NY-ESO-1 tumour associated antigen is a cytoplasmic protein detectable by specific monoclonal antibodies in cell lines and clinical specimens

NY-ESO-1 gene encodes a novel member of the cancer/testis (CT) family of human tumour-associated antigens (TAA). Specific monoclonal antibodies (mAb) have identified the corresponding gene product in lysates of tumour cell lines as a 22 kDa protein but no data are available concerning its intracellular location or distribution within neoplastic tissues. We have generated NY-ESO-1 specific mAbs recognizing the target molecule in cytospin preparations and in sections from clinical tumour specimens. These reagents identify NY-ESO-1 TAA in melanoma cell lines expressing the specific gene as a cytoplasmic protein, sharing the intracellular location of most MAGE TAA. In a series of 12 melanoma specimens, specific staining, limited to neoplastic cells, was detectable in the five cases where NY-ESO-1 gene expression was observed. In two of them over 90% of tumour cells showed evidence of positive staining. Lower percentages of positive neoplastic cells ranging between single cells and 50% were observed in the remaining tumours. These data suggest that active specific immunotherapies targeting NY-ESO-1, alone or in combination with other TAA could be of high clinical relevance in sizeable subgroups of melanoma patients. © 2000 Cancer Research Campaig

Whole blood assessment of antigen specific cellular immune response by real time quantitative PCR: a versatile monitoring and discovery tool

BACKGROUND: Monitoring of cellular immune responses is indispensable in a number of clinical research areas, including microbiology, virology, oncology and autoimmunity. Purification and culture of peripheral blood mononuclear cells and rapid access to specialized equipment are usually required. We developed a whole blood (WB) technique monitoring antigen specific cellular immune response in vaccinated or naturally sensitized individuals. METHODS: WB (300 microl) was incubated at 37 degrees C with specific antigens, in the form of peptides or commercial vaccines for 5-16 hours. Following RNAlater addition to stabilize RNA, the mixture could be stored over one week at room temperature or at 4 degrees C. Total RNA was then extracted, reverse transcribed and amplified in quantitative real-time PCR (qRT-PCR) assays with primers and probes specific for cytokine and/or chemokine genes. RESULTS: Spiking experiments demonstrated that this technique could detect antigen specific cytokine gene expression from 50 cytotoxic T lymphocytes (CTL) diluted in 300 microl WB. Furthermore, the high sensitivity of this method could be confirmed ex-vivo by the successful detection of CD8+ T cell responses against HCMV, EBV and influenza virus derived HLA-A0201 restricted epitopes, which was significantly correlated with specific multimer staining. Importantly, a highly significant (p = 0.000009) correlation between hepatitis B surface antigen (HBsAg) stimulated IL-2 gene expression, as detectable in WB, and specific antibody titers was observed in donors vaccinated against hepatitis B virus (HBV) between six months and twenty years before the tests. To identify additional markers of potential clinical relevance, expression of chemokine genes was also evaluated. Indeed, HBsAg stimulated expression of MIP-1beta (CCL4) gene was highly significantly (p = 0.0006) correlated with specific antibody titers. Moreover, a longitudinal study on response to influenza vaccine demonstrated a significant increase of antigen specific IFN-gamma gene expression two weeks after immunization, declining thereafter, whereas increased IL-2 gene expression was still detectable four months after vaccination. CONCLUSION: This method, easily amenable to automation, might qualify as technology of choice for high throughput screening of immune responses to large panels of antigens from cohorts of donors. Although analysis of cytokine gene expression requires adequate laboratory infrastructure, initial antigen stimulation and storage of test probes can be performed with minimal equipment and time requirements. This might prove important in "field" studies with difficult access to laboratory facilities

The MAGPI Survey: Drivers of kinematic asymmetries in the ionised gas of z∼0.3z\sim0.3 star-forming galaxies

