Regulation of veterinary point-of-care testing in the European Union, the United States of America and Japan.

Point-of-care testing (POCT) is used to detect diseases and other conditions or to monitor therapeutic procedures. In veterinary medicine, POCT not only helps during the prevention, diagnosis and treatment of animal diseases but it also has a direct impact on human health by safeguarding food supplies and preventing zoonoses. Despite its importance, the regulation of the quality, safety and effectiveness of POCT products is rarely discussed. This review reveals that the level of regulatory surveillance of veterinary POCT products in the European Union (EU), the United States of America and Japan is strikingly different, ranging from no regulation (EU) to comprehensive regulation, which is comparable to the procedures for the regulation of human in vitro medical devices (Japan). Details about the licensing procedures in these three locations, discussion of their strengths and weaknesses, and suggestions for possible future development of the regulation of these products are also provided.

Les tests utilisables sur le lieu d'intervention (POCT) permettent de détecter des maladies ou d'autres affections et d'assurer un suivi des procédures thérapeutiques appliquées. En médecine vétérinaire, les POCT apportent un soutien utile lors de la prévention, du diagnostic et du traitement des maladies animales, en plus d'avoir un impact direct sur la santé humaine en participant à la sécurité de l'approvisionnement alimentaire et à la prévention des zoonoses. La réglementation applicable aux produits utilisés pour les POCT, afin d'en garantir la qualité, la sécurité et l'efficacité, fait rarement l'objet de débats, bien qu'il s'agisse d'une question importante. L'examen présenté par les auteurs révèle que le niveau de la surveillance réglementaire exercée sur les produits vétérinaires utilisés pour les POCT dans l'Union européenne (UE), les états-Unis d'Amérique et le Japon est extrêmement variable, allant d'une absence totale de réglementation (UE) à une réglementation exhaustive comparable aux procédures appliquées pour les dispositifs in vitro utilisés en médecine humaine (Japon). Les auteurs décrivent en détail les procédures d'autorisation de mise sur le marché dans ces trois pays, examinent leurs atouts et points faibles respectifs et font quelques propositions pour faire évoluer la réglementation de ces produits à l'avenir.

Las pruebas realizadas en el punto de consulta, o «pruebas de diagnóstico inmediato¼, sirven para detectar enfermedades y otros trastornos o para seguir de cerca los resultados de procedimientos terapéuticos. En medicina veterinaria, estas pruebas no solo ayudan a prevenir, diagnosticar y tratar enfermedades animales, sino que también tienen una influencia directa en la salud humana, en la medida en que permiten proteger los suministros alimentarios y prevenir zoonosis. A pesar de su importancia, rara vez se aborda el tema de la regulación de la calidad, seguridad y eficacia de los productos que contienen pruebas de diagnóstico inmediato. Los autores revelan aquí llamativas disparidades en los regímenes de vigilancia reglamentaria que aplican a las pruebas de diagnóstico veterinario inmediato la Unión Europea (UE), los Estados Unidos de América (EE.UU.) y el Japón, regímenes que van desde la inexistencia de reglamentos (UE) hasta una regulación exhaustiva comparable a los procedimientos aplicados a los dispositivos médicos in vitro (Japón). Los autores también exponen en detalle los procedimientos de obtención de licencia vigentes en estos tres contextos, examinan sus ventajas e inconvenientes y formulan propuestas para el futuro desarrollo de la reglamentación aplicada a los antedichos productos.

Pretreatment and gaseous radiocarbon dating of 40-100 mg archaeological bone.

