Caracterización funcional de zeb1 en células en diferenciación y metastásicas. Functional characterization of zeb1 in differentiating and metastatic cells

IDENTIFICACIÓN ZEB1 (Zinc Finger E-box Binding Homeobox) es un factor de transcripción funcionalmente asociado con la diferenciación de células como miocitos, neuronas, células de sostén y linfocitos T, además de estar involucrado en la Transición Epitelial-Mesenquimatosa (EMT) de los tumores sólidos epiteliales. Aún no se ha revelado en profundidad la participación de ZEB1 en los procesos de proliferación y diferenciación en los que participa. Estamos interesados en los mecanismos de regulación de ZEB1 y los factores que intervienen en los procesos de diferenciación y transformación celular. HIPÓTESIS 1. Las vías de señalamiento regulan el estado de fosforilación y la función de ZEB1 en la célula normal, el cual se desregularía en la célula neoplásica llevando a cambios en la función normal de ZEB1 y consecuentemente a metástasis. 2. IGF-1 es la señal que, en asociación con el supresor de tumores CCN6, juega un rol causal en la regulación de ZEB1 y esto a su vez en la metástasis del cáncer de mama. OBJETIVO GENERAL: establecer el rol funcional de ZEB1, su interrelación con otros factores y su regulación en los procesos de diferenciación y transformación celular. OBJETIVOS ESPECIFICOS (incluye Materiales y Métodos) 1. Estudiar la participación de vías de señalización sobre la función biológica de ZEB1 en células normales y neoplásicas. Analizaremos la participación de señales intracelulares en la fosforilación de ZEB1 por experimentos de ganancia/pérdida de función de la vía (por uso de inhibidores farmacologicos, mutantes silenciadoras y siRNAs), lo cual sera evaluado en EMSAs, ChIP, transfecciones, inmunofluoresc, etc. 2. Estudiar el rol de IGF-1 y CCN6 sobre la expresión y el estado de fosforilación de ZEB1 en tumores mamarios benignos, no invasivos e invasivos y metastatizantes. A) Se estudiará la expresión y localización subcelular de ZEB1 en líneas celulares de cáncer mamario y en xenotransplantes de ratón con variada expresión de CCN6. B) Investigar la relevancia de la fosforilación de ZEB1 mediada por IGF-1 en el EMT por experimentos con ganancia/pérdida de función. RESULTADOS ESPERADOS Esperamos poder delinear la/s vía/s de señalización intracelular que fosforilan ZEB1 y así conocer sobre la regulación del mismo. Podremos establecer algunas bases para entender la biología básica del cáncer de mama e identificar blancos terapéuticos. IMPORTANCIA Un amplio conocimiento de los factores de transcripción y sus vías de señalamiento es necesario para el desarrollo tanto de pruebas diagnósticas como para la identificación de nuevos blancos terapéuticos para neoplasias. De modo que resulta de gran importancia clínica determinar el rol de ZEB1, sus proteínas y vías reguladoras en el proceso de oncogénesis. El desarrollo del proyecto prevé la formación de dos tesistas. Se continuaran colaboraciones con dos grupos extranjeros y se iniciara una tercera. ZEB1 (Zinc Finger E-box Binding Homeobox) is a transcription factor involved in cell differentiation and Epithelial Mesenchymal Transition (EMT) of epithelial tumors. We are interested in the study of mechanisms of regulation (pre and post transcriptional). S.A.1. To investigate post translational mechanisms of ZEB1 regulation in normal and cancer cells. We will analyze the involvement of intracellular signals in phosphorylation of ZEB1 by gain- and lost-of-function experiments. S.A.2. A) To determine the role of IGF-1 signaling and CCN6 in regulating the expression of hypo- and hyperphosphorylated forms of ZEB1 in benign and malignant breast cell lines and in xenograft mouse models by overexpressing and inhibiting CCN6 in breast cancer cells. B) To investigate the relevance of CCN6-mediated ZEB1 phosphorylation to EMT, breast cancer invasion and metastasis. The role of CCN6 on ZEB1 phosphorylation and regulation of E-cadherin, induction of EMT, invasion and metastasis of breast cells will be investigated using gain- and loss-of-function experiments.

