perinueral invasion were significant prognostic factors (P < 0.01), as were the infiltration of lymphocytes and plasma cells in the tumour (P < 0.05). On the other hand, the mitotic indices reflecting the proliferative activity of the tumour cells showed no significant correlation with the prognosis. The results indicate that the prognostic power of the TNM classification can be further increased by assessment of the above special histological features in gastric cancer.


To investigate the underlying mechanisms of carcinogenesis, we have developed a technique to determine the frequency of genetic changes in prostatic carcinoma tissue. We have demonstrated that at a ratio of between 1:4 and 1:9 mutant-normal alleles, the signal from a mutant TP53 allele is not apparent after polymerase chain reaction (PCR) amplification and further direct sequencing or single-strand conformation polymorphism (SSCP) analysis. To bypass this problem, which is inherent in the heterogeneity of the prostate tissue and of the tumour, we selected areas of graded prostate tumours (Gleason score) from cryosectioned preparations and microdissected these cells (20-100 cells). After anionic resin removal of proteins, PCR amplification of TP53 gene exons 5/6 and SSCP analysis, an abnormal SSCP band shift was observed in suspected tumour cells, compared with microdissected stromal cells used as an internal control, while (1) a crude preparation of tissue DNA carrying the tumour did not show any abnormality and (2) immunostaining by a set of monoclonal antibodies against TP53 protein remained negative. Nucleotide sequence analysis of the different bands confirmed the presence of a mutation in the TP53 gene exon 6 position 13,336 in an abnormal band for one specimen, while no mutation was detected in the normal SSCP band. By targeting recognised tumour cells we can find DNA mutations which are undetectable using the standard technique of whole-tissue DNA extraction, particularly in a heterogeneous tumour such as carcinoma of the prostate.


The cytokinetic response of the solid Yoshida sarcoma to hyperthermia was examined at two tumour volumes, 1.0-1.5 ml and 3.0-3.5 ml. The tumour, growing on the feet of rats, was heated at 42 degrees C for 1 h by water-bath immersion. The larger tumour grew more slowly than the smaller one (doubling time 144 h vs 36 h) due to a halving in growth fraction from 67.8 to 39.6% and an increase in cell-loss factor from 59 to 75.9%. Cell cycle and phase times were similar at both volumes. The effect of heat on the population kinetics at both volumes was similar but complex, and involved delayed cell death after up to 10 mitoses. Initial cell killing and blockade of cell-cycle progression (0-24 h) was followed by recovery of proliferation due to recruitment of cells from the non-proliferative compartment, cell cycle and phase times remaining unaltered. From 48 h, the proliferation rate declined progressively, and tumours were completely necrotic 7-8 days after heat. The damaging effects of heat were at least as severe in the large tumours with a low labelling index and small growth fraction as in the smaller tumours with a much larger compartment of proliferating cells and shorter doubling time. The results imply that there may be no simple relationship between proliferative status and thermosensitivity of a tumour, and illustrate the difficulty in predicting tumour response to heat on the basis of cytokinetic studies.


Extraskeletal mesenchymal chondrosarcoma is extremely rare and, in combination with psoriasis, it has never been described before. We report a case of wide resection of an extraskeletal chondrosarcoma of the thigh and reconstruction with a femoral megaprosthesis in a patient with psoriasis vulgaris. Special emphasis has been laid to postoperative wound healing in psoriatic skin which did not show any problems. Psoriasis in patients who should undergo surgery is a controversially discussedproblem and today there are no controlled, prospective studies existing on thistopic . While eA 20-year-old man with a history of Psoriasisvulgaris was presented to our department for resection of an extraosseous mesenchymalchondrosarcoma of the left thigh.The patient recognized a half year before admission a tumour in his leftthigh and suffered of pain over some weeks before he contacted a local hospitalwhere X-rays and anMRI were suspicious for a malignant tumour. Physical examination showed no skindisorders at this point of time. Open biopsy showed an extraosseous mesenchymalchondrosarcoma G3. Tumour size was 18 × 8 × 14 cm. The tumour was localizedclose to the femur in the middle third of the thigh. Tumour staging with CT ofthe chest showed no pathology in the thorax but abdominal CT revealed two intraabdominallymph node metastases besides the left A. and V. iliaca interna. Skeletal scintigraphysupported these findings with high uptake in the middle third of the thigh andthe two lymph nodes.2) and Ifosfamide (8 g/m2) up to 5 months after diagnosis. Curative treatment with a widetumour resection of all contaminated tissue, including the groin (high amputation),was planned but preoperative restaging one month later showed bipulmonal and newinguinal metastases in CT and tumour growth in the thigh. Therefore, curativetreatment seemed to be impossible and it was decided to perform a marginalresection of the tumour and reconstruction of the defect with a cemented diaphysealfemurprosthesis implantation allowing immediate weight bearing and mobilizationof the patient.Chemotherapy with Ifosfamide and Adriamycine was initiated with a good initialtumour response regarding tumour size and lymph node reduction. Four monthslater, a left inguinal and iliacal lymphadenectomy was performed where six metastaseswere removed in another hospital. Chemotherapy was continued afterwards with atotal number of 5 cycles with Adriamycine  crème was initiated and the patient responded well to this treatment. BecauseExtrasceletal mesenchymal chondrosarcoma is extremely rare  and in cWhile some publications indicate that there is no increased risk forinfection or wound healing disturbances after elective surgery in psoriaticskin  and psorAfter operation the patient received a prolonged antibiotic prophylaxisover one week and we could not observe any wound infection or healingcomplications related to the psoriatic skin. A slight necrosis of the skin incentral parts of the wound is not unusual in such operations with widesubcutaneous preparation and has no clinical relevance that causes anintervention.Our case shows that megaprosthesis implantation is possible in patientswith psoriatic lesions in the operation field. There should be applied anadequate psoriasis treatment preoperatively if possible and a prolongedantibiotic prophylaxis seems to be helpful. Whether there is an increasedinfection risk or not remains unclear.


