Radiolabeled fragments of monoclonal antibodies against carcinoembryonic antigen for localization of human colon carcinoma grafted into nude mice.

Four monoclonal antibodies against carcinoembryonic antigen (CEA) have been selected from 32 hybrids that produce antibodies against this antigen, by the criteria of high affinity for CEA and low cross-reactivity with granulocyte glycoprotein(s). The specificity of tumor localization in vivo of the four MAb, and their F(ab')2 and Fab fragments was compared in nude mice bearing grafts of a serially transplanted, CEA-producing, human colon carcinoma. The distribution of radiolabeled MAb and their fragments after intravenous injection was analyzed by direct measurement of radioactivity in tumor and normal organs, as well as by whole-body scanning and by autoradiography of tumor sections. Paired labeling experiments, in which 131I-labeled antibody or fragments and 125I-labeled control IgG are injected simultaneously, were undertaken to determine the relative tumor uptakes of each labeled protein. The tumor antibody uptake divided by that of control IgG defines the specificity index of localization. Tumor antibody uptakes (as compared with the whole mouse), ranging between 7 and 15, and specificity indices ranging between 3.4 and 6.8, were obtained with the four intact MAb at day 4-5 after injection. With F(ab')2 fragments of the four MAb, at day 3, the tumor antibody uptakes ranged between 12 and 24 and the specificity indices between 5.3 and 8.2. With the Fab fragments prepared from the two most promising MAb, the antibody uptakes reached values of 34 and 82 at day 2-3 and the specificity indices were as high as 12 and 19. The scanning results paralleled those obtained by direct measurement of radioactivity. With intact MAb, tumor grafts of 0.5-1 g gave very contrasted positive scans 3 d after injection. Using MAb fragments, tumors of smaller size were detectable earlier. The best results were obtained with Fab fragments of MAb 35, which gave clear detections of tumors weighing only 0.1 g as early as 48 h after injection. Autoradiographs of tumor sections from mice injected with 125I-labeled MAb demonstrated that the radioactivity was localized in the tumor tissues and not in the stromal connective tissue of mouse origin. The highest radioactivity concentration was localized in areas known to contain CEA such as the pseudolumen of glands and the apical side of carcinoma cells. The penetration of radioactivity in the central part of tumor nodules and the pseudolumen appeared to be increased with the use of MAb fragments.

Rendimentos de culturas e alterações químicas do solo tratado com resíduos de curtume e crômio hexavalente

O presente trabalho foi elaborado com o objetivo de avaliar o rendimento das culturas do trigo, alface e rabanete e as alterações químicas de um Latossolo resultantes da adição de resíduos de curtume e de crômio hexavalente. Microparcelas, constituídas por recipientes com 60 L de solo e mantidas em casa de vegetação, receberam os seguintes tratamentos: testemunha; calcário + NPK; lodo do decantador primário + PK; resíduo de rebaixadeira de couro + calcário + NPK; aparas de couro + calcário + NPK; Cr6+ + calcário + NPK ; Cr6+ + calcário + esterco bovino; calcário + esterco bovino. O lodo continha 8,5 g kg-1 de Cr e foi aplicado em dose correspondente a 8,8 t ha-1. O resíduo de rebaixadeira possuía 17,1 g kg-1 Cr e as aparas de couro, 19,4 g kg-1 Cr. Esses resíduos foram aplicados nas doses correspondentes a 4,4 e 3,8 t ha-1. As doses de Cr6+ (K2Cr2O7) e esterco bovino foram de 100 mg kg-1 e 20 t ha-1, respectivamente. A aplicação de lodo elevou o pH do solo de 5,1 para 5,8; os teores de N, de 1,26 para 1,51 g kg-1, e os teores de Ca, de 4,1 para 5,9 cmol c dm-3, proporcionando rendimentos das três culturas equivalentes aos obtidos com a aplicação de calcário + NPK. Os teores de crômio no solo e nas partes vegetativas das culturas nos tratamentos com aplicação dos resíduos de curtume variaram, respectivamente, de 40,7 a 71,2 e de 0,08 a 2,71 mg kg-1, sendo considerados normais. A adição de resíduo de rebaixadeira de couro ou das aparas de couro não reduziu os rendimentos das culturas e não alterou os teores de Cr do solo e das plantas. A adição de Cr6+ provocou um efeito tóxico nas plantas, sendo responsável pela diminuição do rendimento da cultura do trigo em 91 % e da cultura da alface em 86 %, quando comparadas ao tratamento Calcário + NPK. Este efeito foi diminuído com a adição de agentes redutores como o esterco bovino.

Large TCR diversity of virus-specific CD8 T cells provides the mechanistic basis for massive TCR renewal after antigen exposure.