Assessment of Autonomic Function by Phase Rectification of RRInterval Histogram Analysis in Chagas Disease

Background:In chronic Chagas disease (ChD), impairment of cardiac autonomic function bears prognostic implications. Phase‑rectification of RR-interval series isolates the sympathetic, acceleration phase (AC) and parasympathetic, deceleration phase (DC) influences on cardiac autonomic modulation.Objective:This study investigated heart rate variability (HRV) as a function of RR-interval to assess autonomic function in healthy and ChD subjects.Methods:Control (n = 20) and ChD (n = 20) groups were studied. All underwent 60-min head-up tilt table test under ECG recording. Histogram of RR-interval series was calculated, with 100 ms class, ranging from 600–1100 ms. In each class, mean RR-intervals (MNN) and root-mean-squared difference (RMSNN) of consecutive normal RR-intervals that suited a particular class were calculated. Average of all RMSNN values in each class was analyzed as function of MNN, in the whole series (RMSNNT), and in AC (RMSNNAC) and DC (RMSNNDC) phases. Slopes of linear regression lines were compared between groups using Student t-test. Correlation coefficients were tested before comparisons. RMSNN was log-transformed. (α < 0.05).Results:Correlation coefficient was significant in all regressions (p < 0.05). In the control group, RMSNNT, RMSNNAC, and RMSNNDCsignificantly increased linearly with MNN (p < 0.05). In ChD, only RMSNNAC showed significant increase as a function of MNN, whereas RMSNNT and RMSNNDC did not.Conclusion:HRV increases in proportion with the RR-interval in healthy subjects. This behavior is lost in ChD, particularly in the DC phase, indicating cardiac vagal incompetence.

