Variant mouse lymphoma cells with modified response to interferon demonstrate enhanced immunogenicity

 We have previously developed an experimental model for the xenogenization of malignant lymphoma. From highly tumorigenic S49 mouse lymphoma cells that proliferate in suspension culture (designated T-25), we selected variant clones that grew as an adherent monolayer (designated T-25-Adh) and were non-tumorigenic in syngeneic mice. Furthermore, priming of syngeneic hosts with T-25-Adh cells protected them against subsequent challenges with the tumorigenic T-25 cells. Several lines of evidence have indicated that antigens of an endogenous mouse mammary tumor virus (MMTV) are involved in the immunogenicity of T-25-Adh cells. Since interferon (IFN) is known to affect retroviral assembly and maturation on the cell membrane, we have studied the effects of IFN on endogenous MMTV-related structures, as well as on the immunogenicity of T-25-Adh cells. We observed that mouse α and β interferons affect the morphogenesis of intracellular MMTV-related precursors in the immunogenic T-25-Adh cells, but not in tumorigenic T-25 cells. From T-25-Adh cells we selected variants that were either high responders or low responders to the above-mentioned interferon effect. The high-response variants were significantly more protective against tumorigenic T-25 cells than the low-response variants. Involvement of MMTV-related antigens in the immune response of the host to T-25-Adh cells was further suggested by immunoelectron-microscopical analysis, demonstrating that antisera from mice, immunized with T-25-Adh cells, interacted specifically with cell-surface MMTV budding particles. These findings indicate a novel method for xenogenization of lymphoma cells by IFN. Since endogenous retroviruses are present in all tissues of the mouse, this approach might be applicable to a wide variety of tumors. Received: 6 June 1995 / Accepted: 13 March 1997     

Clonal structure and patterns of allozyme diversity in the rare endemic Cycladenia humilis var. jonesii (Apocynaceae)

The rare endemic Cycladenia humilis var. jonesii (Jones cycladenia) has low levels of sexual reproduction. Enzyme electrophoresis was used to explore possible causes of low seed set and high fruit abortion by assessing the clonal structure and genetic diversity in populations. The seven populations studied were composed of multiple, highly interdigitated clones; thus low fruit set is not likely to be due to a scarcity of mates. Genotype frequencies did not differ significantly from Hardy-Weinberg proportions, suggesting that populations are not highly inbred. Jones cycladenia exhibited high levels of genetic diversity at both the population level (A = 1.7; P = 37; He = 0.14) and the taxon level (A = 2.7; P = 60) in comparison to other plants. These data suggest that genetic drift is unlikely to have left this taxon genetically depauperate. Furthermore, we detected little divergence among geographically disjunct populations of Jones cycladenia (FST = 0.10). In comparison, Jones cycladenia populations were highly differentiated from a population of the taxon's close relative, C. h. var. humilis (mean genetic identity = 0.76). Our study suggests that other reasons for low fruit set in Jones cycladenia, such as resource or pollinator limitation, or genetic load, should be explored in future research.     

A model for the impact of herbicide tolerance on the performance of oilseed rape as a volunteer weed

The introduction of genetically modified, herbicide-tolerant, oilseed rape into the agricultural environment will have ramifications beyond weed control of the crop. Herbicide-tolerant rape will undoubtedly become part of established volunteer weed populations that occur in many cereal rotations, but its longevity in these populations and its impact as a weed and contaminant of future oilseed rape crops is uncertain. A life cycle model of volunteer oilseed rape was therefore constructed, incorporating existing information on physiological processes such as emergence pattern, longevity of buried seed, death rates of various structures and flowering and seeding as functions of density. The model was designed to allow interaction with control factors such as harvesting efficiency, herbicide treatment, ploughing and the sequence of crops in the rotation. Many of the physiological parameters (including seed decay rates, fecundity at high density) are uncertain, simply through lack of information in the appropriate context. Other parameters such as harvesting efficiency and herbicide kill rates, are inherently variable in farming. Accordingly, a Monte-Carlo approach, in which the model was run many times with different random realisations of parameter sets, was used to expose factors to which the seedbank was sensitive. Sets of 1250 realisations were compared for each of two extreme conditions: where herbicide could be used according to current intensive farming practice and where it was not an option (representing total herbicide tolerance). Modelled seedbank numbers after 5 yr ranged from 10^-3to 10⁴m^-2, realistic values found in arable soils. The use of herbicide, together with efficient harvesting of seed, clearly has an important suppressive effect on the oilseed rape seedbank, keeping it lower than 10²m^-2(a typical sowing rate) after 5 yr in more than 80% of realisations. In the absence of herbicide, seedbanks were invariably greater, but their absolute value depended strongly on harvesting efficiency and the extent to which high density of plants suppressed fecundity. Analysis of the time series from the simulations showed that the seedbank levels fluctuated by orders in magnitude from year to year in the absence of herbicide use. The sensitivity analysis of the life-cycle model led to the development of a simplified model for the seedbank dynamics. The model shows that the essential features of the dynamics result from an interaction between density-dependent fecundity and the perturbations due to management. Therefore predictions of the effect of herbicide tolerance on seedbank dynamics are highly uncertain without knowledge of the density dependence of fecundity. Furthermore, the sensitivity to management practices suggests that seedbank levels will be substantially more difficult to control if the efficacy of herbicide is compromised. It is concluded that the model and Monte-Carlo approach have many potential uses in exploring the effects of management, cultivar physiology and the nature of the transgenes.     

