Estimation of metabolic flux rates in liver purine catabolism of tumour-bearing mice by computer simulation of radioactive tracer experiments

Mouse hepatocytes from healthy control mice and from Ehrlich ascites tumour-bearing mice were used for tracer-kinetic studies of purine catabolism of liver cells during different periods of tumour growth. The dynamics of the radioactive tracers were modelled mathematically by a system of differential equations. Computer simulations, i.e. direct fitting of numerical solutions of these equations to the observed time-courses of metabolites and specific radioactivites, enables one to estimate unknown kinetic parameters of a simplified model of pathways of hepatic purine catabolism in tumour-bearing mice. There occurred great differences of metabolic flux rates between control hepatocytes, hepatocytes of mice during the proliferating period of tumour growth (6th day after inoculation of the tumour) and hepatocytes of mice during the resting period of tumour growth (12th day after inoculation of the tumour). The final purine degradation of hepatocytes prepared during the proliferating period was lower in comparison with that of control hepatocytes, but it was markedly higher in hepatocytes prepared during the resting period of tumour growth. The changes in hepatocyte purine catabolism during the proliferating period of tumour growth argue for transitions which aim at the maintenance of high purine nucleotide levels in the liver itself rather than for an increased nucleoside and nucleobase supply for the tumour. This suggestion is in accordance with the increased ATP level of the liver during the proliferating phase of tumour growth. The drastic acceleration of the final steps of hepatic purine catabolism forming uric acid and allantoin during the resting period of tumour growth was predominantly due to increased flux rate from xanthosine and guanine in accordance with increased catabolism of monophosphorylated nucleotides.     

Tumour-cell-induced production of tumour necrosis factor by monocytes of gastric cancer patients receiving BCG immunotherapy

Summary Human peripheral blood monocytes cocultured with tumour cells were used as an in vitro model of in situ interactions between tumour-infiltrating macrophages and the tumour. Tumour cells stimulated de novo expression of the human tumour necrosis factor (TNF) gene in monocytes and caused the release of TNF into the culture supernatant. A group of 14 patients with stage IVA gastric cancer receiving adjuvant chemotherapy (5-FU, Adriamycin, mitomycin C: FAM) or immunochemotherapy (BCG+FAM) was investigated for the ability of monocytes to produce TNF in vitro upon stimulation with tumour cells or purified protein derivative of tuberculin (PPD). Patients were followed at biweekly intervals, i.e. before each instillation of BCG epicutaneously over a period of 10 weeks. It was found that monocytes of some patients receiving BCG at the end of the observation period had an enhanced ability to produce TNF following stimulation with tumour cells. In contrast, such production was not substantially altered during the study period in patients on chemotherapy. PPD-induced TNF production was much weaker and was not significantly changed during this observation time. We infer that BCG immunotherapy may induce the subtle changes in some cancer patients that lead to an increased interaction between monocytes and tumour cells and result in enhanced production of cytokine(s) with antitumour properties.     

An analysis of the role of microfilaments in the establishment and maintenance of asymmetry in Caenorhabditis elegans zygotes

Microfilaments are needed to generate asymmetry during the first cell cycle in Caenorhabditis elegans zygotes. To investigate when and how microfilaments participate in this process, we have "pulsed" zygotes with the microfilament inhibitor cytochalasin D (CD) at different times during the cell cycle. We have shown that microfilaments are only required during a narrow time interval approximately three-quarters of the way through the first cell cycle for the manifestations of asymmetry that occur during and subsequent to this interval. When CD treatment spans this critical time interval, pseudocleavage, pronuclear migration, germ-granule segregation (all of which occur during the interval), and movement of the mitotic spindle to an asymmetric position (which occurs later in the cell cycle) are perturbed. In contrast, embryos briefly treated with CD before or after the critical time interval manifest normal asymmetry. Our results suggest that in C. elegans microfilaments participate in the generation of zygotic asymmetry by providing spatial cues and/or serving as a part of the necessary machinery only during a brief period in the first cell cycle, and are not required to maintain asymmetries that have already been established.     