Galaxy gas kinematics are sensitive to the physical processes that contribute to a galaxy's evolution. It is expected that external processes will cause more significant kinematic disturbances in the outer regions, while internal processes will cause more disturbances for the inner regions. Using a subsample of 47 galaxies ($0.27<z<0.36$) from the Middle Ages Galaxy Properties with Integral Field Spectroscopy (MAGPI) survey, we conduct a study into the source of kinematic disturbances by measuring the asymmetry present in the ionised gas line-of-sight velocity maps at the $0.5R_e$ (inner regions) and $1.5R_e$ (outer regions) elliptical annuli. By comparing the inner and outer kinematic asymmetries, we aim to better understand what physical processes are driving the asymmetries in galaxies. We find the local environment plays a role in kinematic disturbance, in agreement with other integral field spectroscopy studies of the local universe, with most asymmetric systems being in close proximity to a more massive neighbour. We do not find evidence suggesting that hosting an Active Galactic Nucleus (AGN) contributes to asymmetry within the inner regions, with some caveats due to emission line modelling. In contrast to previous studies, we do not find evidence that processes leading to asymmetry also enhance star formation in MAGPI galaxies. Finally, we find a weak anti-correlation between stellar mass and asymmetry (ie. high stellar mass galaxies are less asymmetric). We conclude by discussing possible sources driving the asymmetry in the ionised gas, such as disturbances being present in the colder gas phase (either molecular or atomic) prior to the gas being ionised, and non-axisymmetric features (e.g., a bar) being present in the galactic disk. Our results highlight the complex interplay between ionised gas kinematic disturbances and physical processes involved in galaxy evolution.Comment: e.g., 20 pages, 19 figure

The MBHBM Project-I: measurement of the central black hole mass in spiral galaxy NGC 3504 using molecular gas kinematics

We present a dynamical mass measurement of the supermassive black hole (SMBH) in the nearby double-barred spiral galaxy NGC 3504 as part of the Measuring Black Holes in below Milky Way (M sstarf) Mass Galaxies Project. Our analysis is based on Atacama Large Millimeter/submillimeter Array cycle 5 observations of the ${}^{12}\mathrm{CO}(2-1)$ emission line. These observations probe NGC 3504's circumnuclear gas disk (CND). Our dynamical model of the CND simultaneously constrains a black hole (BH) mass of ${1.6}_{-0.4}^{+0.6}\times {10}^{7}$ M ⊙, which is consistent with the empirical BH–galaxy scaling relations and a mass-to-light ratio in the H band of 0.44 ± 0.12 (M ⊙/${L}_{\odot }$). This measurement also relies on our new estimation of the distance to the galaxy of 32.4 ± 2.1 Mpc using the surface brightness fluctuation method, which is much further than the existing distance estimates. Additionally, our observations detect a central deficit in the ${}^{12}\mathrm{CO}(2-1)$ integrated intensity map with a diameter of 6.3 pc at the putative position of the SMBH. However, we find that a dense gas tracer CS(5 − 4) peaks at the galaxy center, filling in the 12CO(2 − 1)-attenuated hole. Holes like this one are observed in other galaxies, and our observations suggest these may be caused by changing excitation conditions rather than a true absence of molecular gas around the nucleus

Tumour antigen expression in hepatocellular carcinoma in a low-endemic western area

Background: Identification of tumour antigens is crucial for the development of vaccination strategies against hepatocellular carcinoma (HCC). Most studies come from eastern-Asia, where hepatitis-B is the main cause of HCC. However, tumour antigen expression is poorly studied in low-endemic, western areas where the aetiology of HCC differs. Methods: We constructed tissue microarrays from resected HCC tissue of 133 patients. Expression of a comprehensive panel of cancer-testis (MAGE-A1, MAGE-A3/4, MAGE-A10, MAGE-C1, MAGE-C2, NY-ESO-1, SSX-2, sperm protein 17), onco-fetal (AFP, Glypican-3) and overexpressed tumour antigens (Annexin-A2, Wilms tumor-1, Survivin, Midkine, MUC-1) was determined by immunohistochemistry. Results: A higher prevalence of MAGE antigens was observed in patients with hepatitis-B. Patients with expression of more tumour antigens in general had better HCC-specific survival (P=0.022). The four tumour antigens with high expression in HCC and no, or weak, expression in surrounding tumour-free-liver tissue, were Annexin-A2, GPC-3, MAGE-C1 and MAGE-C2, expressed in 90, 39, 17 and 20% of HCCs, respectively. Ninety-five percent of HCCs expressed at least one of these four tumour antigens. Interestingly, GPC-3 was associated with SALL-4 expression (P=0.001), an oncofetal transcription factor highly expressed in embryonal stem cells. SALL-4 and GPC-3 expression levels were correlated with vascular invasion, poor differentiation and higher AFP levels before surgery. Moreover, patients who co-expressed higher levels of both GPC-3 and SALL-4 had worse HCC-specific survival (P=0.018). Conclusions: We describe a panel of four tumour antigens with excellent coverage and good tumour specificity in a western area, low-endemic for hepatitis-B. The association between GPC-3 and SALL-4 is a novel finding and suggests that GPC-3 targeting may specifically attack the tumour stem-cell compartment

