Smoking influences the yield of dendritic cells for cancer immunotherapy

Background: Dendritic cell (DC)-based vaccination is considered to be a potentially effective therapeutic strategy against advanced cancer. The aim of this study was to address the smoking history that might affect the preparation of DC vaccines in validated instructional manufacture. Materials and Methods: Data on mature DCs generated from 102 sessions of leukapheresis performed on 92 patients with advanced cancer or sarcoma were retrospectively evaluated and compared in relation to the data between their smoking history and the generation of DCs from these patients. 61 patients with adenocarcinoma, including 7 with lung, 10 with breast, 8 with stomach, 12 with colorectal, and 23 with pancreatic adenocarcinoma were enrolled. Results: The average yield of autologous DCs (15.5 ± 8.3x107) was thought to be dependent on the number of monocytes (124.2 ± 74.1x107) collected by leukapheresis. The average ratio of DCs/apheresed monocytes (DC/aM ratio) was lower in the smoker group (11.1 ± 7.2%) than that in the non-smoker group (17.2 ± 9.3%, p=0.001). The number of DCs and the DC/aM ratio were lower in the patients with gastric and pancreatic cancer than in those with adenocarcinoma of other sites. Conclusions: As cancer therapy moves forward into the field of personaArticlePharmaceutical Regulatory Affairs. 4(1):133 (2015)journal articl

Oscillatory Fractional Brownian Motion and Hierarchical Random Walks

We introduce oscillatory analogues of fractional Brownian motion, sub-fractional Brownian motion and other related long range dependent Gaussian processes, we discuss their properties, and we show how they arise from particle systems with or without branching and with different types of initial conditions, where the individual particle motion is the so-called c-random walk on a hierarchical group. The oscillations are caused by the discrete and ultrametric structure of the hierarchical group, and they become slower as time tends to infinity and faster as time approaches zero. We also give other results to provide an overall picture of the behavior of this kind of systems, emphasizing the new phenomena that are caused by the ultrametric structure as compared with results for analogous models on Euclidean space

Initial low/hard state, multiple jet ejections and X-ray/radio correlations during the outburst of XTE J1859+226

We have studied the 1999 soft X-ray transient outburst of XTE J1859+226 at radio and X-ray wavelengths. The event was characterised by strong variability in the disc, corona and jet - in particular, a number of radio flares (ejections) took place and seemed well-correlated with hard X-ray events. Apparently unusual for the `canonical soft' X-ray transient, there was an initial period of low/hard state behaviour during the rise from quiescence but prior to the peak of the main outburst - we show that not only could this initial low/hard state be an ubiquitous feature of soft X-ray transient outbursts but that it could also be extremely important in our study of outburst mechanisms.Comment: 12 pages, Accepted for publication in MNRA

Broad-band X-ray spectral evolution of GX 339-4 during a state transition

We report on X-ray and soft gamma-ray observations of the black-hole candidate GX 339-4 during its 2007 outburst, performed with the RXTE and INTEGRAL satellites. The hardness-intensity diagram of all RXTE/PCA data combined shows a q-shaped track similar to that observed in previous outbursts.The evolution in the diagram suggested that a transition from hard-intermediate state to soft-intermediate state occurred, simultaneously with INTEGRAL observations performed in March. The transition is confirmed by the timing analysis presented in this work, which reveals that a weak type-A quasi-periodic oscillation (QPO) replaces a strong type-C QPO. At the same time, spectral analysis shows that the flux of the high-energy component shows a significant decrease in its flux. However, we observe a delay (roughly one day) between variations of the spectral parameters of the high-energy component and changes in the flux and timing properties. The changes in the high-energy component can be explained either in terms the high-energy cut-off or in terms of a variations in the reflection component. We compare our results with those from a similar transition during the 2004 outburst of GX 339-4.Comment: 8 pages, 6 figures, accepted for publication in MNRAS Main Journa

Increased S-nitrosylation and proteasomal degradation of caspase-3 during infection contribute to the persistence of adherent invasive escherichia coli (AIEC) in immune cells

Adherent invasive Escherichia coli (AIEC) have been implicated as a causative agent of Crohn's disease (CD) due to their isolation from the intestines of CD sufferers and their ability to persist in macrophages inducing granulomas. The rapid intracellular multiplication of AIEC sets it apart from other enteric pathogens such as Salmonella Typhimurium which after limited replication induce programmed cell death (PCD). Understanding the response of infected cells to the increased AIEC bacterial load and associated metabolic stress may offer insights into AIEC pathogenesis and its association with CD. Here we show that AIEC persistence within macrophages and dendritic cells is facilitated by increased proteasomal degradation of caspase-3. In addition S-nitrosylation of pro- and active forms of caspase-3, which can inhibit the enzymes activity, is increased in AIEC infected macrophages. This S-nitrosylated caspase-3 was seen to accumulate upon inhibition of the proteasome indicating an additional role for S-nitrosylation in inducing caspase-3 degradation in a manner independent of ubiquitination. In addition to the autophagic genetic defects that are linked to CD, this delay in apoptosis mediated in AIEC infected cells through increased degradation of caspase-3, may be an essential factor in its prolonged persistence in CD patients

