A PRESEN��A DA TEOLOGIA SACRAMENTAL ZUINGLIANA NO PRESBITERIANISMO BRASILEIRO E SUAS INFLU��NCIAS LIT��RGICAS. UMA PERSPECTIVA ECUM��NICA

Ulrico Zu��nglio, que iniciou a Reforma protestante na Su����a, rompeu com uma concep����o antiga dos sacramentos, n��o os considerando meios de gra��a. Os sacramentos do batismo e da Ceia do Senhor, mantidos pelo reformador, recebem novos enfoques, em particular a Ceia, ao ser considerada apenas uma recorda����o do sacrif��cio de Cristo na cruz. A ruptura deu-se tamb��m na dissocia����o da Ceia do Senhor do Dia do Senhor. Jo��o Calvino, reformador de Genebra e considerado o principal te��logo da chamada Tradi����o Reformada, manteve a interpreta����o dos sacramentos como meios de gra��a e defendeu, embora sem sucesso, a celebra����o dominical da Ceia do Senhor. A tese demonstra a presen��a dominante da concep����o de Zu��nglio nas denomina����es presbiterianas brasileiras e suas implica����es lit��rgicas, a despeito de essas igrejas considerarem-se calvinistas. Inclui um estudo da media����o norte-americana na teologia dos sacramentos, tendo em vista que o presbiterianismo implantou-se definitivamente no Brasil, no s��culo XIX, atrav��s de mission��rios procedentes dos Estados Unidos. Esses mission��rios consideraram a popula����o cat��lica brasileira como principal alvo de evangeliza����o, praticando o rebatismo de conversos, pr��tica esta que se tornaria usual no presbiterianismo brasileiro. Aspectos da liturgia de igrejas presbiterianas brasileiras, como a celebra����o da Ceia do Senhor de modo informal e mesmo, por vezes, com omiss��o da Ora����o Eucar��stica ou das palavras da institui����o do sacramento, a presen��a ou n��o de elementos cl��ssicos do culto, como o Credo Apost��lico ou Niceno e a Ora����o do Senhor, constitu��ram-se objeto da presente pesquisa in loco. Algumas tentativas de recupera����o de elementos da tradi����o lit��rgica encontram resist��ncias no presbiterianismo brasileiro, seja em raz��o de uma identidade negativa , o anticatolicismo, seja pelo atual contexto cultural de relativiza����o de valores em detrimento de marcos hist��ricos de identidade crist��. A pesquisa constata que a heran��a teol��gica zuingliana, em cujo desenvolvimento intervieram novos fatores, relaciona-se a dificuldades de inser����o das principais igrejas presbiterianas brasileiras no movimento ecum��nico do s��culo XX.(AU)

A PRESEN��A DA TEOLOGIA SACRAMENTAL ZUINGLIANA NO PRESBITERIANISMO BRASILEIRO E SUAS INFLU��NCIAS LIT��RGICAS. UMA PERSPECTIVA ECUM��NICA

Ulrico Zu��nglio, que iniciou a Reforma protestante na Su����a, rompeu com uma concep����o antiga dos sacramentos, n��o os considerando meios de gra��a. Os sacramentos do batismo e da Ceia do Senhor, mantidos pelo reformador, recebem novos enfoques, em particular a Ceia, ao ser considerada apenas uma recorda����o do sacrif��cio de Cristo na cruz. A ruptura deu-se tamb��m na dissocia����o da Ceia do Senhor do Dia do Senhor. Jo��o Calvino, reformador de Genebra e considerado o principal te��logo da chamada Tradi����o Reformada, manteve a interpreta����o dos sacramentos como meios de gra��a e defendeu, embora sem sucesso, a celebra����o dominical da Ceia do Senhor. A tese demonstra a presen��a dominante da concep����o de Zu��nglio nas denomina����es presbiterianas brasileiras e suas implica����es lit��rgicas, a despeito de essas igrejas considerarem-se calvinistas. Inclui um estudo da media����o norte-americana na teologia dos sacramentos, tendo em vista que o presbiterianismo implantou-se definitivamente no Brasil, no s��culo XIX, atrav��s de mission��rios procedentes dos Estados Unidos. Esses mission��rios consideraram a popula����o cat��lica brasileira como principal alvo de evangeliza����o, praticando o rebatismo de conversos, pr��tica esta que se tornaria usual no presbiterianismo brasileiro. Aspectos da liturgia de igrejas presbiterianas brasileiras, como a celebra����o da Ceia do Senhor de modo informal e mesmo, por vezes, com omiss��o da Ora����o Eucar��stica ou das palavras da institui����o do sacramento, a presen��a ou n��o de elementos cl��ssicos do culto, como o Credo Apost��lico ou Niceno e a Ora����o do Senhor, constitu��ram-se objeto da presente pesquisa in loco. Algumas tentativas de recupera����o de elementos da tradi����o lit��rgica encontram resist��ncias no presbiterianismo brasileiro, seja em raz��o de uma identidade negativa , o anticatolicismo, seja pelo atual contexto cultural de relativiza����o de valores em detrimento de marcos hist��ricos de identidade crist��. A pesquisa constata que a heran��a teol��gica zuingliana, em cujo desenvolvimento intervieram novos fatores, relaciona-se a dificuldades de inser����o das principais igrejas presbiterianas brasileiras no movimento ecum��nico do s��culo XX.(AU)