Steady flow of a viscous incompressible fluid in an unbounded funnel-shaped domain.

In this paper we consider a domain which is tube-like at one exit to infinity and the halfspace at the other side. We prove existence of steady motions for the Navier Stokes problem and for the case in which the fluid is moving through a porous medium at rest filling the domain. The flux is arbitrary. We describe the asymptotic behavior in the halfspace

Approach to assess a fast welding simulation in an industrial environment - application for an automotive welded part

Fusion welding processes are widely used for joining metal structures, such as pipes, ships, and cars. In general, these joining processes offer a very good compromise between reliability, safety, cost and maintenance which are important issues in the current economical context. The negative heat effects of welding, i.e. distortions and residual stresses of the welded parts, are well known and many researches in this field have already been done in the last decades in order to minimize them. On the experimental side, many sophisticated procedures have become state of the art to deal with this problem. On the computational side, the improvement of the simulation algorithms and the computing power enables the simulations of many physical phenomena occurring during the welding process. The implementation of welding simulation techniques is nevertheless not an easy task and often associated with expert knowledge which hinders their global application in an industrial environment. This paper is focused on the industrial requirements of a welding simulation software with special respect to the needs of the automotive industry. The necessary information to run a welding simulation and the expectations of a weld specialist without deep knowledge in numerical methods are investigated. These expectations are tested on an automotive welded assembly with a commercially available welding simulation software designed especially for the needs of the automotive industry. A welding experiment is done and the measured temperature distributions and distortions serve as reference to validate the simulation results. The result quality of the simulations of temperature fields and distortions is in best agreement with experimental data. The workflow is well adapted for the considered industrial requirements and the time-tosolution as well as the computational costs are acceptable, whereas the efficient calibration of the heat input model is still a point which will be further investigated in current and future research works

Microstructure of ionic liquid (EAN)-rich and oil-rich microemulsions studied by SANS

In a previous study we investigated the phase behavior of microemulsions consisting of the ionic liquid ethylammonium nitrate (EAN), an n-alkane and a nonionic alkyl polyglycolether (CiEj). We found the same general trends as for the aqueous counterparts, i.e. a transition from an oil-in-EAN microemulsion via a bicontinuous microemulsion to an EAN-in-oil microemulsion with increasing temperature. However, unlike what happens in the corresponding aqueous systems, in EAN-in-oil microemulsions only a very small amount of EAN was detected by NMR-measurements. This is why we investigated the phase behavior and microstructure of EAN-rich n-dodecane-in-EAN microemulsions and oil-rich EAN-in-n-octane microemulsions. We found that the ionic liquid emulsification failure boundary has an extraordinarily small slope, which suggests that the amphiphilic film loses its ability to solubilize EAN with an increase in temperature by only a few degrees. The analysis of the small angle neutron scattering (SANS) curves unambiguously shows that this behavior is due to the fact that the EAN molecules form a substructure with a characteristic length scale of approximate to 8 angstrom inside the EAN-in-oil droplets. In more detail, the analysis of the SANS data with the GIFT method revealed a transition from spherical to cylindrical structures approaching the respective critical endpoint temperatures. By using the respective form factors and combining them with a Gaussian spatial intensity distribution to account for the EAN sub-structure we were able to describe the scattering curves nearly quantitatively