Estudos experimentais sôbre a origem do milho

1) It may seem rather strange that, in spite of the efforts of a considerable number of scientists, the problem of the origin of indian corn or maize still has remained an open question. There are no fossil remains or archaeological relics except those which are quite identical with types still existing. (Fig. 1). The main difficulty in finding the wild ancestor- which may still exist - results from the fact that it has been somewhat difficult to decide what it should be like and also where to look for it. 2) There is no need to discuss the literature since an excellent review has recently been published by MANGELSDORF and REEVES (1939). It may be sufficient to state that there are basically two hypotheses, that of ST. HILAIRE (1829) who considered Brazilian pod corn as the nearest relative of wild corn still existing, and that of ASCHERSON (1875) who considered Euchlaena from Central America as the wild ancestor of corn. Later hypotheses represent or variants of these two hypotheses or of other concepts, howewer generally with neither disproving their predecessors nor showing why the new hypotheses were better than the older ones. Since nearly all possible combinations of ideas have thus been put forward, it har- dly seems possible to find something theoretically new, while it is essential first to produce new facts. 3) The studies about the origin of maize received a new impulse from MANGELSDORF and REEVES'S experimental work on both Zea-Tripsacum and Zea-Euchlaena hybrids. Independently I started experiments in 1937 with the hope that new results might be obtained when using South American material. Having lost priority in some respects I decided to withold publication untill now, when I can put forward more concise ideas about the origin of maize, based on a new experimental reconstruction of the "wild type". 4) The two main aspects of MANGELSDORF and REEVES hypothesis are discussed. We agree with the authors that ST. HILAIRE's theory is probably correct in so far as the tunicata gene is a wild type relic gene, but cannot accept the reconstruction of wild corn as a homozygous pod corn with a hermaphroditic tassel. As shown experimentally (Fig. 2-3) these tassels have their central spike transformed into a terminal, many rowed ear with a flexible rachis, while possessing at the same time the lateral ear. Thus no explanation is given of the origin of the corn ear, which is the main feature of cultivated corn (BRIEGER, 1943). The second part of the hypothesis referring to the origin of Euchlaena from corn, inverting thus ASCHERSON's theory, cannot be accepted for several reasons, stated in some detail. The data at hand justify only the conclusion that both genera, Euchlaena and Zea, are related, and there is as little proof for considering the former as ancestor of the latter as there is for the new inverse theory. 5) The analysis of indigenous corn, which will be published in detail by BRIEGER and CUTLER, showed several very primitive characters, but no type was found which was in all characters sufficiently primitive. A genetical analysis of Paulista Pod Corn showed that it contains the same gene as other tunicates, in the IV chromosome, the segregation being complicated by a new gametophyte factor Ga3. The full results of this analysis shall be published elsewhere. (BRIEGER). Selection experiments with Paulista Pod Corn showed that no approximation to a wild ancestor may be obtained when limiting the studies to pure corn. Thus it seemed necessary to substitute "domesticated" by "wild type" modifiers, and the only means for achieving this substitution are hybridizations with Euchlaena. These hybrids have now been analysed init fourth generation, including backcrosses, and, again, the full data will be published elsewhere, by BRIEGER and ADDISON. In one present publication three forms obtained will be described only, which represent an approximation to wild type corn. 6) Before entering howewer into detail, some arguments against ST. HILAIRE's theory must be mentioned. The premendelian argument, referring to the instability of this character, is explained by the fact that all fertile pod corn plants are heterozygous for the dominant Tu factor. But the sterility of the homozygous TuTu, which phenotypically cannot be identified, is still unexplained. The most important argument against the acceptance of the Tunicata faetor as wild type relic gene was removed recently by CUTLER (not yet published) who showed that this type has been preserved for centuries by the Bolivian indians as a mystical "medicine". 7) The main botanical requirements for transforming the corn ear into a wild type structure are stated, and alternative solutions given. One series of these characters are found in Tripsacum and Euchlaena : 2 rows on opposite sides of the rachis, protection of the grains by scales, fragility of the rachis. There remains the other alternative : 4 rows, possibly forming double rows of female and male spikelets, protection of kernels by their glumes, separation of grains at their base from the cob which is thin and flexible. 8) Three successive stages in the reconstruction of wild corn, obtained experimentally, are discussed and illustrated, all characterized by the presence of the Tu gene. a) The structure of the Fl hybrids has already been described in 1943. The main features of the Tunicata hybrids (Fig. -8), when compared with non-tunicate hybrids (Fig. 5-6), consist in the absence of scaly protections, the fragility of the rachis and finally the differentiation of the double rows into one male and one female spikelet. As has been pointed out, these characters represent new phenotypic effects of the tunicate factor which do not appear in the presence of pure maize modifiers. b) The next step was observed among the first backcross to teosinte (Fig. 9). As shown in the photography, Fig. 9D, the features are essencially those of the Fl plants, except that the rachis is more teosinte like, with longer internodes, irregular four-row-arrangement and a complete fragility on the nodes. c) In the next generation a completely new type appeared (Fig. 10) which resembles neither corn nor teosinte, mainly in consequence of one character: the rachis is thin and flexible and not fragile, while the grains have an abscission layer at the base, The medium sized, pointed, brownish and hard granis are protected by their well developed corneous glumes. This last form may not yet be the nearest approach to a wild grass, and I shall try in further experiments to introduce other changes such as an increase of fertile flowers per spikelet, the reduction of difference between terminal and lateral inflorescences, etc.. But the nature of the atavistic reversion is alveadwy such that it alters considerably our expectation when looking for a still existing wild ancestor of corn. 9) The next step in our deductions must now consist in an reversion of our question. We must now explain how we may obtain domesticated corn, starting from a hypothetical wild plant, similar to type c. Of the several changes which must have been necessary to attract the attention of the Indians, the following two seem to me the most important: the disappearance of all abscission layers and the reduction of the glumes. This may have been brought about by an accumulation of mutations. But it seems much more probable to assume that some crossing with a tripsacoid grass or even with Tripsacum australe may have been responsible. In such a cross, the two types of abscission layer would be counterbalanced as shown by the Flhybrids of corn, Tripsacum and Euchlaena. Furthermore in later generations a.tu-allele of Tripsacum may become homozygous and substitute the wild tunicate factor of corn. The hypothesis of a hybrid origin of cultivated corn is not completely new, but has been discussed already by HARSHBERGER and COLLINS. Our hypothesis differs from that of MANGELSDORF and REEVES who assume that crosses with Tripsacum are responsible only for some features of Central and North American corn. 10) The following arguments give indirects evidence in support of our hypothesis: a) Several characters have been observed in indigenous corn from the central region of South America, which may be interpreted as "tripsacoid". b) Equally "zeoid" characters seem to be present in Tripsacum australe of central South-America. c) A system of unbalanced factors, combined by the in-tergeneric cross, may be responsible for the sterility of the wild type tunicata factor when homozygous, a result of the action of modifiers, brought in from Tripsacum together with the tuallele. d) The hybrid theory may explain satisfactorily the presence of so many lethals and semilethals, responsible for the phenomenon of inbreeding in cultivated corn. It must be emphasized that corn does not possess any efficient mechanism to prevent crossing and which could explain the accumulation of these mutants during the evolutionary process. Teosinte which'has about the same mechanism of sexual reproduction has not accumulated such genes, nor self-sterile plants in spite of their pronounced preference for crossing. 11) The second most important step in domestication must have consisted in transforming a four rowed ear into an ear with many rows. The fusion theory, recently revived byLANGHAM is rejected. What happened evidently, just as in succulent pXants (Cactus) or in cones os Gymnosperms, is that there has been a change in phyllotaxy and a symmetry of longitudinal rows superimposed on the original spiral arrangement. 12) The geographical distribution of indigenous corn in South America has been discussed. So far, we may distinguish three zones. The most primitive corn appears in the central lowlands of what I call the Central Triangle of South America: east of the Andies, south of the Amazone-Basin, Northwest of a line formed by the rivers São Prancisco-Paraná and including the Paraguay-Basin. The uniformity of the types found in this extremely large zone is astonishing (BRIEGER and CUTLER). To the west, there is the well known Andian region, characterized by a large number of extremely diverse types from small pop corn to large Cuszco, from soft starch to modified sweet corn, from large cylindrical ears to small round ears, etc.. The third region extends along the atlantic coast in the east, from the Caribean Sea to the Argentine, and is characterized by Cateto, an orange hard flint corn. The Andean types must have been obtained very early, and undoubtedly are the result of the intense Inca agriculture. The Cateto type may be obtained easily by crosses, for instance, of "São Paulo Pointed Pop" to some orange soft corn of the central region. The relation of these three South American zones to Central and North America are not discussed, and it seems essential first to study the intermediate region of Ecuador, Colombia and Venezuela. The geograprical distribution of chromosome knobs is rapidly discussed; but it seems that no conclusions can be drawn before a large number of Tripsacum species has been analysed.