Tree Harvest in an Experimental Sand Ecosystem: Plant Effects on Nutrient Dynamics and Solute Generation

The hydrochemical signatures of forested ecosystems are known to be determined by a time-variant combination of physical-hydrologic, geochemical, and biologic processes. We studied subsurface potassium (K), calcium (Ca), and nitrate (NO₃) in an experimental red -pine mesocosm to determine how trees affect the behavior of these nutrients in soil water, both during growth and after a harvest disturbance. Solution chemistry was monitored for 2 years at the end of a 15-year period of tree growth, and then for 3 more years after harvest and removal of aboveground biomass. Concentrations were characterized by three distinct temporal patterns that we ascribe to changes in solute generation mechanisms. Prior to harvest, K soil-water concentrations were relatively uniform with depth, whereas Ca soil-water concentrations doubled with depth. Nitrate concentrations were below detection in soil water and discharge (drainage) water. Plant uptake and water/nutrient cycling exerted strong control during this interval. During the 1st year after harvest, K concentrations tripled in shallow soil water, relative to preharvest levels, and showed a strong seasonal peak in discharge that mimicked soil temperature. Summer soil temperatures and annual water flux also increased. Decomposition of labile litter, with complete nitrogen (N) immobilization, characterized this interval. In the third interval (years 2 and 3 after harvest), decomposition shifted from N to carbon (C) limitation, and Ca and NO₃ concentrations in discharge spiked to nearly 200 and 400 μM, respectively. Relatively stable ionic strength and carbonate chemistry in discharge, throughout the study period, indicate that carbonic-acid weathering was sustained by belowground decomposition long after the harvest. This stable chemical weathering regime, along with the persistence of N limitation for a long period after disturbance, may be characteristic of early-phase primary-successional systems.     

Cell kinetic response of an experimental tumour to irradiation. Use of the stathmokinetic method

The cell kinetic perturbations following irradiation (20 Gy) were studied by combining the metaphase arrest method using vincristine with histological indices of cell death. The metaphase arrest method yielded remarkably constant values of rate of entry into mitosis (rM) of around 24 new cells/1,000 cells/hour during a 7 day period in which there was no tumour growth. The time course of cell death as indicated by changes in the pyknotic index during this period may reflect the processes of reoxygenation and repopulation known to influence the results of fractionated radiotherapy.     

Timing of cotyledon damage affects growth and flowering in mature plants

Although the effects of herbivory on plant fitness are strongly linked to age, we understand little about how the timing of herbivory at the seedling stage affects growth and reproduction for plants that survive attack. In this study, we subjected six north-western European, dicotyledonous grassland species (Leontodon autumnalis, Leontodon hispidus, Plantago lanceolata, Plantago major, Trifolium pratense and Trifolium repens) to cotyledon removal at 7, 14 and 21 d old. We monitored subsequent growth and flowering (number of inflorescences recorded, and time taken for first flowers to open) over a 107 d period. Cotyledon removal reduced growth during establishment (35 d) for all species, and a further three exhibited reduced growth at maturity. Four species developed fewer inflorescences, or had delayed flowering after cotyledon removal. Although early damage (7 d old) had the greatest long-term effect on plant performance, responses varied according to the age at which the damage occurred and the species involved. Our results illustrate how growth and flowering into the mature phase is affected by cotyledon damage during different stages of seedling ontogeny, and we highlight the ways in which ontogenetic variation in seedling tolerance of tissue loss might impact upon plant fitness in mature plant communities.     

Can artificial plant beds be used to enhance macroinvertebrate food resources for perch (<Emphasis Type="Italic">Perca fluviatilis</Emphasis> L.) during the initial phase of lake restoration by cyprinid removal?