Radiocarbon dating archaeological bone typically requires 300-1000 mg material using standard protocols. We report the results of reducing sample size at both the pretreatment and 14C measurement stages for eight archaeological bones spanning the radiocarbon timescale at different levels of preservation. We adapted our standard collagen extraction protocol specifically for <100 mg bone material. Collagen was extracted at least twice (from 37-100 mg material) from each bone. Collagen aliquots containing <100 µg carbon were measured in replicate using the gas ion source of the AixMICADAS. The effect of sample size reduction in the EA-GIS-AMS system was explored by measuring 14C of collagen containing either ca. 30 µg carbon or ca. 90 µg carbon. The gas dates were compared to standard-sized graphite dates extracted from large amounts (500-700 mg) of bone material pretreated with our standard protocol. The results reported here demonstrate that we are able to reproduce accurate radiocarbon dates from <100 mg archaeological bone material back to 40,000 BP.

[Carbon accumulation in soils of forest and bog ecosystems of southern Valdai in the Holocene].

Carbon stocks and accumulation rates in humus and peat horizons of the contiguous soil series of forest and bog ecosystems have been studied in the Central Forest State Biosphere Reserve (CFSBR, Tver region). Upland soil types (soddy podzolic, brown, and white podzolic) have been compared to waterlogged (peaty gley podzolic and peaty gley) and bog soils differing in trophic status, including those of raised, transitional, and lowland bogs. The results show that carbon stocks in mineral soils are many times smaller than in waterlogged soils and an order of magnitude smaller than in bog soils. Mineral and bog soils are characterized by similar rates of carbon accumulation averaged over the entire period of their existence. The highest rate of carbon accumulation has been noted for the soils of waterlogged habitats, although this process may be periodically disturbed by fires and other stress influences.

Unusual activity of the Sun during recent decades compared to the previous 11,000 years.

Direct observations of sunspot numbers are available for the past four centuries, but longer time series are required, for example, for the identification of a possible solar influence on climate and for testing models of the solar dynamo. Here we report a reconstruction of the sunspot number covering the past 11,400 years, based on dendrochronologically dated radiocarbon concentrations. We combine physics-based models for each of the processes connecting the radiocarbon concentration with sunspot number. According to our reconstruction, the level of solar activity during the past 70 years is exceptional, and the previous period of equally high activity occurred more than 8,000 years ago. We find that during the past 11,400 years the Sun spent only of the order of 10% of the time at a similarly high level of magnetic activity and almost all of the earlier high-activity periods were shorter than the present episode. Although the rarity of the current episode of high average sunspot numbers may indicate that the Sun has contributed to the unusual climate change during the twentieth century, we point out that solar variability is unlikely to have been the dominant cause of the strong warming during the past three decades.

Case Report. Maxillary sinus infection due to Emericella nidulans.

Fungal infections of the maxillary sinus are frequently caused by Aspergillus species, particularly A. fumigatus. In otherwise healthy persons there is an association with overfilling of dental root canals, when zinc-containing filling materials were used. Below, a maxillary sinus aspergilloma is reported in a young immunocompetent female patient caused by Aspergillus (Emericella) nidulans.

Anatolian tree rings and a new chronology for the east Mediterranean Bronze-Iron Ages.

We report an extensive program of high-precision radiocarbon dating to establish the best date for a floating 1599-year Anatolian tree ring chronology that spans the later third millennium B.C. through the earlier first millennium B.C. This chronology is directly associated with a number of key sites and ancient personages. A previously suggested dating is withdrawn and is replaced by a robust new date fix 22 (+4 or -7) years earlier. These new radiocarbon wiggle-matched dates offer a unique independent resource for establishing the precise chronology of the ancient Near East and Aegean and help resolve, among others, a long-standing debate in favor of the so-called Middle Mesopotamian chronology.

Regional 14CO2 offsets in the troposphere: magnitude, mechanisms, and consequences.

Radiocarbon dating methods typically assume that there are no significant tropospheric (14)CO(2) gradients within the low- to mid-latitude zone of the Northern Hemisphere. Comparison of tree ring (14)C data from southern Germany and Anatolia supports this assumption in general but also documents episodes of significant short-term regional (14)CO(2) offsets. We suggest that the offset is caused by an enhanced seasonal (14)CO(2) cycle, with seasonally peaked flux of stratospheric (14)C into the troposphere during periods of low solar magnetic activity, coinciding with substantial atmospheric cooling. Short-term episodes of regional (14)CO(2) offsets are important to palaeoclimate studies and to high-resolution archaeological dating.

