Antibody-targeted superantigens are potent inducers of tumor-infiltrating T lymphocytes in vivo.

Recruitment of antigen-specific tumor-infiltrating lymphocytes (TILs) is a major goal for immunotherapy of malignant tumours. We now describe that T-cell-activating superantigens targeted to a tumor by monoclonal antibodies induced large numbers of pseudospecific TILs and eradication of micrometastases. As a model for tumor micrometastases, syngeneic B16 melanoma cells transfected with the human colon carcinoma antigen C215 were injected intravenously into C57BL/6 mice and therapy with an anti-C215 Fab fragment-staphylococcal enterotoxin A (C215Fab-SEA) fusion protein reacting with the C215 antigen was initiated when visible lung metastases were established. More than 90% reduction of the number of lung metastases was observed when mice carrying 5-day-old established lung metastases were treated with C215Fab-SEA. The antitumor effect of C215Fab-SEA was shown to be T-cell-dependent since no therapeutic effect was seen in T-cell-deficient nude mice. Depletion of T-cell subsets by injection of monoclonal antibody demonstrated that CD8+ cells were the most prominent effector cells although some contribution from CD4+ cells was also noted. C215Fab-SEA treatment induced massive tumor infiltration of CD4+ and CD8+ T cells, while only scattered T cells were observed in untreated tumors. SEA treatment alone induced a slight general inflammatory response in the lung parenchyme, but no specific accumulation of T cells was seen in the tumor. TILs induced by C215Fab-SEA were mainly CD8+ but a substantial number of CD4+ cells were also present. Immunohistochemical analysis showed strong production of the tumoricidal cytokines tumor necrosis factor alpha and interferon gamma in the tumor. Thus, the C215Fab-SEA fusion protein targets effector T lymphocytes to established tumors in vivo and provokes a strong local antitumor immune response.

Scaling in geology: landforms and earthquakes.

Landforms and earthquakes appear to be extremely complex; yet, there is order in the complexity. Both satisfy fractal statistics in a variety of ways. A basic question is whether the fractal behavior is due to scale invariance or is the signature of a broadly applicable class of physical processes. Both landscape evolution and regional seismicity appear to be examples of self-organized critical phenomena. A variety of statistical models have been proposed to model landforms, including diffusion-limited aggregation, self-avoiding percolation, and cellular automata. Many authors have studied the behavior of multiple slider-block models, both in terms of the rupture of a fault to generate an earthquake and in terms of the interactions between faults associated with regional seismicity. The slider-block models exhibit a remarkably rich spectrum of behavior; two slider blocks can exhibit low-order chaotic behavior. Large numbers of slider blocks clearly exhibit self-organized critical behavior.

Efficient high-resolution genetic mapping of mouse interspersed repetitive sequence PCR products, toward integrated genetic and physical mapping of the mouse genome.

The ability to carry out high-resolution genetic mapping at high throughput in the mouse is a critical rate-limiting step in the generation of genetically anchored contigs in physical mapping projects and the mapping of genetic loci for complex traits. To address this need, we have developed an efficient, high-resolution, large-scale genome mapping system. This system is based on the identification of polymorphic DNA sites between mouse strains by using interspersed repetitive sequence (IRS) PCR. Individual cloned IRS PCR products are hybridized to a DNA array of IRS PCR products derived from the DNA of individual mice segregating DNA sequences from the two parent strains. Since gel electrophoresis is not required, large numbers of samples can be genotyped in parallel. By using this approach, we have mapped > 450 polymorphic probes with filters containing the DNA of up to 517 backcross mice, potentially allowing resolution of 0.14 centimorgan. This approach also carries the potential for a high degree of efficiency in the integration of physical and genetic maps, since pooled DNAs representing libraries of yeast artificial chromosomes or other physical representations of the mouse genome can be addressed by hybridization of filter representations of the IRS PCR products of such libraries.

A simple p53 functional assay for screening cell lines, blood, and tumors.

Mutations in the p53 gene are implicated in the pathogenesis of half of all human tumors. We have developed a simple functional assay for p53 mutation in which human p53 expressed in Saccharomyces cerevisiae activates transcription of the ADE2 gene. Consequently, yeast colonies containing wild-type p53 are white and colonies containing mutant p53 are red. Since this assay tests the critical biological function of p53, it can distinguish inactivating mutations from functionally silent mutations. By combining this approach with gap repair techniques in which unpurified p53 reverse transcription-PCR products are cloned by homologous recombination in vivo it is possible to screen large numbers of samples and multiple clones per sample for biologically important mutations. This means that mutations can be detected in tumor specimens contaminated with large amounts of normal tissue. In addition, the assay detects temperature-sensitive mutants, which give pink colonies. We show here that this form of p53 functional assay can be used rapidly to detect germline mutations in blood samples, somatic mutations in tumors, and mutations in cell lines.

