PRECLINICAL RATIONALE FOR THE USE OF THE AKT INHIBITOR PERIFOSINE IN COMBINATION WITH THE MULTIKINASE INHIBITOR SORAFENIB IN HODGKIN LYMPHOMA

INTRODUCTION: A significant proportion of Hodgkin lymphoma (HL) patients refractory to first-line chemotherapy or relapsing after autologous transplantation are not cured with currently available treatments and require new treatments. The PI3K/AKT and RAF/MEK/ERK pathways are constitutively activated in the majority of HL. These pathways can be targeted using the AKT inhibitor perifosine (\uc6terna Zentaris GmBH, Germany, EU), and the RAF/MEK/ERK inhibitor sorafenib (Nexavar\uae, Bayer, Germany, EU). We hypothesized that perifosine in combination with sorafenib might have a therapeutic activity in HL by overcoming the cytoprotective and anti-apoptotic effects of PI3K/Akt and RAF/MEK/ERK pathways. Since preclinical evidence supporting the anti-lymphoma effects of the perifosine/sorafenib combination are still lacking, the present study aimed at investigating in vitro and in vivo the activity and mechanism(s) of action of this two-drug combination. METHODS: Three HL cell lines (HD-MyZ, L-540 and HDLM-2) were used to investigate the effects of perifosine and sorafenib using in vitro assays analyzing cell growth, cell cycle distribution, gene expression profiling (GEP), and apoptosis. Western blotting (WB) experiments were performed to determine whether the two-drug combination affected MAPK and PI3K/AKT pathways as well as apoptosis. Additionally, the antitumor efficacy and mechanism of action of perifosine/sorafenib combination were investigated in vivo in nonobese diabetic/severe combined immune-deficient (NOD/SCID) mice using tumor growth rates and survival as endpoints. RESULTS: While perifosine and sorafenib as single agents exerted a limited activity against HL cells, exposure of HD-MyZ and L-540 cell lines, but not HDLM-2 cells, to perifosine/sorafenib combination resulted in synergistic cell growth inhibition (40% to 80%) and cell cycle arrest. Upon perifosine/sorafenib exposure, L-540 cell line showed significant levels of apoptosis (up to 70%, P 64.0001) associated with severe mitochondrial dysfunction (cytochrome c, apoptosis-inducing factor release and marked conformational change of Bax accompanied by membrane translocation). Apoptosis induced by perifosine/sorafenib combination did not result in processing of caspase-8, -9, -3, or cleavage of PARP, and was not reversed by the pan-caspase inhibitor Z-VADfmk, supporting a caspase-independent mechanism of apoptosis. In responsive cell lines, WB analysis showed that anti-proliferative events were associated with dephosphorylation of MAPK and PI3K/Akt pathways. GEP analysis of HD-MyZ and L-540 cell lines, but not HDLM-2 cells indicated that perifosine/sorafenib treatment induced upregulation of genes involved in amino acid metabolism and downregulation of genes regulating cell cycle, DNA replication and cell death. In addition, in responsive cell lines, perifosine/sorafenib combination strikingly induced the expression of tribbles homologues 3 (TRIB3) both in vitro and in vivo. Silencing of TRIB3 prevented cell growth reduction induced by perifosine/sorafenib treatment. In vivo, the combined perifosine/sorafenib treatment significantly increased the median survival of NOD/SCID mice xenografted with HD-MyZ cell line as compared to controls (81 vs 45 days, P 64.0001) as well as mice receiving perifosine alone (49 days, P 64.03) or sorafenib alone (54 days, P 64.007). In addition, perifosine/sorafenib treatment had no effect on HDLM-2 nodules, but significantly reduced L-540 nodules with 50% tumor growth inhibition (P 64.0001), compared to controls. In mice bearing subcutaneous nodules generated by HD-MyZ and L-540 cell lines but not HDLM-2 cell line, perifosine/sorafenib treatment induced significantly increased levels of apoptosis (2- to 2.5-fold, P 64.0001) and necrosis (2- to 8-fold, P 64.0001), as compared to controls or treatment with single agents. CONCLUSIONS: Perifosine/sorafenib combination resulted in potent anti-HL activity both in vitro and in vivo. These results warrant clinical evaluation in HL patients

