Distinct EpCAM-Positive stem cell niches are engaged in chronic and neoplastic liver diseases

In normal human livers, EpCAMpos cells are mostly restricted in two distinct niches, which are (i) the bile ductules and (ii) the mucous glands present inside the wall of large intrahepatic bile ducts (the so-called peribiliary glands). These EpCAMpos cell niches have been proven to harbor stem/progenitor cells with great importance in liver and biliary tree regeneration and in the pathophysiology of human diseases. The EpCAMpos progenitor cells within bile ductules are engaged in driving regenerative processes in chronic diseases affecting hepatocytes or interlobular bile ducts. The EpCAMpos population within peribiliary glands is activated when regenerative needs are finalized to repair large intra- or extra-hepatic bile ducts affected by chronic pathologies, including primary sclerosing cholangitis and ischemia-induced cholangiopathies after orthotopic liver transplantation. Finally, the presence of distinct EpCAMpos cell populations may explain the histological and molecular heterogeneity characterizing cholangiocarcinoma, based on the concept of multiple candidate cells of origin. This review aimed to describe the precise anatomical distribution of EpCAMpos populations within the liver and the biliary tree and to discuss their contribution in the pathophysiology of human liver diseases, as well as their potential role in regenerative medicine of the liver

Toxicity and activity of docetaxel in anthracycline-pretreated breast cancer patients: a phase II study

: Docetaxel has proven effective in advanced breast cancer. Myelosuppression and cumulative fluid retention syndrome are troublesome, potentially avoidable toxicities. In this consecutive cohort study, docetaxel (100 mg/m2 by 1 hour i.v. infusion, q3 weeks) activity and toxicity was explored in 56 anthracycline-pretreated patients (eligible: 55: median age: 51 years [range: 28-68 years]; median performance status: 0 [range: 0-3]) with metastatic breast cancer, using two different granulocyte colony-stimulating factor and steroid pre- and postmedication schedules. Twenty-nine patients (group A) received a 5-day oral prednisone premedication, and 26 (group B) received 4-day low-dose i.m. dexamethasone; group B patients also received prophylactic granulocyte colony-stimulating factor. All patients were evaluable for toxicity and 53 for response. Prophylactic granulocyte colony-stimulating factor significantly lowered the incidence of grade III-IV neutropenia and neutropenic fever (p = 0.0001 and 0.01, respectively). The incidence of moderate-severe fluid retention syndrome was lower in patients receiving i.m. dexamethasone (p = 0.08). Overall response rate was 53% (4 complete responses/24 partial responses, 95% confidence interval 39.4-66.2%); 32% have stable disease and 15% progressive disease. In 21 anthracycline-refractory/resistant patients, as well as in 10 paclitaxel-pretreated patients, the overall response rate was 50%. Docetaxel is highly active in anthracycline- and paclitaxel-pretreated metastatic breast cancer, with manageable toxicity. Optimal use of both granulocyte colony-stimulating factor support and steroid premedication deserves further investigation

The FXR agonist obeticholic acid inhibits the cancerogenic potential of human cholangiocarcinoma

Cholangiocarcinoma (CCA) is an aggressive cancer with high resistance to chemotherapeutics. CCA is enriched in cancer stem cells, which correlate with aggressiveness and prognosis. FXR, a member of the metabolic nuclear receptor family, is markedly down-regulated in human CCA. Our aim was to evaluate, in primary cultures of human intrahepatic CCA (iCCA), the effects of the FXR agonist obeticholic acid (OCA), a semisynthetic bile acid derivative, on their cancerogenic potential. Primary human iCCA cell cultures were prepared from surgical specimens of mucinous or mixed iCCA subtypes. Increasing concentrations (0–2.5 μM) of OCA were added to culture media and, after 3–10 days, effects on proliferation (MTS assay, cell population doubling time), apoptosis (annexin V-FITC/propidium iodide), cell migration and invasion (wound healing response and Matrigel invasion assay), and cancerogenic potential (spheroid formation, clonogenic assay, colony formation capacity) were evaluated. Results: FXR gene expression was downregulated (RT-qPCR) in iCCA cells vs normal human biliary tree stem cells (p < 0.05) and in mucinous iCCA vs mixed iCCA cells (p < 0.05) but was upregulated by addition of OCA. OCA significantly (p < 0.05) inhibited proliferation of both mucinous and mixed iCCA cells, starting at a concentration as low as 0.05 μM. Also, CDCA (but not UDCA) inhibited cell proliferation, although to a much lower extent than OCA, consistent with its different affinity for FXR. OCA significantly induced apoptosis of both iCCA subtypes and decreased their in vitro cancerogenic potential, as evaluated by impairment of colony and spheroid formation capacity and delayed wound healing and Matrigel invasion. In general, these effects were more evident in mixed than mucinous iCCA cells. When tested together with Gemcitabine and Cisplatin, OCA potentiated the anti-proliferative and pro-apoptotic effects of these chemotherapeutics, but mainly in mixed iCCA cells. OCA abolished the capacity of both mucinous and mixed iCCA cells to form colonies when administered together with Gemcitabine and Cisplatin. In subcutaneous xenografts of mixed iCCA cells, OCA alone or combined with Gemcitabine or Cisplatin markedly reduced the tumor size after 5 weeks of treatment by inducing necrosis of tumor mass and inhibiting cell proliferation. In conclusion, FXR is down-regulated in iCCA cells, and its activation by OCA results in anti-cancerogenic effects against mucinous and mixed iCCA cells, both in vitro and in vivo. The effects of OCA predominated in mixed iCCA cells, consistent with the lower aggressiveness and the higher FXR expression in this CCA subtype. These results, showing the FXR-mediated capacity of OCA to inhibit cholangiocarcinogenesis, represent the basis for testing OCA in clinical trials of CCA patients

