"Pues doy Fe de que son tenidos como tales" : Prácticas y discursos legitimantes en el intento de conformación de una posible nobleza rioplatense

A mediados del siglo XVIII los grandes comerciantes de distintos espacios hispanoamericanos, acumulan suficientes caudales que les permiten comprar títulos de nobleza, distinciones o formar mayorazgos que relumbren sus nombres y perpetúen sus bienes adquiridos. Este proceso es mayormente evidente en los espacios mexicanos y peruanos; pero no se conocen casos concretos para el espacio rioplatense. Como planteó José Torre Revello, esto no implica que los comerciantes rioplatenses no intentasen ennoblecerse. El presente estudio de caso detalla como Don Vicente de Azcuénaga intenta fundar un mayorazgo en la ciudad de Buenos Aires a favor de su primogénito Miguel. A través de este estudio basado en las "probanzas" se puede observar como la familia Azcuénaga pretende resaltar su nombre frente al resto de sus contemporáneos, pero las relaciones entre padre e hijo nos conducen a la vez a replantearnos interrogantes referentes a las tradiciones de acumulación y conservación de patrimonios

Nuevo enlace ferrovial interprovincial Santa Fe-Entre Ríos : Una oportunidad de escala regional

La provincia de Santa Fe, Argentina, se encuentra en una localización estratégica. La potencialidad de la Hidrovía Paraná-Paraguay, los corredores bioceánicos viales y la red existente de trazados ferroviarios le confieren gran dinamismo a su integración económica, social, cultural y política, no sólo hacia el interior del propio territorio, sino también en relación a las demás provincias y más allá de los confines nacionales. La región capital, cuyo núcleo es la ciudad de Santa Fe, se encuentra caracterizada por factores realmente dinámicos: el riesgo hídrico que es intrínseco del área, la intensificación de los flujos económicos pasantes, los procesos de concentración demográfica y la creciente interdependencia entre ciudades, como es el casode Santa Fe y Paraná (capital de la vecina provincia de Entre Ríos), bajo un progresivo proceso de metropolización binuclear. Estos factores, sumados a la escasa cantidad de conexiones físicas sobre el sistema fluvial del río Paraná, han instalado la creciente necesidad de contar con un nuevo enlace interprovincial, adaptado a una hipótesis de reactivación ferroviaria. El proyecto se encuentra en fase preliminar. La cuestión principal gira en torno a la decisión de su localización específica, que deberá considerar el profundo efecto transformador propio de una obra civil de gran calibre, tanto en relación a la plataforma natural como al sistema de asentamientos humanos. También sus alcances territoriales y el impacto potencial en la micro, meso y macroescala. El propósito de la investigación reside en profundizar sobre las dimensiones involucradas por el proyecto (técnica, social, económica, ambiental, de movilidad), en la búsqueda de una toma de posición que permita echar luz sobre los escenarios más beneficiosos y/o menos desfavorables, en relación a las numerosas propuestas de localización que se encuentran actualmente en discusión. El resultado es una matriz analítica basada en variables cuantitativas y cualitativas, que permite una evaluación integral de las propuestas en función de considerar, en síntesis, el grado de impacto sobre la plataforma natural sustentante, sus capacidades para revertir las problemáticas territoriales actuales, y finalmente sus posibilidades para generar nuevos ejes de desarrollo en la región o bien potenciar los existentes. Se concluye que análisis preliminares de tipo pluridimensional son necesarios para someter a discusión, como instancia previa a estudios específicos de factibilidad y viabilidad, puesto que permiten una visualización integral de las variables intervinientes, marcando el camino hacia su adecuada ponderación. Palabras clave: enlace, multimodalidad, región, transformaciones

Nuevo enlace ferrovial interprovincial Santa Fe-Entre Ríos : Una oportunidad de escala regional

