The asialoglycoprotein receptor in human hepatocellular carcinomas: its expression on proliferating cells

The expression of the asialoglycoprotein receptor (ASGP-R) on human hepatocellular carcinoma (HCC) cells might be exploited to reduce the extrahepatic toxicity of DNA synthesis inhibitors by their conjugation with galactosyl-terminating peptides. In the present study we first assessed the frequency of ASGP-R expression in 60 HCCs. Secondly, we investigated whether the receptor was maintained on the plasma membranes of DNA synthesizing cancer cells. Needle biopsies of HCC were evaluated. Diagnosis and grading of HCC were performed on routine haematoxylin and eosin-stained sections according to Edmondson and Steiner (1953). Thirty-five tumours were grade I and II and were classified as well differentiated, while 25 tumours were grade III and IV and were classified as poorly differentiated. Sections from formalin-fixed, paraffin-embedded samples were incubated, after antigen retrieval, with an anti-ASGP-R monoclonal antibody revealed by secondary biotinylated antibody and streptavidin–biotin–peroxidase–diaminobenzidine reaction. A clear immunolabelling of plasma membranes of HCC cells was observed in 28 out of 35 (80%) well differentiated (grade I and II) and in five out of 25 (20%) poorly differentiated (grade III and IV) HCCs. The presence of the ASGP-R on the surface of DNA synthesizing cancer cells was also investigated after in vitro bromodeoxyuridine (BrdU) labelling of HCC samples by immunohistochemical visualization of both the ASGP-R and incorporated BrdU on the same section. The results obtained clearly demonstrated that DNA synthesizing cancer cells expressed the ASGP-R on their surface. The presence of ASGP-R on cell plasma membrane in the majority of differentiated HCCs and its maintenance on proliferating cells encourages studies in order to restrict the action of the inhibitors of DNA synthesis of HCC cells by their conjugation with galactosyl-terminating carriers internalized through this receptor. © 1999 Cancer Research Campaig

Hercules X-1: Empirical Models of UV Emission Lines

The UV emission lines of Hercules X-1, resolved with the HST GHRS and STIS, can be divided into broad (FWHM 750 km/s) and narrow (FWHM 150 km/s) components. The broad lines can be unambiguously identified with emission from an accretion disk which rotates prograde with the orbit. The narrow lines, previously identified with the X-ray illuminated atmosphere of the companion star, are blueshifted at both phi=0.2 and phi=0.8 and the line flux at phi=0.2 is 0.2 of the flux at phi=0.8. Line ratio diagnostics show that the density of the narrow line region is log n=13.4+/-0.2 and the temperature is T=1.0+/-0.2x10^5 K. The symmetry of the eclipse ingress suggests that the line emission on the surface of the disk is left-right symmetric relative to the orbit. Model fits to the O V, Si IV, and He II line profiles agree with this result, but fits to the N V lines suggest that the receding side of the disk is brighter. We note that there are narrow absorption components in the N V lines with blueshifts of 500 km/s.Comment: To be published in the Astrophysical Journa

Influence of tumor microenvironment and fibroblast population plasticity on melanoma growth, therapy resistance and immunoescape

Cutaneous melanoma (CM) tissue represents a network constituted by cancer cells and tumor microenvironment (TME). A key feature of CM is the high structural and cellular plasticity of TME, allowing its evolution with disease and adaptation to cancer cell and environmental alter-ations. In particular, during melanoma development and progression each component of TME by interacting with each other and with cancer cells is subjected to dramatic structural and cellular modifications. These alterations affect extracellular matrix (ECM) remodelling, phenotypic profile of stromal cells, cancer growth and therapeutic response. The stromal fibroblast populations of the TME include normal fibroblasts and melanoma‐associated fibroblasts (MAFs) that are highly abun-dant and flexible cell types interacting with melanoma and stromal cells and differently influencing CM outcomes. The shift from the normal microenvironment to TME and from normal fibroblasts to MAFs deeply sustains CM growth. Hence, in this article we review the features of the normal mi-croenvironment and TME and describe the phenotypic plasticity of normal dermal fibroblasts and MAFs, highlighting their roles in normal skin homeostasis and TME regulation. Moreover, we dis-cuss the influence of MAFs and their secretory profiles on TME remodelling, melanoma progres-sion, targeted therapy resistance and immunosurveillance, highlighting the cellular interactions, the signalling pathways and molecules involved in these processes

The 1999 Hercules X-1 Anomalous Low State

A failed main-on in the 35d cycle of Her X-1 was observed with the Rossi X-Ray Timing Explorer (RXTE) on 1999 April 26. Exceptions to the normal 35d cycle have been seen only twice before; in 1983 and again 1993. We present timing and spectral results of this latest Anomalous Low State (ALS) along with comparisons to the main-on and normal low states. Pulsations were observed in the 3-18 keV band with a fractional RMS variation of (0.037+-0.003). Spectral analysis indicates that the ALS spectrum has the same shape as the main-on but is modified by heavy absorption and scattering. We find that 70% of the observed emission has passed through a cold absorber (N_H=5.0x10^{23}cm^{-2}). This partially absorbing spectral fit can be applied to the normal low state with similar results. We find that the ALS observations may be interpreted as a decrease in inclination of the accretion disk causing the central X-Ray source to be obscured over the entire 35d cycle.Comment: revised text, 6 revised figures, accepted for publication in Ap