A Fair Day's Wages: Liberty, Legality, and Liability Among Denver's Day Laborers

Day laborers occupy an essential position in Denver’s booming construction industry. Day laborers make up a highly flexible, highly effective workforce able to respond to market changes. For day laborers, informal day-labor gathering points provide increased control over working hours and employee-employer relationships when compared to traditional wage labor. Still, recent legislation and policies around irregular migration has forced large numbers of workers who may have benefited from the stability of full-time regular employment into the informal sector. The day laborers’ flexibility also exposes them to employers constantly inventing ways to deny them the wages and benefits they are owed. Despite changes in Colorado law in attempts to strengthen workers’ recourse against their employers, and despite social and individual tactics day laborers employ to mitigate their vulnerability, systematic structural, symbolic, and everyday violence continue to advantage employers.

Composición de la dieta y estrategia alimentaria de cinco especies de peces bentónicos de la costa de Cullera (España)

This paper provides data on diet and feeding habits of five benthic fish species (Torpedo torpedo (Linnaeus,1758), Mullus surmuletus (Linnaeus, 1758), Uranoscopus scaber (Linnaeus,1758), Scorpaena scrofa (Linnaeus,1758) and Synaptura lusitanica (Capello,1868)) common in the artisanal fisheries in the Cullera coast (Mediterranean sea – Spain) and T.torpedo, U.scaber y S. lusitánica feeding habits are almost unknown. T. torpedo preferred small preys like fishes, polychaetes and molluscs, these preys were feed in small portions. M. surmuletus showed the highest feeding dynamic, consuming small prey in large numbers like crustaceans (brachyura and amphypoda). U. scaber had similar feeding habits, but the numbers of preys in the stomach were lower. The principal preys were fishes, crustaceans, molluscs and polychaetes. S.scrofa ate larger prey items such as fish, followed by crustaceans and molluscs. Finally S. lusitánica had a high vacuity index, feeding polychaetes as the most important prey in their diet. Feeding strategy indicates a specialization of T.torpedo, S.scrofa and S. lusitanica; conversely M. surmuletus and U. scaber were generalized species.

Research-action Methodology between Computer Science and Educational Psychology: a Multidisciplinary Approach

Computer science studies possess a strong multidisciplinary aptitude since most graduates do their professional work outside of a computing environment, in close collaboration with professionals from many different areas. However, the training offered in computer science studies lacks that multidisciplinary factor, focusing more on purely technical aspects. In this paper we present a novel experience where computer studies and educational psychology find a common ground and realistic working through laboratory practices. Specifically, the work enables students of computer science education the development of diagnosis support systems, with artificial intelligence techniques, which could then be used for future educational psychologists. The applications developed by computer science students are the creation of a model for the diagnosis of pervasive developmental disorders (PDD), sometimes also commonly called the autism spectrum disorders (ASD). The complexity of this diagnosis, not only by the exclusive characteristics of every person who suffers from it, but also by the large numbers of variables involved in it, requires very strong and close interdisciplinary participation. This work demonstrates that it is possible to intervene in a curricular perspective, in the university, to promote the development of interpersonal skills. What can be shown, in this way, is a methodology for interdisciplinary practices design and a guide for monitoring and evaluation. The results are very encouraging since we obtained significant differences in academic achievement between students who attended a course using the new methodology and those who did not use it.

Influence of tree hollow characteristics on the diversity of saproxylic insect guilds in Iberian Mediterranean woodlands

Saproxylic diversity assessment is a major goal for conservation strategies in woodlands and it should consider woodland composition and configuration at site and tree level as key modelling factors. However, in Mediterranean woodlands little is known about the relation with the environmental factors that structure their assemblages, especially those linked to tree hollow microhabitats. We assessed the diversity of Syrphidae (Diptera) and Coleoptera saproxylic guilds that co-occurred in tree hollows located in three different Iberian Mediterranean woodlands in the Cabañeros National Park (Spain). Furthermore, we evaluated how differences in tree hollow microenvironmental variables (understood as the physical and biotic characteristics of a hollow and tree individual) influenced saproxylic guild diversity both within and among woodland sites. We found that woodland sites that provided greater heterogeneity of trees and hollow microhabitats determined higher saproxylic guild diversity. Nevertheless, certain species or even complete guilds can be favoured in woodlands where some hollow microhabitats predominate as a consequence of historical tree management. In general, hollow volume was the main determining factor for saproxylic guild richness and abundance in woodland sites, and large hollow volume was usually related to higher diversity, which highlighted the importance of multi-habitat hollow trees. Moreover, saproxylic guilds also responded to other different microenvironmental variables, which indicated different ecological preferences among guilds. The conservation of saproxylic insects in Iberian Mediterranean areas must be addressed to protect woodland sites that provide high diversity and large numbers of tree hollow microhabitats, and practices to enhance microhabitat heterogeneity should even be encouraged.

