Patient doses in CT examinations in Switzerland: implementation of national diagnostic reference levels.

Diagnostic reference levels (DRLs) were established for 21 indication-based CT examinations for adults in Switzerland. One hundred and seventy-nine of 225 computed tomography (CT) scanners operated in hospitals and private radiology institutes were audited on-site and patient doses were collected. For each CT scanner, a correction factor was calculated expressing the deviation of the measured weighted computed tomography dose index (CTDI) to the nominal weighted CTDI as displayed on the workstation. Patient doses were corrected by this factor providing a realistic basis for establishing national DRLs. Results showed large variations in doses between different radiology departments in Switzerland, especially for examinations of the petrous bone, pelvis, lower limbs and heart. This indicates that the concept of DRLs has not yet been correctly applied for CT examinations in clinical routine. A close collaboration of all stakeholders is mandatory to assure an effective radiation protection of patients. On-site audits will be intensified to further establish the concept of DRLs in Switzerland.

The CD4-like molecule LAG-3, biology and therapeutic applications.

IMPORTANCE OF THE FIELD: Promising immunotherapeutic agents targeting co-stimulatory pathways are currently being tested in clinical trials. One player in this array of regulatory pathways is the LAG-3/MHC class II axis. The lymphocyte activation gene-3 (LAG-3) is a negative co-stimulatory receptor that modulates T cell homeostasis, proliferation and activation. A recombinant soluble dimeric form of LAG-3 (sLAG-3-Ig, IMP321) shows adjuvant properties and enhances immunogenicity of tumor vaccines. Recent clinical trials produced encouraging results, especially when the human dimeric soluble form of LAG-3 (hLAG-3-Ig) was used in combination with chemotherapy. AREAS COVERED IN THIS REVIEW: The biological relevance of LAG-3 in vivo. Pre-clinical data demonstrating adjuvant properties, as well as the improvement of tumor immunity by sLAG-3-Ig. Recent advances in the clinical development of the therapeutic reagent IMP321, hLAG-3-Ig, for cancer treatment. WHAT THE READER WILL GAIN: This review summarizes preclinical and clinical data on the biological functions of LAG-3. TAKE HOME MESSAGE: The LAG-3 inhibitory pathway is attracting attention, in the light of recent studies demonstrating its role in T cell unresponsiveness, and Treg function after chronic antigen stimulation. As a soluble recombinant dimer, the sLAG-3-Ig protein acts as an adjuvant for therapeutic induction of T cell responses, and may be beneficial to cancer patients when used in combination therapies.

Nouvelles techniques de stimulation cérébrale: perspectives thérapeutiques en psychiatrie [Novel brain stimulation techniques: therapeutic perspectives in psychiatry]

Recent advances have allowed the development of new physical techniques in neurology and psychiatry, such as Transcranial Magnetic Stimulation (TMS), Vagus Nerve Stimulation (VNS), and Deep Brain Stimulation (DBS). These techniques are already recognized as therapeutic approaches in several late stage refractory neurological disorders (Parkinson's disease, tremor, epilepsy), and currently investigated in psychiatric conditions, refractory to medical treatment (obsessive-compulsive disorder, resistant major depression). In Paralell, these new techniques offer a new window to understand the neurobiology of human behavior.

Produtividade e bienalidade da produção de cafezais adensados, sob diferentes doses de N e K

O objetivo deste trabalho foi avaliar o efeito da adubação com N e K sobre a produtividade e a bienalidade de produção de cafeeiros em plantio adensado, em estudo de longa duração. Foram avaliados dois experimentos, na região da Zona da Mata de Minas Gerais, conduzidos em blocos ao acaso, com parcelas subdivididas no tempo. No primeiro experimento, realizado no espaçamento de 1,5x0,7 m (9.523 plantas por hectare), foram avaliados os efeitos de sete doses de N + K2O na proporção 1:1 (0, 200, 400, 600, 800, 1.000 e 1.200 kg ha‑1 por ano), durante oito safras (2001/2002 a 2008/2009). Como o fertilizante utilizado tinha proporção 1:1, para obter a dose utilizada de cada elemento (N ou K2O), deve-se dividir a dose total por dois. O segundo experimento foi conduzido no espaçamento de 2,5x0,6 m (6.666 plantas por hectare), em arranjo fatorial, com cinco doses de N (0, 150, 300, 450 e 600 kg ha‑1 por ano) e cinco doses de K2O (0, 150, 300, 450 e 600 kg ha‑1 por ano), durante seis safras (2005/2006 a 2010/2011). As doses associadas à máxima produtividade de café beneficiado variaram de 424 a 560 kg ha‑1, para N, e de 21 a 338 kg ha‑1, para K2O. A adubação com N reduz os efeitos da bienalidade da produção do cafeeiro, e a adubação com K, em anos de baixa produtividade, favorece a recuperação da produtividade do cafeeiro no ano seguinte.

