bluuebunny/crossref_metadata_2025 huggingface dataset:
Bill Litigation and the Observance of Sundays and Major Festivals in the Court of King's Bench in the Fifteenth Century
Authors: Jenks Susanne
Published: February 2008

The attitudes of medieval people toward Sundays and Holy Days have always been of interest to historians. They have been studied from at least five different perspectives. Max Levy, for instance, explained how Sunday developed from a day that commemorated Christ's resurrection, but was originally a working day (dies dominica), to a day of worship, contemplation, and rest. Initially no (servile) work was allowed, but exceptions were accepted because of necessity, like harvest work, or because of good intentions, like concern for the common good or for a pious cause. Others looked at the stance of the Church, analyzing the protests against the non-observance of Holy Days as well as the objections raised to the observance of Holy Days from the clergy or from laymen, or concentrated mainly on the work ban and its implications for working life in the Middle Ages. Willard and Haskett studied the observance of Sundays and Holy Days in government departments like the Lower Exchequer or the Chancery to see to what extent the working of the English government was affected. Legal historians, however, have not shown much interest in how the courts observed Sundays and Holy Days, mainly because everything seemed to have been settled since the late thirteenth century. Paul Brand recently stated that it had “clearly become the general practice by the second half of the reign of Edward I for the Common Bench and King's Bench not to sit on Sundays,” or on All Saints Day (1 November), All Souls Day (2 November), the feast of the Purification (2 February), Ascensiontide, and the feast of the Nativity of St. John the Baptist (24 June), all of which fell within term time.
The Jewish Question in the Genre System of Dostoevskii's Diary of a Writer and the Problem of the Authorial Image
Authors: Vassena Raffaella
Published: February 2008

The second edition of theDiary of a Writer(1876-1877) marked a crucial point in Fedor Dostoevskii's literary career: in spite of critics' attacks, many “ordinary” readers were overwhelmed by the author's charisma and began writing to Dostoevskii from different parts of Russia, expressing their views on the moral, social, and political issues dealt with in theDiary. Such success was also guaranteed by the original rhetorical and genre system of theDiary of a Writer, which, wisely modulated and addressed, aimed to involve readers and persuade them to share the author's beliefs. Raffaella Vassena explores the case of the article “The Jewish Question” in the issue of March 1877, where Dostoevskii's rhetoric actually failed to bring about what he had intended. By concentrating on new archival materials, Vassena investigates the reasons for this failure and submits a new perspective on the controversial question of Dostoevskii's attitude toward Jews.
Apports et perspectives de la technologie LiDAR aéroporté pour la pratique forestière | Contributions and perspectives of airborne LiDAR technology for forestry
Authors: Gachet Gilles , Junod Pascal
Published: February 2008

LiDAR is an optical remote sensing system that determines the distance to an object or to a surface by measuring the time delay between the transmission of a pulse and the detection of the reflected signal. A wide variety of applications arise from this technology which facilitate a better knowledge of land use. In forestry, airborne LiDAR systems provide, in addition to an accurate model of the relief, a 3D representation of the forest structure at various scales. This contribution presents the foundations of LiDAR technology and illustrates several forestry applications: underneath vegetation topography, canopy height and texture, vegetation delineation and stand delimitation. This contribution also outlines the potential of airborne laser scanning for forestry.
Solvent‐impregnated resins as an in situ product recovery tool for phenol recovery from Pseudomonas putida S12TPL fermentations
Authors: van den Berg Corjan , Wierckx Nick , Vente Johan , Bussmann Paul , de Bont Jan , van der Wielen Luuk
Published: February 2008

The sustainable production of fine/bulk chemicals is often hampered by product toxicity and inhibition to the producing micro‐organisms. Consequently, the product must be removed from the micro‐organisms' environment. To achieve this, so‐called solvent‐impregnated resins (SIRs) as well as commercial resins have been added to a Pseudomonas putida S12TPL fermentation that produces phenol as a model compound from glucose. The SIRs contained an ionic liquid which extracts phenol effectively. It was observed that the addition of these particles resulted in an increased phenol production of more than a fourfold while the commercial resin (XAD‐4) which is widely used in aromatic removal from aqueous phases, only gave a 2.5‐fold increase in volumetric production. Biotechnol. Bioeng. 2008;100: 466–472. © 2008 Wiley Periodicals, Inc.
Multiplex bacterial growth monitoring in 24‐well microplates using a dual optical sensor for dissolved oxygen and pH
Authors: Kocincová Anna S. , Nagl Stefan , Arain Sarina , Krause Christian , Borisov Sergey M. , Arnold Matthias , Wolfbeis Otto S.
Published: February 2008

Non‐invasive, simultaneous optical monitoring of oxygen and pH during bacterial cultivation in 24‐well microplates is presented using an integrated dual sensor for dissolved oxygen and pH values. The dual sensor is based on oxygen‐sensitive organosilica microparticles and pH‐sensitive microbeads from a polymethacrylate derivative embedded into a polyurethane hydrogel. The readout is based on a phase‐domain fluorescence lifetime‐based method referred to as modified frequency domain dual lifetime referencing using a commercially available detector system for 24‐well microplates. The sensor was used for monitoring the growth of Pseudomonas putida bacterial cultures. The method is suitable for parallelized, miniaturized bioprocessing, and cell‐based high‐throughput screening applications. Biotechnol. Bioeng. 2008;100: 430–438. © 2008 Wiley Periodicals, Inc.
In situ protein folding and activation in bacterial inclusion bodies
Authors: Gonzalez‐Montalban Nuria , Natalello Antonino , García‐Fruitós Elena , Villaverde Antonio , Doglia Silvia Maria
Published: February 2008