Peribiliary glands are key in regeneration of the human biliary epithelium after severe bile duct injury

Peribiliary glands (PBG) are a source of stem/progenitor cells organized in a cellular network encircling large bile ducts. Severe cholangiopathy with loss of luminal biliary epithelium has been proposed to activate PBG, resulting in cell proliferation and differentiation to restore biliary epithelial integrity. However, formal evidence for this concept in human livers is lacking. We, therefore, developed a novel ex vivo model using precision-cut slices of extrahepatic human bile ducts obtained from discarded donor livers, providing an intact anatomical organization of cell structures, to study spatiotemporal differentiation and migration of PBG cells after severe biliary injury. Post-ischemic bile duct slices were incubated in oxygenated culture medium for up to a week. At baseline, severe tissue injury was evident with loss of luminal epithelial lining and mural stroma necrosis. In contrast, PBG remained relatively well preserved and different reactions of PBG were noted, including PBG dilatation, cell proliferation and maturation. Proliferation of PBG cells increased after 24 h of oxygenated incubation, reaching a peak after 72 h. Proliferation of PBG cells was paralleled by a reduction in PBG apoptosis and differentiation from a primitive and pluripotent (Nanog+/Sox9+) to a mature (CFTR+/secretin receptor+) and activated phenotype (increased expression of HIF-1α, Glut-1, and VEGF-A). Migration of proliferating PBG cells in our ex vivo model was unorganized, but resulted in generation of epithelial monolayers at stromal surfaces. CONCLUSION: Human PBG contain biliary progenitor cells and are able to respond to bile duct epithelial loss with proliferation, differentiation, and maturation to restore epithelial integrity. The ex vivo spatiotemporal behaviour of human PBG cells provides evidence for a pivotal role of PBG in biliary regeneration after severe injury. This article is protected by copyright. All rights reserved

Multiperiodicity in the newly discovered mid-late Be star V2104 Cygni

We obtained the first long, homogenous time-series of V2104Cyg, consisting of 679 datapoints, with the uvbybeta photometers of Sierra Nevada and San Pedro Martir Observatories with the aim to detect and subsequently interpret the intrinsic frequencies of this previously unstudied variable star, which turned out to be a Be star. We try to figure out its place among the variable B stars on the upper Main Sequence. In order to obtain additional information on physical parameters we collected a few spectra with the ELODIE and FIES instruments. We searched for frequencies in the uvby passbands using 2 different frequency analysis methods and used the S/N>4 criterion to select the significant periodicities. We obtained an estimate of the physical parameters of the underlying B star of spectral type between B5 and B7, by correcting for the presence of a circumstellar disk, using a formalism based on the strenght of the Halpha line emission. We detected 3 independent frequencies with amplitudes below 0.01mag, f1 = 4.7126 c/d, f2 = 2.2342 c/d and f3 = 4.671 c/d, and discovered that V2104Cyg is a Be star. The fast rotation (vsini=290+/-10 km/s, and 27<i<45) hampered the investigation of the associated pulsational parameters l. Nevertheless, the most plausible explanation for the observed variability of this mid-late type Be star is a non-radial pulsation model. This paper is based on observations obtained at the Observatorio Astronomico Nacional San Pedro Martir (Mexico), Observatorio de Sierra Nevada (Spain), Observatoire de Haute Provence (France), and on observations made with the Nordic Optical Telescope, Observatorio Roque de los Muchachos, La Palma, Spain.Comment: 7 pages, 4 figures, A&A accepte

Activation of Fas/FasL pathway and the role of c-FLIP in primary culture of human cholangiocarcinoma cells