Cell adhesion and the integrin-linked kinase regulate the LEF-1 and ��-catenin signaling pathways

The integrin-linked kinase (ILK) is an ankyrin repeat containing serine-threonine protein kinase that can interact directly with the cytoplasmic domains of the ��1 and ��3 integrin subunits and whose kinase activity is modulated by cell���extracellular matrix interactions. Overexpression of constitutively active ILK results in loss of cell���cell adhesion, anchorage-independent growth, and tumorigenicity in nude mice. We now show that modest overexpression of ILK in intestinal epithelial cells as well as in mammary epithelial cells results in an invasive phenotype concomitant with a down-regulation of E-cadherin expression, translocation of ��-catenin to the nucleus, formation of a complex between ��-catenin and the high mobility group transcription factor, LEF-1, and transcriptional activation by this LEF-1/��-catenin complex. We also find that LEF-1 protein expression is rapidly modulated by cell detachment from the extracellular matrix, and that LEF-1 protein levels are constitutively up-regulated at ILK overexpression. These effects are specific for ILK, because transformation by activated H-ras or v-src oncogenes do not result in the activation of LEF-1/��-catenin. The results demonstrate that the oncogenic properties of ILK involve activation of the LEF-1/��-catenin signaling pathway, and also suggest ILK-mediated cross-talk between cell���matrix interactions and cell���cell adhesion as well as components of the Wnt signaling pathway.

Substantial narrowing of the Niemann���Pick C candidate interval by yeast artificial chromosome���complementation

Niemann���Pick disease type C (NP-C) is an autosomal recessive lipidosis linked to chromosome 18q11���12, characterized by lysosomal accumulation of unesterified cholesterol and delayed induction of cholesterol-mediated homeostatic responses. This cellular phenotype is identifiable cytologically by filipin staining and biochemically by measurement of low-density lipoprotein-derived cholesterol esterification. The mutant Chinese hamster ovary cell line (CT60), which displays the NP-C cellular phenotype, was used as the recipient for a complementation assay after somatic cell fusions with normal and NP-C murine cells suggested that this Chinese hamster ovary cell line carries an alteration(s) in the hamster homolog(s) of NP-C. To narrow rapidly the candidate interval for NP-C, three overlapping yeast artificial chromosomes (YACs) spanning the 1 centimorgan human NP-C interval were introduced stably into CT60 cells and analyzed for correction of the cellular phenotype. Only YAC 911D5 complemented the NP-C phenotype, as evidenced by cytological and biochemical analyses, whereas no complementation was obtained from the other two YACs within the interval or from a YAC derived from chromosome 7. Fluorescent in situ hybridization indicated that YAC 911D5 was integrated at a single site per CT60 genome. These data substantially narrow the NP-C critical interval and should greatly simplify the identification of the gene responsible in mouse and man. This is the first demonstration of YAC complementation as a valuable adjunct strategy for positional cloning of a human gene.

In vivo antagonism of a T cell response by an endogenously expressed ligand

3.L2 T cell receptor transgenic T cells are activated by the 64���76 peptide of the mouse hemoglobin d �� chain [Hb(64���76)], and their response is antagonized by the position 72 alanine substitution of this peptide (A72). To test the effect of this altered peptide ligand (APL) on 3.L2 T cell function in vivo, a transgene expressing A72 in major histocompatibility complex II positive cells (A72tg) has been introduced into mice. We demonstrate that 3.L2 T cells, when transferred to A72tg+ mice show a dramatically reduced proliferative response to Hb(64���76). Identical decreased responses were observed using T cells that developed in either A72tg+ or A72tg��� hosts. This affect was not attributable to diminished precursor frequency, anergy, or competition for binding to I-Ek molecules. These results unequivocally demonstrate in vivo antagonism by an endogenous APL and characterize a class of self-peptides that, although inefficient in causing deletion in the thymus, effectively modulate T cell responses in the periphery.