Recent observations indicate that bacterial inclusion bodies formed in absence of the main chaperone DnaK result largely enriched in functional, properly folded recombinant proteins. Unfortunately, the molecular basis of this intriguing fact, with obvious biotechnological interest, remains unsolved. We have explored here two non‐excluding physiological mechanisms that could account for this observation, namely selective removal of inactive polypeptides from inclusion bodies or in situ functional activation of the embedded proteins. By combining structural and functional analysis, we have not observed any preferential selection of inactive and misfolded protein species by the dissagregating machinery during inclusion body disintegration. Instead, our data strongly support that folding intermediates aggregated as inclusion bodies could complete their natural folding process once deposited in protein clusters, which conduces to significant functional activation. In addition, in situ folding and protein activation in inclusion bodies is negatively regulated by the chaperone DnaK. Biotechnol. Bioeng. 2008;100: 797–802. © 2008 Wiley Periodicals, Inc.
Growth and storage processes in aerobic granules grown on soybean wastewater
Authors: Ni Bing‐Jie , Yu Han‐Qing
Published: February 2008

Aerobic granules in a sequencing batch reactor (SBR) are subjected to alternative feast and famine conditions, and are able to take up carbon substrate in wastewater rapidly and to store it as intracellular storage products when the substrate is in excess. This phenomenon could not be described by the widely used activated sludge model No.3 (ASM3). In this work, taking adsorption process, microbial maintenance, and substrate diffusion into account, the simultaneous growth and storage processes occurring in an aerobic‐granule‐based SBR are investigated with experimental and modeling approaches. A new model is established and successfully validated with the experimental results of an SBR fed with soybean‐processing wastewater. Simulation results show that our approach is appropriate for elucidating the fates of major model components. Comparison between ASM3 and the model established in this work demonstrates that the latter is better to describe the substrate removal mechanisms and simultaneous growth and storage processes in aerobic granules. Biotechnol. Bioeng. 2008;100: 664–672. © 2008 Wiley Periodicals, Inc.
Characterization of mechanical properties of transgenic tobacco roots expressing a recombinant monoclonal antibody against tooth decay
Authors: Hassan Sally , Liu Wei , Ma Julian K‐C. , Thomas Colin R. , Keshavarz‐Moore Eli
Published: February 2008

In this article, we describe a new approach that allows the determination of the magnitude of force required to break single plant roots. Roots were taken from transgenic tobacco plants, expressing a secreted monoclonal antibody. They were divided into four key developmental stages. A novel micromanipulation technique was used to pull to breakage, single tobacco roots in buffer in order to determine their breaking force. A characteristic uniform step‐wise increase in the force up to a peak force for breakage was observed. The mean breaking force and mean work done were 101mN and 97µJ per root respectively. However, there was a significant increase in breaking force from the youngest white roots to the oldest, dark red–brown roots. We speculate that this was due to increasing lignin deposition with root stage of development (shown by phloroglucinol staining). No significant differences between fresh root mass, original root length, or mean root diameter for any of the root categories were found, displaying their uniformity, which would be beneficial for bioprocessing. In addition, no significant difference in antibody yield from the different root categories was found. These data show that it is possible to characterise the force requirements for root breakage and should assist in the optimisation of recombinant protein extraction from these roots. Biotechnol. Bioeng. 2008;100: 803–809. © 2008 Wiley Periodicals, Inc.
Hypervideo application on an experimental control system as an approach to education
Authors: Debevc Matjaž , Šafarič Riko , Golob Marjan
Published: February 2008

Hypervideo, as an interactive tool with links within video frames, is becoming widely used in multimedia presentations for e‐learning applications. Its concept of rich multimedia presentation together with temporal based link objects, gives a chance for use in engineering education courses, where many practical work and experiments are needed. In the article we present a strategy for the development and navigation of hypervideo application, to be used in engineering education. Example of magnetic suspension system experiment was built as hypervideo application. The system usability was tested with two methods, software usability measurement inventory (SUMI) evaluation and general approach to usability engineering. Test results have shown that hypervideo increases the users' motivation to work and helps students recognize, organize and present specific information. By applying some of the proposed improvements in future development, hypervideo could be a significant tool in the future of engineering education. © 2008 Wiley Periodicals, Inc. Comput Appl Eng Educ 16: 31–44, 2008; Published online in Wiley InterScience (www.interscience.wiley.com); DOI 10.1002/cae.20116
Chemistry and the Missing Era of Evolution
Authors: Cairns‐Smith A. Graham
Published: February 2008

The Darwinian evolution of life on earth depends utterly on complex molecular machinery, which, it seems, could only have arisen through a Darwinian evolution. The “RNA world” idea reduces this paradox, but requires a geochemically implausible supply of RNA monomers. A pre‐RNA era of natural selection is implied. I suggest that originally this was based on inorganic materials that came to replicate permutations with specific (e.g. catalytic) effects.
A novel modular probe base design
Authors: Behr Volker C. , Kaufmann Ilja , Haase Axel , Jakob Peter M.
Published: February 2008