Intrahepatic cholangiocarcinoma (iCCA) represents a heterogeneous group of malignancies emerging from the biliary tree, often in the context of chronic bile ducts inflammation. The immunological features of iCCA cells and their capability to control the lymphocytes response have not yet been investigated. The aims of the present study were to evaluate the interaction between iCCA cells and human peripheral blood mononuclear cells (PBMCs) and the role of Fas/FasL in modulating T-cells and NK-cells response after direct co-culture. iCCA cells express high levels of Fas and FasL that increase after co-culture with PBMCs inducing apoptosis in CD4(+), CD8(+) T-cells and in CD56(+) NK-cells. In vitro, c-FLIP is expressed in iCCA cells and the co-culture with PBMCs induces an increase of c-FLIP in both iCCA cells and biliary tree stem cells. This c-FLIP increase does not trigger the caspase cascade, thus hindering apoptotis of iCCA cells which, instead, underwent proliferation. The increased expression of Fas, FasL and c-FLIP is confirmed in situ, in human CCA and in primary sclerosing cholangitis. In conclusion our data indicated that iCCA cells have immune-modulatory properties by which they induce apoptosis of T and NK cells, via Fas/FasL pathway, and escape inflammatory response by up-regulating c-FLIP system

The double-mode nature of the HADS star GSC 00144-03031 and the Petersen diagram of the class

The double--mode pulsation of GSC 00144-03031 has been detected when searching for COROT targets. A very large dataset composed of 4722 photometric measurements was collected at six observatories in Europe and America. There is no hint of the excitation of additional modes (down to 0.6 mmag) and therefore GSC 00144-03031 seems to be a pure double--mode pulsator, with a very short fundamental radial mode (P=84 min). From Stromgren photometry and evolutionary tracks it appears to be a Pop. I star with M=1.75 solar masses, located in the middle of the instability strip, close to the Zero--Age Main Sequence. We also discovered other new double--mode pulsators in the databases of large--scale projects: OGLE BW2_V142, OGLE BW1_V207, ASAS3 094303-1707.3, ASAS3 000116-6037.0, NSVS 3234596 and NSVS 3324715. An observational Petersen diagram is presented and explained by means of new models. A common sequence connecting Pop. I stars from the shortest to the longest periods is proposed and the spreads in the period ratios are ascribed to different metallicities (at the shortest periods) and to different masses (at the longest ones). The paper is based on data collected at S. Pedro Martir and Sierra Nevada Observatories and on the contributions from several amateur astronomers.Comment: 9 pages, 5 pages, accepted for publication in Astronomy and Astrophysic

Ground-based astrometry calibrated by Gaia DR1: new perspectives in asteroid orbit determination

Context. The Gaia Data Release 1 (GDR1) is a first, important step on the path of evolution of astrometric accuracy towards a much improved situation. Although asteroids are not present in GDR1, this intermediate release already impacts asteroid astrometry. Aims. Our goal is to investigate how the GDR1 can change the approach to a few typical problems, including the determination of orbits from short-arc astrometry, the exploitation of stellar occultations, and the impact risk assessment. Methods.We employ optimised asteroid orbit determination tools, and study the resulting orbit accuracy and post-fit residuals. For this goal, we use selected ground-based asteroid astrometry, and occultation events observed in the past. All measurements are calibrated by using GDR1 stars. Results. We show that, by adopting GDR1, very short measurement arcs can already provide interesting orbital solutions, capable of correctly identifying near-Earth asteroids (NEAs) and providing a much more accurate risk rating. We also demonstrate that occultations, previously used to derive asteroid size and shapes, now reach a new level of accuracy at which they can be fruitfully used to obtain astrometry at the level of accuracy of Gaia star positions

The Italian Registry for Primary Immunodeficiencies (Italian Primary Immunodeficiency Network; IPINet): Twenty Years of Experience (1999&#8211;2019)

Primary immunodeficiencies (PIDs) are heterogeneous disorders, characterized by variable clinical and immunological features. National PID registries offer useful insights on the epidemiology, diagnosis, and natural history of these disorders. In 1999, the Italian network for primary immunodeficiencies (IPINet) was established. We report on data collected from the IPINet registry after 20\ua0years of activity. A total of 3352 pediatric and adult patients affected with PIDs are registered in the database. In Italy, a regional distribution trend of PID diagnosis was observed. Based on the updated IUIS classification of 2019, PID distribution in Italy showed that predominantly antibody deficiencies account for the majority of cases (63%), followed by combined immunodeficiencies with associated or syndromic features (22.5%). The overall age at diagnosis was younger for male patients. The minimal prevalence of PIDs in Italy resulted in 5.1 per 100.000 habitants. Mortality was similar to other European registries (4.2%). Immunoglobulin replacement treatment was prescribed to less than one third of the patient cohort. Collectively, this is the first comprehensive description of the PID epidemiology in Italy