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

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

Caracterização etnopedológica de Planossolos utilizados em cerâmica artesanal no Agreste Paraibano

Técnicas adaptadas da etnociência clássica foram utilizadas para descrever e avaliar os conhecimentos de um grupo de artesãos camponeses do Agreste Paraibano sobre alguns solos que eles usam como recurso cerâmico. Cinco perfis de solo foram descritos por agrônomos-pesquisadores (abordagem eticista) e por camponeses (abordagem emicista), em locais onde a população local extrai material para cerâmica. Amostras coletadas em ambas as abordagens foram usadas para caracterização morfológica e analítica desses solos. Os camponeses pesquisados foram capazes de distinguir, identificar e nomear diversos materiais de solo, arranjados em estratos ao longo dos perfis de solo, de modo comparável ao arranjo dos horizontes pedogenéticos. A visão, o tato e o paladar foram empregados pelos artesãos na avaliação da qualidade do solo para cerâmica. Quatro perfis descritos junto às fontes de material cerâmico foram classificados como Planossolo Nátrico e um como Planossolo Háplico. A realização de pesquisas etnopedológicas em diferentes contextos sociais e pedológicos pode contribuir para o avanço da ciência do solo, sendo também uma oportunidade para melhor compreender as formas camponesas de conhecimento e manejo de solos.

[Lettre autographe signée de Giuseppina Verdi adressée à Escudier, Rome, 20 janvier 1859]

En français, post-scriptum de Giuseppe Verdi

[Lettre autographe signée de Giuseppina Verdi adressée à Escudier, 24 décembre 1859]

Post-scriptum de Giuseppe Verdi

Role of peroxynitrite in the redox regulation of cell signal transduction pathways.

Peroxynitrite is a potent oxidant and nitrating species formed from the reaction between the free radicals nitric oxide and superoxide. An excessive formation of peroxynitrite represents an important mechanism contributing to cell death and dysfunction in multiple cardiovascular pathologies, such as myocardial infarction, heart failure and atherosclerosis. Whereas initial works focused on direct oxidative biomolecular damage as the main route of peroxynitrite toxicity, more recent evidence, mainly obtained in vitro, indicates that peroxynitrite also behaves as a potent modulator of various cell signal transduction pathways. Due to its ability to nitrate tyrosine residues, peroxynitrite affects cellular processes dependent on tyrosine phosphorylation. Peroxynitrite also exerts complex effects on the activity of various kinases and phosphatases, resulting in the up- or downregulation of signalling cascades, in a concentration- and cell-dependent manner. Such roles of peroxynitrite in the redox regulation of key signalling pathways for cardiovascular homeostasis, including protein kinase B and C, the MAP kinases, Nuclear Factor Kappa B, as well as signalling dependent on insulin and the sympatho-adrenergic system are presented in detail in this review.

PEASANT AND SCIENTIFIC KNOWLEDGE ON PLANOSOLS AS A SOURCE OF MATERIALS IN THE MAKING OF NON-INDUSTRIAL POTTERY

Ethnopedological studies have mainly focused on agricultural land uses and associated practices. Nevertheless, peasant and indigenous populations use soil and land resources for a number of additional purposes, including pottery. In the present study, we describe and analyze folk knowledge related to the use of soils in non-industrial pottery making by peasant potters, in the municipality of Altinho, Pernambuco State, semiarid region at Brazil. Ethnoscientific techniques were used to record local knowledge, with an emphasis on describing the soil materials recognized by the potters, the properties they used to identify those soil materials, and the criteria employed by them to differentiate and relate such materials. The potters recognized three categories of soil materials: “terra” (earth), “barro” (clay) and, “piçarro” (soft rock). The multi-layered arrangement of these materials within the soil profiles was similar to the arrangement of the soil horizon described by formal pedologists. “Barro vermelho” (red clay) was considered by potters as the principal ceramic resource. The potters followed morphological and utilitarian criteria in distinguishing the different soil materials. Soils from all of these sites were sodium-affected Alfisols and correspond to Typic Albaqualf and Typic Natraqualf in the Soil Taxonomy (Soil Survey Staff, 2010).

Jouer au théâtre. Le rythme de l'expression

Qu'est-ce qui se joue entre des personnes qui écrivent un même livre, entre un maître et des élèves, entre un élève et ses maîtres, entre un dispositif et une performance, entre les joueurs d'une pièce de théâtre, entre les acteurs d'une photographie ? En explorant chacune de ces questions, ce livre est tout à la fois une manière de jouer et de répondre collectivement à la question : comment s'accorde-t-on quand on joue ? Lorsque des acteurs singuliers font quelque chose ensemble, ils ne font pas tous exactement la même chose. On cherche un accord pratique : on se donne la réplique.