Comportamento dos cromossômios na meiose de Euryophthalmus rufipennis Laporte (Hemiptera Pyrrhocoridae)

In this paper an account is given of the principal facts observer in the meiosis of Euryophthalmus rufipennis Laporte which afford some evidence in favour of the view held by the present writer in earlier publications regarding the existence of two terminal kinetochores in Hem ip ter an chromosomes as well as the transverse division of the chromosomes. Spermatogonial mitosis - From the beginning of prophase until metaphase nothing worthy of special reference was observed. At anaphase, on the contrary, the behavior of the chromosomes deserves our best attention. Indeed, the chromoso- mes, as soon as they begin to move, they show both ends pronouncedly turned toward the poles to which they are connected by chromosomal fibres. So a premature and remarkable bending of the chromosomes not yet found in any other species of Hemiptera and even of Homoptera points strongly to terminally localized kinetochores. The explanation proposed by HUGHES-SCHRADER and RIS for Nautococcus and by RIS for Tamalia, whose chromosomes first become bent late in anaphase do not apply to chromosomes which initiate anaphase movement already turned toward the corresponding pole. In the other hand, the variety of positions assumed by the anaphase chromosomes of Euryophthalmus with regard to one another speaks conclusively against the idea of diffuse spindle attachments. First meiotic division - Corresponding to the beginning of the story of the primary spermatocytes cells are found with the nucleus entirelly filled with leptonema threads. Nuclei with thin and thick threads have been considered as being in the zygotente phase. At the pachytene stage the bivalents are formed by two parallel strands clearly separated by a narrow space. The preceding phases differ in nothing from the corresponding orthodox ones, pairing being undoubtedly of the parasynaptic type. Formation of tetrads - When the nuclei coming from the diffuse stage can be again understood the chromosomes reappear as thick threads formed by two filaments intimately united except for a short median segment. Becoming progressively shorter and thicker the bivalents sometimes unite their extremities forming ring-shaped figures. Generally, however, this does not happen and the bivalents give origin to more or less condensed characteristic Hemipteran tetrads, bent at the weak median region. The lateral duplicity of the tetrads is evident. At metaphase the tetrads are still bent and are connected with both poles by their ends. The ring-shaped diakinesis tetrads open themselves out before metaphase, showing in this way that were not chiasmata that held their ends together. Anaphase proceeds as expected. If we consider the median region of the tetrads as being terminalized chiasmata, then the chromosomes are provided with a single terminal kinetochore. But this it not the case. A critical analysis of the story of the bivalents before and after the diffuse stage points to the conclusion that they are continuous throughout their whole length. Thence the chromosomes are considered as having a kinetochore at each end. Orientation - There are some evidences that Hemipteran chromosomes are connected by chiasmata. If this is true, the orientation of the tetrads may be understood in the following manner: Chiasmata being hindered to scape by the terminal kinetochores accumulate at the ends of the tetrads, where condensation begins. Repulsion at the centric ends being prevented by chiasmata the tetrads orient themselves as if they were provided with a single kinetochore at each extremity, taking a position parallelly to the spindle axis. Anaphase separation - Anaphase separation is consequently due to a transverse division of the chromosomes. Telophase and secund meiotic division - At telophase the kinetochore repeli one another following the moving apart of the centosomes, the chiasmata slip toward the acentric extremities and the chromosomes rotate in order to arrange themselves parallelly to the axis of the new spindle. Separation is therefore throughout the pairing plane. Origin of the dicentricity of the chromosomes - Dicentricity of the chromosomes is ascribed to the division of the kinetochore of the chromosomes reaching the poles followed by separation and distension of the chromatids which remain fused at the acentric ends giving thus origin to terminally dicentric iso-chromosomes. Thence, the transverse division of the chromosomes, that is, a division through a plane perpendicular to the plane of pairing, actually corresponds to a longitudinal division realized in the preceding generation. Inactive and active kinetochores - Chromosomes carrying inactive kinetochore is not capable of orientation and active anaphasic movements. The heterochromosome of Diactor bilineatus in the division of the secondary spermatocytes is justly in this case, standing without fibrilar connection with the poles anywhere in the cell, while the autosomes are moving regularly. The heterochromosome of Euryophthalmus, on the contrary, having its kinetochores perfectly active ,is correctly oriented in the plane of the equator together with the autosomes and shows terminal chromosomal connection with both poles. Being attracted with equal strength by two opposite poles it cannot decide to the one way or the other remaining motionless in the equator until some secondary causes (as for instances a slight functional difference between the kinetochores) intervene to break the state of equilibrium. When Yiothing interferes to aide the heterochromosome in choosing its way it distends itself between the autosomal plates forming a fusiform bridge which sometimes finishes by being broken. Ordinarily, however, the bulky part of the heterochromosome passes to one pole. Spindle fibers and kinetic activity of chromosomal fragments - The kinetochore is considered as the unique part of the chromosome capable of being influenced by other kinetochore or by the poles. Under such influence the kinetochore would be stimulated or activited and would elaborate a sort of impulse which would run toward the ends. In this respect the chromosome may be compared to a neüròn, the cell being represented by the kinetochore and the axon by the body of the chromosome. Due to the action of the kinetochore the entire chromosome becomes also activated for performing its kinetic function. Nothing is known at present about the nature of this activation. We can however assume that some active chemical substance like those produced by the neuron and transferred to the effector passes from the kinetochore to the body of the chromosome runing down to the ends. And, like an axon which continues to transmit an impulse after the stimulating agent has suspended its action, so may the chromosome show some residual kinetic activity even after having lost its kinetochore. This is another explanation for the kinetic behavior of acentric chromosomal fragmehs. In the orthodox monocentric chromosomes the kinetic activity is greater at the kinetochore, that is, at the place of origin of the active substance than at any other place. In chromosomes provided with a kinetochore at each end the entire body may become active enough to produce chromosomal fibers. This is probably due to a more or less uniform distribution and concentration of the active substance coming simultaneously from both extremities of the chromosome.