Peritoneal macrophages from Sarcoma-180-bearing mice against Staphylococcus aureus were studied to determine the in vitro phagocytic capacities. When the phagocytic system was opsonized with normal mouse serum, macrophage phagocytic activity increased markedly soon after tumour graft and then returned to normal. A new antiphagocytic factor was detected in the serum of tumour-bearers soon after tumour implantation. This factor was of tumour origin, stable even at a temperature of 56 degrees C for 30 min and non-dialysable. Peptone-starch-induced macrophages were less sensitive to the factor than their unstimulated counterpart.


Antibody was detected by membrane immunofluorescence tests in sera of rats bearing an ascitic variant of a transplanted hepatoma, and in concentrated cell-free ascitic fluid. Ascites hepatoma cells were also shown to have immunoglobulin, possibly tumour specific antibody, bound to their surface. The kinetics of antibody responses to ascites hepatoma and hepatoma cells from solid tumours were compared: both tumour types gave positive reactions by the third day after implantation; antibody was present throughout subsequent tumour growth with the ascites whereas antibody was not detected after tumour became palpable in rats injected with hepatoma cells from solid tumour. Antibody responses to ascites tumour were investigated in rats bearing solid hepatoma tumour. Subcutaneous hepatoma did not influence the antibody response to ascites, but rats bearing intraperitoneal tumours showed a diminished serum antibody response to ascitic hepatoma.


The combination of hyperthermia and the hypoxic cell radiosensitizer Ro-07-0582 has been investigated on the EMT6 tumour implanted into the legs of BALB/c mice. Treatments at a drug dose of 1 mg/g over a range of waterbath temperatures from 37 to 45 degrees C are described. The surviving clonogenic fraction following treatment was assayed in vitro. Measurements of intra-tumour temperature have been made, and shown to be better correlated with the cytocidal effect on the tumour than the waterbath temperature. No significant effect of Ro-07-0582 was observed at 37 degrees C. However, marked cytotoxicity due to the drug was seen at intra-tumour temperatures above 42-5 degrees C for 1 h. These were in addition to the cytocidal effect of the hyperthermia. The results are discussed in relation to the distribution of temperatures and hypoxic cell populations throughout the tumour.