Electronic structure of a neutral oxygen vacancy in SrTiO3

The electronic structure of an isolated oxygen vacancy in SrTiO3 has been investigated with a variety of ab initio quantum mechanical approaches. In particular we compared pure density functional theory (DFT) approaches with the Hartree-Fock method, and with hybrid methods where the exchange term is treated in a mixed way. Both local cluster models and periodic calculations with large supercells containing up to 80 atoms have been performed. Both diamagnetic (singlet state) and paramagnetic (triplet state) solutions have been considered. We found that the formation of an O vacancy is accompanied by the transfer of two electrons to the 3d(z2) orbitals of the two Ti atoms along the Ti-Vac-Ti axis. The two electrons are spin coupled and the ground state is diamagnetic. New states associated with the defect center appear in the gap just below the conduction band edge. The formation energy computed with respect to an isolated oxygen atom in the triplet state is 9.4 eV.

Development of Elispot and CFSE flow cytometric assays for detecting beryllium sensitivity

Background: Beryllium sensitization (BeS) is caused by exposure to beryllium in the workplace and may progress to chronic beryllium disease (CBD). This granulomatous lung disorder mimicks sarcoidosis clinically, but is characterized by beryllium specific CD4+ T-cells immune response. BeS is classically detected by beryllium lymphocyte proliferation test (BeLPT), but this assay requires radioactivity and is not very sensitive. In the context of a study aiming to evaluate if CBD patients are misdiagnosed as sarcoidosis patients in Switzerland, we developed EliSpot and CFSE beryllium flow cytometric test. Methods: 23 patients considered as having sarcoidosis (n = 21), CBD (n = 1) and possible CBD (n = 1) were enrolled. Elispot was performed using plate covered with gamma-IFN mAb. Cells were added to wells and incubated overnight at 37 °C with medium (neg ctrl), SEB (pos ctrl) or BeSO4 at 1, 10 and 100 microM. Anti-IFN-gamma biotinylated mAb were added and spots were visualized using streptavidinhorseradish peroxidase and AEC substrate reagent. Results were reported as spot forming unit (SFU). For Beryllium specific CFSE flow cytometry analysis, CFSE labelled cells were cultured in the presence of SEB and 1, 10 or 100 microM BeSO4. Unstimulated CFSE labeled cells were defined as controls. The cells were incubated for 6 days at 37 °C and 5% CO2. Surface labelling of T-lymphocytes and vivid as control of cells viability was performed at the time of harvest. Results: Using EliSpot technology, we were able to detect a BeS in 1/23 enrolled patients with a mean of 780 SFU (cut off value at 50 SFU). This positive result was confirmed using different concentration of BeSO4. Among the 23 patients tested, 22 showed negative results with EliSpot. Using CFSE flow cytometry, 1/7 tested patients showed a positive result with a beryllium specific CD4+ count around 30% versus 45% for SEB stimulation as positif control and 0.6 % for negative control. This patient was the one with a positive EliSpot assay. Conclusions: The preliminary data demonstrated the feasibility of Elispot and CFSE flow cytometry to detect BeS. The patient with a beryllium specific positive EliSpot and CFSE flow cytometry result had been exposed to beryllium at her workplace 20 years ago and is still regularly controlled for her pulmonary status. A positive BeLPT had already been described in 2001 in France for this patient. Further validation of these techniques are in progress.

Dominant TNF-α(+) Mycobacterium tuberculosis-specific CD4(+) T cell responses discriminate between latent infection and active disease.

Rapid diagnosis of active Mycobacterium tuberculosis (Mtb) infection remains a clinical and laboratory challenge. We have analyzed the cytokine profile (interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2)) of Mtb-specific T cells by polychromatic flow cytometry. We studied Mtb-specific CD4(+) T cell responses in subjects with latent Mtb infection and active tuberculosis disease. The results showed substantial increase in the proportion of single-positive TNF-α Mtb-specific CD4(+) T cells in subjects with active disease, and this parameter was the strongest predictor of diagnosis of active disease versus latent infection. We validated the use of this parameter in a cohort of 101 subjects with tuberculosis diagnosis unknown to the investigator. The sensitivity and specificity of the flow cytometry-based assay were 67% and 92%, respectively, the positive predictive value was 80% and the negative predictive value was 92.4%. Therefore, the proportion of single-positive TNF-α Mtb-specific CD4(+) T cells is a new tool for the rapid diagnosis of active tuberculosis disease.