La provincia de Santa Fe, Argentina, se encuentra en una localización estratégica. La potencialidad de la Hidrovía Paraná-Paraguay, los corredores bioceánicos viales y la red existente de trazados ferroviarios le confieren gran dinamismo a su integración económica, social, cultural y política, no sólo hacia el interior del propio territorio, sino también en relación a las demás provincias y más allá de los confines nacionales. La región capital, cuyo núcleo es la ciudad de Santa Fe, se encuentra caracterizada por factores realmente dinámicos: el riesgo hídrico que es intrínseco del área, la intensificación de los flujos económicos pasantes, los procesos de concentración demográfica y la creciente interdependencia entre ciudades, como es el casode Santa Fe y Paraná (capital de la vecina provincia de Entre Ríos), bajo un progresivo proceso de metropolización binuclear. Estos factores, sumados a la escasa cantidad de conexiones físicas sobre el sistema fluvial del río Paraná, han instalado la creciente necesidad de contar con un nuevo enlace interprovincial, adaptado a una hipótesis de reactivación ferroviaria. El proyecto se encuentra en fase preliminar. La cuestión principal gira en torno a la decisión de su localización específica, que deberá considerar el profundo efecto transformador propio de una obra civil de gran calibre, tanto en relación a la plataforma natural como al sistema de asentamientos humanos. También sus alcances territoriales y el impacto potencial en la micro, meso y macroescala. El propósito de la investigación reside en profundizar sobre las dimensiones involucradas por el proyecto (técnica, social, económica, ambiental, de movilidad), en la búsqueda de una toma de posición que permita echar luz sobre los escenarios más beneficiosos y/o menos desfavorables, en relación a las numerosas propuestas de localización que se encuentran actualmente en discusión. El resultado es una matriz analítica basada en variables cuantitativas y cualitativas, que permite una evaluación integral de las propuestas en función de considerar, en síntesis, el grado de impacto sobre la plataforma natural sustentante, sus capacidades para revertir las problemáticas territoriales actuales, y finalmente sus posibilidades para generar nuevos ejes de desarrollo en la región o bien potenciar los existentes. Se concluye que análisis preliminares de tipo pluridimensional son necesarios para someter a discusión, como instancia previa a estudios específicos de factibilidad y viabilidad, puesto que permiten una visualización integral de las variables intervinientes, marcando el camino hacia su adecuada ponderación. Palabras clave: enlace, multimodalidad, región, transformaciones

"Pues doy Fe de que son tenidos como tales" : Prácticas y discursos legitimantes en el intento de conformación de una posible nobleza rioplatense

A mediados del siglo XVIII los grandes comerciantes de distintos espacios hispanoamericanos, acumulan suficientes caudales que les permiten comprar títulos de nobleza, distinciones o formar mayorazgos que relumbren sus nombres y perpetúen sus bienes adquiridos. Este proceso es mayormente evidente en los espacios mexicanos y peruanos; pero no se conocen casos concretos para el espacio rioplatense. Como planteó José Torre Revello, esto no implica que los comerciantes rioplatenses no intentasen ennoblecerse. El presente estudio de caso detalla como Don Vicente de Azcuénaga intenta fundar un mayorazgo en la ciudad de Buenos Aires a favor de su primogénito Miguel. A través de este estudio basado en las "probanzas" se puede observar como la familia Azcuénaga pretende resaltar su nombre frente al resto de sus contemporáneos, pero las relaciones entre padre e hijo nos conducen a la vez a replantearnos interrogantes referentes a las tradiciones de acumulación y conservación de patrimonios

Direct visualization of antigen-specific T cells: HTLV-1 Tax11–19- specific CD8+ T cells are activated in peripheral blood and accumulate in cerebrospinal fluid from HAM/TSP patients