Tumour markers are important in the diagnosis and monitoring of many tumours. This study tested the hypothesis that an oncofoetal protein, foetal haemoglobin (HbF) is a potential tumour marker in embryonic tumours, useful for management. An immunohistochemical investigation of HbF blood cell (Fc) distribution was carried out in tumours and in bone marrow samples from 83 children and 13 adults with various embryonic tumours (blastomas), and in bone marrow samples of 24 leukaemia patients. In the three, main blastoma types, nephroblastoma (Wilms' tumour), neuroblastoma and retinoblastoma, where all the patients, except two, were children, around 80% of the tumour samples had Fc within proliferating blood vessels and spaces between tumour cells. In parallel, clusters of Fc, mostly F-erythroblasts (Feb), were distributed in the bone marrow of some of those patients and in the bone marrow of 79% of the leukaemia patients. Foetal haemoglobin, as well as being a potential prognostic cancer marker, is a potential indicator of DNA hypomethylation implicated in the development of these tumours, as well as in others previously noted for the presence of HbF. Foetal haemoglobin in most cases is not a product of the tumour cells. Foetal haemoglobin expression by the blood cells surrounding the tumour appears to be induced under the conditions of malignancy. Defining these conditions might contribute to the understanding of carcinogenesis at its basic levels. In this respect, we note that DNA hypomethylation (DNhpom) at CpG dinucleotides (cytosine nucleotide found adjacent to guanine nucleotide) is an example of an epigenetic process implicated in the promotion of carcinogenesis  are common in the predominant kinds of solid tumours as well as in haematological malignancies . In prelogenesis , as wellpression . For exapression . By thatpression . By estas of DNA . Since tThe aim of this study was to investigate the presence of F-cells in blastoma-related tumours by the use of immunohistochemistry. In this way, conditions inducing HbF expression can be related locally to the tumour tissue, rather than to the whole organism, as in our study of colorectal tumours, where we detected local development of F-cells (Our study included Wilms' tumour (nephroblastoma), neuroblastoma, retinoblastoma, childhood leukaemia, rhabdomyosarcoma and others, all of which originate from primordial embryonic cells, including the embryonic neural crest as the presumed source of neuroblastoma, Ewing's sarcoma, medulloblastoma and rhabdomyosarcoma. All have similar morphology, with uniform, primitive embryonic, blastoma cells. Some of them are reported to share the same oncogenes, as WT1 in Wilms' tumour (The study was conducted in two institutions: (1) Pathology Group, Institute of Cell and Molecular Sciences, The Royal London Hospital, England and (2) Department of Histology and Embryology, Poznan University of Medical Sciences, Poland. The programme of research including studies on archival and stored materials was approved by The East London and City Health Authority Research Ethics Committee and by the Poznan University of Medical Sciences, Department of Paediatric Oncology, Haematology and Transplantology, Ethics Committee.We used the peroxidase avidin–biotin method with an affinity purified anti-HbF (n=24), leukaemia (n=24), retinoblastoma (n=18), neuroblastoma (n=17), rhabdomyosarcoma (n=14), medulloblastoma (n=11), Ewing's sarcoma (n=2), hepatoblastoma, neurofibroma and glioblastoma (n=1 for each). All these patients, except those with leukaemia and medulloblastoma, were predominantly children.The study included examination of histological tumour and bone marrow specimens of the following diseases: nephroblastoma  comprised mature erythrocytes (Fer) and nucleated erythroblasts (Feb), which could sometimes be binucleated or in mitosis. Foetal haemoglobin blood cells were identified and were distributed in blood vessels or free in tumour tissues. In the blood vessels their concentrations were 0–100%. They could be distributed either randomly in the blood vessels, in most of the cases not exceeding 50% of the total blood cells , or at 1The scarce cases treated by chemotherapy prior to our examination were noted in the tables of results.The results are summarized in The results are summarized in Some of the Feb were binucleated or in mitosis. We noted here the high rate of proliferating Fc in regenerating blood vessels and in free cell clusters The results are summarized in All specimens were from tumour in the cerebellum. As described in A list of Fc positive and negative cases is shown in One tumour specimen, from the temporal bone of a 60-year-old man, had Fc distributed at 10–50% in part of the blood vessels and in necrotic areas. About 1/4 of these Fc were Feb. Another sacral tumour from a 2-year-old boy had a few fine blood vessels with 100% Fer.In a tumour specimen from a cyst in the leg of 1-year-old boy, we observed Fc cells infiltrating among tumour cells and in adjacent necrotic areas, where some were Feb, as shown in Tumour specimen from the brain of a 52-year-old woman consisted of blastoma-like cells infiltrated by Fer. The adjacent brain tissue was free of Fc.In a bone marrow specimen from a 2-year-old girl, we observed Fc, most of them Feb, distributed as foci and as single cells.As described in the results, F-cells (Fc) were distributed in various patterns inside the embryonic tumour tissues. Foetal haemoglobin blood cells probably originate in the bone marrow and, delivered by circulation, were found randomly at 0–50% in the main blood vessels . HoweverAnother aspect of the abundance of F-cells inside these tumours is related to their embryonic characteristics. Some of these tumours mimic normal embryonic development, such as three-phase nephroblastoma, containing mesenchyme, tubular and glomerular elements, or neuroblastoma cells, which may differentiate to ganglion cells or neural cells. It is therefore possible that these tumours are programmed for autonomous foetal haematopoiesis by F-cell precursors. This hypothesis is supported by the finding of HbF erythroblasts (Feb), some of them in mitosis, in all of these embryonic tumours, most prominently in retinoblastoma and nephroblastoma. Another support for Fc development in tumour tissue comes from the finding of Fc, including Feb, in proliferating blood vessels. These Fc were found only in the tumour and not in other adjacent tissues like normal kidney tissue adjacent to nephroblastoma , or braiIt seems thus that local regeneration rather than transportation through the general circulation is the source of Fc among tumour cells, and in proliferating blood vessels inside the tumours.


NO concentration was estimated indirectly from serum nitrite. The ANOVA test showed a significant difference in serum VEGF165 concentrations between tumour stages (P < 0.001). A striking relationship was found between serum NO levels and tumour stage (P < 0.001). A significant difference was also seen between healthy individuals and patients with stage 1 disease. The present study suggested that large tumour burden was associated with significantly increased levels of VEGF165 and NO. © 1999 Cancer Research CampaignThe importance of tumour angiogenesis in the process of tumour growth and progression in solid tumours has been widely accepted. Among many angiogenic factors, vascular endothelial growth factor (VEGF) has been shown to play a major role in the development and dissemination of the malignant tumours. Nitric oxide (NO) production was also observed in solid tumour tissues. NO has been reported to play an important role for the mitogenic effect of VEGF in the angiogenic process. However, little is known about the correlation between VEGF and NO in circulating levels. Therefore, we investigated serum VEGF and NO concentrations in human gastric cancers as well as healthy individuals, and examined the influence of tumour stage on circulating level of VEGF. The study consisted of 11 healthy individuals and 37 patients with primary gastric cancer who did not receive any prior therapy. Patients were categorized into four groups according to TNM classification. The level of VEGF