Commercially available probe bases often impose limitations for their use with different setups. The goal of this work was to design and construct a universally applicable probe base to be used in radio frequency (rf) applications such as NMR imaging and spectroscopy. The concept is to support several arbitrarily combinable rf channels as well as additional devices, for example, a climate chamber for plants, an animal handling system, a perfusion device, or further specialized equipment. The concept and a prototype of such a probe base were successfully developed and are presented in this work. © 2008 Wiley Periodicals, Inc. Concepts Magn Reson Part B (Magn Reson Engineering) 33B: 55–61, 2008
A comparison of survival and disease‐specific survival in surgically resected, lymph node‐positive bronchioloalveolar carcinoma versus nonsmall cell lung cancer
Authors: Varlotto John M. , Flickinger John C. , Recht Abram , Nikolov Margaret C. , DeCamp Malcolm M.
Published: February 2008

BACKGROUND

The objective of this study was to assess whether disease‐specific survival (DSS) and overall survival (OS) differed among patients who had N1 and N2 bronchioloalveolar carcinoma (BAC) compared with patients who had non‐BAC nonsmall cell lung cancer (NSCLC).

METHODS

The Surveillance, Epidemiology, and End Results (SEER) Program database from 1992 to 2002 contained 684 patients with BAC and 9809 patients with non‐BAC NSCLC who had N1/N2 tumors and who underwent a definitive surgical procedure. OS and DSS rates were compared according to potential prognostic factors, including the use of a matched‐pair analysis.

RESULTS

The BAC patients with either pathologic N1 or N2 lymph node status were significantly more likely to be women, and nonblack/nonwhite race, but significantly less likely to have poorly differentiated or undifferentiated tumors than patients with non‐BAC cancers with comparable lymph node status. The median follow‐up of all patients was 29 months. There was a significant difference in DSS between patients with N2 BAC and non‐BAC cancers, but not for patients with N1 disease. There was a nonsignificant trend toward longer OS for patients with N2 BAC compared with non‐BAC cancers.

CONCLUSIONS

Patients with lymph node‐positive BAC had distinctly different patient and tumor characteristics than patients with lymph node‐positive non‐BAC NSCLC. Because DSS appears to be better for patients with N2 BAC, they may not benefit as much from adjuvant therapy as patients with non‐BAC NSCLC. Cancer 2008. ©2008 American Cancer Society.
External validation of IMP3 expression as an independent prognostic marker for metastatic progression and death for patients with clear cell renal cell carcinoma
Authors: Hoffmann Nathan E. , Sheinin Yuri , Lohse Christine M. , Parker Alexander S. , Leibovich Bradley C. , Jiang Zhong , Kwon Eugene D.
Published: February 2008

BACKGROUND

High‐quality external validation studies have recently been highlighted to be of paramount importance for proper translation of prognostic markers into the clinical setting. To that end, the authors examined associations of the insulin‐like growth factor‐II mRNA binding protein, IMP3, with clinical and pathologic features of clear cell renal cell carcinoma (ccRCC) in an independent cohort of patients to validate recent work showing IMP3 as a prognostic marker for RCC progression and death.

METHODS

The authors studied 716 consecutive tumor specimens from patients treated with surgery at the study institution for unilateral, sporadic, noncystic ccRCC between 1990 and 1999. Tissues were stained and scored for IMP3 expression and these expression levels were correlated with clinical and pathologic features as well as clinical outcomes including progression to distant metastases and death from RCC.

RESULTS

It was observed that 213 ccRCC specimens (29.8%) expressed tumor cell IMP3. IMP3 expression was associated with advanced stage and grade of primary tumors as well as other adverse features including coagulative tumor necrosis and sarcomatoid differentiation. After multivariate adjustment for ccRCC prognostic features, positive IMP3 expression was still found to be associated with a 42% increase in the risk of death from RCC (hazards ratio [HR], 1.42; P = .024). Among the subset of patients with clinically localized disease, positive IMP3 expression was associated with a nearly 5‐fold increased risk of distant metastases (HR, 4.71; P < .001).

CONCLUSIONS

Using a large and independent cohort of ccRCC patients, the authors confirmed that tumor cell IMP3 expression represents an independent predictor of aggressive ccRCC tumor behavior. Cancer 2008. ©2008 American Cancer Society.
Symptom burden after autologous stem cell transplantation for multiple myeloma
Authors: Campagnaro Erica , Saliba Rima , Giralt Sergio , Roden Linda , Mendoza Floralyn , Aleman Ana , Cleeland Charles , Weber Donna , Brown Jane , Anderson Karen O.
Published: February 2008

BACKGROUND

Multiple myeloma (MM) is the most common indication for high‐dose chemotherapy with autologous stem cell transplantation (ASCT) in the U.S. and can be associated with substantial morbidity. Thorough assessment and understanding of symptoms and risk factors for symptom development after ASCT are logical first steps toward developing strategies aimed at reducing the symptom burden associated with this procedure.

METHODS

The authors performed a prospective evaluation of symptom burden among 64 patients with myeloma who underwent ASCT. Symptom data were collected using the M. D. Anderson Symptom Inventory (MDASI) at 4 time points: baseline, the day of stem cell infusion (Day 0), nadir of counts, and Day 30. Univariate analysis was performed to correlate pretransplantation variables with post‐transplantation symptom burden at these time points.

RESULTS

MDASI scores increased significantly throughout transplantation, with most patients returning to baseline by Day 30 after the procedure. Patients with the highest MDASI scores at baseline had the highest MDASI scores at nadir (P = .02). Patients with prolonged time to transplantation and women had a trend toward higher nadir global symptom severity scores. These groups, as well as patients aged >60 years, had a trend toward higher nadir interference scores.