Human T lymphotropic virus type 1 (HTLV-1) -associated myelopathy/tropic spastic paraparesis is a demyelinating inflammatory neurologic disease associated with HTLV-1 infection. HTLV-1 Tax11–19-specific cytotoxic T cells have been isolated from HLA-A2-positive patients. We have used a peptide-loaded soluble HLA-A2–Ig complex to directly visualize HTLV-1 Tax11–19-specific T cells from peripheral blood and cerebrospinal fluid without in vitro stimulation. Five of six HTLV-1-associated myelopathy/tropic spastic paraparesis patients carried a significant number (up to 13.87%) of CD8+ lymphocytes specific for the HTLV-1 Tax11–19 peptide in their peripheral blood, which were not found in healthy controls. Simultaneous comparison of peripheral blood and cerebrospinal fluid from one patient revealed 2.5-fold more Tax11–19-specific T cells in the cerebrospinal fluid (23.7% vs. 9.4% in peripheral blood lymphocyte). Tax11–19-specific T cells were seen consistently over a 9-yr time course in one patient as far as 19 yrs after the onset of clinical symptoms. Further analysis of HTLV-1 Tax11–19-specific CD8+ T lymphocytes in HAM/TSP patients showed different expression patterns of activation markers, intracellular TNF-α and γ-interferon depending on the severity of the disease. Thus, visualization of antigen-specific T cells demonstrates that HTLV-1 Tax11–19-specific CD8+ T cells are activated, persist during the chronic phase of the disease, and accumulate in cerebrospinal fluid, showing their pivotal role in the pathogenesis of this neurologic disease.

Fe-catalyzed cleavage of the α subunit of Na/K-ATPase: Evidence for conformation-sensitive interactions between cytoplasmic domains

Incubation of Na/K-ATPase with ascorbate plus H2O2 produces specific cleavage of the α subunit. Five fragments with intact C termini and complementary fragments with intact N termini were observed. The β subunit is not cleaved. Cleavages depend on the presence of contaminant or added Fe2+ ions, as inferred by suppression of cleavages with nonspecific metal complexants (histidine, EDTA, phenanthroline) or the Fe3+-specific complexant desferrioxamine, or acceleration of cleavages by addition of low concentrations of Fe2+ but not of other heavy metal ions. Na/K-ATPase is inactivated in addition to cleavage, and both effects are insensitive to OH⋅ radical scavengers. Cleavages are sensitive to conformation. In low ionic strength media (E2) or media containing Rb ions [E2(Rb)], cleavage is much faster than in high ionic strength media (E1) or media containing Na ions (E1Na). N-terminal fragments and two C-terminal fragments (N-terminals E214 and V712) have been identified by amino acid sequencing. Approximate positions of other cleavages were determined with specific antibodies. The results suggest that Fe2+ (or Fe3+) ions bind with high affinity at the cytoplasmic surface and catalyze cleavages of peptide bonds close to the Fe2+ (or Fe3+) ion. Thus, cleavage patterns can provide information on spatial organization of the polypeptide chain. We propose that highly conserved regions of the α subunit, within the minor and major cytoplasmic loops, interact in the E2 or E2(Rb) conformations but move apart in the E1 or E1Na conformations. We discuss implications of domain interactions for the energy transduction mechanism. Fe-catalyzed cleavages may be applicable to other P-type pumps or membrane proteins.

Expression of HLA class I-specific inhibitory natural killer cell receptors in HIV-specific cytolytic T lymphocytes: Impairment of specific cytolytic functions

Human T lymphocytes have been shown to express inhibitory natural killer cell receptors (NKR), which can down-regulate T cell antigen receptor-mediated T cell function, including cytolytic activity. In the present study, we demonstrate that CD3+NKR+ cells can be identified in HIV-infected patients. HIV-specific cytolytic activity was analyzed in five patients in whom autologous lymphoblastoid B cell lines could be derived as a source of autologous target cells. Phytohemagglutinin-activated T cell populations that had been cultured in interleukin 2 displayed HIV-specific cytotoxic T lymphocyte (CTL) activity against HIV env, gag, pol, and nef in 3 of 5 patients. Addition of anti-NKR mAb of IgM isotype could increase the specific CTL activity. Moreover, in one additional patient, HIV-specific CTL activity was undetectable; however, after addition of anti-NKR mAb such CTL activity appeared de novo. Similar results were obtained by analysis of CD3+NKR+ clones derived from two patients. These data provide direct evidence that CD3+NKR+ cells may include antigen (HIV)-specific CTLs and that mAb-mediated masking of inhibitory NKR may revert the down-regulation of CTL function.