Slices of human prostatic adenocarcinoma obtained by transurethral resection were maintained in organ culture for 4 days. Preservation of histological appearance was good with little evidence of necrosis within the viable tissue. Slices of tumour cultured in the presence of testosterone showed a morphological change to a more differentiated type of neoplasm whereas explants cultured in the absence of steroid hormone, or with stilboestrol diphosphate, showed no change. In the case of a relatively anaplastic tumour, testosterone produced a significant increase in the number of mitotic figures seen.


Xenografts originated from human tumours offer the most appropriate research material for in vivo experimental research. However, primary human breast carcinomas are difficult to grow when transplanted in athymic mice: tumour take is less than 15%. Recently, we have achieved 60% tumour take by injecting tumour cell suspensions mixed with Matrigel. Human breast xenografts originated from primary breast carcinoma also frequently show the potential to metastasize spontaneously. In the present study, we generated a human breast carcinoma xenograft line (UISO-BCA-NMT-18) that shows 100% tumorigenicity and 80-100% lung metastasis when transplanted s.c. in athymic mice. We have studied in detail the characteristics of the xenograft and the patient's tumour from which the xenograft line originated. Both the xenograft and the patient's tumour showed intense staining for mutant p53 nuclear protein, and high expression of U-PA, PAI and u-PAR. In vivo growth of the xenograft is stimulated by exogenous supplementation of oestrogen. This xenograft is continuously growing in mice and has shown 80-100% metastasis for the last three successive in vivo passages. This well-characterized, oestrogen-responsive, metastatic breast carcinoma xenograft line will provide excellent research material for metastasis-related research.


The effect of 17 beta-oestradiol on a "receptor positive" and on a "receptor negative" human breast carcinoma grown in nude mice was studied. Experimental growth data were used to determine the effect on tumour growth. Flow cytometric DNA analysis (FCM) performed on tumour tissue obtained by sequential fine-needle aspirations was used to estimate the effect on the cell cycle. In the receptor-positive breast carcinoma, oestradiol induced complete tumour regression and characteristic cell cycle changes. In the receptor-negative breast carcinoma, no changes in tumour growth and cell cycle distribution could be demonstrated following the treatment. The results indicate that the oestradiol-induced cell kill could be explained to some extent by the induction of polyploid cells, which eventually die. Since the cell cycle changes monitored by FCM in the receptor-positive breast carcinoma appeared prior to any reduction in the tumour size, the results suggest that FCM may prove a valuable method in the early detection of tumour response to hormone treatment in human breast cancer.


Strain BALB/c female mice bearing syngeneic implants of 2 mammary adenocarcinomata were treated with MER, x-irradiation or both. MER was administered either subcutaneously at a site contralateral to the neoplastic growth or both into such a site and directly at the tumour location. None of the treatments effected cures but many of the treated animals survived significantly longer than did the saline injected controls. There was no evidence that introduction of MER into, or directly adjacent to, a tumour is a generally more efficacious route of administration than application at only a distal site and there was, indeed, the strong contrary impression that distal treatment alone bestowed survival protection more often and to a greater extent. In no instance was there a shortening of survival time following administration of MER at a location away from the tumour implant.


These investigations included research on the effects of macrophage–derived eicosanoids (cycloxygenase and lipoxygenase derivates of arachidonic acid) and of monokines such as tumour necrosis factor-α, interleukin-1 and granulocyte–monocyte–macrophage–colony stimulating factor) and of lymphocyte products: interleukins and interferons. The investigations yielded information on the effects of various factors on macrophage and T-cell activation in vitro, determination of direct anti-tumour properties on animal and human tumour cells in vitro and on therapeutic effectiveness in tumour-bearing individuals either alone or in combination with other therapeutic factors and their production by tumour cells. During recent years much effort has been dedicated towards the use of the tumour cells transfected with cytokine genes in the preparation of cancer vaccines. Cycloxygenase products (prostaglandins) were usually assumed to inhibit expression of anti-tumour activity by macrophages and an increase in their production in cancer patients was considered as a poor prognostic index. Lipoxygenase (leukotrienes) products were assumed to exhibit antitumour activity and to induce production of IL-1 by macrophages. Interleukins 2, 4, 6, 7, 12 and the interferons were extensively tested for their therapeutic effectiveness in experimental tumour models and in cancer clinical trials. The general conclusion on the use of cell-free mediators for cancer immunotherapy is that much still has to be done in order to assure effective and reproducible therapeutic effectiveness for routine use in the treatment of human neoplasia.The description of a cell-free soluble anti-tumour factor by Carswell