Restoration of shallow lakes to a clear-water state, often characterized by high submerged macrophyte cover and a high proportion of piscivores such as perch, Perca fluviatilis L., frequently involves removal of a large proportion of the zoobenthivorous fish, such as bream, Abramis brama L., and roach, Rutilus rutilus L. (i.e. biomanipulation). However, establishment of submerged macrophytes is often delayed following fish removal. This is unfortunate because plant beds typically host high densities of the macroinvertebrates constituting the diet of small perch and thus help perch to go through the bottleneck from feeding on macroinvertebrates to feeding on fish. Establishment of artificial plant beds may be a useful tool to enhance macroinvertebrate population growth and thus food resources for small perch until the natural plants have established. To investigate this restoration option, we studied during two growing seasons (June–October) the composition and abundance of the macroinvertebrate community in artificial plant beds installed in shallow Lake Væng (Denmark) comprising the initial phase of a biomanipulation effort by fish removal. Lake areas with artificial plant beds exhibited substantially higher macroinvertebrate densities than the lake bottom. This suggests that artificial plant beds may be used as feeding grounds for small perch, similarly to the well-known refuge effect for zooplankton against fish predation. In this way, artificial plant beds could help maintain a clear-water state during the transient period when natural submerged vegetation is not yet established in the lake.     

Modelling the early growth of ductal carcinoma in situ of the breast

The growth of a tumour in a rigid walled cylindrical duct is examined in order to model the initial stages of tumour cell expansion in ductal carcinoma in situ (DCIS) of the breast. A nutrient-limited growth model is formulated, in which cell movement is described by a Stokes flow constitutive relation. The effects on the shape of the tumour boundary of the material properties (i.e. the viscosity) and the extent to which the cells adhere to the duct wall are studied using numerical and asymptotic methods. It is shown how stable, non-planar, interface configurations result and that, during these initial stages, before the duct wall has been breached, few cells die and a nutrient-rich model is usually sufficient to capture the behaviour. Finally, we discuss the relevance of this approach to DCIS and suggest possible avenues for further work. Send offprint requests to:S.J. Franks at Centre of Mathematical Medicine.     

Spontaneous regression of tumours. Possible cross reactivity of autoantibodies against carbonic anhydrase I

Spontaneous tumour regression in patients after high dose therapy and autologous stem cell transplantation or patients with standard therapy is accompanied with the presence of high titers autoantibodies against carbonic anhydrase I (CA I). The concomitant presence of aplastic anaemia-like syndrome in these patients points to parallel bone marrow suppression during this period. It seems that CA I, an ‘obscure’ enzyme, does not have any significant physiological role in humans. One possible explanation points to the fact that autoantibodies against CA I may target another antigen(s) which is(are) important in tumour growth as well as in normal haematopoiesis. One of the candidates for such a target is the DNA polymerase theta.     

Oncogenes, anti-oncogenes and the immune response to cancer: a mathematical model.

We develop a mathematical model for the initial growth of a tumour after a mutation in which either an oncogene is expressed or an anti-oncogene (i.e. tumour suppressor gene) is lost. Our model incorporates mitotic control by several biochemicals, with quite different regulatory characteristics, and we consider mutations affecting the cellular response to these control mechanisms. Our mathematical representation of these mutations reflects the current understanding of the roles of oncogenes and anti-oncogenes in controlling cell proliferation. Numerical solutions of our model, for biologically relevant parameter values, show that the different types of mutations have quite different effects. Mutations affecting the cell response to chemical regulators, or resulting in autonomy from such regulators, cause an advancing wave of tumour cells and a receding wave of normal cells. By contrast, mutations affecting the production of a mitotic regulator cause a slow localized increase in the numbers of both normal and mutant cells. We extend our model to investigate the possible effects of an immune response to cancer by including a first order removal of mutant cells. When this removal rate exceeds a critical value, the immune system can suppress tumour growth; we derive an expression for this critical value as a function of the parameters characterizing the mutation. Our results suggest that the effectiveness of the immune response after an oncogenic mutation depends crucially on the way in which the mutation affects the biochemical control of cell division.     