Synergistic up-regulation of the myeloid-specific promoter for the macrophage colony-stimulating factor receptor by AML1 and the t(8;21) fusion protein may contribute to leukemogenesis.

AML1 is involved in the (8;21) translocation, associated with acute myelogenous leukemia (AML)-type M2, which results in the production of the AML1-ETO fusion protein: the amino-terminal 177 amino acids of AML1 and the carboxyl-terminal 575 amino acids of ETO. The mechanism by which AML1-ETO accomplishes leukemic transformation is unknown; however, AML1-ETO interferes with AML1 transactivation of such AML1 targets as the T-cell receptor beta enhancer and the granulocyte-macrophage colony-stimulating factor promoter. Herein, we explored the effect of AML1-ETO on regulation of a myeloid-specific AML1 target, the macrophage colony-stimulating factor (M-CSF) receptor promoter. We found that AML1-ETO and AML1 work synergistically to transactivate the M-CSF receptor promoter, thus exhibiting a different activity than previously described. Truncation mutants within the ETO portion of AML1-ETO revealed the region of ETO necessary for the cooperativity between AML1 and AML1-ETO lies between amino acids 347 and 540. Endogenous M-CSF receptor expression was examined in Kasumi-1 cells, derived from a patient with AML-M2 t(8;21) and the promonocytic cell line U937. Kasumi-1 cells exhibited a significantly higher level of M-CSF receptor expression than U937 cells. Bone marrow from patients with AML-M2 t(8;21) also exhibited a higher level of expression of M-CSF receptor compared with normal controls. The upregulation of M-CSF receptor expression by AML1-ETO may contribute to the development of a leukemic state in these patients.

Correction in trans for Fabry disease: expression, secretion and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector.

Fabry disease is an X-linked metabolic disorder due to a deficiency of alpha-galactosidase A (alpha-gal A; EC 3.2.1.22). Patients accumulate glycosphingolipids with terminal alpha-galactosyl residues that come from intracellular synthesis, circulating metabolites, or from the biodegradation Of senescent cells. Patients eventually succumb to renal, cardio-, or cerebrovascular disease. No specific therapy exists. One possible approach to ameliorating this disorder is to target corrective gene transfer therapy to circulating hematopoietic cells. Toward this end, an amphotropic virus-producer cell line has been developed that produces a high titer (>10(6) i.p. per ml) recombinant retrovirus constructed to transduce and correct target cells. Virus-producer cells also demonstrate expression of large amounts of both intracellular and secreted alpha-gal A. To examine the utility of this therapeutic vector, skin fibroblasts from Fabry patients were corrected for the metabolic defect by infection with this recombinant virus and secreted enzyme was observed. Furthermore, the secreted enzyme was found to be taken up by uncorrected cells in a mannose-6-phosphate receptor-dependent manner. In related experiments, immortalized B cell lines from Fabry patients, created as a hematologic delivery test system, were transduced. As with the fibroblasts, transduced patient B cell lines demonstrated both endogenous enzyme correction and a small amount of secretion together with uptake by uncorrected cells. These studies demonstrate that endogenous metabolic correction in transduced cells, combined with secretion, may provide a continuous source of corrective material in trans to unmodified patient bystander cells (metabolic cooperativity).

Detection of JC virus DNA sequence and expression of the viral oncoprotein, tumor antigen, in brain of immunocompetent patient with oligoastrocytoma.