Differential Expression and Functions of Cortical Myosin IIA and IIB Isotypes during Meiotic Maturation, Fertilization, and Mitosis in Mouse Oocytes and Embryos

To explore the role of nonmuscle myosin II isoforms during mouse gametogenesis, fertilization, and early development, localization and microinjection studies were performed using monospecific antibodies to myosin IIA and IIB isotypes. Each myosin II antibody recognizes a 205-kDa protein in oocytes, but not mature sperm. Myosin IIA and IIB demonstrate differential expression during meiotic maturation and following fertilization: only the IIA isoform detects metaphase spindles or accumulates in the mitotic cleavage furrow. In the unfertilized oocyte, both myosin isoforms are polarized in the cortex directly overlying the metaphase-arrested second meiotic spindle. Cortical polarization is altered after spindle disassembly with Colcemid: the scattered meiotic chromosomes initiate myosin IIA and microfilament assemble in the vicinity of each chromosome mass. During sperm incorporation, both myosin II isotypes concentrate in the second polar body cleavage furrow and the sperm incorporation cone. In functional experiments, the microinjection of myosin IIA antibody disrupts meiotic maturation to metaphase II arrest, probably through depletion of spindle-associated myosin IIA protein and antibody binding to chromosome surfaces. Conversely, the microinjection of myosin IIB antibody blocks microfilament-directed chromosome scattering in Colcemid-treated mature oocytes, suggesting a role in mediating chromosome���cortical actomyosin interactions. Neither myosin II antibody, alone or coinjected, blocks second polar body formation, in vitro fertilization, or cytokinesis. Finally, microinjection of a nonphosphorylatable 20-kDa regulatory myosin light chain specifically blocks sperm incorporation cone disassembly and impedes cell cycle progression, suggesting that interference with myosin II phosphorylation influences fertilization. Thus, conventional myosins break cortical symmetry in oocytes by participating in eccentric meiotic spindle positioning, sperm incorporation cone dynamics, and cytokinesis. Although murine sperm do not express myosin II, different myosin II isotypes may have distinct roles during early embryonic development.

Division of Labor among the ��6��4 Integrin, ��1 Integrins, and an E3 Laminin Receptor to Signal Morphogenesis and ��-Casein Expression in Mammary Epithelial Cells

Contact of cultured mammary epithelial cells with the basement membrane protein laminin induces multiple responses, including cell shape changes, growth arrest, and, in the presence of prolactin, transcription of the milk protein ��-casein. We sought to identify the specific laminin receptor(s) mediating the multiple cell responses to laminin. Using assays with clonal mammary epithelial cells, we reveal distinct functions for the ��6��4 integrin, ��1 integrins, and an E3 laminin receptor. Signals from laminin for ��-casein expression were inhibited in the presence of function-blocking antibodies against both the ��6 and ��1 integrin subunits and by the laminin E3 fragment. The ��6-blocking antibody perturbed signals mediated by the ��6��4 integrin, and the ��1-blocking antibody perturbed signals mediated by another integrin, the �� subunit(s) of which remains to be determined. Neither ��6- nor ��1-blocking antibodies perturbed the cell shape changes resulting from cell exposure to laminin. However, the E3 laminin fragment and heparin both inhibited cell shape changes induced by laminin, thereby implicating an E3 laminin receptor in this function. These results elucidate the multiplicity of cell-extracellular matrix interactions required to integrate cell structure and signaling and ultimately permit normal cell function.