CONCLUSIONS

ASCT for MM was associated with significant but reversible symptom burden during the first 30 days, and the baseline symptom burden was the most important predictor of symptom burden after transplantation. The MDASI was useful as a tool for following the symptom burden associated with ASCT and may be used to evaluate interventions aimed at reducing transplantation‐related morbidity in these patients. Cancer 2008. ©2008 American Cancer Society.
Oncogenic properties and prognostic implications of the ubiquitin ligase Skp2 in cancer
Authors: Hershko Dan D.
Published: February 2008

The expression of Skp2, the ubiquitin ligase subunit that targets p27Kip1 for degradation, is commonly overexpressed in human cancers. p27Kip1 is a negative regulator of the cell cycle that plays an important role in tumor suppression. Loss of p27Kip1 secondary to enhanced ubiquitin‐mediated degradation results in uncontrolled proliferation and promotes tumor progression. In the present study the prognostic implications of Skp2 are reviewed and the mechanisms that regulate its expression in different human cancers. A review and analysis of the English literature was undertaken. Overexpression of Skp2 mRNA and protein levels was observed in many aggressive cancers and was commonly associated with down‐regulation of p27Kip1 levels and loss of tumor differentiation. Skp2 is an independent prognostic marker for disease‐free and overall survival and may provide additional predictive information to that provided by p27Kip1 alone. Targeting Skp2 in experimental models resulted in up‐regulation of p27Kip1 and arrested cellular proliferation. Alterations in Skp2 expression have profound effects on cancer progression and may serve as an accurate and independent prognostic marker. Thus, determination of levels of Skp2 and p27Kip1 by readily available immunohistochemical studies may be a useful tool in the assessment of prognosis, especially in patients with intermediate disease, and may potentially assist in the planning of adjuvant therapy. Skp2 may be an attractive target for the development of novel interventional therapy. Cancer 2008. © 2008 American Cancer Society.
Expression of cyclic guanosine monophosphate‐dependent protein kinase in metastatic colon carcinoma cells blocks tumor angiogenesis
Authors: Kwon In‐Kiu , Schoenlein Patricia V. , Delk Jennifer , Liu Kebin , Thangaraju Muthusmy , Dulin Nickolai O. , Ganapathy Vadivel , Berger Franklin G. , Browning Darren D.
Published: February 2008

BACKGROUND

Type 1 cyclic guanosine monophosphate (cGMP)‐dependent protein kinase (PKG) reportedly has exhibited antitumor properties, and its expression is down‐regulated in many tumors.

METHODS

The authors recently demonstrated that PKG re‐expression in metastatic colon carcinoma cells results in decreased tumorigenesis: In the current study, they addressed that mechanism.

RESULTS

Over‐expression of PKG in SW620 cells produced smaller, more apoptotic subcutaneous tumors in athymic mice, but the observed effect of PKG expression on growth and apoptosis in vitro was minimal. Closer examination of the subcutaneous xenografts revealed highly vascular tumors produced by the parental SW620 cells, which contrasted greatly with the PKG‐expressing tumors, in which cell growth was limited to “islands” surrounding CD31‐positive cells. The idea that PKG expression was associated with reduced tumor angiogenesis was supported by decreased levels of vascular endothelial growth factor in these tumors compared with tumors that were derived from parental SW620 cells. Investigation of potential mechanisms revealed that PKG expression was associated with reduced levels of β‐catenin compared with parental cells. Moreover, this effect of exogenous PKG on β‐catenin expression in SW620 cells also occurred in vitro, where the decrease was associated with reduced T‐cell factor‐dependent transcription.

CONCLUSIONS

Together the findings indicated that PKG down‐regulation in colon cancer cells is important for optimal tumor angiogenesis and that regulation of β‐catenin expression may be important to this process. Cancer 2008. © 2008 American Cancer Society.
Liver disease induced by radioembolization of liver tumors
Authors: Sangro Bruno , Gil‐Alzugaray Belen , Rodriguez Javier , Sola Iosu , Martinez‐Cuesta Antonio , Viudez Antonio , Chopitea Ana , Iñarrairaegui Mercedes , Arbizu Javier , Bilbao Jose I.
Published: February 2008

BACKGROUND

To the authors' knowledge, liver damage after liver radioembolization with yttrium90‐labeled microspheres has never been studied specifically.

METHODS

Using a complete set of data recorded prospectively among all patients without previous chronic liver disease treated by radioembolization at the authors' institution from September 2003 to July 2006, patterns of liver damage were identified and possible risk factors were analyzed.

RESULTS

In all, 20% of patients developed a distinct clinical picture that appeared 4 to 8 weeks after treatment and was characterized by jaundice and ascites. Veno‐occlusive disease was the histologic hallmark observed in the most severe cases. This form of sinusoidal obstruction syndrome was not observed among patients who never received chemotherapy or in those in whom a single hepatic lobe was treated. Relevant to treatment planning, a possible risk factor was a higher treatment dose in relation to the targeted liver volume. A transjugular intrahepatic stent shunt improved liver function in 2 patients with impending liver failure, although 1 of them eventually died from it.