We describe molecular and clinical findings in an immunocompetent patient with an oligoastrocytoma and the concomitant presence of the human papovavirus, JC virus (JCV), which is the etiologic agent of the subacute, debilitating demyelinating disease, progressive multifocal leukoencephalopathy. Histologic review revealed a glial neoplasm consisting primarily of a moderately cellular oligodendroglioma with distinct areas of a fibrillary astrocytoma. Immunohistochemical analysis revealed nuclear staining of tumor cells with antibodies against the viral oncoprotein [tumor antigen (T antigen)], the proliferation marker (Ki67), and the cellular proliferation regulator (p53). Using primers specific to the JCV control region, PCR yielded amplified DNA that was identical to the control region of the Mad-4 strain of the virus. PCR analysis demonstrated the presence of the genome for the viral oncoprotein, T antigen, and results from primer extension studies revealed synthesis of the viral early RNA for T antigen in the tumor tissues. The presence of viral T antigen in the tumor tissue was further demonstrated by immunoblot assay. To our knowledge, this is the first report of the presence of JCV DNA, RNA, and T antigen in tissue in which viral T antigen is localized to tumor cell nuclei and suggests the possible association of JCV with some glial neoplasms.

Peroxynitrite disables the tyrosine phosphorylation regulatory mechanism: Lymphocyte-specific tyrosine kinase fails to phosphorylate nitrated cdc2(6-20)NH2 peptide.

To determine if nitration of tyrosine residues by peroxynitrite (PN), which can be generated endogenously, can disrupt the phosphorylation of tyrosine residues in proteins involved in cell signaling networks, we studied the effect of PN-promoted nitration of tyrosine residues in a pentadecameric peptide, cdc2(6-20)NH2, on the ability of the peptide to be phosphorylated. cdc2(6-20)NH2 corresponds to the tyrosine phosphorylation site of p34cdc2 kinase, which is phosphorylated by lck kinase (lymphocyte-specific tyrosine kinase, p56lck). PN nitrates both Tyr-15 and Tyr-19 of the peptide in phosphate buffer (pH 7.5) at 37 degrees C. Nitration of Tyr-15. which is the phosphorylated amino acid residue, inhibits completely the phosphorylation of the peptide. The nitration reaction is enhanced by either Fe(III)EDTA or Cu(II)-Zn(II)-superoxide dismutase (Cu,Zn-SOD). The kinetic data are consistent with the view that reactions of Fe(111)EDTA or Cu,Zn-SOD with the cis form of PN yield complexes in which PN decomposes more slowly to form N02+, the nitrating agent. Thus, the nitration efficiency of PN is enhanced. These results are discussed from the point of view that PN-promoted nitration will result in permanent impairment of cyclic cascades that control signal transduction processes and regulate cell cycles.

T-cell epitope analysis using subtracted expression libraries (TEASEL): application to a 38-kDA autoantigen recognized by T cells from an insulin-dependent diabetic patient.

Studies on circulating T cells and antibodies in newly diagnosed type 1 diabetic patients and rodent models of autoimmune diabetes suggest that beta-cell membrane proteins of 38 kDa may be important molecular targets of autoimmune attack. Biochemical approaches to the isolation and identification of the 38-kDa autoantigen have been hampered by the restricted availability of islet tissue and the low abundance of the protein. A procedure of epitope analysis for CD4+ T cells using subtracted expression libraries (TEASEL) was developed and used to clone a 70-amino acid pancreatic beta-cell peptide incorporating an epitope recognized by a 38-kDa-reactive CD4+ T-cell clone (1C6) isolated from a human diabetic patient. The minimal epitope was mapped to a 10-amino acid synthetic peptide containing a DR1 consensus binding motif. Data base searches did not reveal the identity of the protein, though a weak homology to the bacterial superantigens SEA (Streptococcus pyogenes exotoxin A) and SEB (Staphylococcus aureus enterotoxin B) (23% identity) was evident. The TEASEL procedure might be used to identify epitopes of other autoantigens recognized by CD4+ T cells in diabetes as well as be more generally applicable to the study low-abundance autoantigens in other tissue-specific autoimmune diseases.

Human neoplasms elicit multiple specific immune responses in the autologous host.