Biparental Inheritance of ��-Tubulin during Human Fertilization: Molecular Reconstitution of Functional Zygotic Centrosomes in Inseminated Human Oocytes and in Cell-free Extracts Nucleated by Human Sperm

Human sperm centrosome reconstitution and the parental contributions to the zygotic centrosome are examined in mammalian zygotes and after exposure of spermatozoa to Xenopus laevis cell-free extracts. The presence and inheritance of the conserved centrosomal constituents ��-tubulin, centrin, and MPM-2 (which detects phosphorylated epitopes) are traced, as is the sperm microtubule-nucleating capability on reconstituted centrosomes. ��-Tubulin is biparentally inherited in humans (maternal >> than paternal): Western blots detect the presence of paternal ��-tubulin. Recruitment of maternal ��-tubulin to the sperm centrosome occurs after sperm incorporation in vivo or exposure to cell-free extract, especially after sperm ���priming��� induced by disulfide bond reduction. Centrin is found in the proximal sperm centrosomal region, demonstrates expected calcium sensitivity, but appears absent from the zygotic centrosome after sperm incorporation or exposure to extracts. Sperm centrosome phosphorylation is detected after exposure of primed sperm to egg extracts as well as during the early stages of sperm incorporation after fertilization. Finally, centrosome reconstitution in cell-free extracts permits sperm aster microtubule assembly in vitro. Collectively, these results support a model of a blended zygotic centrosome composed of maternal constituents attracted to an introduced paternal template after insemination.

E-cadherin induces mesenchymal-to-epithelial transition in human ovarian surface epithelium

Ovarian carcinomas are thought to arise in the ovarian surface epithelium (OSE). Although this tissue forms a simple epithelial covering on the ovarian surface, OSE cells exhibit some mesenchymal characteristics and contain little or no E-cadherin. However, E-cadherin is present in metaplastic OSE cells that resemble the more complex epithelia of the oviduct, endometrium and endocervix, and in primary epithelial ovarian carcinomas. To determine whether E-cadherin was a cause or consequence of OSE metaplasia, we expressed this cell-adhesion molecule in simian virus 40-immortalized OSE cells. In these cells the exogenous E-cadherin, all three catenins, and F-actin localized at sites of cell���cell contact, indicating the formation of functional adherens junctions. Unlike the parent OSE cell line, which had undergone a typical mesenchymal transformation in culture, E-cadherin-expressing cells contained cytokeratins and the tight-junction protein occludin. They also formed cobblestone monolayers in two-dimensional culture and simple epithelia in three-dimensional culture that produced CA125 and shed it into the culture medium. CA125 is a normal epithelial-differentiation product of the oviduct, endometrium, and endocervix, but not of normal OSE. It is also a tumor antigen that is produced by ovarian neoplasms and by metaplastic OSE. Thus, E-cadherin restored some normal characteristics of OSE, such as keratin, and it also induced epithelial-differentiation markers associated with weakly preneoplastic, metaplastic OSE and OSE-derived primary carcinomas. The results suggest an unexpected role for E-cadherin in ovarian neoplastic progression.

Comparative evaluation of the antitumor activity of antiangiogenic proteins delivered by gene transfer

Although the systemic administration of a number of different gene products has been shown to result in the inhibition of angiogenesis and tumor growth in different animal tumor models, the relative potency of those gene products has not been studied rigorously. To address this issue, recombinant adenoviruses encoding angiostatin, endostatin, and the ligand-binding ectodomains of the vascular endothelial growth factor receptors Flk1, Flt1, and neuropilin were generated and used to systemically deliver the different gene products in several different preexisting murine tumor models. Single i.v. injections of viruses encoding soluble forms of Flk1 or Flt1 resulted in ���80% inhibition of preexisting tumor growth in murine models involving both murine (Lewis lung carcinoma, T241 fibrosarcoma) and human (BxPC3 pancreatic carcinoma) tumors. In contrast, adenoviruses encoding angiostatin, endostatin, or neuropilin were significantly less effective. A strong correlation was observed between the effects of the different viruses on tumor growth and the activity of the viruses in the inhibition of corneal micropocket angiogenesis. These data underscore the need for comparative analyses of different therapeutic approaches that target tumor angiogenesis and provide a rationale for the selection of specific antiangiogenic gene products as lead candidates for use in gene therapy approaches aimed at the treatment of malignant and ocular disorders.

Purification and Characterization of Phosphoribulokinase from the Marine Chromophytic Alga Heterosigma carterae1

In this study we characterized phosphoribulokinase (PRK, EC 2.7.1.19) from the eukaryotic marine chromophyte Heterosigma carterae. Serial column chromatography resulted in approximately 300-fold purification of the enzyme. A polypeptide of 53 kD was identified as PRK by sequencing the amino terminus of the protein. This protein represents one of the largest composite monomers identified to date for any PRK. The native holoenzyme demonstrated by flow performance liquid chromatography a molecular mass of 214 �� 12.6 kD, suggesting a tetrameric structure for this catalyst. Because H. carterae PRK activity was insensitive to NADH but was stimulated by dithiothreitol, it appears that the enzyme may require a thioredoxin/ferredoxin rather than a metabolite mode of regulation. Kinetic analysis of this enzyme demonstrated Michaelis constant values of ribulose-5-phosphate (226 ��m) and ATP (208 ��m), respectively. In summary, H. carterae PRK is unique with respect to holoenzyme structure and function, and thus may represent an alternative evolutionary pathway in Calvin-cycle kinase development.