Effect of flour and pasta debris on larval development of Rhyzopertha dominica

The development of Rhyzopertha dominica (F.) larvae on different types of flour and on pasta debris was evaluated under laboratory condition. Tests were carried out on wheat flour, semolina, wheat bran, corn meal, corn starch, rice flour, buckwheat flour, chickpea flour, almond flour, chestnut flour, potato starch, and pasta debris. Two layers of flour or pasta debris, 3 and 6 mm high, were placed in Petri dishes and 20 first instar larvae were put into the middle of the substrate. Four replicates were carried out for each combination of substrate, particle size, and thickness of the layer. The emergence of adults was assessed daily. Chestnut flour and semolina permitted the development of larvae in a period of time similar to that observed in cereal kernels while chickpea flour and wheat flour were less appropriate food; in fact, only 25% of larvae underwent complete development into adults. In the case of corn meal, rice flour, and wheat bran, only 5% of adults emerged, while larvae were unable to develop on corn starch and potato starch. Although R. dominica is a well-known pest of pasta, pasta debris was less suitable for the development of larvae, as few adults emerged and a longer development time was observed. A three-millimeter layer of suitable food was sufficient for the development of larvae, although a higher number of adults was observed in the 6 mm layer

Physical properties of the jet from DG Tauri on sub-arcsecond scales with HST/STIS

We derive the physical properties at the base of the jet from DG Tau both along and across the flow and as a function of velocity. We analysed seven optical spectra of the DG Tau jet, taken with the Hubble Space Telescope Imaging Spectrograph. The spectra were obtained by placing a long-slit parallel to the jet axis and stepping it across the jet width. The resulting position-velocity diagrams in optical forbidden emission lines allowed access to plasma conditions via calculation of emission line ratios. We find at the base of the jet high electron density, $n_e \sim$ 10$^5$, and very low ionisation, $x_e \sim 0.02-0.05$, which combine to give a total density up to $n_H \sim$ 3 10$^6$. This analysis confirms previous reports of variations in plasma parameters along the jet, (i.e. decrease in density by several orders of magnitude, increase of $x_e$ from 0.05 to a plateau at 0.7 downstream at 2$''$ from the star). Furthermore, a spatial coincidence is revealed between sharp gradients in the total density and supersonic velocity jumps. This strongly suggests that the emission is caused by shock excitation. The position-velocity diagrams indicate the presence of both fast accelerating gas and slower, less collimated material. We derive the mass outflow rate, $\dot{M}_j$, in the blue-shifted lobe in different velocity channels, that contribute to a total of $\dot{M}_j \sim$ 8 $\pm$ 4 10$^{-9}$ M$_\odot$ yr$^{-1}$. We estimate that a symmetric bipolar jet would transport at the low and intermediate velocities probed by rotation measurements, an angular momentum flux of $\dot{L}_j \sim$ 2.9 $\pm$ 1.5 10$^{-6}$ M$_\odot$ yr$^{-1}$ AU km s$^{-1}$. The derived properties of the DG Tau jet are demonstrated to be consistent with magneto-centrifugal theory. However, non-stationary modelling is required in order to explain all of the features revealed at high resolution.Comment: 16 pages, 18 figure

A produção orgânica no município de Cacoal, RO: uma análise da dinâmica econômica e o desenvolvimento sustentável.