Persistent solar influence on North Atlantic climate during the Holocene.

Surface winds and surface ocean hydrography in the subpolar North Atlantic appear to have been influenced by variations in solar output through the entire Holocene. The evidence comes from a close correlation between inferred changes in production rates of the cosmogenic nuclides carbon-14 and beryllium-10 and centennial to millennial time scale changes in proxies of drift ice measured in deep-sea sediment cores. A solar forcing mechanism therefore may underlie at least the Holocene segment of the North Atlantic's "1500-year" cycle. The surface hydrographic changes may have affected production of North Atlantic Deep Water, potentially providing an additional mechanism for amplifying the solar signals and transmitting them globally.

The vasoconstrictor effect of 8-epi prostaglandin F2alpha in the hypoxic rat heart.

1. 8-epi prostaglandin (PG) F2alpha, a vasoconstrictor isoprostane, is synthesized under conditions of oxidative stress. This study was undertaken to investigate the vasoconstrictor effect of 8-epi PGF2alpha in the coronary circulation before and after a period of oxidative stress. 2. The effects of the isoprostane 8-epi PGF2alpha and the thromboxane mimetic U46619 were compared in the isolated rat heart perfused in the Langendorff mode at a constant pressure of 80 mmHg. 3. In normal hearts U46619 caused a dose-related reduction in coronary flow (ED50 4.7+/-2.2 nmol). In contrast, 8-epi PGF2alpha had no effect. 4. After reducing perfusion pressure to 20 mmHg for 30 min and reperfusing at 80 mmHg, the dose-response curve to U46619 was unaffected. In contrast, 8-epi PGF2alpha caused a dose-dependent drop in coronary flow (ED50 52.6+/-12.7 nmol), producing a similar maximal reduction to U46619. 5. Similarly, after perfusion with xanthine and xanthine oxidase for either 15 or 30 min there was little change in the response to U46619 in comparison to control hearts. In contrast, 8-epi PGF2alpha caused a reduction in coronary flow similar to that produced by U46619, the magnitude of the response being related to the length of xanthine/xanthine oxidase perfusion. 6. Responses to both U46619 and 8-epi PGF2alpha after xanthine/xanthine oxidase perfusion were blocked by the selective thromboxane receptor antagonist SQ29548 10(-7) M. 7. These results show that oxidative stress in the isolated perfused rat heart reveals a potent vasoconstrictor effect of the isoprostane 8-epi PGF2alpha by an action on the thromboxane receptor. 8. The data also suggest that, since 8-epi PGF2alpha is a partial agonist at the thromboxane receptor, thromboxane receptor reserve is increased by oxidative stress.

Actions of 8 epi prostaglandin F2alpha on isolated rat aorta.

8-epi prostaglandin F2alpha(8-epi PGF2alpha) contracted rat thoracic aorta rings in a concentration-dependent manner in the presence or absence of functional endothelium [median effective concentration (EC50) values, 455+/-52 and 268+/-34 nM, respectively; Student's t test; p=0.006]. U46619 was a more potent agonist with or without functional endothelium (EC50 values, 6.8+/-1.6 and 4.5+/-1.0 nM, respectively). SQ29548 [a thromboxane (TP)-receptor antagonist] inhibited contractions to both 8-epi PGF2alpha and U46619 in a competitive manner, with mean pA2 values of 8.3 and 7.9, respectively. 8-Epi PGF2alpha had a further contractile effect in vessels that had been contracted with noradrenaline and had been shown to possess a functional endothelium. Inhibition of thromboxane synthesis with OKY-046 or blockade of endothelin receptors with bosentan had no effect on responses to 8-epi PGF2alpha or U46619. Preincubation with 8-epi PGF2alpha or noradrenaline shifted the concentration-response curves to U46619 upward at low concentrations of U46619 with no significant change in EC50 values or maximal responses. Reduction of TP-receptor number in rat aorta with dithiothreitol caused a concentration-dependent inhibition of responses to both U46619 and 8-epi PGF2alpha, with no effect on maximal responses and or on the responses to U46619 after the preincubation with 8-epi PGF2alpha. These results indicate that 8-epi PGF2alpha is a potent vasoconstrictor in the rat aorta and are suggestive of an action of 8-epi PGF2alpha at the TP receptor.