Moderately High Temperatures Inhibit Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase (Rubisco) Activase-Mediated Activation of Rubisco1

We tested the hypothesis that light activation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is inhibited by moderately elevated temperature through an effect on Rubisco activase. When cotton (Gossypium hirsutum L.) or wheat (Triticum aestivum L.) leaf tissue was exposed to increasing temperatures in the light, activation of Rubisco was inhibited above 35 and 30��C, respectively, and the relative inhibition was greater for wheat than for cotton. The temperature-induced inhibition of Rubisco activation was fully reversible at temperatures below 40��C. In contrast to activation state, total Rubisco activity was not affected by temperatures as high as 45��C. Nonphotochemical fluorescence quenching increased at temperatures that inhibited Rubisco activation, consistent with inhibition of Calvin cycle activity. Initial and maximal chlorophyll fluorescence were not significantly altered until temperatures exceeded 40��C. Thus, electron transport, as measured by Chl fluorescence, appeared to be more stable to moderately elevated temperatures than Rubisco activation. Western-blot analysis revealed the formation of high-molecular-weight aggregates of activase at temperatures above 40��C for both wheat and cotton when inhibition of Rubisco activation was irreversible. Physical perturbation of other soluble stromal enzymes, including Rubisco, phosphoribulokinase, and glutamine synthetase, was not detected at the elevated temperatures. Our evidence indicates that moderately elevated temperatures inhibit light activation of Rubisco via a direct effect on Rubisco activase.

A nuclear gene of eubacterial origin in Euglena gracilis reflects cryptic endosymbioses during protist evolution.

Genes for glycolytic and Calvin-cycle glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of higher eukaryotes derive from ancient gene duplications which occurred in eubacterial genomes; both were transferred to the nucleus during the course of endosymbiosis. We have cloned cDNAs encoding chloroplast and cytosolic GAPDH from the early-branching photosynthetic protist Euglena gracilis and have determined the structure of its nuclear gene for cytosolic GAPDH. The gene contains four introns which possess unusual secondary structures, do not obey the GT-AG rule, and are flanked by 2- to 3-bp direct repeats. A gene phylogeny for these sequences in the context of eubacterial homologues indicates that euglenozoa, like higher eukaryotes, have obtained their GAPDH genes from eubacteria via endosymbiotic (organelle-to-nucleus) gene transfer. The data further suggest that the early-branching protists Giardia lamblia and Entamoeba histolytica--which lack mitochondria--and portions of the trypanosome lineage have acquired GAPDH genes from eubacterial donors which did not ultimately give rise to contemporary membrane-bound organelles. Evidence that "cryptic" (possibly ephemeral) endosymbioses during evolution may have entailed successful gene transfer is preserved in protist nuclear gene sequences.

The Pursuit of Balanced Income An Economic Perspective on Wealth

The purpose of this thesis is to determine if households with moderate incomes have the opportunity to acquire pecuniary wealth. In an environment of increasing globalization, labor-eliminating technology, and an overburdened entitlement system, the position of wealth is necessary for households with a penchant for early retirement and particularly for those that will inevitably succumb to involuntary retirement. In these writings, qualitative and quantitative descriptions of wealth are offered; two microeconomic metrics are introduced to identify baseline wealth and to gauge proximity to this value; and strategies are devised to close the gap between these metrics. The construct of the economic intertemporal household budget was used as the basis for this research whereby conventional measures of wealth, i.e. net worth and liquid net worth, have been coupled with historical and empirical household data to define and extrapolate wealth and wealth proximity metrics. The major finding is wealth is a dynamic variable contingent upon consumption level rather than income or saving levels. And because households exercise a greater degree of autonomy over consumption, moderate earners are afforded the same opportunities as high earners. The position of wealth is necessary because it sustains consumption in the event of an unwelcomed retirement and it can assuage reliance on entitlement programs. For the majority of households, wealth is a matter of choice.