Assessment of tumour response to chemotherapy is important when assessing efficacy of treatment and comparing differing therapeutic regimens. Percentage hepatic replacement (PHR) is commonly used to assess response to treatment of colorectal hepatic metastases. PHR is dependent not only on tumour volume, but also on hepatic parenchymal volume. The effect of tumour growth on hepatic parenchymal volume is unclear but is of importance owing to its effect on PHR. We assessed tumour and hepatic parenchymal weights in an animal tumour model using dissection, and tumour and hepatic parenchymal volumes in patients with colorectal hepatic metastases using CT scanning, in order to establish how hepatic parenchyma varied with change in metastasis size. There was no significant correlation between tumour and liver parenchyma in either the animal model  or the patient study . This suggests that hepatic parenchymal volume was preserved in the presence of increasing tumour volume. In a further study of computerised tomographic (CT) scans before and after treatment in patients whose tumours either responded to chemotherapy or continued to grow, change in PHR (median proportion of PHR change = 0.40) significantly (P = 0.04) underestimated the change in tumour volume (median proportion of tumour volume change = 0.56), particularly at higher (> 400 ml) volumes. There was good correlation between change in tumour volume and WHO criteria in assigning patients to tumour growth, stable disease or tumour response categories. This study suggests that, in clinical trials comparing colorectal liver metastasis treatments, metastasis volume and not PHR should be used to assess extent of disease and the effect of treatment.


However, they formed aggressive tumours upon co-implantation with a ‘foreign body’, i.e. a gelatin sponge, and the rate of tumour take ranged from 8% to 58% among QR clones. The enhanced tumorigenicity was due to host cell-mediated reaction to the gelatin sponge (inflammation). Immunoblot analysis and enzyme activity assay revealed a significant inverse correlation between the frequencies of tumour formation by QR clones and the levels of manganese superoxide dismutase  and glutathione peroxidase  in the respective tumour clones. Electron spin resonance (ESR) revealed that superoxide-scavenging ability of cell lysates of the QR clone with high level of Mn-SOD was significantly higher than that with low level of the antioxidative enzyme in the presence of potassium cyanide, an inhibitor for copper–zinc superoxide dismutase (CuZn-SOD) (P<0.001). Minisatellite mutation (MSM) induced by the inflammatory cells in tumour cells were investigated by DNA fingerprint analysis after QR clones had been co-cultured with gelatin-sponge-reactive cells. The MSM rate was significantly higher in the subclones with low levels of Mn-SOD and GPχ (P<0.05) than in the subclones with high levels of both enzymes. The MSM of the subclones with low levels of both enzymes was inhibited in the presence of mannitol, a hydroxyl radical scavenger. The content of 8-hydroxydeoxyguanosine (8-OHdG) by which the cellular DNA damage caused by active oxygen species can be assessed was significantly low in the tumours arising from the QR clone with high levels of Mn-SOD and GPχ even if the clone had been co-implanted with gelatin sponge, compared with the arising tumour from the QR clone with low levels of those antioxidative enzymes (P<0.001). In contrast, CuZn-SOD and catalase levels in the six QR clones did not have any correlation with tumour progression parameters. These results suggest that tumour progression is accelerated by inflammation-induced active oxygen species particularly accompanied with declined levels of intracellular antioxidative enzymes in tumour cells. © 1999 Cancer Research CampaignWe isolated six clones of weakly tumorigenic fibrosarcoma (QR) from the tumorigenic clone BMT-11 cl-9. The QR clones were unable to grow in normal C57BL/6 mice when injected s.c. (1 × 10


The growth of tumours in guinea-pigs was observed for 20 weeks after placing them on various doses of vitamin C. Complete tumour regression occurred in 55% of those animals receiving 0-3 mg/kg/day ascorbic acid, whereas animals given 10 mg/kg/day showed tumour inhibition but no regression. In contrast, tumours in animals maintained on 1 g/kg/day ascorbic acid grew without sign of retardation. When increased amounts of ascorbic acid were restored to the diet of scorbutic tumour-bearing animals, tumours which had not regressed responded with enhanced growth. Likewise, animals previously maintained on 10 mg/kg ascorbic acid responded in turn to the additional vitamin with enhanced tumour growth. In contrast, all tumour-bearing animals maintained on 1 g/kh ascorbic acid died within 3 weeks when this dose was replaced with 0-3 mg/kg.


A mouse monoclonal antibody (LICR-LON/HT13) has been developed to a cell-surface antigen carried on a human germ-cell tumour xenograft (HX39). After radioiodination, the antibody localized in vivo preferentially in xenografted tumours as opposed to normal mouse tissue, whereas tumor uptake did not occur with normal mouse IgG or nonspecific monoclonal IgG. This selective localization could be abolished by simultaneous injection of an excess of the unlabelled LICR-LON/HT13. The kinetics of and factors influencing localization have been examined. Tumour weight was important in that the smaller the tumour the better the localization. LICR-LON/HT13 was found to localize also in other xenografted germ-cell tumours, but not in non-germ-cell tumour xenografts. Thus monoclonal antibodies are capable of selective in vivo localization of human tumours in an animal model, and their clinical value should now be assessed.