Ex vivo analysis of virus-specific CD8 T cell populations by anchored PCR has shown that the CD8 TCR repertoire was less oligoclonal (seven to nine clonotypes per individual epitope) than previously thought. In the current study, TCR diversity was investigated by assessing both the overall TCR β-chain variable regions usage as well as the CDR3 regions in ex vivo-isolated CMV- and EBV-specific CD8 T cells from 27 healthy donors. The average number of clonotypes specific to most single viral epitopes comprised between 14 and 77. Changes in the CD8 TCR repertoire were also longitudinally assessed under conditions of HIV-1 chronic infection (i.e., in patients with suppressed virus replication and after treatment interruption and Ag re-exposure). The results showed that a large renewal (≤80%) of the TRB repertoire occurred after Ag re-exposure and was eventually associated with an increased T cell recognition functional avidity. These results demonstrate that the global CD8 TCR repertoire is much more diverse (≤9-fold) than previously estimated and provide the mechanistic basis for supporting massive repertoire renewal during chronic virus infection and Ag re-exposure.

Mudanças da fertilidade do solo e crescimento de um povoamento de Eucalyptus grandis fertilizado com biossólido

Antes da recomendação em larga escala de biossólido em plantações florestais, é preciso compreender seus efeitos no solo e na planta. Assim, a fertilidade do solo, o estado nutricional e o crescimento de um povoamento de Eucalyptus grandis fertilizado com biossólido foram avaliados em um experimento na Estação Experimental de Ciências Florestais de Itatinga (SP), ESALQ/USP. O delineamento experimental foi o de blocos casualizados, com quatro blocos e nove tratamentos: (1) Testemunha; (2) Adubação mineral; (3) 5 t ha-1 de bios. + K; (4) 10 t ha-1 de bios. + K; (5) 10 t ha-1 de bios.; (6) 10 t ha-1 de bios. + K + P; (7) 15 t ha-1 de bios. + K; (8) 20 t ha-1 de bios. + K, e (9) 40 t ha-1 de bios. + K. Foram analisadas quimicamente amostras de solo (camadas de 0-5, 5-10 e 10-20 cm) e de folhas. A produção de madeira foi avaliada por meio da colheita e pesagem de árvores. Até 32 meses após a aplicação do biossólido, 36 meses pós-plantio, constataram-se aumentos do pH, dos teores de C orgânico, de P-resina e de Ca trocável nas três camadas, diretamente associados às doses de biossólido aplicadas. Os teores de S-SO4(2-) e K trocável diminuíram 13 meses após a aplicação do biossólido e, 19 meses depois, os teores estavam aumentados. O Al trocável diminuiu com o aumento das doses de biossólido, nas três camadas amostradas. A aplicação de biossólido influiu positivamente na nutrição das plantas, proporcionando uma produção de madeira igual à obtida no tratamento que só recebeu adubação mineral (1,5 t ha-1 de calcário dolomítico e, em kg ha-1, 98 de N, 79,5 de P2O5, 165 de K2O, 1,3 de B e 1,2 de Zn), quando a dose de biossólido foi equivalente a 12 t ha-1.

Fluorine-19 magnetic resonance angiography of the mouse.

PURPOSE: To implement and characterize a fluorine-19 ((19)F) magnetic resonance imaging (MRI) technique and to test the hypothesis that the (19)F MRI signal in steady state after intravenous injection of a perfluoro-15-crown-5 ether (PCE) emulsion may be exploited for angiography in a pre-clinical in vivo animal study. MATERIALS AND METHODS: In vitro at 9.4T, the detection limit of the PCE emulsion at a scan time of 10 min/slice was determined, after which the T(1) and T(2) of PCE in venous blood were measured. Permission from the local animal use committee was obtained for all animal experiments. 12 µl/g of PCE emulsion was intravenously injected in 11 mice. Gradient echo (1)H and (19)F images were obtained at identical anatomical levels. Signal-to-noise (SNR) and contrast-to-noise (CNR) ratios were determined for 33 vessels in both the (19)F and (1)H images, which was followed by vessel tracking to determine the vessel conspicuity for both modalities. RESULTS: In vitro, the detection limit was ∼400 µM, while the (19)F T(1) and T(2) were 1350±40 and 25±2 ms. The (19)F MR angiograms selectively visualized the vasculature (and the liver parenchyma over time) while precisely coregistering with the (1)H images. Due to the lower SNR of (19)F compared to (1)H (17±8 vs. 83±49, p<0.001), the (19)F CNR was also lower at 15±8 vs. 52±35 (p<0.001). Vessel tracking demonstrated a significantly higher vessel sharpness in the (19)F images (66±11 vs. 56±12, p = 0.002). CONCLUSION: (19)F magnetic resonance angiography of intravenously administered perfluorocarbon emulsions is feasible for a selective and exclusive visualization of the vasculature in vivo.

