Physical considerations on discrepancies in target volume delineation.

BACKGROUND AND PURPOSE: To compare the delineations and interpretations of target volumes by physicians in different radio-oncology centers. MATERIALS AND METHODS: Eleven Swiss radio-oncology centers delineated volumes according to ICRU 50 recommendations for one prostate and one head and neck case. In order to evaluate the consistency of the volume delineations, the following parameters were determined: 1) the target volumes (GTV, CTV and manually expanded PTV) and their extensions in the three main axes and 2) the correlation of the volume delineated by each pair of centers using the ratio of the intersection to the union (called proximity index). RESULTS: The delineated prostate volume was 105+/-55cm(3) for the CTV and 218+/-44cm(3) for the PTV. The delineated head and neck volume was 46+/-15cm(3) for the GTV, 327+/-154cm(3) for the CTV and 528+/-106cm(3) for the PTV. The mean proximity index for the prostate case was 0.50+/-0.13 for the CTV and 0.57+/-0.11 for the PTV. The proximity index for the head and neck case was 0.45+/-0.09 for the GTV, 0.42+/-0.13 for the CTV and 0.59+/-0.06 for the PTV. CONCLUSIONS: Large discrepancies between all the delineated target volumes were observed. There was an inverse relationship between the CTV volume and the margin between CTV and PTV, leading to less discrepancies in the PTV than is the CTV delineations. There was more spread in the sagittal and frontal planes due to CT pixel anisotropy, which suggests that radiation oncologists should delineate the target volumes not only in the transverse plane, but also in the sagittal and frontal planes to improve the delineation by allowing a consistency check.

Analyse der Hepatitis-C-Situation bei den drogenkonsumierenden Personen in der Schweiz

Die Analyse der Hepatitis-C-Situation bei den drogenkonsumierenden Personen in der Schweiz beruht auf verschiedenen Methoden: Analyse der publizierten und grauen Literatur zu diesem Thema, Sekundäranalyse verfügbarer Daten, Befragung nationaler Fachpersonen, Befragung drogenkonsumierender Personen (DU), Fallstudien in vier Kantonen (Zürich, Waadt, Aargau, Wallis), Internetumfrage bei den Einrichtungen, die DU stationär oder ambulant behandeln, sowie bei den niederschwelligen Einrichtungen zur Risikominderung. In den vergangenen zwei Jahrzehnten erreichte die Zahl der neu gemeldeten Infektionen mit dem Hepatitis-C-Virus (HCV) Ende der 90er-Jahre eine Spitze mit mehr als 2500 Fällen pro Jahr, wobei mehr als 30 % dieser Fälle Personen betrafen, die intravenös Drogen konsumieren (IDU). Die Zahl neu gemeldeter Fälle sank anschliessend bis auf 1500 Fälle im Jahr 2011 und stieg dann wieder auf mehr als 1700 Fälle im Jahr 2013 an. Der Anteil der IDU belief sich auf etwas mehr 20 % (436 Fälle, kein Wiederanstieg der Anzahl neuer Fälle in dieser Bevölkerungsgruppe). Die für die Gesamtbevölkerung der Schweiz geschätzte Prävalenz der HCV-Infektionen bewegt sich je nach Autor zwischen 0,7 % und 1,75 %. Bei Männern, die Sex mit Männern haben (MSM), ist dieser Wert etwas höher (2 %). Bei Staatsangehörigen aus Ländern mit einer höheren Prävalenz (Afrika südlich der Sahara, bestimmte Länder des Nahen Ostens und Südostasiens) ist die Prävalenzrate vermutlich erhöht; es sind jedoch keine Werte bekannt. Auch in Gefängnissen ist die HCV-Prävalenz höher als in der Gesamtbevölkerung, da DU und Staatsangehörige aus Ländern mit höherer Prävalenz stärker vertreten sind (zwischen 5 und 10 %). Es wird davon ausgegangen, dass es bei etwa zwei Drittel der Fälle zu einem chronischen Krankheitsverlauf kommt.