Tumour cells from 7 patients with ovarian carcinoma and from 22 different human tumour xenografts representing a wide range of histological sub-types have been examined for multicellular spheroid forming ability. Spheroid formation was limited to cells derived from xenografts. Of the 22 lines tested, 5 formed spheroids capable of growth in isolation. There was no clear relationship between histological type and spheroid-forming ability. The plating efficiency of tumour cells obtained from spheroids was always greater than for the cells obtained from the dissociated tumour of origin and was in some cases as much as 6-fold greater. Spheroid growth was nearly exponential for 4 cell lines. Volume growth delay was used to investigate the activity of melphalan, adriamycin, the Vinca alkaloids, CCNU and cisplatin. Differences between lines in drug response broadly reflected patient and in vivo xenograft response.


Hexadecafluorinated zinc phthalocyanine (ZnPcF16), an analogue of zinc phthalocyanine (ZnPc) in which all hydrogen atoms have been substituted by fluorine, was prepared as a single isomeric product via the condensation of tetrafluorophthalonitrile with zinc acetate. Fluorination renders the ZnPc soluble in most common solvents. The photodynamic properties and pharmacokinetics of the ZnPcF16 were evaluated in EMT-6 tumour-bearing Balb/c mice using 65Zn-radiolabelled analogues. Both dyes, administered i.v. at 1 mumol kg-1 as Cremophor emulsions, revealed good tumour uptake [approximately 8-9 per cent of the injected dose per g tissue (%IDg-1)] at 24 h post injection (p.i.), with the fluorinated dye reaching higher concentrations (approximately 11%IDg-1) at 48 h p.i. and subsequently higher tumour-blood ratios due to rapid blood clearance. ZnPcF16 at a dose of 5 mumol kg-1 (4.3 mg kg-1) induced complete tumour regression after phototherapy . At a dose of 2 mumol kg-1 and phototherapy at 24 h p.i., the tumour volume doubling time increased to 11 days vs 6 days for the control tumours. A similar tumour growth delay was observed when phototherapy was conducted at 48 h or 72 h after dye injection implying that tumour response correlates with tumour dye concentrations rather than serum concentrations. As a result of its low solubility, the administered dose of ZnPc was limited to 1 mumol kg-1 and at this drug level significant tumour response was only observed when the dye was solubilised as the pyridinium salt. Isolation of the neoplastic cells after in vivo dye administration and in vitro exposure to red light followed by a colony formation assay showed that the ZnPcF16 exhibited a 1-2 order of magnitude higher potential for direct cell killing as compared with Photofrin and about a five times lower efficiency than ZnPc. However, all three photosensitisers induced complete occlusion of tumour vasculature immediately after PDT, suggesting that tumour regression mainly resulted from vascular stasis. The ZnPcF16 offers several advantages over ZnPc for clinical applications, including improved solubility in most solvents, resulting in facilitated drug formation, favourable pharmacokinetics as well as the potential use in fluorine magnetic resonance (F-MR) imaging.


Nine patients receiving the CRC Phase II study schedule of temozolomide were assessed with [18F]fluorodeoxyglucose ([18F]FDG) dynamic positron emission tomography (PET) scans prior to and 14 days after treatment with temozolomide given as 750–1000 mg m−2over 5 days every 28 days. Tumour MRGlu was calculated and compared with objective response at 8 weeks. Pretreatment MRGlu was higher in responders than non-responders. The responding patient group had a greater than 25% reduction in MRGlu in regions of high focal tumour uptake (HFU). Whole tumour changes in MRGlu did not correlate with response. Percentage change in HFU standardized uptake value (SUV) did discriminate the responding from the non-responding patients, but not as well as with MRGlu. Large differences also occurred in the normal brain SUV following treatment. Thus, MRGlu appeared to be a more sensitive discriminator of response than the simplified static SUV analysis. Changes in MRGlu may reflect the degree of cell kill following chemotherapy and so may provide an objective, quantitative subclinical measure of response to therapy. © 2000 Cancer Research CampaignQuantitation of metabolic changes in tumours may provide an objective measure of clinical and subclinical response to anticancer therapy. This pilot study assesses the value of quantitation of metabolic rate of glucose (MRGlu) measured in mmol min