A transient variable 6 Hz QPO from GX 339-4

We report the results of an observation with the Rossi X-ray Timing Explorer of the black hole candidate GX 339-4 during its 2002/2003 outburst. This observation took place during a spectral transition from the hard to the soft state. A strong (6% rms) transient quasi-periodic oscillation (QPO) appears suddenly in the power density spectrum during this observation. The QPO centroid is ~6 Hz, but it varies significantly between 5 and 7 Hz with a characteristic time scale of ~10 seconds, correlated with the 2-30 keV count rate. The appearance of the QPO is related to spectral hardening of the flux, due to a change in the relative contribution of the soft and hard spectral components. We compare this peculiar behavior with results from other systems that show similar low frequency QPO peaks, and discuss the results in terms of possible theoretical models for QPO production.Comment: Author list corrected, small typos fixe

Endocannabinoids Generated by Ca2+ or by Metabotropic Glutamate Receptors Appear to Arise from Different Pools of Diacylglycerol Lipase

The identity and subcellular sources of endocannabinoids (eCBs) will shape their ability to affect synaptic transmission and, ultimately, behavior. Recent discoveries support the conclusion that 2-arachidonoyl glycerol, 2-AG, is the major signaling eCB, however, some important issues remain open. 2-AG can be synthesized by a mechanism that is strictly Ca2+-dependent, and another that is initiated by G-protein coupled receptors (GPCRs) and facilitated by Ca2+. An important question is whether or not the 2-AG in these cases is synthesized by the same pool of diacylglycerol lipase alpha (DAGLα). Using whole-cell voltage-clamp techniques in CA1 pyramidal cells in acute in vitro rat hippocampal slices, we investigated two mechanistically distinct eCB-mediated responses to address this issue. We now report that pharmacological inhibitors of DGLα have quantitatively different effects on eCB-mediated responses triggered by different stimuli, suggesting that functional, and perhaps physical, distinctions among pools of DAGLα exist

A biophysical model of endocannabinoid-mediated short term depression in hippocampal inhibition

Memories are believed to be represented in the synaptic pathways of vastly interconnected networks of neurons. The plasticity of synapses, that is, their strengthening and weakening depending on neuronal activity, is believed to be the basis of learning and establishing memories. An increasing number of studies indicate that endocannabinoids have a widespread action on brain function through modulation of synap–tic transmission and plasticity. Recent experimental studies have characterised the role of endocannabinoids in mediating both short- and long-term synaptic plasticity in various brain regions including the hippocampus, a brain region strongly associated with cognitive functions, such as learning and memory. Here, we present a biophysically plausible model of cannabinoid retrograde signalling at the synaptic level and investigate how this signalling mediates depolarisation induced suppression of inhibition (DSI), a prominent form of shortterm synaptic depression in inhibitory transmission in hippocampus. The model successfully captures many of the key characteristics of DSI in the hippocampus, as observed experimentally, with a minimal yet sufficient mathematical description of the major signalling molecules and cascades involved. More specifically, this model serves as a framework to test hypotheses on the factors determining the variability of DSI and investigate under which conditions it can be evoked. The model reveals the frequency and duration bands in which the post-synaptic cell can be sufficiently stimulated to elicit DSI. Moreover, the model provides key insights on how the state of the inhibitory cell modulates DSI according to its firing rate and relative timing to the post-synaptic activation. Thus, it provides concrete suggestions to further investigate experimentally how DSI modulates and is modulated by neuronal activity in the brain. Importantly, this model serves as a stepping stone for future deciphering of the role of endocannabinoids in synaptic transmission as a feedback mechanism both at synaptic and network level

Tetrahydrobiopterin modulates ubiquitin conjugation to UBC13/UBE2N and proteasome activity by S-nitrosation

Nitric Oxide (NO) is an intracellular signalling mediator, which affects many biological processes via the posttranslational modification of proteins through S-nitrosation. The availability of NO and NOS-derived reactive oxygen species (ROS) from enzymatic uncoupling are determined by the NO synthase cofactor Tetrahydrobiopterin (BH4). Here, using a global proteomics “biotin-switch” approach, we identified components of the ubiquitin-proteasome system to be altered via BH4-dependent NO signalling by protein S-nitrosation. We show S-nitrosation of ubiquitin conjugating E2 enzymes, in particular the catalytic residue C87 of UBC13/UBE2N, leading to impaired polyubiquitylation by interfering with the formation of UBC13~Ub thioester intermediates. In addition, proteasome cleavage activity in cells also seems to be altered by S-nitrosation, correlating with the modification of cysteine residues within the 19S regulatory particle and catalytic subunits of the 20S complex. Our results highlight the widespread impact of BH4 on downstream cellular signalling as evidenced by the effect of a perturbed BH4-dependent NO-Redox balance on critical processes within the ubiquitin-proteasome system (UPS). These studies thereby uncover a novel aspect of NO associated modulation of cellular homeostasis