Population genetic structure of the gynodioecious Thymus vulgaris L. (Labiatae) in southern France

In a self-compatible gynodioecious species, the abundance of female plants (which are obligate outcrossers) relative to hermaphrodites (which may self and outcross) may be a critical factor influencing genetic diversity and population structure. In the gynodioecious Thymus vulgaris L., female frequency varies from 5 to 95%, providing a suitable model to examine this issue. In this study, we use allozyme markers to (1) evaluate the relationship between female frequency, genetic diversity and population structure, (2) determine whether females and hermaphrodites vary in heterozygote deficiency and (3) examine whether other factors such as plant density are related to heterozygote deficiency. Twenty three natural populations, with female frequencies ranging from 11 to 92%, were sampled in and around the St-Martin-de-Londres basin in southern France. Based on four polymorphic allozyme loci, we found no significant correlation between female frequency and heterozygote deficiency. A significant (P < 0.05) F_IS value over loci and over populations of 0.11 was detected. The F_IS value per population showed a significant heterozygote deficiency in 11 of the 23 populations. However, no significant difference between female and hermaphrodite F_IS values was found. A significant heterozygote deficiency only occurred in populations of intermediate density. There was little differentiation among populations (F_ST = 0.038) nor among subpopulations within each population. The significant F_IS values are thus mostly due to inbreeding effects. The lack of a correlation between F_IS values and female frequency may be due to outcrossing in hermaphrodites and/or restoration of male fertility which may occur to a greater extent at low female frequency. The similarity of female and hermaphrodite F_IS values indicates that females may occasion high levels of biparental inbreeding.     

Annual Abundances, Diversity and Growth Potential of Gymnamoebae in a Shallow Freshwater Pond

ABSTRACT. Abundance and diversity of gymnamoebae were assessed at monthly intervals for one year in the sediment surface layer of a shallow, highly productive, freshwater pond located in northeastern United States using laboratory culture enumeration techniques. The range in total abundance was 81/ml in January to 1,568/ml in June (spring peak) and 1,813/ml in October (autumn peak). The latter is equivalent to over 1.8 million per liter, one of the highest values reported for gymnamoebae from a freshwater pond. Maximum diversity (H = 3.3) occurred in April, July, and August. The growth potential of the gymnamoebae was determined in laboratory cultures (20° C) containing only the pond water (control culture) and pond water emended with malt-yeast extract and glucose (nutrient culture) using pond water collected in December. The total abundance of gymanmoebae increased from 363/ml in the initial pond water sample to approximately 1,800/ml in each of the culture conditions in the first week. Therafter, abundances reached 5,678/ml (control culture) and 8,016/ml (nutrient culture) by the third week indicating the vigorous growth potential from winter standing stock.     

Assessment and comparison of the Marennes-Oléron Bay (France) and Carlingford Lough (Ireland) carrying capacity with ecosystem models

Based on the individual growth, food limitation,population renewal through seeding, and individualmarketable size, a theoretical model of the culturedspecies population dynamics was used to assess thecarrying capacity of an ecosystem. It gave adome-shape curve relating the annual production andthe standing stock under the assumption of individual growth limited by the available food inan ecosystem. It also showed the influence ofmortality rate and marketable size on this curve andwas introduced as a means to explore the globalproperties resulting from the interactions betweenthe ecophysiology of the reared species and theenvironment at the ecosystem level. In a second step, an ecosystem model was built to assess the carryingcapacity of Marennes-Oléron bay, the mostimportant shellfish culture site in France, with astanding stock of Crassostrea gigas around 100000tonnes fresh weight (FW) and an annual production of30000 tonnes FW. The ecosystem model focused on theoyster growth rate and considered the interactionbetween food availability, residence time of thewater, oyster ecophysiology and number ofindividuals. It included a spatial discretization ofthe bay (box design) based on a hydrodynamic model,and the nitrogen or carbon cycling betweenphytoplankton, cultured oysters, and detritus. Fromsimulations of the oyster growth with differentseeding values, a curve relating the total annualproduction and the standing stock was obtained. Thiscurve exhibited a dome shape with a maximumproduction corresponding to an optimum standingstock. The model predicted a maximum annualproduction of 45000 tonnes FW for a standing stockaround 115000 tonnes FW. The prediction confirmedsome results obtained empirically in the case ofMarennes-Oléron bay and the results of thetheoretical model. Results were compared with thoseobtained in Carlingford Lough (Ireland) using asimilar ecosystem model. Carlingford Lough is asmall intertidal bay where the same species iscultured at a reduced scale, with current biomassless than 500 tonnes FW. The model showed that thestanding stock can be increased from 200 tonnes FWto approximately 1500 tonnes FW before any decreaseof the production. This revised version was published online in August 2006 with corrections to the Cover Date.     