CD44 is a marker of tumour-initiating cells and is upregulated in invasive breast carcinoma; however, its role in the cancer progression is unknown. Here, we show that antibody-mediated CD44-targeting in human breast cancer xenografts (HBCx) significantly reduces tumour growth and that this effect is associated to induction of growth-inhibiting factors. Moreover, treatment with this antibody prevents tumour relapse after chemotherapy-induced remission in a basal-like HBCx. In vivo CD44 targeting by siRNA and by specific antibody results in anti-tumor activity in colon cancer xenografts and human acute leukaemia xenografts, respectively  and maintained in specific pathogen-free conditions. Their care and housing were in accordance with institutional guidelines as put forth by the French Ethical Committee  under the supervision of an authorised investigator (MF Poupon).4 to 2 × 106 Da) was provided by Javenech Cie .The murine anti-CD44 monoclonal antibody (P245) and Hermes3 were supplied by Laurence Boumsell  and David Naor, respectively, and the isotype control (IgG1) was from Sigma-Aldrich . Adriamycin (Doxorubicin) and cyclophosphamide (Endoxan) were purchased from Teva Pharmaceuticals  and Baxter , respectively. Adriamycin and cyclophosphamide combination was designated as AC. Hyaluronan . After washing, cells were labelled with FITC-conjugated P245 anti-CD44 mAb (1 μg per 105 cells) or IgG1 . The proportion of CD44+ cells was measured by flow cytometry relative to isotype-matched controls, using a FACSvantage . Each measurement was done on 5,000 cells. Dead cells were excluded using 7-AAD.CD44 earlier . The freCD44 mRNA expression was analysed by nested RT–PCR using human specific intron-spanning primers  from mRNA of tumours or fat pad tissues frozen in liquid nitrogen and extracted as described earlier .The differences between median Δ+ cells  in the P245 mAb-treated group was 70% at day 28 (P<0.01) in both xenografts, with HBCx-3 tumour growth being fully stabilized as early as day 5, whereas HBCx-8 was strongly inhibited from days 15–20 , a treatment commonly used for this breast cancer subtype  induces a complete regression of all tumours by day 35, both by local palpation and histological analysis of the injected fat pads + cells: one in every 107 cells) , whereas injections of IgG1 isotype control had no effect. These results support the idea that CD44+ surviving cells play a role in tumour recurrences.We next addressed the role of CD447 cells) , showinganalysis . Small ianalysis . To test+ cells play an important role in tumour growth-driving mechanisms. This effect was associated with the activation of cytokine gene expression, suggesting that these cytokines may be involved in the P245-mediated inhibition of tumour growth   . As HA btuzumab) . This re+ cancer cells, suggesting that a subpopulation of CD44+ tumour cells survives in spite of the macroscopic complete tumour remission induced by chemotherapy. The fact that P245 treatment during tumour remission inhibits tumour recurrence suggests that CD44+ cells play a direct role in the tumour recurrence, providing an interesting target and a useful model to finalise breast cancer therapies associations.In breast cancer treatment, combined chemotherapy and radiotherapy can induce disease remission; however, tumour recurrences are frequent and responsible for treatment failures. Such recurrences are due to the persistence and proliferation of chemoresistant breast cancer cells. This is particularly true for basal-like breast cancers, which are sensitive to anthracyclines and taxanes combinations, but have higher risk of recurrence after therapy and worse survival rates . By usin+ breast cancer cells inhibits tumour growth and post-chemotherapy tumour recurrence in both ER+ and basal-like breast cancer. In addition, we show that the effect of P245 mAb was associated with induction of cytokines known to have anti-proliferative activity. The finding that CD44+ cells play a key role in chemoresistant breast cancer recurrences provides an innovative strategy to improve breast cancer treatment.In conclusion, we show here for the first time that targeting CD44


This paper gives a brief survey of the use of algebraic rewriting systems for modellingand simulating various biological processes, particularly at the cellular level.


Clonal interaction between three subpopulations of Ehrlich carcinoma were studied during growth as mixed solid tumours and as ascites tumours in immune-incompetent nude NMRI mice. The tumour cell lines differed in DNA content as determined by DNA flow cytometry (FCM). Tumour growth was evaluated by tumour growth curves including calculation of tumour volume doubling times, tumour weight on day 14, cell cycle times (per cent labelled mitoses) and cell cycle distributions (FCM). Two subpopulations (E1.15 and E1.95) showed nearly identical growth characteristics during both solid and ascites tumour growth. The third subpopulation (E1.80) grew more slowly. FCM on fine-needle tumour aspirates was used to determine the relative proportions of the cell populations in mixed solid tumours in which E1.95 showed a growth-dominating effect on E1.15. No such effect was demonstrated during single-cell tumour growth in ascitic fluid in which the cells had no intimate contact. Ascitic fluid from E1.95-bearing animals or radiation-killed E1.95 cells had no effect on the growth of E1.15, and no remote effect was seen when the two cell lines were growing in opposite flanks. This indicates that only viable E1.95 cells in close in vivo contact were able to induce growth inhibition of the E1.15 subpopulation. Both the E1.95 and the E1.15 cells dominated the E1.80 cells, but in these cases cell kinetic differences may have played a role as the E1.95 and the E1.15 lines grew faster than the E1.80. The E1.80 cell line had no dominating effect on the E1.15 or E1.95. It is concluded that non-immunologically mediated cellular dominance in heterogeneous tumours may contribute to the evolution of these tumours and may be involved in fundamental tumour biological phenomena.