CONCLUSIONS

Radioembolization of liver tumors, particularly after antineoplastic chemotherapy, may result in an uncommon but potentially life‐threatening form of hepatic sinusoidal obstruction syndrome that presents clinically with jaundice and ascites. Cancer 2008. © 2008 American Cancer Society.
Direct and indirect connections between cochlear root neurons and facial motor neurons: Pathways underlying the acoustic pinna reflex in the albino rat
Authors: Horta‐Júnior José de Anchieta C. , López Dolores E. , Alvarez‐Morujo Antonio Jesús , Bittencourt Jackson C.
Published: February 2008

Cochlear root neurons (CRNs) are involved in the acoustic startle reflex, which is widely used in behavioral models of sensorimotor integration. A short‐latency component of this reflex, the auricular reflex, promotes pinna movements in response to unexpected loud sounds. However, the pathway involved in the auricular component of the startle reflex is not well understood. We hypothesized that the auricular reflex is mediated by direct and indirect inputs from CRNs to the motoneurons responsible for pinna movement, which are located in the medial subnucleus of the facial motor nucleus (Mot7). To assess whether there is a direct connection between CRNs and auricular motoneurons in the rat, two neuronal tracers were used in conjunction: biotinylated dextran amine, which was injected into the cochlear nerve root, and Fluoro‐Gold, which was injected into the levator auris longus muscle. Under light microscopy, close appositions were observed between axon terminals of CRNs and auricular motoneurons. The presence of direct synaptic contact was confirmed at the ultrastructural level. To confirm the indirect connection, biotinylated dextran amine was injected into the auditory‐responsive portion of the caudal pontine reticular nucleus, which receives direct input from CRNs. The results confirm that the caudal pontine reticular nucleus also targets the Mot7 and that its terminals are concentrated in the medial subnucleus. Therefore, it is likely that CRNs innervate auricular motoneurons both directly and indirectly, suggesting that these connections participate in the rapid auricular reflex that accompanies the acoustic startle reflex. J. Comp. Neurol. 507:1763–1779, 2008. © 2008 Wiley‐Liss, Inc.
An adaptive variational multiscale method for convection–diffusion problems
Authors: Larson Mats G. , Målqvist Axel
Published: February 2008

The adaptive variational multiscale method is an extension of the variational multiscale method where the fine‐scale part of the solution is approximated by a sum of numerically computed solutions to localized subgrid problems. Furthermore, the crucial discretization parameters are chosen automatically by an adaptive algorithm based on a posteriori error estimates. This method has been developed for diffusion‐dominated problems and applied to multiscale problems that arise in oil reservoir simulation. In this paper, we extend the method to convection–diffusion problems. We present a duality based a posteriori error representation formula and an adaptive algorithm that tunes the fine‐scale mesh size and the patch sizes of the local problems. Numerical results show rapid convergence of the adaptive algorithm. Copyright © 2008 John Wiley & Sons, Ltd.
Effects of ampholyte concentration on protein behavior in on‐chip isoelectric focusing
Authors: Shim Jaesool , Dutta Prashanta , Ivory Cornelius F.
Published: February 2008

The effects of mobility corrections on carrier ampholytes are studied at various ampholyte concentrations to understand protein behavior during IEF. IEF simulations are conducted in the presence of 25 biprotic carrier ampholytes within a pH range of 6–9 after applying the Onsager–Debye–Hückel correction to the carrier ampholytes. Two model proteins with ten charge states but without ionic strength corrections are allowed to focus under an electric field of 300 V/cm in a 1 cm long channel. The IEF simulation results show that higher ionic strengths (50 – 100 mM) cause significant changes in the transient movement as well as the final focused profiles of both ampholytes and proteins. The time required for a single, well‐defined peak to form increases with ionic strength when Onsager corrections are applied to the carrier ampholytes. For a particular ampholyte concentration, the space‐averaged conductivity does not change during the final focusing stage, but the magnitude of space averaged conductivity is different for different ampholyte concentration. The simulation results also reveal that at steady‐state ionic strength profiles remain flat throughout the channel except at the locations of proteins where a significant change in ampholyte concentration is obtained.
The cyclophilin inhibitor Debio-025 shows potent anti–hepatitis C effect in patients coinfected with hepatitis C and human immunodeficiency virus
Authors: Flisiak Robert , Horban Andrzej , Gallay Philippe , Bobardt Michael , Selvarajah Suganya , Wiercinska-Drapalo Alicja , Siwak Ewa , Cielniak Iwona , Higersberger Jozef , Kierkus Jarek , Aeschlimann Christian , Grosgurin Pierre , Nicolas-Métral Valérie , Dumont Jean-Maurice , Porchet Hervé , Crabbé Raf , Scalfaro Pietro
Published: February 2008

Debio-025 is an oral cyclophilin (Cyp) inhibitor with potent anti–hepatitis C virus activity

in vitro

. Its effect on viral load as well as its influence on intracellular Cyp levels was investigated in a randomized, double-blind, placebo-controlled study. Mean hepatitis C viral load decreased significantly by 3.6 log10 after a 14-day oral treatment with 1200 mg twice daily (

P

< 0.0001) with an effect against the 3 genotypes (1, 3, and 4) represented in the study. In addition, the absence of viral rebound during treatment indicates that Debio-025 has a high barrier for the selection of resistance. In Debio-025–treated patients, cyclophilin B (CypB) levels in peripheral blood mononuclear cells decreased from 67 ± 6 (standard error) ng/mg protein (baseline) to 5 ± 1 ng/mg protein at day 15 (

P

< 0.01).