Treating high blood pressure: is reaching the target more important than the means? No, the means are important.

All major antihypertensive drug classes i.e. diuretics, beta-blockers, calcium antagonists and blockers of the renin-angiotensin system have been shown to effectively lower blood pressure and hence to reduce cardiovascular outcomes in hypertensive patients. These drugs decrease cardiovascular complications in hypertension essentially because they reduce systemic blood pressure. Nevertheless, there is growing evidence that the extent of the benefits differed between drug classes suggesting that the various classes of antihypertensive agents are not equivalent in their ability to protect against target organ damages and cardiovascular and renal endpoints. More recently, evidence has also accumulated to demonstrate that even combination therapies are not equally effective in reducing the occurrence of cardiovascular complications in hypertension. These recent observations suggest that the means to lower blood pressure are as important as the achieved target blood pressure in the management of hypertensive patients.

The p63 target HBP1 is required for skin differentiation and stratification.

Genetic experiments established that p63 is crucial for the development and maintenance of pluristratified epithelia. In the RNA interference (RNAi) screening for targets of p63 in keratinocytes, we identified the transcription factor, High Mobility Group (HMG) box protein 1 (HBP1). HBP1 is an HMG-containing repressor transiently induced during differentiation of several cell lineages. We investigated the relationship between the two factors: using RNAi, overexpression, chromatin immunoprecipitations and transient transfections with reporter constructs, we established that HBP1 is directly repressed by p63. This was further confirmed in vivo by evaluating expression in p63 knockout mice and in transgenics expressing p63 in basal keratinocytes. Consistent with these findings, expression of HBP1 increases upon differentiation of primary keratinocytes and HaCaT cells in culture, and it is higher in the upper layers of human skin. Inactivation of HBP1 by RNAi prevents differentiation of keratinocytes and stratification of organotypic skin cultures. Finally, we analyzed the keratinocyte transcriptomes after HBP1 RNAi; in addition to repression of growth-promoting genes, unexpected activation of differentiation genes was uncovered, coexisting with repression of other genes involved in epithelial cornification. Our data indicate that suppression of HBP1 is part of the growth-promoting strategy of p63 in the lower layers of epidermis and that HBP1 temporally coordinates expression of genes involved in stratification, leading to the formation of the skin barrier.

Improved HIV-1 RNA quantitation by COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, v2.0 using a novel dual-target approach.

BACKGROUND: HIV-1 RNA viral load is a key parameter for reliable treatment monitoring of HIV-1 infection. Accurate HIV-1 RNA quantitation can be impaired by primer and probe sequence polymorphisms as a result of tremendous genetic diversity and ongoing evolution of HIV-1. A novel dual HIV-1 target amplification approach was realized in the quantitative COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, v2.0 (HIV-1 TaqMan test v2.0) to cope with the high genetic diversity of the virus. OBJECTIVES AND STUDY DESIGN: The performance of the new assay was evaluated for sensitivity, dynamic range, precision, subtype inclusivity, diagnostic and analytical specificity, interfering substances, and correlation with the COBAS AmpliPrep/COBAS TaqMan HIV-1 (HIV-1 TaqMan test v1.0) predecessor test in patients specimens. RESULTS: The new assay demonstrated a sensitivity of 20 copies/mL, a linear measuring range of 20-10,000,000 copies/mL, with a lower limit of quantitation of 20 copies/mL. HIV-1 Group M subtypes and HIV-1 Group O were quantified within +/-0.3 log(10) of the assigned titers. Specificity was 100% in 660 tested specimens, no cross reactivity was found for 15 pathogens nor any interference for endogenous substances or 29 drugs. Good comparability with the predecessor assay was demonstrated in 82 positive patient samples. In selected clinical samples 35/66 specimens were found underquantitated in the predecessor assay; all were quantitated correctly in the new assay. CONCLUSIONS: The dual-target approach for the HIV-1 TaqMan test v2.0 enables superior HIV-1 Group M subtype coverage including HIV-1 Group O detection. Correct quantitation of specimens underquantitated in the HIV-1 TaqMan test v1.0 test was demonstrated.