Absence of LKM-1 antibody reactivity in autoimmune and hepatitis-C-related chronic liver disease in Sweden. Swedish Internal Medicine Liver club.

BACKGROUND: Type-2 autoimmune hepatitis is a subgroup of chronic hepatitis characterized by the presence of liver/kidney microsomal autoantibodies type 1 (LKM-1). A frequent association with chronic hepatitis C suggests that hepatitis virus might trigger autoimmune reactivity. LKM-1-positive chronic hepatitis is not uncommon in southern Europe but is rarely seen in the USA and the UK. The prevalence in Scandinavia is hitherto unknown. METHODS: We used an automated prototype LKM-1 immunometry-based assay (IMx) to detect LKM-1 antibodies in sera from 350 Swedish patients with chronic liver diseases (100 with primary biliary cirrhosis, 80 with primary sclerosing cholangitis, 100 with hepatitis C, and 70 patients with various forms of chronic hepatitis, including 36 autoimmune cases), and from 17 children with autoimmune hepatitis. Sera reactive in the IMx assay were subjected to immunofluorescence testing. RESULTS: No clearly LKM-reactive sera were detected. Serum samples from 29 patients were borderline reactive in the IMx assay but tested negative in the confirmatory immunofluorescence test. Positive tests in the former assay were likely caused by reactivity against microsomal antigens other than LKM-1/cytochrome P450IID6. CONCLUSIONS: LKM-1-positive type-2 autoimmune hepatitis is very rare in Sweden. Furthermore, chronic hepatitis C did not trigger this type of autoimmune reactivity in our patients, probably owing to genetic insusceptibility.

cea5, a structurally divergent member of the murine carcinoembryonic antigen gene family, is exclusively expressed during early placental development in trophoblast giant cells.

The carcinoembryonic antigen (CEA) gene family encodes a large family of glycoproteins. Some are probably involved in the homeostasis/development of epithelial cells and granulocyte activation, while others e.g. the pregnancy-specific glycoproteins, are expressed in the placenta and are essential for a positive outcome of pregnancy. In this paper, we have characterized cea5, a member of the murine CEA gene family. RNase protection and in situ hybridization analyses revealed that Cea5 mRNA is exclusively synthesized in primary and secondary trophoblast giant cells of the placenta only during early stages of development. Full-length Cea5 cDNA was obtained by a reverse transcription-polymerase chain reaction using day 10.5 post-coitum placental RNA. The 1.6-kilobase pair (kb) Cea5 mRNA encodes a secreted glycoprotein with a predicted size of 30 kDa. It is composed of a leader peptide (L), one immunoglobulin (Ig) variable or N, and one Ig constant-like or A domain. This domain organization is unique within the human and murine CEA families. Two overlapping cosmid clones covering 54 kb of the cea5 gene locus were mapped. cea5 consists of three exons (L, N, A/3'-untranslated region exon) located within a 4-kb region. rnCGM2, the rat cea5 counterpart, exhibits the same restricted expression pattern. This together with their exceptional conservation within the rat and murine CEA families and their absence from the human CEA family suggests that cea5 and rnCGM2 are of functional importance for rodent placental development.

Coordinate expression of splice variants of the murine pregnancy-specific glycoprotein (PSG) gene family during placental development.