Conclusion:

Debio-025 induced a strong drop in CypB levels, coinciding with the decrease in hepatitis C viral load. These are the first preliminary human data supporting the hypothesis that CypB may play an important role in hepatitis C virus replication and that Cyp inhibition is a valid target for the development of anti–hepatitis C drugs.
Defective DNA strand break repair causes chromosomal instability and accelerates liver carcinogenesis in mice
Authors: Teoh Narci C. , Dan Yock Young , Swisshelm Karen , Lehman Stacey , Wright Jocelyn H. , Haque Jamil , Gu Yansong , Fausto Nelson
Published: February 2008

Chromosomal instability is a characteristic feature of hepatocellular carcinoma (HCC) but its origin and role in liver carcinogenesis are undefined. We tested whether a defect in the nonhomologous end-joining (NHEJ) DNA repair gene

Ku70

was associated with chromosomal abnormalities and enhanced liver carcinogenesis. Male Ku70 NHEJ-deficient (Ku70−/−), heterozygote (Ku70 +/−), and wild-type (WT) mice were injected with diethylnitrosamine (DEN), a liver carcinogen, at age 15 days. Animals were killed at 3, 6, and 9 months for assessment of tumorigenesis and hepatocellular proliferation. For karyotype analysis, primary liver tumor cell cultures were prepared from HCCs arising in Ku70 mice of all genotypes. Compared to WT littermates, Ku70−/− mice injected with DEN displayed accelerated HCC development. Ku70−/− HCCs harbored clonal increases in numerical and structural aberrations of chromosomes 4, 5, 7, 8, 10, 14, and 19, many of which recapitulated the spectrum of equivalent chromosomal abnormalities observed in human HCC. Ku70−/− HCCs showed high proliferative activity with increased cyclin D1 and proliferating cell nuclear antigen expression, Aurora A kinase activity, enhanced ataxia telangiectasia mutated kinase and ubiquitination, and loss of p53 via proteasomal degradation, features which closely resemble those of human HCC.

Conclusion:

These findings demonstrate that defects in the NHEJ DNA repair pathway may participate in the disruption of cell cycle checkpoints leading to chromosomal instability and accelerated development of HCC.
Loss of orphan receptor small heterodimer partner sensitizes mice to liver injury from obstructive cholestasis
Authors: Park Young Joo , Qatanani Mohammed , Chua Steven S. , LaRey Jennifer L. , Johnson Stacy A. , Watanabe Mitsuhiro , Moore David D. , Lee Yoon Kwang
Published: February 2008

The orphan nuclear hormone receptor small heterodimer partner (SHP) regulates the expression of several genes involved in bile acid homeostasis in the liver. Because bile acid toxicity is a major source of liver injury in cholestatic disease, we explored the role of SHP in liver damage induced by common bile duct ligation (BDL).

Shp

−/− mice show increased sensitivity in this model of acute obstructive cholestasis, with greater numbers of bile infarcts and higher mortality than wild-type C57BL/6 mice. This increased sensitivity could not be accounted for by differences in expression of bile acid homeostatic genes 2 or 5 days after BDL. Instead, higher basal expression of such genes, including the key biosynthetic enzyme cholesterol 7α hydroxylase (Cyp7A1) and the bile salt export pump, is associated with both an increase in bile flow prior to BDL and an increase in acute liver damage at only 1.5 hours after BDL in

Shp

−/− mice, as shown by bile infarcts. At 3 hours, Cyp7A1 expression still remained elevated in

Shp

−/− with respect to wild-type mice, and the hepatic and serum bile acid levels and total hepatobiliary bile acid pool were significantly increased. The increased sensitivity of mice lacking SHP contrasts with the decreased sensitivity of mice lacking the farnesoid X receptor (FXR; nuclear receptor subfamily 1, group H, member 4) to BDL, which has been associated with decreased intraductal pressure and fewer bile infarcts.

Conclusion:

We propose that differences in acute responses to BDL, particularly the early formation of bile infarcts, are a primary determinant of the differences in longer term sensitivity of the

Fxr

−/− and

Shp

−/− mice to acute obstructive cholestasis.
Vascular biology and pathobiology of the liver: Report of a single-topic symposium
Authors: Iwakiri Yasuko , Grisham Matthew , Shah Vijay
Published: February 2008

Portal hypertension and its complications account for the majority of morbidity and mortality that occurs in patients with cirrhosis. In addition to portal hypertension, a number of other vascular syndromes are also of great importance, especially the ischemia-reperfusion (IR) injury. With the identification of major vascular defects that could account for many of the clinical sequelae of these syndromes, the liver vasculature field has now integrated very closely with the broader vascular biology discipline. In that spirit, the Henry and Lillian Stratton Basic Research Single Topic Conference was held on the topic of Vascular Biology and Pathobiology of the Liver. The course took place approximately 10 years after the first American Association for the Study of Liver Disease (AASLD)-sponsored conference on this topic that occurred in Reston, Virginia. The conference initiated with an introduction to basic vascular cell signaling and then explored vascular biology specifically as it relates to liver cells. Subsequently, specific disease syndromes were discussed in more detail including portal hypertension and IR injury. Finally, clinical and translational sessions focused on emerging therapies and technologies to treat vascular diseases of the liver.
TIS21 negatively regulates hepatocarcinogenesis by disruption of cyclin B1–Forkhead box M1 regulation loop
Authors: Park Tae Jun , Kim Ji Yeon , Oh Paul S. , Kang So Young , Kim Bong Wan , Wang Hee Jung , Song Kye Yong , Kim Hyoung Chin , Lim In Kyoung
Published: February 2008