Danos do complexo broca-podridão à produtividade e à qualidade da cana-de-açúcar fertirrigada com doses de nitrogênio

O objetivo deste trabalho foi avaliar o efeito da fertirrigação nitrogenada sobre os danos causados pelo complexo broca-podridão à produtividade e à qualidade da cana-de-açúcar. O experimento foi realizado em campo, em delineamento de blocos ao acaso, com quatro repetições e cinco doses de N (0, 50, 100, 150 e 200 kg ha-1), aplicadas por irrigação por gotejamento. Quantificaram-se as seguintes variáveis: número de orifícios causados por Diatraea saccharalis (Lepidoptera: Crambidae); número de internódios infectados com a podridão-vermelha; percentagem de incidência de podridão-vermelha; produtividade de colmos; e teor de açúcar. Em laboratório, foram avaliados a atratividade e o consumo de fragmentos de colmos por lagartas de quarto instar, em testes com ou sem chance de escolha. A irrigação nitrogenada aumentou a incidência de D. saccharalis e de podridão-vermelha. O número de orifícios causados pelas lagartas correlacionou-se à ocorrência de podridão-vermelha. A percentagem de açúcar diminuiu com o aumento do número de internódios danificados pela podridão-vermelha. As doses de N não afetaram a atratividade dos fragmentos de colmos de cana-de-açúcar; no entanto, maiores doses de N fertilizante aumentaram o consumo destes fragmentos por lagartas de D. saccharalis. Apesar dos danos à qualidade, a adubação nitrogenada aumentou expressivamente a produtividade de colmos e de açúcar.

Nutrição e produtividade da nogueira-macadâmia em função de doses de nitrogênio

O objetivo deste trabalho foi avaliar a influência de doses de nitrogênio na nutrição mineral e na produtividade da nogueira-macadâmia (Macadamia integrifolia). O experimento foi conduzido durante três anos agrícolas, em Jaboticabal, SP. Utilizou-se o delineamento experimental de blocos ao acaso, com quatro repetições. Foram avaliadas cinco doses de N (0, 50, 100, 150 e 200 kg ha-1 por ano), aplicadas na forma de ureia. O aumento nas doses de N aumentou o teor do nutriente nas folhas e a produtividade de nozes e amêndoas. Os teores de Ca, Mg e S diminuíram e os de Fe e Mg aumentaram linearmente com a adubação nitrogenada. A produtividade de nozes e de amêndoas correlacionou-se positivamente com o teor de N nas folhas, e a faixa de 14 a 18 g kg-1 foi a que possibilitou as maiores produtividades. A dose de 150 kg ha-1 de N por ano proporciona maior produtividade de noz, sem reduzir a taxa de recuperação de amêndoas.

Non-viral ocular gene therapy: potential ocular therapeutic avenues.

Non-viral vectors for potential gene replacement and therapy have been developed in order to overcome the drawbacks of viral vectors. The diversity of non-viral vectors allows for a wide range of various products, flexibility of application, ease of use, low-cost of production and enhanced "genomic" safety. Using non-viral strategies, oligonucleotides (ODNs) can be delivered naked (less efficient) or entrapped in cationic lipids, polymers or peptides forming slow release delivery systems, which can be adapted according to the organ targeted and the therapy purposes. Tissue and cell internalization can be further enhanced by changing by physical or chemical means. Moreover, a specific vector can be selected according to disease course and intensity of manifestations fulfilling specific requirements such as the duration of drug release and its level along with cells and tissues specific targeting. From accumulating knowledge and experience, it appears that combination of several non-viral techniques may increase the efficacy and ensure the safety of these evolving and interesting gene therapy strategies.