Informe sobre les X Jornades d'Arxivística de Catalunya: "Els arxivers com (ens) comuniquem?" (Terrassa, 19, 20 i 21 de maig de 2005)

Com ja és costum des de fa vint anys (1985-2005), pel maig l'Associació d'Arxivers de Catalunya (AAC) convoca els seus associats i la comunitat arxivística en general a les Jornades d'Arxivística de Catalunya. Enguany han estat les desenes i amb el títol "Els arxivers com (ens) comuniquem?" s'han dut a terme els dies 19, 20 i 21 de maig a la ciutat de Terrassa. L'objectiu de les X Jornades ha estat doble: en primer lloc, s'ha volgut reflexionar sobre les estratègies de què disposen els arxivers per fer difusió de la seva activitat professional ("com comuniquem") i a continuació s'ha produït el debat sobre la comunicació intraprofessional dels arxivers ("com ens comuniquem").

Informe sobre les VII Jornadas Españolas de Documentación, FESABID 2000 (Bilbao, 19-21 d'octubre de 2000)

Aquestes jornades espanyoles de documentació tenen lloc cada dos anys en una localitat diferent d'Espanya, i són organitzades conjuntament per FESABID (Federación Española de Sociedades de Archivística, Biblioteconomía, Documentación y Museística) i l'associació professional de la comunitat autònoma on se celebren. En aquesta ocasió, Bilbao fou la ciutat amfitriona, i ALDEE (Artxibozain, Liburuzain eta Dokumentazainen Euskal Elkartea), una de les associacions professionals que existeixen al País Basc, participà en l'organització

Analysis of viral- and tumor antigen-specific CD4 T cell responses in healthy donors and melanoma patients

Résumé Des tentatives pour développer des traitements anti-cancéreux basés sur l'utilisation d'antigènes tumoraux ont commencé il y a plus de 10 ans. Depuis quelques années, un certain intérêt s'est portée sur une sous-population particulière des cellules du système immunitaire, les lymphocytes T CD4. Ces cellules jouent un rôle central dans les réponses immunitaires tant contre les virus que contre les cellules tumorales. Comme d'autres lymphocytes T, ces cellules sont activées de manière spécifique en reconnaissant un morceau d'antigène, appelé peptide. Ces peptides proviennent soit de protéines des cellules de l'hôte, soit des protéines étrangères (virus ou bactéries) soit de cellules transformées (cellules tumorales) et sont présentés aux lymphocytes T par des molécules du soi appelées CMH (complexe majeur d'histocompatibilité). Dans le cas des lymphocytes T CD4, ces molécules sont plus précisément des molécules du CMH de classe II (CMH II). Mis à part l'intérêt porté aux réponses médiées par les lymphocytes T cytotoxiques, un intérêt croissant pour les lymphocytes T CD4 s'est développé à cause de la place centrale qu'occupent ces cellules dans les réponses immunitaires. L'identification d'épitopes présentés par des molécules du CMH de classe II dérivés d'un grand nombre d'antigènes tumoraux, ainsi que le développement de techniques permettant de suivre les réponses immunitaires, offre des opportunités pour étudier de manière quantitative et qualitative les lymphocytes T CD4 spécifiques pour un antigène particulier chez des patients cancéreux. De plus, ces épitopes permettent d'induire des réponses médiées par les lymphocytes T CD4 et CD8 chez ces mêmes patients. Dans ce travail, notre premier but était de valider l'utilisation de multimères formés par des complexes peptide:molécules de CMH de class II (pCMH II) pour quantifier la réponse des cellules T CD4 dirigée contre l'épitope HA307-319 dérivé de la protéine hémaglutinine du virus de la grippe et présenté par HLA-DRB1*0401. En analysant des échantillons provenant de volontaires sains ayant reçus un vaccin contre la grippe, nous avons pu démontrer une expansion et une activation transitoires des lymphocytes T CD4 spécifiques pour le peptide HA307-319 après vaccination. De plus, les multimères pCMH II nous ont permis d'analyser plus en détails hétérogénéité des cellules T CD4 spécifiques pour le peptide HA307-319 présents dans le sang périphérique d'individus sains. Par la suite, notre but a été d'analyser les réponses des lymphocytes T CD4 spécifiques pour l'antigène Melan-A chez des patients atteints de mélanome métastatique. Nous avons tout d'abord démontré la présence de cellules T CD4 spécifiques pour l'épitope Melan-A51-73, présenté par HLA-DRBl*0401, qui avait déjà été préalablement décrit. Ensuite, nous avons décrit et caractérisé 2 nouveaux peptides issus de Melan-A qui sont présentés aux cellules T CD4 par différentes molécules du CMH de clans II. Des cellules spécifiques pour ces deux épitopes ont été trouvées chez 9/ 16 patients analysés. De plus, des multimères pCMH II chargés avec un des épitopes nous ont permis de détecter ex vivo des lymphocytes T CD4 spécifiques pour Melan-A dans le sang périphérique d'un patient atteint de mélanome. Mis ensemble, tous ces résultats suggèrent une potentielle utilisation des multimères pCMH II pour analyser en détail les lymphocytes T CD4 spécifiques d'antigènes définis. Cependant, le suivi ex vivo de telles cellules ne semble être possible que dans des cas bien particuliers. Néanmoins, les nouveaux épitopes issus de Melan-A et présentés par des molécules du CMH de classe II que nous avons décrits dans cette étude aideront à étudier plus en détails les lymphocytes T CD4 spécifiques pour Melan-A chez des patients atteints de mélanome, un sujet d'étude sur lequel peu de résultats sont à ce jour disponibles. Summary Attempts to develop cancer vaccines based on molecularly defined tumorassociated antigens were initiated more than 10 years ago. Apart from CTLmediated anti-tumor immunity, interests are. now focused on CD4 T cells that are central players of immune responses. The identification of MHC class-II-restricted epitopes from numerous tumor antigens together with the development of monitoring tools offers the opportunity to quantitatively and qualitatively study antigen-specific CD4 T lymphocytes in cancer patients and to induce both CTL and T helper responses in cancer patients. In this work, we first aimed at validating the use of peptide:MHC class II complex (pMHC II) multimers to quantitate the CD4 T cell response against the hemagglutinin-derived epitope HAso~-si9 from influenza virus presented by HLA-DRBl*0401. By analysing samples from healthy volunteers vaccinated with ananti-influenza vaccine, we could demonstrate a transient expansion and activation of HA-specific CD4 T cells after treatment. Moreover, pMHC II multimers helped us to study the heterogeneity of HAspecific CD4 T cells found in peripheral blood of healthy individuals. Then, we aimed to analyse Melan-A-specific CD4 T cell responses in metastatic melanoma patients. We first demonstrated the presence of CD4 T cells specific for the previously described Melan-A51_73 epitope presented by HLA-DRB 1 *0401 in peripheral blood of those patients. Second, we described and characterised 2 new Melan-A-derived peptides that are presented by different MHC II molecules to CD4 T cells. Specific cells for these epitopes were found in 9/ 16 rnelánoma patients analysed. In addition, pMHC II multimers loaded with one of the two epitopes allowed us to detect ex vivo Melan-A-specific CD4 T cells in peripheral blood of a melanoma patient. Together, these results suggest a potential use of pMHC II multimers in analysing in detail antigen-specific CD4 T cells. However, ex vivo monitoring of such cells will be possible only in particular conditions. Nevertheless, the new Melan-A-derived MHC II-restricted epitopes described here will help to study in more detail Melan-A-specific CD4 T cells in melanoma patients, a field where only scarce data are available.