Adjustment to target capital, finance and growth

Does financial development result in capital being reallocated more rapidly to industries where it is most productive? We argue that if this was the case, financially developed countries should see faster growth in industries with investment opportunities due to global demand and productivity shifts. Testing this cross-industry cross-country growth implication requires proxies for (latent) global industry investment opportunities. We show that tests relying only on data from specific (benchmark) countries may yield spurious evidence for or against the hypothesis. We therefore develop an alternative approach that combines benchmark-country proxies with a proxy that does not reflect opportunities specific to a country or level of financial development. Our empirical results yield clear support for the capital reallocation hypothesis.

Which inflation to target? A small open economy with sticky wages indexed to past inflation

In a closed economy context there is common agreement on price inflation stabilization being one of the objects of monetary policy. Moving to an open economy context gives rise to the coexistence of two measures of inflation: domestic inflation (DI) and consumer price inflation (CPI). Which one of the two measures should be the target variable? This is the question addressed in this paper. In particular, I use a small open economy model to show that once sticky wages indexed to past CPI inflation are introduced, a complete inward looking monetary policy is no more optimal. I first, derive a loss function from a secondorder approximation of the utility function and then, I compute the fully optimalmonetary policy under commitment. Then, I use the optimal monetary policy as a benchmark to compare the performance of different monetary policy rules. The main result is that once a positive degree of indexation is introduced in the model the rule performing better (among the Taylor type rules considered) is the one targeting wage inflation and CPI inflation. Moreover this rule delivers results very close to the one obtained under the fully optimal monetary policy with commitment.

Glycogen synthase 2 is a novel target gene of peroxisome proliferator-activated receptors.

Glycogen synthase 2 (Gys-2) is the ratelimiting enzyme in the storage of glycogen in liver and adipose tissue, yet little is known about regulation of Gys-2 transcription. The peroxisome proliferator-activated receptors (PPARs) are transcription factors involved in the regulation of lipid and glucose metabolism and might be hypothesized to govern glycogen synthesis as well. Here, we show that Gys-2 is a direct target gene of PPARalpha, PPARbeta/delta and PPARgamma. Expression of Gys-2 is significantly reduced in adipose tissue of PPARalpha-/-, PPARbeta/delta-/- and PPARgamma+/- mice. Furthermore, synthetic PPARbeta/delta, and gamma agonists markedly up-regulate Gys-2 mRNA and protein expression in mouse 3T3-L1 adipocytes. In liver, PPARalpha deletion leads to decreased glycogen levels in the refed state, which is paralleled by decreased expression of Gys-2 in fasted and refed state. Two putative PPAR response elements (PPREs) were identified in the mouse Gys-2 gene: one in the upstream promoter (DR-1prom) and one in intron 1 (DR-1int). It is shown that DR-1int is the response element for PPARs, while DR-1prom is the response element for Hepatic Nuclear Factor 4 alpha (HNF4alpha). In adipose tissue, which does not express HNF4alpha, DR-1prom is occupied by PPARbeta/delta and PPARgamma, yet binding does not translate into transcriptional activation of Gys-2. Overall, we conclude that mouse Gys-2 is a novel PPAR target gene and that transactivation by PPARs and HNF4alpha is mediated by two distinct response elements.