The human and murine pregnancy-specific glycoprotein (PSG) gene families encode a large number of closely related proteins which are abundantly expressed in the fetal trophoblast and secreted into the maternal circulation. Although the presence of a well conserved tripeptide sequence His or Arg-Gly-Asp or Glu or Lys (H/RGD/E/K) similar to the RGD motif found in extracellular matrix proteins hints towards a possible interaction with integrin-type receptors, the function of this group of proteins related to the carcinoembryonic antigen family is still unknown. It is also not clear whether the various members of the PSG family exert the same function. Here we describe the cloning of two splice variants of Cea4 (Cea4a, Cea4b), a murine PSG family member, which lacks the RGD-related consensus motif. Cea4a, like most of the other rodent PSG members, is composed of three immunoglobulin (Ig) variable-like domains (N1-N3) and and one Ig constant-like domain (A). In contrast, Cea4b lacks the N2 domain (N1N3A), demonstrating for the first time that PSG isoforms produced by alternative splicing also exist in mice. The mRNAs coding for Cea4a and Cea4b exhibit the same expression kinetics during placental development as found for two other murine PSGs, Cea2 and Cea3, which contain the RGD-like motif. Expression starts after day 12.5 of embryonic development (E12.5) and maximum steady-state levels are reached around E15.5-E17.5 as determined by RNase protection analyses. At E17.5, PSG transcripts can be detected exclusively in the spongiotrophoblast of the placenta. In addition, PCR analyses revealed that Cea2, Cea3, and Cea4 transcripts are also found in RNA from a pool of embryos (E12-E15) but are absent from a number of adult tissues tested (kidney, lung, testis, ovary, liver, brain, thymus, heart, spleen). These results indicate that the various PSG isoforms exert their function(s) at the same time during placental and embryonic development.

Synchronized Terrestrial-Atmospheric Deglacial Records Around the North Atlantic

On the basis of synchronization of three carbon-14 (14C)-dated lacustrine sequences from Sweden with tree ring and ice core records, the absolute age of the Younger Dryas-Preboreal climatic shift was determined to be 11,450 to 11,390 +/- 80 years before the present. A 150-year-long cooling in the early Preboreal, associated with rising Delta14C values, is evident in all records and indicates an ocean ventilation change. This cooling is similar to earlier deglacial coolings, and box-model calculations suggest that they all may have been the result of increased freshwater forcing that inhibited the strength of the North Atlantic heat conveyor, although the Younger Dryas may have begun as an anomalous meltwater event.

Coronary artery constriction by the isoprostane 8-epi prostaglandin F2 alpha.

1. This study was undertaken to compare the effects of 8-epi prostaglandin F2 alpha (8-epi PGF2 alpha) to those of prostaglandin F2 alpha (PGF2 alpha) and U46619, a thromboxane mimetic, on ovine, bovine and porcine coronary arteries. 2. 8-epi PGF2 alpha constricted porcine and bovine coronary arteries in a concentration-dependent manner with EC50 values of 689.0 +/- 229.3 and 1361.0 +/- 272.3 nM, respectively, but had no effect on ovine coronary arteries. 3. U46619 was a potent vasoconstrictor of porcine, ovine and bovine coronary arteries with EC50 values of 33.0 +/- 23.5, 373.3 +/- 69.7 and 254.1 +/- 134.3 nM, respectively. Emax values were significantly greater than those obtained with 8-epi PGF2 alpha. 4. PGF2 alpha constricted procine and bovine coronary arteries in a concentration-dependent manner with EC50 values of 1631.0 +/- 207.6 and 3644.0 +/- 344.8 nM, respectively, but had no effect on ovine coronary arteries. 5. Concentration-dependent constriction to U46619 in porcine coronary arteries was competitively inhibited by SQ29548 (10(-8) M to 10(-7) M) and BM13505 (10(-8) M to 10(-6) M) with no decrease in maximal responses. 6. Concentration-dependent constriction to 8-epi PGF2 alpha in porcine coronary arteries was inhibited in a concentration-dependent manner by SQ29548 (10(-8) M to 10(-7) M) and BM13505 (10(-8) M to 10(-6) M). However, the inhibition was associated with a decrease in maximal response. 7. Maximal responses of porcine coronary artery to U46619 (1 microM) and 8-epi PGF2 alpha (30 microM) were inhibited in a concentration-dependent manner by SQ29548 with IC50 values 99 +/- 12.36 nM and 46.5 +/- 18.67 nM, respectively. 8. Although ovine coronary arteries did not constrict to 8-epi PGF2 alpha pre-incubation of these vessels with 8-epi PGF2 alpha caused a rightward shift of the U46619 response curve in a concentration-dependent manner. 9. Pre-incubation of porcine coronary arteries with 8-epi PGF2 alpha competitively inhibited responses to U46619 with a Schild slope of 0.99 and a pA2 of 6.13. 10. We conclude that 8-epi PGF2 alpha is a vasoconstrictor within porcine and bovine coronary arteries, with a potency approximately twice that of PGF2 alpha but 5-20 times lower than U46619. The data suggest that 8-epi PGF2 alpha is acting as a partial agonist on the TP-receptor in the coronary vasculature.