The Second Meeting of the British Society for Immunology Tumour Immunology Affinity Group (TIAG) took place at King's College  on 17-18 June 1997 and brought together over 100 tumour immunologists from the UK and abroad. In contrast to previous meetings the focus of the meeting was on the role of adhesion in immunosurveillance and tumour dissemination. In addition, recent achievements in the areas of chemokines, cytotoxic T-lymphocyte (CTL) and natural killer (NK) cells, co-stimulation, gene and adoptive immunotherapy were also addressed. The purpose of this report is to outline current trends in tumour immunology.


The tumour used, designated MT1, is a more radiosensitive form of the anaplastic MT tumour previously described. No explanation for the increased radiosensitivity was found, but it was shown not to be due to infection or to a change in immunological status, growth rate or histology. The sensitivity has remained constant throughout the present work. No cytotoxicity in the tumour was observed when 1 mg/g body weight of Ro-07-0582 was injected immediately after a single dose of X-rays; indeed a small protective effect was seen. A radiosensitization enhancement of 1-5 was achieved with a relatively low drug dose of Ro-07-0582 in a 5F/4d fractionated regime. The interval between the injection of a low dose of Ro-07-0582 and the start of irradiation was found to be critical, the optimum interval being 45-60 min. The subsequent incidence of distant metastases was not increased by the use of Ro-07-0582 at the time of "primary" tumour irradiation.


The therapeutic effects of intraperitoneal topotecan, a water-soluble camptothecin analogue, were investigated in two models of human ovarian carcinoma xenografted intraperitoneally into nude mice: the IGROV-1 tumour, which originated from an untreated patient, and the A2780 tumour, selected for resistance in vitro to cisplatin (A2780DDP). In IGROV-1 tumour-bearing mice, the optimal dose (10 mg kg-1) of topotecan, given intraperitioneally every 4 days for four occasions markedly increased survival time over control mice (300 T/C%) and cured 4/9 mice, and such effects were not achieved by any of the clinically available drugs tested, i.e. cisplatin carboplatin and doxorubicin delivered intraperitonally according to their optimal doses and schedules. In the treatment of A2780DDP tumour-bearing mice, topotecan was very effective since, at dose levels of 6.6 and 10 mg kg-1 every 4 days for four occasions, 15/18 mice survived more than 100 days, and most of them (12/15) were found to be tumour free. The high responsiveness of this tumour to topotecan might be related to the elevated expression of the target enzyme topoisomerase I. From these results, intraperitoneal treatment with topotecan appears to be a promising approach in the therapy of refractory ovarian cancer confined to the peritoneal cavity.


The anti-tumour activity of MEN 11467 was observed both with s.c. or intravenous treatments and was partially reverted by the concomitant administration of exogenous SP. Doxorubicin did not show any activity in controlling U373 MG growth in this in vivo model. A novel therapeutic approach to treat malignant gliomas with tachykinin NK1receptor antagonists is suggested by these findings. © 2000 Cancer Research CampaignAstrocytes harbour functional receptors to many neurotransmitters, including substance P (SP), an undecapeptide belonging to the tachykinin family of peptide transmitters. SP activates malignant glial cells to induce cytokine release and proliferation, both responses being relevant for tumour progression. In tumours developed in nude mice transplanted subcutaneously (s.c.) to U373 MG human glioma cells, the presence of SP was observed at immunohistochemistry. Although the administration of exogenous SP did not significantly affect the size or development of U373 MG xenograft, a role of SP in supporting glioma progression in vivo was highlighted by the tumour growth inhibition induced by highly specific and selective human tachykinin NK


A hypothesis is proposed that tumour lysis may be an important factor affecting blood levels of CEA. This has been explored in an experimental study with a model tumour system, consisting of immune-deprived mice bearing human CEA-producing tumours. Using agents such as irradiation, chemotherapeutic drugs, diphtheria toxin and techniques such as cryosurgery, it has been shown that tumour lysis is important when it is both rapid and extensive. The extent to which this may occur in patients remains uncertain, except in rare instances of dramatic response of malignant disease to treatment.


Cell survival and the cytotoxic effects of the drugs on the tumour cells were determined by using a double-headed gamma counter to measure the retention of the 125I label.As a more sensitive measure of cell survival, 125I retention).Both AraC and AraCTP, either as the free drugs or liposomally associated, had no effects on the tumour. Due to the lack of response of tumour cells to these drugs, further studies were initiated with free and liposomally associated methotrexate (MTX), a drug to which the cells were known to be sensitive. It was found that the liposomally associated MTX, at a 5-10-fold lower dose than the free drug, was (a) more effective in prolonging the survival of tumour-bearing mice and (b) as effective as the free drug in killing tumour cells (as measured by the In vivo MTX was more effective in the liposomally associated form, whereas liposomally entrapped AraC and AraCTP were ineffective. It is proposed that in vivo liposomally associated drugs may be acting not by actively localizing in the tumour cells, but by the liposomes providing a slow-release drug depot, improving the pharmacokinetic properties of MTX.