Mechanisms of interaction of therapeutic nanoparticles with cells

Nanoparticles (NPs) have gained a lot of interest in recent years due to their huge potential for applications in industry and medicine. Their unique properties offer a large number of attractive possibilities in the biomedical field, providing innovative tools for diagnosis of diseases and for novel therapies. Nevertheless, a deep understanding of their interactions with living tissues and the knowledge about their possible effects in the human body are necessary for the safe use of nanoparticulate formulations. The aim of this PhD project was to study in detail the interactions of therapeutic NPs with living cells, including cellular uptake and release, cellular localization and transport across the cell layers. Moreover, the effects of NPs on the cellular metabolic processes were determined using adapted in vitro assays. We evaluated the biological effect of several NPs potentially used in the biomedical field, including titanium dioxide (Ti02) NPs, 2-sized fluorescent silica NPs, ultrasmall superparamagnetic iron oxide (USPIO) NPs, either uncoated or coated with oleic acid or with polyvinylamine (aminoPVA) and poly(lactic-co-glycolic acid) - polyethylene-oxide (PLGA-PEO) NPs. We have found that the NPs were internalized by the cells, depending on their size, chemical composition, surface coating and also depending on the cell line considered. The uptake of aminoPVA-coated USPIO NPs by endothelial cells was enhanced in the presence of an external magnetic field. None of the tested USPIO NPs and silica NPs was transported across confluent kidney cell layers or brain endothelial cell layers, even in the presence of a magnetic field. However, in an original endothelium-glioblastoma barrier model which was developed, uncoated USPIO NPs were directly transferred from endothelial cells to glioblastoma cells. Following uptake, Ti02 NPs and uncoated USPIO NPs were released by the kidney cells, but not by the endothelial cells. Furthermore, these NPs induced an oxidative stress and autophagy in brain endothelial cells, possibly associated with their enhanced agglomeration in cell medium. A significant DNA damage was found in brain endothelial cells after their exposure to TiO2NPs. Altogether these results extend the existing knowledge about the effects of NPs on living cells with regard to their physicochemical characteristics and provide interesting tools for further investigation. The development of the in vitro toxicological assays with a special consideration for risk evaluation aims to reduce the use of animal experiments. -Les nanoparticules (NPs) présentent beaucoup d'intérêt dans le domaine biomédical et industriel. Leurs propriétés uniques offrent un grand nombre de possibilités de solutions innovantes pour le diagnostique et la thérapie. Cependant, pour un usage sûr des NPs il est nécessaire d'acquérir une connaissance approfondie des mécanismes d'interactions des NPs avec les tissus vivants et de leur effets sur le corps humain. Le but de ce projet de thèse était d'étudier en détail les mécanismes d'interactions de NPs thérapeutiques avec des cellules vivantes, en particulier les mécanismes d'internalisation cellulaire et leur subséquente sécrétion par les cellules, leur localisation cellulaire, leur transport à travers des couches cellulaires, et l'évaluation des effets de NPs sur le métabolisme cellulaire, en adaptant les méthodes existante d'évaluation cyto-toxico logique s in vitro. Pour ces expériences, les effets biologiques de nanoparticules d'intérêt thérapeutique, telles que des NPs d'oxyde de titane (TiO2), des NPs fluorescents de silicate de 2 tailles différentes, des NPs, d'oxyde de fer super-para-magnétiques ultra-petites (USPIO), soit non- enrobées soit enrobées d'acide oléique ou de polyvinylamine (aminoPVA), et des NPs d'acide poly(lactique-co-glycolique)-polyethylene-oxide (PLGA-PEO) ont été évalués. Les résultats ont démontré que les NPs sont internalisées par les cellules en fonction de leur taille, composition chimique, enrobage de surface, et également du type de cellules utilisées. L'internalisation cellulaire des USPIO NPs a été augmentée en présence d'un aimant externe. Aucune des NPs de fer et de silicate n'a été transportée à travers des couches de cellules épithéliales du rein ou endothéliales du cerveau, même en présence d'un aimant. Cependant, en développant un modèle original de barrière endothélium-glioblastome, un transfert direct de NPs d'oxyde de fer de cellule endothéliale à cellule de glioblastome a été démontré. A la suite de leur internalisation les NPs d'oxyde de fer et de titane sont relâchées par des cellules épithéliales du rein, mais pas des cellules endothéliales du cerveau. Dans les cellules endothéliales du cerveau ces NPs induisent en fonction de leur état d'agglomération un stress oxydatif et des mécanismes d'autophagie, ainsi que des dommages à l'ADN des cellules exposées aux NPs d'oxyde de titane. En conclusion, les résultats obtenus élargissent les connaissances sur les effets exercés par des NPs sur des cellules vivantes et ont permis de développer les outils expérimentaux pour étudier ces effets in vitro, réduisant ainsi le recours à des expériences sur animaux.