Estudos sobre a alimentação mineral do cafeeiro: VI Efeitos das deficiências de micronutrientes em Coffea arabica L. var. Mundo Novo cultivado em solução nutritiva

Young coffee plants (Coffea arabica L., var. Mundo Novo) were grown in nutrient solution purified from micronutrients contaminants by the method of MUNNS & JOHNSON (1960). All plants, except those in the control treatment, wer given all macronutrients and all micronutrients except one which was omitted in order to induce its shortage. Symptoms of deficiency were obtained for all known micronutrients but chlorine. Measurements, observations and chemical analysis of leaves allowed the following main conclusions to be drawn. 1. The relative influence of micronutrients in growth-measured by the fresh weight of the entire plant - was as follows: -Fe -Zn -Cu -Mo -Mn complete = -B = -CI. that is: the omission of iron from the nutrient solution caused the severest reduction in growth; lack of B and Cl had no effect. 2. Symptoms of deficiency of B, Fe, Mn, and Zn were found to be in good agreement with those in the literature. Effects of Cu and Mo shortage, however, had not been described so far: In the case of the Cu-deficient plants, the younger leaves were distorted, having an "S" shape, due probably to lack of growth of the veins; they lost their green color and developed rather large, necrotic patches near the margins. When molybdenum was omitted from the nutrient solution yellow spots develop near the margen of subterminal (fully mature) leaves; they became necrotic; there was a characteristic downward curling of the leaf blade along the mid rib so that the opposite edges touched each other underneath. 3. The levels of micronutrients found in normal and deficient leaves are given in Table 4. It is hoped that those values will serve as a basis of judgement of micronutrient contents found in leaves of field grown plants.