Baroreflex responsiveness is maintained during isometric exercise in humans

The simultaneous rise in heart rate and arterial pressure during isometric handgrip exercise suggests that arterial baroreflex control may be altered. We applied incremental intensities of neck suction and pressure to nine healthy young men to alter carotid sinus transmural pressure. Carotid stimuli were delivered during 1) supine control, 2) "anticipation" of beginning exercise, and 3) handgrip (20% of maximum voluntary contraction). Anticipation was a quiet period, immediately preceding the beginning of handgrip, when no muscular work was being performed. Compared with control, the R-R interval prolongation and mean arterial pressure decline provoked by carotid stimuli were decreased during the anticipation period. These data suggest that influences from higher central neural locations may alter baroreflex function. Furthermore, we derived stimulus-response curves relating carotid sinus transmural pressure to changes in R-R interval and mean arterial pressure. These curves were shifted during handgrip; however, calculated regression slopes were not changed from control. The data indicate that isometric handgrip exercise has a specific influence on human carotid baroreflex control of arterial pressure and heart period: baroreflex function curves are shifted rightward during handgrip, whereas baroreflex sensitivity is unchanged. Furthermore, central neural influences may be partially involved in these alterations.     

The epidermal-growth-control theory of stem elongation: an old and a new perspective

The botanist G. Kraus postulated in 1867 that the peripheral cell layers determine the rate of organ elongation based on the observation that the separated outer and inner tissues of growing stems spontaneously change their lengths upon isolation from each other. Here, we summarize the modern version of this classical concept, the "epidermal-growth-control" or "tensile skin" theory of stem elongation. First, we present newly acquired data from sunflower hypocotyls, which demonstrate that the expansion of the isolated inner tissues is not an experimental artefact, as recently claimed, but rather the result of metabolism-independent cell elongation caused by the removal of the growth-controlling peripheral walls. Second, we present data showing that auxin-induced elongation of excised stem segments is attributable to the loosening of the thick epidermal walls, which provides additional evidence for the "epidermal-growth-control concept". Third, we show that the cuticle of aerial organs can be thin and mechanically weak in seedlings raised at high humidity, but thick and mechanically important for organs growing under relatively dry air conditions. Finally, we present a modified model of the "tensile skin-theory" that draws attention to the mechanical and physiological roles of (a) the thickened, helicoidal outer cell walls, (b) the mechanical constraint of a cuticle, and (c) the interactions among outer and inner cell layers as growth is coordinated by hormonal signals.     

Timing in the life histories of insects

This paper presents a heuristic model illustrating some major problems in analyzing seasonal life histories of multigeneration insects. The concept of the critical interval is introduced and defined as the age classes that survive at the end of a period of population growth. These conclusions follow from the results: The optimal age for occupying a habitat depends upon the duration of the habitat as well as the life history of the insect. Two positions of the initial age distribution may give local maxima for fitness. The critical interval should often include the youngest age classes to maximize fitness while the optimal position of the initial age distribution may be at a much older age. In this case, conflicts arise between the positions of the critical interval at the end of one growing period and the initial age distribution at the start of the next. The length of the critical interval that maximizes fitness in a particular environment may be relatively small in which case mortality at the end of a growing period may be high and timing would appear to be poor even though fitness is maximized. In this model, optimum generation lengths exist which are not the shortest attainable. Finally, the length of time that a habitat remains suitable influences all of the above results and must be taken into account in analyzing the adaptedness of life history traits.     