Informe del World Library and Information Congress: 73rd IFLA General Conference and Council (Durban, Sud-àfrica, 19-23 d'agost de 2007)

This year, IFLA's World Library and Information Congress (WLIC) was held in Durban, South Africa, under the title ¿Libraries for the future: progress, development and partnerships¿. The association thus continued its policy of holding the event in different continents: Buenos Aires (South America) 2004, Oslo (Europe) 2005, Seoul (Asia) 2006, Durban (Africa) 2007, Quebec (North America) 2008, Milan (Europe) 2009 and Brisbane (Oceania) 2010.

Biochemical analysis of the major vaccinia virus envelope antigen.

The major envelope antigen of vaccinia virus is an acylated protein of M(r) 37,000 (p37K) which is required for the formation of extracellular enveloped virions (EEV). Despite its important role in the wrapping process, p37K has not been studied in much detail. In order to better characterize this protein we have undertaken a detailed biochemical analysis. Sodium carbonate treatment showed that p37K is tightly bound to the viral envelope. Its resistance to proteinase K digestion indicates that it is not exposed on the surface of EEV but lines the inner side of the envelope. Since p37K does not contain a signal peptide characteristic of most membrane proteins, we examined the possibility that the protein acquires its membrane affinity through the addition of fatty acids. Indeed, Triton X-114 phase partitioning experiments demonstrated that p37K is hydrophobic when acylated, but hydrophilic in the absence of fatty acids. Three other viral proteins have been shown to be required for virus envelopment and release from the host cell and we therefore tested whether p37K interacts with viral proteins. In EEV and in absence of reducing agents, an 80-kDa complex reacting with an anti-37K antiserum was found. Analysis of this complex showed that it most likely consists of a p37K homodimer. Interestingly, only a small amount of p37K occurs as a complex, most of it is present in the viral envelope as monomers.