The rediscovery of Stenogeocoris horvathi Montandon (Heteroptera, Geocoridae) in Argentina

ABSTRACT Stenogeocoris Montandon, 1913 was described to include S. horvathi based in one specimen from Córdoba Province, Argentina. The type specimen is lost and there are no records or additional material since the Montandon's description; thus, the identity of this taxon has remained uncertain until now. In this contribution, we redescribe the genus Stenogeocoris and the species S. horvathi, based on male and female specimens, including characters from the male genitalia, and compare Stenogeocoris with the other Neotropical genera

Polarization in the chilean party system: changes and continuities, 1990-1999

The focus of this thesis is the evolution of programmatic polarization in the post-authoritarian Chilean party system at the elite level. It shows the distance/proximity between parties located along the left-right ideological continuum on three sets of issues. The paper demonstrates that important changes have taken place in the meaning of the right and, especially, left poles. This implies convergence on socio-economic issues between parties, but persistence of differences on religious-value issues, and on issues related to the authoritarian/democratic cleavage. Distance between the poles has been reduced, and as a result the center has lost its own political space. In addition, the paper shows that the pattern followed by programmatic polarization at the elite level is explained by the authoritarian experience, the institutional framework, and socio-economic transformations. Together with this factors, the degree of negotiability of the issues and the cross-cutting nature of the cleavages have also shaped polarization.

Where Has the Old Debate Gone? Realism, Institutionalism, and IR Theory

This paper resorts to the contribution of the science philosopher Gerald Holton to map some of the IR arguments and debates in an unconventional and more insightful way. From this starting point, it is sustained that the formerly all-pervading neorealism-neoinstitutionalism debate has lost its appeal and is attracting less and less interest among scholars. It does not structure the approach of the theoretically-oriented authors any more; at least, not with the habitual intensity. More specifically, we defend that the neo-neo rapprochement, even if it could have demonstrated that international cooperation is possible and relevant in a Realist world, it has also impoverished theoretical debate by hiding some of the most significant issues that preoccupied classical transnationalists. Hence, some authors appear to be trying to rescue some of these arguments in an analytical and systematic fashion, opening up a theoretical querelle that may be the next one to pay attention to.

Trypanosoma cruzi: antigen-receptor mediated endocytosis of antibody

Trypanomastigote forms of Trypanosoma cruzi were derived from tissue culture and incubated with immune and non-immune human sera. All immune sera showed high titers of specific humoral antibodies of the IgM or the IgG type. Agglutination and swelling of parasites were observed after incubation at 37ºC, but many trypomastigotes remained free-swimming in the sera for two to three days. The quantitiy of immune serum capable of lysing a maximum of 10 x 10 [raised to the power of 6] sensitized red cells was not capable of lysing 4 x 10 [raised to the power of 3] tripomastigotes. Typically, the parasites underwent cyclical changes with the formation of clumps of amastigotes and the appearance of epimastigote forms. Multiplication of the parasites was observed in immune sera. Further, the infectivity of the parasites to susceptible mice was not lost. All sera used produced similar general effects on the growth of the parasite. The antibody bound to T. cruzi appeard to enter cells by antigen-receptor mediated endocytosis. The ferritin-conjugated antibody was internalized and delivered to phagolysosomes where they might be completely degraded to amino-acids. This seemed to be a coupled process by which the immunoglobulin is first bound to specific parasite surface receptor and then rapidly endocytosed by the cell.

Cultured human fetal astrocytes can be induced by interferon-gamma to express HLA-DR.

Interferon-gamma (IFN-gamma) modulates the expression of Class II major histocompatibility antigens (MHC), thus providing a potential regulatory mechanism for local immune reactivity in the context of MHC-restricted antigen presentation. Within the central nervous system (CNS), the expression of MHC Class II antigens has been demonstrated on human reactive astrocytes and glioma cells. In order to investigate the modulation of HLA-DR on normal astrocytes, two cell lines were grown from a 20-week-old fetal brain. In situ none of the fetal brain cells expressed HLA-DR as determined by immunohistology on frozen tissue sections. The two cell lines, FB I and FB II, expressed GFAP indicating their astrocytic origin. FB I was HLA-DR negative at the first tissue culture passages, but could be induced to express HLA-DR when treated with 500 U/ml IFN-gamma. FB II was spontaneously HLA-DR positive in the early passages, lost the expression of this antigen after 11 passages and could also be induced to express HLA-DR by IFN-gamma. The induction of HLA-DR expression was demonstrated both by a binding RIA and by immunoprecipitation using a monoclonal antibody (MAB) directed against a monomorphic determinant of HLA-DR. The HLA-DR alloantigens were determined on FB II cells after IFN-gamma treatment, by immunofluorescence and by cytotoxicity assays, and were shown to be DR4, DR6, Drw52, DRw53 and DQwl. These results show that human fetal astrocytes can be induced to express HLA-DR by IFN-gamma in vitro and support the concept that astrocytes may function as antigen-presenting cells.

Donor-derived aspergillosis from use of a solid organ recipient as a multiorgan donor.