Expression of cDNA libraries from human melanoma, renal cancer, astrocytoma, and Hodgkin disease in Escherichia coli and screening for clones reactive with high-titer IgG antibodies in autologous patient serum lead to the discovery of at least four antigens with a restricted expression pattern in each tumor. Besides antigens known to elicit T-cell responses, such as MAGE-1 and tyrosinase, numerous additional antigens that were overexpressed or specifically expressed in tumors of the same type were identified. Sequence analyses suggest that many of these molecules, besides being the target of a specific immune response, might be of relevance for tumor growth. Antibodies to a given antigen were usually confined to patients with the same tumor type. The unexpected frequency of human tumor antigens, which can be readily defined at the molecular level by the serological analysis of autologous tumor cDNA expression cloning, indicates that human neoplasms elicit multiple specific immune responses in the autologous host and provides diagnostic and therapeutic approaches to human cancer.

Anti-melanoma antibodies from melanoma patients immunized with genetically modified autologous tumor cells: selection of specific antibodies from single-chain Fv fusion phage libraries.

Fusion phage libraries expressing single-chain Fv antibodies were constructed from the peripheral blood lymphocytes of two melanoma patients who had been immunized with autologous melanoma cells transduced the gamma-interferon gene to enhance immunogenicity, in a trial conducted at another institution. Anti-melanoma antibodies were selected from each library by panning the phage against live cultures of the autologous tumor. After two or three rounds of panning, clones of the phage were tested by ELISA for binding to the autologous tumor cells; > 90% of the clones tested showed a strong ELISA reaction, demonstrating the effectiveness of the panning procedure for selecting antimelanoma antibodies. The panned phage population was extensively absorbed against normal melanocytes to enrich for antibodies that react with melanoma cells but not with melanocytes. The unabsorbed phage were cloned, and the specificities of the expressed antibodies were individually tested by ELISA with a panel of cultured human cells. The first tests were done with normal endothelial and fibroblast cells to identify antibodies that do not react, or react weakly, with two normal cell types, indicating some degree of specificity for melanoma cells. The proportion of phage clones expressing such antibodies was approximately 1%. Those phage were further tested by ELISA with melanocytes, several melanoma lines, and eight other tumor lines, including a glioma line derived from glial cells that share a common lineage with melanocytes. The ELISA tests identified three classes of anti-melanoma antibodies, as follows: (i) a melanoma-specific class that reacts almost exclusively with the melanoma lines; (ii) a tumor-specific class that reacts with melanoma and other tumor lines but does not react with the normal melanocyte, endothelial and fibroblast cells; and (iii) a lineage-specific class that reacts with the melanoma lines, melanocytes, and the glioma line but does not react with the other lines. These are rare classes from the immunized patients' repertoires of anti-melanoma antibodies, most of which are relatively nonspecific anti-self antibodies. The melanoma-specific class was isolated from one patient, and the lineage-specific class was isolated from the other patient, indicating that different patients can have markedly different responses to the same immunization protocol. The procedures described here can be used to screen the antibody repertoire of any person with cancer, providing access to an enormous untapped pool of human monoclonal anti-tumor antibodies with clinical and research potential.

Persistent high frequency of human immunodeficiency virus-specific cytotoxic T cells in peripheral blood of infected donors.

Human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTLs) are thought to play a major role in the immune response to HIV infection. The HIV-specific CTL response is much stronger than previously documented in an infectious disease, yet estimates of CTL frequency derived from limiting-dilution analysis (LDA) are relatively low and comparable to other viral infections. Here we show that individual CTL clones specific for peptides from HIV gag and pol gene products are present at high levels in the peripheral blood of three infected patients and that individual CTL clones may represent between 0.2% and 1% of T cells. Previous LDA in one donor had shown a frequency of CTL precursors of 1/8000, suggesting that LDA may underestimate CTL effector frequency. In some donors individual CTL clones persisted in vivo for at least 5 years. In contrast, in one patient there was a switch in CTL usage suggesting that different populations of CTLs can be recruited during infection. These data imply strong stimulation of CTLs, potentially leading some clones to exhaustion.