Concentration time courses of troponin and myosin subunits after acute myocardial infarction.

BACKGROUND: As a result of the limited sensitivity and specificity of creatine kinase and lactate dehydrogenase (LDH) as well as their isoenzymes, there is increasing interest in the use of cardiac contractile proteins for the diagnosis of acute myocardial infarction (AMI) and myocardial damage. METHODS: This study compared the release of creatine kinase, creatine kinase MB, myoglobin, cardiac troponin I (cTnI), cardiac troponin T (cTnT), cardiac myosin light chain-1 (cMLC-1), and beta-type myosin heavy chains (bMHC) in serial blood samples from 13 patients (10 men, three women; median age 54 years, range 40-74 years) with first-time AMI (11 Q-wave, two non-Q-wave AMI; three anterior and 10 inferior wall AMI). All but one patient received intravenous thrombolytic treatment. RESULTS: Myoglobin was the first marker to increase in blood after AMI and showed the earliest peak levels, whereas bMHC increased latest, showing the latest peak levels. cTnI and cTnT increased significantly earlier than cMLC-1 and bMHC. cTnI and cTnT increased and reached peak levels parallel to each other, but the latter tended to stay increased longer. cTnT time courses were biphasic in the majority of AMI patients, unlike cTnI time courses. cMLC-1 release was mostly biphasic. cMLC-1 allows diagnosis during the acute phase as well as several days after the onset of AMI. The time courses of bMHC were usually monophasic. Its delayed appearance makes it useful for the diagnosis of remote infarction. In contrast to cTnI and cTnT, cMLC-1 and bMHC time courses were not significantly influenced by early reperfusion. CONCLUSION: Our results demonstrate the impact of the intracellular compartmentation of an intramyocardial protein (cytosolic, structurally bound, or structurally bound with soluble pool) on its concentration time course after AMI, particularly on the rapidity of its release.

Measurement of liver microsomal cytochrome p450 (CYP2D6) activity using [O-methyl-14C]dextromethorphan.

The activity of human liver microsomal cytochrome P4502D6 (CYP2D6) is readily estimated by following the O-demethylation of [O-methyl-14C]dextromethorphan. The basis of the assay is the quantitative measurement of [14C]formaldehyde (0.05-4.0 microM) after addition of NaOH to the microsomal incubates and extraction with methylene chloride. The assay is relatively simple, sensitive (limit of detection is approximately 5.0 pmol HCHO/h/mg microsomal protein) and does not require the use of HPLC or an internal standard. Formation of radiolabeled formaldehyde in human liver microsomes is linear for 20 min, up to a final protein concentration of 1.0 mg/ml. Furthermore, the O-demethylase activity in a panel of microsomes prepared from a series of human livers was significantly correlated with the immunochemically determined levels of CYP2D6 protein (r = 0.925, p < 0.001), and was inhibited (> 89%) by quinidine and lobeline. In addition, [O-methyl-14C]-dextromethorphan O-demethylation was exclusively catalyzed by cDNA-expressed CYP2D6 in microsomes prepared from human B-lymphoblast cells. The method is suitable for rapid screening of compounds as potential CYP2D6 cosubstrates and/or inhibitors.