The growing need for organs and the scarcity of donors has resulted in an increased use of extended criteria donors. We report a case where a recipient of a cardiac graft was used as an organ donor. Death of the recipient occurred 9 days after transplantation and was attributed to presumed cerebral hemorrhage, which post mortem was diagnosed as invasive aspergillosis of the brain. One recipient of a kidney transplant lost the graft due to infection with Aspergillus fumigatus, whereas prompt initiation of therapy successfully prevented disseminated aspergillosis in the other recipients. Despite the pressure to extend the use of organs by lowering the acceptance criteria, organs should only be accepted if the cause of death of the donors is unequivocally explained.

Stem Cell Transplantation for Retinal Degenerations

Within the last few years, several reports have revealed that cell transplantation can be an effective way to replace lost neurons in the central nervous system (CNS) of patients affected with neurodegenerative diseases. Concerning the retina, the concept that newborn photoreceptors can integrate the retina and restore some visual functions was univocally demonstrated recently in the mouse eye (MacLaren et al. 2006) and remains to be achieved in human. These results pave the way to a standard approach in regenerative medicine aiming to replace lost photoreceptors. With the discovery of stem cells a great hope has appeared towards elaborating protocols to generate adequate cells to restore visual function in different retinal degeneration processes. Retinal stem cells (RSCs) are good candidates to repair the retina and are present throughout the retina development, including adulthood. However, neonatal mouse RSCs derived from the radial glia population have a different potential to proliferate and differentiate in comparison to adult RSCs. Moreover, we observed that adult mouse RSCs, depending on the culture conditions, have a marked tendency to transform, whereas neonatal RSCs show subtle chromosome abnormalities only after extensive expansion. These characteristics should help to identify the optimal cell source and culture conditions for cell transplantation studies. These results will be discussed in light of other studies using RSCs as well as embryonic stem cells. Another important factor to consider is the host environment, which plays a crucial role for cell integration and which was poorly studied in the normal and the diseased retina. Nonetheless, important results were recently generated to reconsider cell transplantation strategy. Perspectives to enhance cell integration by manipulating the environment will also be presented.

Intraseasonal variation in immune defence, body mass and hematocrit in adult house martins Delichon urbica

The intensity of parasite infections often increases during the reproductive season of the host as a result of parasite reproduction, increased parasite transmission and increased host susceptibility. We report within-individual variation in immune parameters, hematocrit and body mass in adult house martins Delichon urbica rearing nestlings in nests experimentally infested with house martin bugs Oeciacus hirundinis and birds rearing nestlings in initially parasite-free nests. From first to second broods body mass and hematocrit of breeding adult house martins decreased. In contrast leucocytes and immunoglobulins became more abundant. When their nests were infested with ectoparasites adults lost more weight compared with birds raising nestlings in nests treated with pyrethrin, whereas the decrease in hematocrit was more pronounced during infection with blood parasites. Neither experimental infestation with house martin bugs nor blood parasites had a significant effect on the amount of immune defences.

Albuterol delivery in an in vitro pediatric ventilator lung model: comparison of jet, ultrasonic, and mesh nebulizers.

OBJECTIVE: : To determine the influence of nebulizer types and nebulization modes on bronchodilator delivery in a mechanically ventilated pediatric lung model. DESIGN: : In vitro, laboratory study. SETTING: : Research laboratory of a university hospital. INTERVENTIONS: : Using albuterol as a marker, three nebulizer types (jet nebulizer, ultrasonic nebulizer, and vibrating-mesh nebulizer) were tested in three nebulization modes in a nonhumidified bench model mimicking the ventilatory pattern of a 10-kg infant. The amounts of albuterol deposited on the inspiratory filters (inhaled drug) at the end of the endotracheal tube, on the expiratory filters, and remaining in the nebulizers or in the ventilator circuit were determined. Particle size distribution of the nebulizers was also measured. MEASUREMENTS AND MAIN RESULTS: : The inhaled drug was 2.8% ± 0.5% for the jet nebulizer, 10.5% ± 2.3% for the ultrasonic nebulizer, and 5.4% ± 2.7% for the vibrating-mesh nebulizer in intermittent nebulization during the inspiratory phase (p < 0.01). The most efficient nebulizer was the vibrating-mesh nebulizer in continuous nebulization (13.3% ± 4.6%, p < 0.01). Depending on the nebulizers, a variable but important part of albuterol was observed as remaining in the nebulizers (jet and ultrasonic nebulizers), or being expired or lost in the ventilator circuit (all nebulizers). Only small particles (range 2.39-2.70 µm) reached the end of the endotracheal tube. CONCLUSIONS: : Important differences between nebulizer types and nebulization modes were seen for albuterol deposition at the end of the endotracheal tube in an in vitro pediatric ventilator-lung model. New aerosol devices, such as ultrasonic and vibrating-mesh nebulizers, were more efficient than the jet nebulizer.