Retinal degeneration progression changes lentiviral vector cell targeting in the retina

Ce travail de thèse a été réalisé au sein de l'Unité de Thérapie Génique et Biologie des Cellules Souches de l'Hôpital Jules- Gonin dans le Service d'Ophtalmologie de l'Université de Lausanne. Ce laboratoire recherche des solutions thérapeutiques pour des maladies dégénératives et incurables de la rétine comme les rétinites pigmentaires (RP). Ayant déjà montré certains résultats dans le domaine, la thérapie génique a été notre outil pour ce travail. Cette méthode se base sur le principe de remplacer un gène déficient par sa copie normale, en transportant celle-ci au coeur même du noyau par un vecteur. Il existe à l'heure actuelle différents vecteurs. Un des plus efficaces est un vecteur viral non-réplicatif : le lentivirus, dérivé de HIV-1. Celui-ci a la capacité d'intégrer le génome de la cellule cible, lui conférant ainsi un nouveau matériel génétique. Notre but a été d'établir le tropisme du lentivirus dans une rétine en dégénérescence. Ce lentivirus est connu pour transduire efficacement les cellules de l'épithélium pigmentaire rétinien dans l'oeil adulte sain, ainsi que celles de la neurorétine, mais ce, uniquement durant le développement. On sait aussi que le vecteur lentiviral présente un tropisme différent selon les enveloppes dont il est muni ; par exemple, le lentivirus avec une enveloppe Mokola est connu pour transduire les cellules gliales du système nerveux central. La rétine qui dégénère montre quant à elle des changements de sa structure qui pourraient influencer la diffusion du vecteur et/ou son tropisme. Le postulat de base a été le suivant : chez l'adulte, la transduction des neurones de la rétine via le lentivirus pourrait être facilitée par l'altération de la membrane limitante externe induite par la dégénérescence (meilleure pénétrance du virus). D'un point de vue technique, nous avons utilisé deux types distincts de modèles murins de dégénérescence rétinienne : des souris Balb/C soumises à une dose toxique de lumière et les souris Rhodopsin knockout, animaux génétiquement modifiés. Comme vecteur viral, nous avons employé deux différents pseudotypes de lentivirus (caractérisés par les enveloppes virales) avec différents promoteurs (séquence d'ADN qui initie la transduction et confère la spécificité d'expression d'un gène). En changeant l'enveloppe et le promoteur, nous avons essayé de trouver la meilleure combinaison pour augmenter l'affinité du vecteur vis-à-vis des photorécepteurs d'abord, puis vis-à-vis d'autres cellules de la rétine. Nos résultats ont montré que la membrane limitante externe est effectivement altérée chez les deux modèles de dégénérescence, mais que cette modification ne favorise pas la transduction des photorécepteurs lorsqu'on utilise un vecteur lentiviral contenant une enveloppe VSVG et un promoteur photorécepteur-spécifique ou ubiquitaire. En effet, une forte réaction gliale a été observée. Par contre, en utilisant le lentivirus avec une enveloppe Mokola et un promoteur ubiquitaire, nous avons constaté une très bonne transduction au niveau des cellules de Millier dans la rétine en dégénérescence, phénomène non observé chez les souris sauvages. Ce travail a donc permis de trouver un vecteur viral efficace pour atteindre et transduire les cellules de Miiller, ceci seulement pendant la dégénérescence de la rétine. Ces cellules, une fois transduites, pourraient être utilisées pour sécréter dans la rétine des agents thérapeutiques tels que des facteurs neurotrophiques pour soutenir la survie des photorécepteurs ou des facteurs anti-angiogéniques pour prévenir la néo-vascularisation lors de diabète ou de dégénérescence maculaire liée à l'âge. - In normal mice, the lentiviral vector (LV) is very efficient to target the RPE cells, but transduces retinal neurons well only during development. In the present study, the tropism of LV has been investigated in the degenerating retina of mice, knowing that the retina structure changes during degeneration. We postulated that the viral transduction would be increased by the alteration of the iuter limiting membrane (OLM). Two different LV pseudotypes were tested using the VSVG arid the Mokola envelopes, as well as two animal models of retinal degeneration: light-damaged Balb-C and Rhodopsin knockout (Rho-/-) mice. After light damage, the OLM is altered and no significant increase of the number of transduced photoreceptors can be obtained with a LV-VSVG-Rhop-GFP vector. In the Rho-/- mice, an altération of the OLM was also observed, but the possibility of transducing photoreceptors was decreased, probably by ongoing gliosis. The use of a ubiquitous promoter allows better photoreceptor transduction, suggesting that photoreceptór-specific promoter activity change during late stages of photoreceptor degeneration. However, the number of targeted photoreceptors remains low. In contrast, LV pseudotyped with the tfokola envelope allows a wide dispersion of the ctor into the retina (corresponding to the injection bleb) with preferential targeting of Muller cells, a situation Mc\ does ot occur in the wild- type retina. Mokola-pseudotyped lentiviral vectors may serve to engineer these glial cells to deliver secreted therapeutic factors to a diseased area of the retina.