Local colonization and extinction of field layer plants in a deciduous forest and their dependence upon life history features

Abstract. In this study we used species inventory data collected in 1970 and 1993 from 132 plots in a 14.5ha deciduous forest to examine local extinction and colonization processes among 45 field layer species. Local colonization rate was positively related to both seed size and seed dispersal features. Local extinction rate was negatively related to seed size. Growth form (clonal vs. non-clonal) and presence of a seed bank were not found to be associated with local dynamics. Despite an overall constancy in species composition during this period, plants exhibited a considerable mobility among the 132 plots. This pattern conformed to a suggested ‘carousel model’ of species mobility in grasslands. A tentative suggestion is that this mobility acts on a comparatively broader spatio-temporal scale in deciduous forests as compared with grasslands. Additional data are presented indicating that species abundance (frequency) among field layer plants in deciduous forest communities is consistent among forest fragments, and when comparing local and regional scales. The main conclusion is that life history features of the seed dispersal/recruitment phase, particularly seed size, are causally associated with abundance patterns at least at a local scale, but possibly also on a broader regional scale.     

Structure of population,grouping patterns and density of forest elephants in north-west Congo

The structure of the population of forest elephants visiting a clearing in north-west Congo was studied over 8 months. Out of 3314 sightings, 629 elephants were identified (including 64%, of adults). The sex ratio was about 1:1. The number of offspring per female was 1.12. Solitary elephants (91.4%, of males) made up 35% of the population. Of females, 93.6% were grouped. Groups (mean size 3.5) included females and offspring (3.4), males and females (5.0), or males only (2.3). The return rate of elephants at the clearing suggested that up to 1900 individuals have visited the clearing. This confirms that elephant densities are especially high in north Congo.     

DOES COSMOPOLITANISM RESULT FROM OVERCONSERVATIVE SYSTEMATICS? A CASE STUDY USING THE MARINE SPONGE CHONDRILLA NUCULA

The sponge species Chondrilla nucula has a simple morphology and a very wide geographical distribution. To verify whether the latter might be an artifact of the former, samples of this species were collected across 10,000 km of its range, in the Mediterranean, the Caribbean, and the southwestern atlantic. The classical (spicule morphology) and molecular (allozymes) systematic approaches were compared, to try to define the geographic limits between populations and detect possible cryptic species. We found five distinct genetic forms within C. nucula that sometimes showed morphological homogeneity and other times plasticity. The difference in size of spicules could not be related to the clear-cut genetic differences, suggesting that the use of spicule sizes for sponge systematics should be reappraised. The population of one of the genetic forms along 3000 km of the Brazilian coast was highly structured (F_ST = 0.21; N_em = 0.96). Our results reject the null hypothesis of cosmopolitanism of C. nucula and indicate that the putative worldwide distribution of some marine sponges, and possibly many other benthic invertebrates, may be the result of overly conservative systematics. Cryptic species appear to be particularly prevalent when genera are well defined but species are characterized by only a few morphological characters.     

Population dynamics of Mus minutoides and Steatomys pratensis (Muridae: Rodentia) in a subtropical grasslandin Swaziland

The population dynamics of Mus minutoides and Steatomys pratensis are virtually unknown. These two species were live-trapped over a 12-month period in a subtropical grassland in Swaziland. Numbers of M. minutoides were relatively high in winter, declined in spring and the population disappeared in summer and autumn. By contrast, numbers of S. pratensis increased gradually from winter to summer and reached a peak in autumn. There were no differences between the mean weights of male and female M. minutoides and S. pratensis. There were, however, seasonal differences in the mean weight of male S. pratensis, with highest weights recorded in summer. Individuals of both species came into breeding condition in spring (October–November). Reproduction had ceased by the end of autumn (April–May). Monthly survival rates of M. minutoides were highest in winter, but did not vary seasonally in S. pratensis. Burning had a pronounced effect on the distribution of S. pratensis. Steatomys pratensis individuals selected recently burnt but revegetated areas over unburnt areas. The effect of burning on the M. minutoides population is difficult to assess, as this species disappeared shortly after the fire.