Morphometric redefinition of the component chronospecies of the Globorotalia conoidea - G. inflata lineage in DSDP Hole 29-284

Phylo-zonations (or lineage-zonations) are based upon morphological changes within individual evolutionary lineages. These zonations, although potentially of use for stratigraphic subdivision and correlation, often suffer from a lack of quantitative exactness in the definitions of chronospecies. Thus exact reproducibility is hindered for stratigraphic determinations. The potential of morphometrically defined phylo-zonations is demonstrated on a temperate South Pacific Late Cenozoic lineage of planktonic foraminifera (Globorotalia conoidea through intermediate forms to Globorotalia inflata in DSDP Site 284) exhibiting phyletic gradualism. Our sampling interval is about 0.1 m.y. during the last 8 m.y. Changes in the number of chambers in the final whorl, test conicalness, percentage of keeled forms, and test roundness or inflatedness, are used to quantitatively define the following five chronospecies: G. conoidea (Late Miocene; 6.1->8.3 m.y.), G. conomiozea (latest Miocene ; 5.3-6.1 m.y.), G. puncticulata sphericomiozea (earliest Pliocene; 4.5-5.3 m.y.), G. puncticulata puncticulata (Early-Middle Pliocene; 2.9-4.5 m.y.), and G. inflata (Late Pliocene-Quaternary; 0-2.9 m.y.). This phylo-zonation is directly applicable to temperate cool subtropical Southern Hemisphere areas where the evolution took place (Kennett, 1967, 1973; Scott, 1979). It is still not known if the lineage occurs elsewhere; thus the applicability of the phylo-zonation over broader areas is still uncertain. Trends in general size and aperture shape seem to be climatically controlled, and thus may be only of local stratigraphic utility. The practical applications of morphometric phylo-zonation for stratigraphy is to a large extent dependent upon the amount of time and effort required to statistically define the trends. Experiments with large numbers of subsamples from this lineage demonstrate that accurate stratigraphic determinations are possible from measurements on only 15 specimens per sample, except for those very close to chronospecies boundaries.

Pleistocene planktonic foraminifera at ODP Site 128-798

ODP Site 798 on the Oki Ridge in the Southern Japan Sea yielded the first continuous and well-preserved record of Pleistocene planktonic foraminifers in the Northwestern Pacific Ocean region. Quantitative analysis of planktonic foraminifers completed for 122 samples from the 200-m-thick Pleistocene section cored at ODP Site 798 provides a proxy record of variations in sea-surface temperature, productivity, and circulation during the past 1.6 m.y. in an area beneath the track of the Tsushima Current. Faunal census data allow recognition of five distinct assemblages: (1) type A assemblages dominated by sinistrally coiling forms of Neogloboquadrina pachyderma representing polar-subpolar surface temperatures, (2) type B assemblages dominated by Globigerina bulloides and thought to represent periods of increased surface productivity and upwelling, (3) type C assemblages marked by significant abundances of dextrally coiling forms of N. pachyderma thought to represent the warm transitional waters of the Tsushima Current, (4) type D assemblages distinguished by relatively high percentages of dextral N. pachyderma and Globorotalia inflata that also represent warmer surface temperatures and increased flow of the Tsushima Current, and (5) type E assemblages marked by relatively large numbers of the delicate species Globigerina quinqueloba and Globigerinita spp., indicative of exceptional preservation conditions and/or episodic high production of these taxa. Early and middle Pleistocene coiling patterns of Neogloboquadrina pachyderma at Site 798 can be correlated with Pleistocene coiling trends and planktonic foraminiferal datums established in the onshore Oga Peninsula sequence of Northern Honshu and open-ocean N. pachyderma coiling dominance shifts in the North Pacific region. A sustained early Pleistocene warm period recognized in both the Oga Peninsula sequence and the Northern Pacific can clearly be recognized at Site 798. In addition, the late Pleistocene planktonic foraminiferal record at Site 798 shows good correlation with glaciation/deglaciation events for the Northern Hemisphere as delineated by oxygen isotopes and represents the first detailed analysis of Pleistocene sea-surface temperature changes in the Northwestern Pacific Ocean region.