The parties' voices in a therapeutic interview

Drawing on a dialogical approach inspired by Bakhtin, we start from the assumptionthat a concrete discussion is an intermingling between dialogue in praesentia and dialogue inabsentia, and we refer to the notion of 'enunciative positioning' to account for the variousrelations that a speaker may express towards the voices that he or she invokes. Our data arebased on a first therapeutic interview between a therapist, a mother and a child in a counselingcenter for children and adolescents. We identify the various voices invoked in this interviewand show that three levels of discursive process were involved: (a) the speakers invokedabsent speakers; (b) at the same time they developed their own discourse on the basis of theirinterlocutors' discourse which (c) itself drew on absent speakers or voices. We highlight thevarious discursive processes through which the speakers integrate their own voice into absentvoices, or integrate a distant voice so that it loses its property of being a distant (andborrowed) voice. As a theoretical and methodological contribution to dialogism, our resultsshow that absent voices and their specific intermingling with hic et nunc exchanges were amajor resource for therapeutic changes.

Treatment deviating from guidelines does not influence status epilepticus prognosis.

Status epilepticus (SE) prognosis is related to nonmodifiable factors (age, etiology), but the exact role of drug treatment is unclear. This study was undertaken to address the prognostic role of treatment adherence to guidelines (TAG). We prospectively studied over 26 months a cohort of adults with incident SE (excluding postanoxic). TAG was assessed in terms of drug doses (± 30 % of recommendations) and medication sequence; its prognostic impact on mortality and return to baseline conditions was adjusted for etiology, SE severity [Status Epilepticus Severity Score (STESS)], and comorbidities. Of 225 patients, 26 (12 %) died and 82 (36 %) were discharged with a new handicap; TAG was observed in 142 (63 %). On univariate analysis, age, etiology, SE severity, and comorbidities were significantly related to outcome, while TAG was associated with neither outcome nor likelihood of SE control. Logistic regression for mortality identified etiology [odds ratio (OR) 18.8, 95 % confidence interval (CI) 4.3-82.8] and SE severity (STESS ≥ 3; OR 1.7, 95 % CI 1.2-2.4) as independent predictors, and for lack of return to baseline, again etiology (OR 7.4, 95 % CI 3.9-14.0) and STESS ≥ 3 (OR 1.7, 95 % CI 1.4-2.2). Similar results were found for the subgroup of 116 patients with generalized-convulsive SE. Receiver operator characteristic (ROC) analyses confirmed that TAG did not improve outcome prediction. This study of a large SE cohort suggests that treatment adherence to recommendations using current medications seems to play a negligible prognostic role (class III), confirming the importance of the biological background. Awaiting further treatment trials, it appears mandatory to apply resources towards identification of new therapeutic approaches.

A pilot study of the efficacy of IL1 blockade by anakinra in acute calcific periarthritis of the rotator cuff

Background/Purpose: Calcific periarthritis of rotator cuff can induce acute and severe shoulder pain and is accompnied by signs of acute inflammation. The calcific deposits are composed of calcium phosphate crystals such as hydroxyapatite or basic calcium phosphate. These crystals stimulate the production and release of IL1b from macrophages, in an analogous manner to MSU and CPPD crystals. As IL1 blockade is effective in reducing signs and symptoms of inflammation in acute gout, we performed a pilot study to study if it is also effective in calcific periarthritis Methods: 5 consecutive patients were included (mean age: 62, 3 females, 2 males) between March 2011 and March 2012. Symptoms of acute shoulder pain at rest had to be present for _7 days before inclusion, associated with limitation of shoulder mobility and the presence on calcification in the rotator cuff by conventional radiography. None of the patients had responded to at least 48 hours of high doses of NSAIDs. Exclusion criteria included no corticosteroid therapy in the last 2 weeks and the exclusion of other rheumatologic or infectious diseases- .Clinical evaluation consisted of patient assessment of pain (total, rest and activity) by VAS (100mm scale) at days 0, 1, 3, 15, 42 and clinical examination of shoulder mobility at days 0, 3, 15. ESR and CRP were measured at days 0, 3. Plain radiographs were performed at days 0 and 15 and an ultrasound examination (including Doppler) was performed at days 0, 3, 15. Anakinra 100mg daily was administered for 3 consecutive days after the first evaluation (day 0). Rescue analgesics were allowed and recorded. Results: At inclusion, all patients had severe shoulder pain: mean (SD) VAS day pain of 72mm (_25mm), mean VAS night pain of 96 (_ 5) and impaired shoulder mobility. CRP was elevated in all of them (mean of 3X). Treatment with anakinra lead to rapid relief of pain in all patients, starting already on the first night following the first injection. The reduction of VAS pain was particularly striking for rest pain: mean (SD) VAS of 4mm (_ 5) at day 1 and this response was maintained for the 5 patients at the end of the three injections without any need of rescue medication. Mean rest VAS was 6 (_8) at day 3. The effect on day pain was less spectacular: mean (SD) VAS at D1 of 30 (_ 18), at D3 of 27 (_ 11). Shoulder mobility also improved and the CRP normalized in 4 of 5 patients at day 3. At day 42, 4 of 5 the patients were still totally asymptomatic. On X rays and US, the calcifications were reduced in size: mean maximal diameter of 21 mm at day 0 to 12 mm at day 15, but did not disappear in any patient. The main change on US was a significant and rapid (at day 3) reduction of Doppler activity around the calcification. Conclusion: This pilot open study suggests that IL-1_ inhibition may be an interesting therapeutic approach in acute calcific periarthritis, especially in patients who have not responded adequately to NSAIDs. The effect on pain seems to be more rapid (within a few hours) than steroid injection although a randomized controlled study needs to be performed to confirm this observation.