Does Bt maize cultivation affect the non-target insect community in the agro ecosystem?

ABSTRACT The cultivation of genetically modified crops in Brazil has led to the need to assess the impacts of this technology on non-target species. Under field conditions, the potential effect on insect biodiversity was evaluated by comparing a homogeneous corn field with conventional and transgenic maize, expressing different Bt proteins in seven counties of Minas Gerais, Brazil. The richness pattern of non-target insect species, secondary pests and natural enemies were observed. The results do not support the hypothesis that Bt protein affects insect biodiversity. The richness and diversity data of insects studied were dependent on the location and other factors, such as the use of insecticides, which may be a major factor where they are used.

Potential target antigens for immunotherapy in human pancreatic cancer.

BACKGROUND: To be effective and selective, immunotherapy ideally targets specifically tumor cells and spares normal tissues. Identification of tumor specific antigens is a prerequisite to establish an effective immunotherapy. Still very little is known about the expression of tumor-related antigens in pancreatic neoplasms. Cancer Testis antigens (CT) are antigens shared by a variety of malignant tumors, but not by normal tissues with the exception of germ cells in testis. Restricted expression in neoplastic tissues and inherent immunogenic features make CT antigens ideal for use in immunotherapy. We analyzed the expression of a selected panel of nine CT antigens that have been proven to elicit an efficient immunogenic response in other malignancies. In addition we analyzed the expression of HERV-K-MEL, an immunogenic antigen of viral origin. METHODS: Pancreatic adenocarcinoma tumor samples (n=130) were obtained intraoperatively, control tissues (n=23) were collected from cadaveric donor and from patients with chronic pancreatitis. Tumor-associated antigen expression of MAGE-A1, MAGE-A3, MAGE-A4, MAGE-A10, LAGE-1, NY-ESO-1, SCP-1, SSX-2, SSX-4 and HERV-K-MEL was assessed by PCR. Sequencing of PCR products were performed to assess the expression of SSX-4 in neoplastic and normal pancreatic tissues. RESULTS: Three of 10 tested antigens were expressed in over 10% of malignant pancreatic tissue samples. SSX-4 was found positive in 30% of cases, SCP-1 in 19% and HERV-K-MEL in 23% of cases. No expression of CT antigens was found in non-malignant pancreatic tissue with the exception of SSX-4 and and SSX-2. CONCLUSIONS: Fifty two percentage of the analyzed tissues expressed at least one CT antigen. The concomitant expression of SSX-4 in both malignant and non-malignant pancreatic tissue is a new finding which may raise concerns for immunotherapy. However, HERV-K-MEL is expressed with a relatively high prevalence and may be a candidate for specific immunotherapy in a large subgroup of pancreatic cancer patients. This study advocates the analysis of patients with regard to their immunogenic profile before the onset of antigen-specific immunotherapy.

Maladies cardiovasculaires: une cible de prévention pour contrecarrer les effets de l'évolution démographique [Cardiovascular diseases: a target for prevention to thwart the effects of population aging]

The prevalence of cardiovascular diseases is high in the old age. These conditions have a negative impact on the quality of life and are associated with a high risk of disability. A marked increase in the number of affected individuals is likely, in coming decades, with population aging. Primary cardiovascular prevention, but also an early recognition of subclinical heart diseases and secondary and tertiary prevention will be of up most importance for individuals (quality of life) and for societies (burden of functional impairments) as the baby-boom generation reaches retirement age.