Modelling the Role of Angiogenesis and Vasculogenesis in Solid Tumour Growth

Recent experimental evidence suggests that vasculogenesis may play an important role in tumour vascularisation. While angiogenesis involves the proliferation and migration of endothelial cells (ECs) in pre-existing vessels, vasculogenesis involves the mobilisation of bone-marrow-derived endothelial progenitor cells (EPCs) into the bloodstream. Once blood-borne, EPCs home in on the tumour site, where subsequently they may differentiate into ECs and form vascular structures. In this paper, we develop a mathematical model, formulated as a system of nonlinear ordinary differential equations (ODEs), which describes vascular tumour growth with both angiogenesis and vasculogenesis contributing to vessel formation. Submodels describing exclusively angiogenic and exclusively vasculogenic tumours are shown to exhibit similar growth dynamics. In each case, there are three possible scenarios: the tumour remains in an avascular steady state, the tumour evolves to a vascular equilibrium, or unbounded vascular growth occurs. Analysis of the full model reveals that these three behaviours persist when angiogenesis and vasculogenesis act simultaneously. However, when both vascularisation mechanisms are active, the tumour growth rate may increase, causing the tumour to evolve to a larger equilibrium size or to expand uncontrollably. Alternatively, the growth rate may be left unaffected, which occurs if either vascularisation process alone is able to keep pace with the demands of the growing tumour. To clarify further the effects of vasculogenesis, the full model is also used to compare possible treatment strategies, including chemotherapy and antiangiogenic therapies aimed at suppressing vascularisation. This investigation highlights how, dependent on model parameter values, targeting both ECs and EPCs may be necessary in order to effectively reduce tumour vasculature and inhibit tumour growth.     

The application of ultrasound in neuroendoscopic procedures: first results with the new tool "NECUP-2"

In this paper, our experience with originally constructed Neurosurgical Endoscopic Contact Ultrasound Probe "NECUP-2" in neuroendoscopy is reported. Between June 1997 and June 2007, 132 neuroendoscopic procedures have been performed: 102 endoscopic thrid ventriculostomies (ETV), 15 arachnoid cysts and 5 intraventricular tumours operations. The "NECUP-2" was applied effectively in all cases in which blunt perforation was not possible: 38/102 ETY, 10/10 septostomies, 15/15 arachnoid cysts. In five cases of intraventricular tumours, neuroendoscopic procedure was combined with open microsurgery for tumour removal with preservation of vascular structures. There were no "NECUP-2" related complications. Of postoperative complications, we had liquorrhea (9 patients), and symptoms of meningitis (6 patients). In the follow-up period (6 months to 6 years), we had a patency rate of 80% (50/63 patients). All patients improved in clinical status. According to the first results, it seems that ultrasonic contact probe NECUP-2 presents a new device in neurosurgical armamentarium that can be used in various fields of neurosurgery. With minimal and controlled lesion that is produced at the tip of the probe, it can be used in highly demanding operations such as third ventriculostomy and tumour resection.     

Exercise inhibits progressive growth of the Morris hepatoma 7777 in male and female rats

Male and female rats were either trained to swim for a 6-week period or they remained sedentary. Rats were implanted with Morris hepatoma 7777 after 3 weeks of swimming and were sacrificed after a further 3 weeks. Exercised rats of both sexes showed a significant reduction in tumour weight at sacrifice, compared with sedentary controls (p less than 0.01). Similarly, when rats were first implanted with tumours and then placed on an exercise program of 3 weeks duration, tumour growth was also reduced (p less than 0.05). These results suggest that the tumour may be sensitive to exercise at more than one point in its development. Tumour growth was inhibited to a similar extent whether the total swimming time was 10, 20, or 30 h over the 3-week period. Although sedentary, tumour-bearing rats were anorexic; both male and female rats showed significant improvement of appetite during the period of tumour growth, in response to exercise. Tumour implantation was associated with significant losses of whole body and muscle protein. The progression of this wasting was not significantly altered by exercise.     