Precursor-specific requirements for SecA, SecB, and delta muH+ during protein export of Escherichia coli.

We compare translocation into inside-out plasma membrane vesicles (INV) of the in vitro synthesized outer membrane proteins LamB and OmpA and the periplasmic protein Skp of Escherichia coli and demonstrate a precursor-specific dependence on the export factors SecA, SecB, and the proton-motive force (delta mu H+). A partial reduction in soluble SecA caused a 50% decrease in translocation of preLamB. In contrast, removal of INV-bound SecA by urea extraction was required to see a decrease in translocation of preOmpA and preSkp, with 8% of preSkp still being translocated into urea-treated INV. Translocation of the three precursors into INV showed a corresponding differential sensitivity toward dissipation of delta mu H+ following removal of the F1-ATPase from the INV. While depletion of both F1 and SecA or simply lowering of the reaction temperature resulted in an inhibition of complete transmembrane translocation, it interfered less severely with signal sequence cleavage, indicating the formation of translocation intermediates under these conditions. The relative amounts of intermediate obtained were also different for the three preproteins correlating a low requirement for SecA and delta mu H+ with a facilitated initiation of translocation. Whereas preSkp was translocated independently of SecB, preLamB was not even targeted to the INV in its absence. Functional targeting of preOmpA required the presence of SecB during incubation of the precursor with INV and not during its synthesis. SecB, exogenously added during the period of synthesis, did not prevent the formation of translocation-incompetent preLamB. The latter results are consistent with an important targeting function of SecB, which so far has mostly been described as a molecular chaperone. The findings are discussed with respect to current models of bacterial protein export usually derived from the analysis of a single precursor.

Human carcinoembryonic antigen cDNA expressed in rat carcinoma cells can function as target antigen for tumor localization of antibodies in nude rats and as rejection antigen in syngeneic rats.

We have tried to develop a new model consisting of rats transplanted with syngeneic colon carcinoma PROb cells transfected with cDNA coding for the carcinoembryonic antigen (CEA), the human tumor marker most commonly used as target for MAbs. The antigenic density of the 4 CEA-expressing clones selected for a precise characterization ranged from 5 x 10(4) to 1 x 10(6) CEA molecules per cell. In all clones the CEA was shown to be attached to the membrane by a phosphatidylinositol (PI) anchor. Using a panel of radiolabeled MAbs directed against the 5 major epitopes described on the CEA molecule, we showed that all these CEA epitopes were expressed by the 4 transfectants. Southern-blot analysis showed that the entire CEA cDNA was present in the transfectants. Western-blot analysis, however, showed that the size of the CEA expressed by the 4 transfectants was slightly smaller than that of CEA produced by 2 reference human colon-carcinoma cell lines. Two clones, expressing 1 x 10(5) and 1 x 10(6) CEA molecules per cell, respectively, were grafted s.c. in nude mice and rats. Injection of radiolabeled anti-CEA F(ab')2 fragments into these animals showed specific tumor localization with the highest percentages of injected doses for the transfectants expressing the highest CEA level. When grafted into immunocompetent syngeneic BDIX rats, the CEA-expressing clones induced a strong antibody response against CEA and tumor rejections in a majority of the animals. Although the analysis of the immune response against the CEA-cDNA-transfected carcinoma cells is under investigation, the present results demonstrate that human CEA could function as a rejection antigen when transfected into rat carcinoma cells.