IBtkα Activates the β‐Catenin‐Dependent Transcription of MYC through Ubiquitylation and Proteasomal Degradation of GSK3βin Cancerous B Cells

The IBTK gene encodes the IBtkα protein that is a substrate receptor of E3 ubiquitin ligase, Cullin 3. We have previously reported the pro‐tumorigenic activity of Ibtk in MYC‐dependent B‐ lymphomagenesis observed in Eμ‐myc transgenic mice. Here, we provide mechanistic evidence of the functional interplay between IBtkα and MYC. We show that IBtkα, albeit indirectly, activates the β‐catenin‐dependent transcription of the MYC gene. Of course, IBtkαassociates with GSK3β and promotes its ubiquitylation, which is associated with proteasomal degradation. This event increases the protein level of β‐catenin, a substrate of GSK3β, and results in the transcriptional activation of the MYC and CCND1 target genes of β‐catenin, which are involved in the control of cell division and apoptosis. In particular, we found that in Burkitt’s lymphoma cells, IBtkα silencing triggered the downregulation of both MYC mRNA and protein expression, as well as a strong decrease of cell survival, mainly through the induction of apoptotic events, as assessed by using flow cytometry‐based cell cycle and apoptosis analysis. Collectively, our results shed further light on the complex puzzle of IBtkα interactome and highlight IBtkα as a potential novel therapeutic target to be employed in the strategy for personalized therapy of B cell lymphoma

Probing the outer edge of an accretion disk : a Her X-1 turn-on observed with RXTE

We present the analysis of Rossi X-ray Timing Explorer (RXTE) observations of the turn-on phase of a 35 day cycle of the X-ray binary Her X-1. During the early phases of the turn-on, the energy spectrum is composed of X-rays scattered into the line of sight plus heavily absorbed X-rays. The energy spectra in the 3–17 keV range can be described by a partial covering model, where one of the components is influenced by photoelectric absorption and Thomson scattering in cold material plus an iron emission line at 6.5 keV. In this paper we show the evolution of spectral parameters as well as the evolution of the pulse profile during the turn-on. We describe this evolution using Monte Carlo simulations which self-consistently describe the evolution of the X-ray pulse profile and of the energy spectrum

Stability of the Cyclotron Resonance Scattering Feature in Her X-1 with RXTE

Five observations of the hard X-ray spectrum of Her X-1 from \xte show that the $\sim 41$ keV energy of the cyclotron scattering line is constant within statistics of a few percent per observtion. Comparison with other observations over many years indicates strongly that the centroid energy of this absorption line has increased some time between 1991 and 1993 by 23%, from 34 keV to 41 keV. Moreover, the cutoff energy of the spectral continuum increased at the same time from 16 keV to 20 keV, which is, within the statistical error of 5%, in direct proportion to the centroid. This may be a sign thatboth these characteristics of the spectrum are controlled in the same way by the magnetic field strength in the region of scattering.Comment: 30 page, 5 figure

The expression of inhibitor of bruton's tyrosine kinase gene is progressively up regulated in the clinical course of chronic lymphocytic leukaemia conferring resistance to apoptosis.

Chronic lymphocytic leukaemia (CLL) is the most common B-cell malignancy with a variable clinical outcome. Biomarkers of CLL progression are required for optimising prognosis and therapy. The Inhibitor of Bruton's tyrosine kinase-isoform α (IBTKα) gene encodes a substrate receptor of Cullin 3-dependent E3 ubiquitin ligase, and promotes cell survival in response to the reticulum stress. Searching for novel markers of CLL progression, we analysed the expression of IBTKα in the peripheral blood B-cells of CLL patients, before and after first line therapy causing remission. The expression of IBTKα was significantly increased in disease progression, and decreased in remission after chemotherapy. Consistently with a pro-survival action, RNA interference of IBTKα increased the spontaneous and Fludarabine-induced apoptosis of MEC-1 CLL cells, and impaired the cell cycle of DeFew B-lymphoma cells by promoting the arrest in G0/G1 phase and apoptosis. Consistently, RNA interference of IBTKα up regulated the expression of pro-apoptotic genes, including TNF, CRADD, CASP7, BNIP3 and BIRC3. Our results indicate that IBTKα is a novel marker of CLL progression promoting cell growth and resistance to apoptosis. In this view, IBTKα may represent an attractive cancer drug target for counteracting the therapy-resistance of tumour cells

Insights into Thymus Development and Viral Thymic Infections

T-cell development in the thymus is a complex and highly regulated process, involving a wide variety of cells and molecules which orchestrate thymocyte maturation into either CD4+ or CD8+ single-positive (SP) T cells. Here, we briefly review the process regulating T-cell differentiation, which includes the latest advances in this field. In particular, we highlight how, starting from a pool of hematopoietic stem cells in the bone marrow, the sequential action of transcriptional factors and cytokines dictates the proliferation, restriction of lineage potential, T-cell antigen receptors (TCR) gene rearrangements, and selection events on the T-cell progenitors, ultimately leading to the generation of mature T cells. Moreover, this review discusses paradigmatic examples of viral infections affecting the thymus that, by inducing functional changes within this lymphoid gland, consequently influence the behavior of peripheral mature T-lymphocytes