Foraminifera in sediment cores of the Lomonosov Ridge and Morris Jesup Rise

X-ray fluorescence (XRF) scanning of sediment cores from the Lomonosov Ridge and the Morris Jesup Rise reveals a distinct pattern of Ca intensity peaks through Marine Isotope Stages (MIS) 1 to 7. Downcore of MIS 7, the Ca signal is more irregular and near the detection limit. Virtually all major peaks in Ca coincide with a high abundance of calcareous microfossils; this is particularly conspicuous in the cores from the central Arctic Ocean. However, the recorded Ca signal is generally caused by a combination of biogenic and detrital carbonate, and in areas influenced by input from the Canadian Arctic, detrital carbonates may effectively mask the foraminiferal carbonates. Despite this, there is a strong correlation between XRF-detected Ca content and foraminiferal abundance. We propose that in the Arctic Ocean north of Greenland a common palaeoceanographic mechanism is controlling Ca-rich ice-rafted debris (IRD) and foraminiferal abundance. Previous studies have shown that glacial periods are characterized by foraminfer-barren sediments. This implies that the Ca-rich IRD intervals with abundant foraminifera were most likely deposited during interglacial periods when glaciers left in the Canadian Arctic Archipelago were still active and delivered a large amount of icebergs. At the same time, conditions were favourable for planktic foraminifera, resulting in a strong covariance between these proxies. Therefore, we suggest that the XRF scanner's capability to efficiently map Ca concentrations in sediment cores makes it possible to systematically examine large numbers of cores from different regions to investigate the palaeoceanographic reasons for the calcareous microfossils' spatial and temporal variability.

Distribution of deep-sea meiobenthos in surface sediments off North Carolina

Observations on the ecology and distribution of meiofauna occurring on the outer continental shelf and continental slope at depths from 50 to 2500 m in the region where the Blake Plateau cuts across the North Carolina slope are reported. Total numbers of meiofauna ranged from 151/100 cm**3 of sediment at 400 m to 1196/100 cm**3 of sediment at 250 m. Sediments of the upper region (50-500 m) consisted of medium-sized calcareous sands with relatively low organic carbon contents, while the deeper sediments (600-2500 m) consisted of sandy silts and silts with organic carbon contents 6-10 times that of the shallower sediments. Two basic faunas appear to be present in the areas investigated; a shallow-water fauna extending from 50 to 500 m and a deep-water fauna from 800 to 2500 m. The shallow-water fauna consists of nematodes (the dominant taxon) and relatively large numbers of harpactacoid copepods, ostracods, benthic foraminifera, polychaetes, gastrotrichs and several other groups, while below 500 m only nematodes and foraminifera are present in large numbers, the latter being especially abundant between 800 and 2000 m. A major change in the meiofauna occurs on the Blake Plateau between the depths of approximately 400-500 m and 600-750 m where the composition of the sediment changes from sand to silty sand. From 50 m to 400-500 m gastrotrichs, turbellaria, tardigrades, kinorhynchs, halicarids, hydrozoans, gnathostomulids, lamellibranchs and cumaceans are commonly encountered; these groups are absent below 500 m. In addition, there are significant reductions in the numbers of harpactacoids, ostracods, nemerteans and polychaetes below 500 m. Examination of the nematode population also show faunal differences between the shallower sediments (50-500 m) and the deeper sediments (600-2500 m). High indices of affinity exist among the faunas between 50 and 500 m and among the faunas between 800 and 2500 m; the fauna at 600-750 m represents a transition between these two regions, but it is more closely related to the deep-water fauna. Changes in the distribution of both the total meiofuna and also the nematodes are highly correlated with changes in sediments composition and bottom water temperatures. It is suggested that changes in grain size and accompanying changes in sources of nutrition, which are the results of Gulf Stream and other current activity, are the dominant environmental factors influencing the meiofauna of the area.