A functional and biochemical interaction of TIS21/BTG2/PC3 with Forkhead box M1 (FoxM1), essential transcription factor for hepatocyte regeneration and a master regulator of mitotic gene expression, was explored. Growth of hepatocellular carcinoma (HCC), developed by a single injection of diethylnitrosamine (DEN), was the same in both the TIS21+/+ and TIS21−/− mice until 6 months, whereas it was significantly higher in the TIS21−/− mice at 9 months. Expression of TIS21 was significantly lower in both human and murine HCCs than in the surrounding tissues. Forced expression of TIS21 impaired growth, proliferation, and tumorigenic potential of Huh7 cells. At the mechanistic level, TIS21 inhibited FoxM1 phosphorylation, a required modification for its activation, by reducing cyclin B1–cdk1 activity, examined by

in vitro

kinase assay and FoxM1 mutant analyses. These observations were further confirmed

in vivo

by the reciprocal control of TIS21 expression and FoxM1 phosphorylation in the diethylnitrosamine-induced HCCs and TIS21−/− mouse embryonic fibroblast (MEF), in addition to increased expression of cyclin B1 and cdk1 activity.

Conclusion:

TIS21 negatively regulated hepatocarcinogenesis in part by disruption of the FoxM1–cyclin B1 regulatory loop, thereby inhibiting proliferation of transformed cells developed in mouse and human livers.
Suppressor of cytokine signaling 1 protects mice against concanavalin A–induced hepatitis by inhibiting apoptosis
Authors: Torisu Takehiro , Nakaya Mako , Watanabe Satoko , Hashimoto Masayuki , Yoshida Hideyuki , Chinen Takatoshi , Yoshida Ryoko , Okamoto Fuyuki , Hanada Toshikatsu , Torisu Kumiko , Takaesu Giichi , Kobayashi Takashi , Yasukawa Hideo , Yoshimura Akihiko
Published: February 2008

Acute liver failure is associated with significant mortality. However, the underlying pathophysiological mechanism is not yet fully understood. Suppressor of cytokine signaling-1 (SOCS1), which is a negative-feedback molecule for cytokine signaling, has been shown to be rapidly induced during liver injury. Here, using liver-specific SOCS1-conditional-knockout mice, we demonstrated that SOCS1 deletion in hepatocytes enhanced concanavalin A (ConA)–induced hepatitis, which has been shown to be dependent on activated T and natural killer T (NKT) cells. Although serum cytokine level and NKT cell activation were similar in wild-type (WT) and SOCS1-deficient mice after ConA treatment, proapoptotic signals, including signal transducers and activators of transcription 1 (STAT1) and Jun-terminal kinase (JNK) activation, were enhanced in SOCS1-deficient livers compared with those in WT livers. SOCS1-deficient hepatocytes had higher expression of Fas antigen and were more sensitive to anti-Fas antibody–induced apoptosis than were WT hepatocytes. Furthermore, SOCS1-deficient hepatocytes were more sensitive to tumor necrosis factor (TNF)-α-induced JNK activation and apoptosis. These data indicate that SOCS1 is important to the prevention of hepatocyte apoptosis induced by Fas and TNF-α. In contrast, SOCS1 overexpression in the liver by adenoviral gene transfer prevented ConA-induced liver injury.

Conclusion:

These findings indicate that SOCS1 plays important negative roles in fulminant hepatitis and that forced expression of SOCS1 is therapeutic in preventing hepatitis.
Recombinant factor VIIa for variceal bleeding in patients with advanced cirrhosis: A randomized, controlled trial
Authors: Bosch Jaime , Thabut Dominique , Albillos Agustín , Carbonell Nicolas , Spicak Julius , Massard Julien , D'Amico Gennaro , Lebrec Didier , de Franchis Roberto , Fabricius Sφren , Cai Yan , Bendtsen Flemming
Published: February 2008

A beneficial effect of recombinant activated factor VII (rFVIIa) in Child-Pugh class B and C patients with cirrhosis who have variceal bleeding has been suggested. This randomized controlled trial assessed the efficacy and safety of rFVIIa in patients with advanced cirrhosis and active variceal bleeding. At 31 hospitals in an emergency setting, 256 patients (Child-Pugh > 8; Child-Pugh B = 26%, C = 74%) were randomized equally to: placebo; 600 μg/kg rFVIIa (200 + 4× 100 μg/kg); or 300 μg/kg rFVIIa (200 + 100 μg/kg). Dosing was intravenous at 0, 2, 8, 14, and 20 hours after endoscopy, in addition to standard vasoactive, prophylactic antibiotic, and endoscopic treatment. The primary composite endpoint consisted of failure to control 24-hour bleeding, or failure to prevent rebleeding or death at day 5. Secondary endpoints included adverse events and 42-day mortality. Baseline characteristics were comparable between groups. Administration of rFVIIa had no significant effect on the composite endpoint compared with placebo (

P

= 0.37). There was no significant difference in 5-day mortality between groups; however, 42-day mortality was significantly lower with 600 μg/kg rFVIIa compared with placebo (odds ratio 0.31, 95% confidence interval = 0.13–0.74), and bleeding-related deaths were reduced from 12% (placebo) to 2% (600 μg/kg). A marked heterogeneity in the failure rate in all treatment groups was observed across participating centers. Adverse events, including overall thromboembolic events, were comparable between groups.