A preocupação com a segurança alimentar, qualidade de vida e a questão ambiental vem ao longo do tempo adquirindo uma dinâmica em escala global com novos hábitos alimentares e modos de produção. Com o objetivo verificar o desenvolvimento produtivo e sua viabilidade comercial diante da implantação de sistemas orgânicos na associação dos produtores do município de Cacoal, RO buscou-se analisar e caracterizar as relações comerciais e sociais que se estabelece entre o produtor e consumidor sendo identificados os diferentes cenários como: produção, distribuição e comercialização no município, através do método de pesquisa com a identificação dos principais meios e espaços de comercialização e, a realização de entrevistas com os principais membros da associação e feirante envolvidos no processo. Conclui-se que diante dos obstáculos a serem superados, a produção e a comercialização agroecológica apresenta possibilidades de expansão no município

The phenomenology of depression

The phenomenological method, characterized by the suspension of judgment (epoché), has helped analyzing the subjective experience of patients affected by mental disorders. Psychiatry, dealing with the human being itself in its complexity and unicity, is placed between the biological positivistic attempt, for which the symptoms of mental illness are a mere consequence of brain dysfunctions and the phenomenological-existential approach, inclined to consider the symptoms as meaningful phenomena of the person’s subjective experience. Eugène Minkowski, Ludwig Binswanger, Arthur Tatossian, Kimura Bin, Henri Maldiney and Hubertus Tellenbach are fundamental authors in the phenomenological psychopathology of depression; they described the alterations of the lived time, space, body and others experienced by the depressed. Starting from the main theoretical contributions of the authors, we will focus on the psychopathology and discuss the key themes of clinical depression: guilt, poverty and hypochondriasis. Finally we will focus on the typus melancholicus construct

The effects of competition between larvae of stored-product moths

The competition between Idaea inquinata, Corcyra cephalonica, and Plodia interpunctella on an artificial diet was investigated. The experiments were carried out with eggs laid within 24 hours. In the first experiment, 20 eggs of one species were placed in a ventilated Petri dish with 10 g of diet and 20 eggs of one of the other species; in the second experiment, 20 eggs of one species and, after 7 days, 20 eggs of one of the other species were added. Experiments were carried out at 27\ub11 \ub0C, 70\ub15% R.H. Each experiment was replicated 5 times and the number of days to complete the development and the number of emerged adults were recorded. The number of P. interpunctella adults was not affected by the presence of C. cephalonica or I. inquinata. A delay in the development was only observed when P. interpunctella eggs were added to the medium already colonized by the other species. I. inquinata was the less competitive, as the number of adults decreased and the days to complete the cycle increased. The development of I. inquinata was prevented if P. interpunctella eggs were started at the same time. Larvae of this last species cannibalized eggs of the other two species. I. inquinata was the least competitive due to the longer cycle, the lower mobility and the aggressiveness of the larvae of the other two species that cannibalized its eggs

Egg hatching at different temperatures and relative humidities in Idaea inquinata (Scopoli) (Lepidoptera: Geometridae)

Idaea inquinata (Scopoli) feeds mainly on dried plants, nevertheless, it is also a potential pest of stored products as it is able to develop on cereal products. The few references on the biology of this species do not deal with the influence of temperature and relative humidity on egg hatching. To fill this gap, groups of 100 eggs, 24-48 hours old, were exposed to five constant temperatures (17, 21, 26, 29 and 34±1°C), two relative humidities (35 and 70±5%) and a photoperiod of 0:24 (light:dark); ten tests were carried out. Each test was replicated four times and egg hatching was observed daily. The highest mean number of hatched eggs was observed at 26 and 29±1°C, 70±5% r.h. with 91.5 and 91.0 eggs, respectively. The lowest mean number of hatched eggs was 61.5 observed at 17°C and 70±5% r.h. The mean numbers of hatched eggs, 83.5, 77.5, 78.5 and 79.8 were similar at 21, 26, 29 and 34±1°C, 35±5% r.h., respectively. Eggs hatched between the sixth and the eighth day at all the temperatures tested, except for 17±1°C and 35±5% r.h., where hatching started on the twelfth day. At this temperature, the duration of the hatching period increased with increasing humidity: 11 d at 35% r.h. and 15 d at 70% r.h. Keywords: Egg, Hatching, Temperature, Relative humidity, Rusty wave mot