Competition between polyphosphate and glycogen accumulating organisms in enhanced biological phosphorus removal systems with acetate and propionate as carbon sources

Enhanced biological phosphorus removal (EBPR) is a widely used process for achieving phosphorus removal from wastewater. A potential reason for EBPR failure is the undesirable growth of glycogen accumulating organisms (GAOs), which can compete for carbon sources with the bacterial group responsible for phosphorus removal from wastewater: the polyphosphate accumulating organisms (PAOs). This study investigates the impact of carbon source on EBPR performance and the competition between PAOs and GAOs. Two sequencing batch reactors (SBRs) were operated during a 4-6 month period and fed with a media containing acetate or propionate, respectively, as the sole carbon source. It was found that the acetate fed SBR rarely achieved a high level of phosphorus removal, and that a large portion of the microbial community was comprised of "Candidatus Competibacter phosphatis", a known GAO. The propionate fed SBR, however, achieved stable phosphorus removal throughout the study, apart from one brief disturbance. The bacterial community of the propionate fed SBR was dominated by "Candidatus Accumulibacter phosphatis", a known PAO, and did not contain Competibacter. In a separate experiment, another SBR was seeded with a mixture of PAOs and a group of alphaproteobacterial GAOs, both enriched with propionate as the sole carbon source. Stable EBPR was achieved and the PAO population increased while the GAOs appeared to be out-competed. The results of this paper suggest that propionate may provide PAOs with a selective advantage over GAOs in the PAO-GAO competition, particularly through the minimisation of Competibacter. Propionate may be a more suitable substrate than acetate for enhancing phosphorus removal in EBPR systems.     

A delayed effect of the aquatic parasite <Emphasis Type="Italic">Margaritifera laevis</Emphasis> on the growth of the salmonid host fish <Emphasis Type="Italic">Oncorhynchus masou masou</Emphasis>

Parasitic species often have detrimental effects on host growth and survival. The larvae of the genus Margaritifera (Bivalvia), called glochidia, are specialist parasites of salmonid fishes. Previous studies have reported negligible influences of the parasite on their salmonid hosts at natural infection levels. However, those studies focused mainly on their instantaneous effects (i.e., during the parasitic period). Given the time lag between physiological and somatic responses to pathogen infections, the effect of glochidial infection may become clearer during the post-parasitic period. Here, we examined whether the effect of glochidial infections of Margaritifera laevis on its salmonid host Oncorhynchus masou masou would emerge during the post-parasitic period. We performed a controlled aquarium experiment and monitored fish growth at two time intervals (i.e., parasitic and post-parasitic periods) to test this hypothesis. Consistent with previous observations, the effects of glochidial infection were unclear in the middle of the experiment (day 50; parasitic period). However, even with a natural glochidial load (48 glochidia per fish), we found a significant reduction in growth rates of infected fish in the extended period of the experiment (day 70; post-parasitic period). Our results suggest that examining only instantaneous effects may provide misleading conclusions about mussel–host relationships.     

Irreversible Effects of Cycloheximide During the Early Period of Vaccinia Virus Replication

The presence of cycloheximide, an inhibitor of protein synthesis, during the period 30 to 60 min after vaccinia infection produced an irreversible block in virus replication. In contrast (i) cycloheximide given at earlier or later times, even for prolonged periods, did not prevent continuation of the infectious cycle after removal of the drug, and (ii) treatment with cycloheximide during the first 2 hr did not prevent virus growth when the early stages of replication proceeded more slowly due to infection with a low multiplicity of virus. These findings were interpreted as an indication that protein synthesis is required at a critical time in the virus growth cycle. Under the conditions in which brief cycloheximide treatment prevented virus growth, ribonucleic acid (RNA) synthesis continued at an undiminished rate for at least 2 hr after removal of the drug. Although this RNA appeared identical by polyacrylamide gel electrophoresis to "early" viral messenger RNA, it was not found associated with ribosomes or polyribosomes. Failure to observe viral protein synthesis was consistent with the latter finding. It appeared unlikely that the translational block resulted from inadequate removal of cycloheximide, since the effects of the drug were shown to be reversible at earlier or later times in infection or even at the same time when a lower multiplicity of virus was used. Interference with the normal synthesis of specific viral protein factors required for translation was postulated to explain the results.