Dinoflagellates of multicorer surface sediments

Only very few studies focus on recent calcareous dinoflagellate cyst diversity, geographic distribution and ecology, so that information on the distribution patterns and environmental affinities of individual cyst species is extremely limited. This information is, however, essential if we want to use calcareous dinoflagellate cysts for palaeoenvironmental reconstruction. Surface sediment samples from the generally oligotrophic western equatorial Atlantic Ocean, offshore northeast Brazil, were therefore quantitatively analysed for their calcareous dinoflagellate cyst content, including the calcareous vegetative coccoid Thoracosphaera heimii. Seven calcareous dinoflagellate cyst species/morphotypes and T. heimii were encountered in high concentrations throughout the area. Substantial differences in the distribution patterns were observed. The highest concentrations of cysts are found in sediments of the more oligotrophic, oceanic regions, beyond the influence of Amazon River discharge waters. Dinoflagellates producing calcareous cysts thus appear to be capable of surviving low nutrient concentrations and produce large numbers of cysts in relatively stable and predictable environments affected by minimal seasonality. To test for the environmental affinities of individual species, distribution patterns in surface sediments were compared with temperature, salinity, density and stratification gradients within the upper water column (0-100 m) over different times of the year, using principal components analysis and redundancy analysis. T. heimii and four of the seven encountered cyst species (Sphaerodinella? albatrosiana, two morphotypes of Sphaerodinella? tuberosa and Scrippsiella regalis) relate to these parameters significantly and the variations in the cyst associations appear to be associated with the different surface water currents characterising the area. The results imply that calcareous dinoflagellate cyst distributions can potentially be used to distinguish between different open oceanic environments and they could, therefore, be useful in tracing water mass movements throughout the late Quaternary.

Targeting a complex transcriptome: The construction of the mouse full-length cDNA encyclopedia

We report the construction of the mouse full-length cDNA encyclopedia, the most extensive view of a complex transcriptome, on the basis of preparing and sequencing 246 libraries. Before cloning, cDNAs were enriched in full-length by Cap-Trapper, and in most cases, aggressively subtracted/normalized. We have produced 1,442,236 successful 3'-end sequences clustered into 171,144 groups, from which 60,770 clones were fully sequenced cDNAs annotated in the FANTOM-2 annotation. We have also produced 547,149 5' end reads, which clustered into 124,258 groups. Altogether, these cDNAs were further grouped in 70,000 transcriptional units (TU), which represent the best coverage of a transcriptome so far. By monitoring the extent of normalization/subtraction, we define the tentative equivalent coverage (TEC), which was estimated to be equivalent to >12,000,000 ESTs derived from standard libraries. High coverage explains discrepancies between the very large. numbers of clusters (and TUs) of this project, which also include non-protein-coding RNAs, and the lower gene number estimation of genome annotations. Altogether, S'-end clusters identify regions that are potential promoters for 8637 known genes and S'-end clusters suggest the presence of almost 63,000 transcriptional starting points. An estimate of the frequency of polyadenylation signals suggests that at least half of the singletons in the EST set represent real mRNAs. Clones accounting for about half of the predicted TUs await further sequencing. The continued high-discovery rate suggests that the task of transcriptome discovery is not yet complete.

A macrophage colony-stimulating factor receptor-green fluorescent protein transgene is expressed throughout the mononuclear phagocyte system of the mouse

The c-fms gene encodes the receptor for macrophage colony-stimulating factor (CSF-1). The gene is expressed selectively in the macrophage and trophoblast cell lineages. Previous studies have indicated that sequences in intron 2 control transcript elongation in tissue-specific and regulated expression of c-fms. In humans, an alternative promoter was implicated in expression of the gene in trophoblasts. We show that in mice, c-fms transcripts in trophoblasts initiate from multiple points within the 2-kilobase (kb) region flanking the first coding exon. A reporter gene construct containing 3.5 kb of 5' flanking sequence and the down-stream intron 2 directed expression of enhanced green fluorescent protein (EGFP) to both trophoblasts and macrophages. EGFP was detected in trophoblasts from the earliest stage of implantation examined at embryonic day 7.5. During embryonic development, EGFP highlighted the large numbers of c-fms-positive macrophages, including those that originate from the yolk sac. In adult mice, EGFP location Was consistent with known F4/80-positive macrophage populations, including Langerhans cells of the skin, and permitted convenient sorting of isolated tissue macrophages from disaggregated tissue. Expression of EGFP in transgenic mice was dependent on intron 2 as no lines with detectable EGFP expression were obtained where either all of intron 2 or a conserved enhancer element FIRE (the Fms intronic regulatory element) was removed. We have therefore defined the elements required to generate myeloid- and trophoblast-specific transgenes as well as a model system for the study of mononuclear phagocyte development and function. (C) 2003 by The American Society of Hematology.