Conclusion:

Treatment with rFVIIa had no significant effect on the primary composite endpoint compared with placebo. Therefore, decision on the use of this hemostatic agent in acute variceal bleeding should be carefully considered, because results of this study do not support the routine use of rFVIIa in this setting. Adverse events were comparable across groups.
Racial disparity in liver disease: Biological, cultural, or socioeconomic factors
Authors: Nguyen Geoffrey C. , Thuluvath Paul J.
Published: February 2008

Chronic liver diseases are a major public health issue in the United States, and there are substantial racial disparities in liver cirrhosis–related mortality. Hepatitis C virus (HCV) is the most significant contributing factor in the development of chronic liver disease, complications such as hepatocellular carcinoma, and the need for liver transplantation. In the United States, African Americans have twice the prevalence of HCV seropositivity and develop hepatocellular carcinoma at more than twice the rate as whites. African Americans are, however, less likely to respond to interferon therapy for HCV than are whites and have considerably lower likelihood of receiving liver transplantation, the only definitive therapy for end-stage liver disease. Even among those who undergo transplantation, African Americans have lower 2-year and 5-year graft and patient survival compared to whites. We will review these racial disparities in chronic liver diseases and discuss potential biological, socioeconomic, and cultural contributions. An understanding of their underlying mechanisms is an essential step in implementing measures to mollify racially based inequities in the burden and management of liver disease in an increasingly racially and ethnically diverse population.
Adoptive transfer of CD8+ T cells from transforming growth factor beta receptor type II (dominant negative form) induces autoimmune cholangitis in mice
Authors: Yang Guo-Xiang , Lian Zhe-Xiong , Chuang Ya-Hui , Moritoki Yuki , Lan Ruth Y. , Wakabayashi Kanji , Ansari Aftab A. , Flavell Richard A. , Ridgway William M. , Coppel Ross L. , Tsuneyama Koichi , Mackay Ian R. , Gershwin Eric M.
Published: February 2008

We recently reported that mice with a T cell–restricted expression of a dominant negative form of transforming growth factor β receptor type II (dnTGFβRII) spontaneously develop autoimmune cholangitis that resembles human primary biliary cirrhosis (PBC), including antimitochondrial antibodies (AMAs) and extensive portal CD4+ and CD8+ lymphocytic infiltrates. On the basis of these data, we performed a series of experiments to determine whether the pathology was secondary to direct dnTGFβRII disruption of the liver and/or alternatively the appearance of autoreactive T cells. First, using dnTGFβRIIRag1−/− mice, we noted a normal hepatic and biliary structure. Hence, we performed a rigorous series of adoptive transfer studies, transferring Ly5.1+ unfractionated spleen cell CD4+ or CD8+ T cells from dnTGFβRII mice into B6/Rag−/− (Ly 5.2) recipients. In unmanipulated dnTGFβRII mice, there was a marked increase in CD4+ and CD8+ T cell biliary infiltrates with AMA. Indeed, B6/Rag−/− recipients of dnTGFβRII unfractionated cells develop features of liver disease similar to PBC, suggesting that splenic loss of self-tolerance alone is sufficient to cause disease in this model and therefore that there is no specific abnormality in the biliary targets required for appearance of disease. More importantly, adoptive transfer of CD8+ but not CD4+ T cells into B6/Rag−/− mice led to liver histopathology remarkably similar to PBC, emphasizing a prominent role for CD8 T cell–mediated pathogenesis. In contrast, B6/Rag−/− recipients of CD4+ T cells from dnTGFβRII mice predominantly developed inflammatory bowel disease associated with higher levels of serum interferon γ and tumor necrosis factor α.

Conclusion:

These data suggest that in this model of PBC, autoreactive CD8+ cells destroy bile ducts.
Temporal-spatial activation of apoptosis and epithelial injury in murine experimental biliary atresia
Authors: Erickson Nissa , Mohanty Sujit Kumar , Shivakumar Pranavkumar , Sabla Gregg , Chakraborty Ranajit , Bezerra Jorge A.
Published: February 2008

Biliary atresia is a fibro-inflammatory cholangiopathy that obstructs the extrahepatic bile ducts in young infants. Although the pathogenesis of the disease is undefined, studies in livers from affected children and neonatal mice with experimental biliary atresia have shown increased expression of proapoptosis molecules. Therefore, we hypothesized that apoptosis is a significant mechanism of injury to duct epithelium. To test this hypothesis, we quantified apoptosis using terminal transferase dUTP nick end labeling and active caspase-3 staining in livers and extrahepatic bile ducts from Balb/c mice infected with

Rhesus

rotavirus (RRV) within 24 hours of birth. RRV induced a significant increase in labeled cells in the portal tracts and in epithelial and subepithelial compartments of extrahepatic bile ducts, with onset within 3 days and peaks at 5–10 days. Exploring mechanisms of injury, we found increased biliary expression of caspases 1 and 4 and of interferon-gamma (IFNγ)–related and tumor necrosis factor-alpha (TNFα)–related genes. Using a cholangiocyte cell line, we found that neither IFNγ nor TNFα alone affected cell viability; however, simultaneous exposure to IFNγ and TNFα activated caspase-3 and decreased cell viability. Inhibition of caspase activity blocked apoptosis and restored viability to cultured cholangiocytes. In vivo, administration of the caspase inhibitor IDN-8050 decreased apoptosis in the duct epithelium and the extent of epithelial injury after RRV challenge.

Conclusion:

The biliary epithelium undergoes early activation of apoptosis in a mouse model of biliary atresia. The synergistic role of IFNγ and TNFα in activating caspase-3 in cholangiocytes and the decreased apoptosis following pharmacologic inhibition of caspases support a prominent role for apoptosis in the pathogenesis of experimental biliary atresia.