Les juifs et l'idée de Bildung dans l'Allemagne de culture protestante : chronique d'un mésamour à travers le long XIXe siècle

Des années 1780, quand surgit la question de l'émancipation des juifs, à la Première Guerre mondiale, qui dément l'optimisme de la perfectibilité du genre humain cultivé par la Bildung, le « long » XIXe siècle est la scène sur laquelle se déploient les efforts d'intégration des juifs dans la société et la culture allemandes, où la Bildung, intimement liée à l'esprit du protestantisme allemand qui l'a profondément marquée de son empreinte, tient lieu de médiation. Fil conducteur de ma recherche, la Bildung me permet de montrer en quoi son idéal est devenu un élément constitutif de l'identité des juifs allemands, en même temps qu'il cesse, sous les effets de la nationalisation d'une culture allemande devenue un outil au service d'un peuple particulier, d'être le projet, certes d'une communauté donnée, mais porteur d'universalisme. De fait, tout en adhérant à sa définition originale, les juifs ont su réinterpréter l'idée de Bildung en désamorçant l'alliance entre culture, germanité et nationalisme, afin de construire une nouvelle identité judéo-allemande qui réponde aux enjeux et aux exigences de la modernité ainsi qu'aux évolutions du temps, tout en visant à la reconnaissance des valeurs et du statut du judaïsme. Dans la mesure où cet idéal de la Bildung, sous les coups du nationalisme allemand, a perdu sa portée universelle pour, dans un processus de germanisation, devenir un instrument au service du projet nationaliste, les juifs vont progressivement se voir exclus de la nation allemande, quand bien même ou précisément parce qu'ils se sont identifiés à tel point au projet initial de la Bildung qu'ils en sont devenus les garants. From the 1780s, when the question of the emancipation of the Jews emerged, until World War I-a disappointment for those who were optimistic about cultivating a perfected humanity through Bildung (education)-the "long" nineteenth century is the stage on which the efforts to integrate the Jews into German society and culture took place. In this context, Bildung, which was decidedly bound to and profoundly marked by the German Protestant spirit, served as mediation. The underlying theme of Bildung in my research enables me to show how its ideal became the constitutive element of German Jewish identity. Concurrently, under the effects of the nationalization of German culture that became a tool in the service of a specific folk, the ideal of Bildung ceased to be a project that conveyed universal meaning. In fact, although the Jewish people agreed with its original definition, they succeeded in reinterpreting the idea of Bildung by neutralizing the alliance between culture, being German, and nationalism in order to elaborate a new German-Jewish identity in reply to the challenges and requirements of modernity and the evolution of society while still recognizing the values and status of Judaism. Inasmuch as the ideal of Bildung lost its universal significance for serving the nationalist project under the influence of German nationalism, the Jews were gradually excluded from the German folk, which took place despite, or precisely because, they identified to such an extent with the original aims of Bildung that they became the guarantors for it. Das ,,lange" 19. Jahrhundert bildet die Kulisse der Integrationsbemühungen der Juden in die deutsche Gesellschaft und Kultur, von den 1780er Jahren, als die Frage nach der Judenemanzipation zutage kommt, bis zum Ersten Weltkrieg, der den Optimismus der menschlichen Verbesserungsfahigkeit durch die Bildung widerlegt. Die mit dem Geist des deutschen Protestantismus eng verbundene Bildung dient hier als Mediation. Der rote Faden der Bildung ermöglicht mir zu zeigen, inwiefern ihr Ideal wesentlich für die jüdische Identität geworden ist. Zur gleichen Zeit hat das Bildungsideal, unter der Wirkung der Nationalisierung der deutschen Kultur, die zum Werkzeug eines eigenartigen Volkes gemacht wurde, sein universales Wesen verloren. In der Tat, obwohl die Juden dem ursprünglichen Bildungsideal zustimmten, haben sie die Bildung neu interpretiert, indem sie die Verbindung zwischen Kultur, Germanentum und Nationalismus entschärft und eine neue deutsch-jüdische Identität gebildet haben, die den Herausforderungen und den Ansprüchen der Moderne sowie dem Gesellschaftswandel entsprach und gleichzeitig darauf abzielte, die Werte und den Status des Judentums zu anerkennen. Insoweit, als das Bildungsideal seine universale Geltung unter dem Einfluss des deutschen Nationalismus verloren hat, um den nationalistischen Absichten zu dienen, wurden die Juden nach und nach vom deutschen Volk ausgeschlossen, selbst wenn oder gerade weil sie sich dermassen mit dem ursprünglichen Zweck der Bildung identifiziert haben, dass sie ihre Garanten geworden sind.