Produtividade de grãos e componentes de produção da canola de acordo com fontes e doses de nitrogênio

O objetivo deste trabalho foi avaliar a resposta da canola a fontes e doses de nitrogênio aplicadas na semeadura. O experimento foi conduzido em Latossolo Vermelho distroférrico típico, com textura muito argilosa. Utilizou-se delineamento experimental de blocos ao acaso, em arranjo fatorial 7x2, com sete doses de N em superfície na semeadura (0, 20, 40, 60, 80, 100 e 120 kg ha-1), duas fontes de N (sulfato de amônio e ureia) e quatro repetições. O experimento foi realizado com o híbrido Hyola 61, por dois anos, e foram avaliadas as seguintes variáveis: altura de planta, número de plantas por metro quadrado, massa de matéria seca da parte aérea, massa de síliquas por planta, massa de mil grãos, produtividade de grãos, e teores de proteína e de óleo nos grãos. As variáveis não foram influenciadas pelas fontes de N. A maior produtividade de grãos é alcançada com 88 kg ha-1 de N. Doses crescentes de N aumentam os teores de proteína e diminuem os de óleo nos grãos de canola.

Tumor sensitizing function and mechanism of action of a RasGAP derived peptide

Cancer is the second cause of death after cardio-vascular diseases in economically developed countries. Two of the most commonly used anti-cancer therapies are chemo and radiotherapy. Despite the remarkable advances made in term of delivery and specificity of these two anti-tumor regimens, their toxicity towards healthy tissue remains a limitation. A promising approach to overcome this obstacle would be the utilization of therapeutic peptides that specifically augment the sensitivity of tumoral cells to treatments. Lower therapeutical doses would then be required to kill malignant cells, limiting toxic effects on healthy tissues. It was previously shown in our laboratory that the caspase-3 generated fragment N2 of RasGAP is able to potentiate the genotoxin-induced apoptosis selectively in cancer cells. In this work we show that fragment N2 strictly requires a cytoplasmic localization to deliver its pro-apoptotic effect in genotoxin-treated cancer cells. The tumor sensitizing capacity of fragment N2 was found to reside within the 10 amino acid sequence 317-326. Our laboratory earlier demonstrated that a peptide corresponding to amino acids 317 to 326 of RasGAP fused to the TAT cell permeable moiety, called TAT-RasGAP317.326, is able to sensitize cancer cells, but not normal cells, to genotoxin-induced apoptosis. In the present study we describe the capacity of TAT-RasGAP 317.326 to sensitize tumors to both chemo and radiotherapy in an in vivo mouse model. The molecular mechanism underlying the TAT-RasGAP 317.326-mediated sensitization starts now to be elucidated. We demonstrate that G3BP1, an endoribonuclease binding to amino acids 317-326 of RasGAP, is not involved in the sensitization mechanism. We also provide evidence showing that TAT-RasGAP3 17-326 potentiates the genotoxin-mediated activation of Bax in a tBid-dependent manner. Altogether our results show that TAT-RasGAP 317.326 could be potentially used in cancer therapy as sensitizer, in order to improve the efficacy of chemo and radiotherapy and prolong the life expectancy of cancer patients. Moreover, the understanding of the TAT-RasGAP317.326 mode of action might help to unravel the mechanisms by which cancer cells resist to chemo and radiotherapy and therefore to design more targeted and efficient anti-tumoral strategies.